Effect of Biofertilizer On Growth, Productivity, Quality and Economics of Rainfed Organic Ginger (Zingiber Officinale Rosc.) Bhaisey Cv. in North-Eastern Region of India

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Jour na l of
Agric ult ura l Sc ie nc e
a nd Te chnology A
Volume 3, Number 2, February 2013 (Serial Number 22)
Contents
Research Papers
83
Effect of Biofertilizer on Growth, Productivity, Quality and Economics of Rainfed Organic

Ginger (Zingiber officinale Rosc.) Bhaisey cv. in North-Eastern Region of India
Nongmaithem Jyotsna, Mainak Ghosh, Dulal Chandra Ghosh, Wahengbam Ingo Meitei and Jagadish
Timsina
99
Research on the Soil Carbon Storage of Alpine Grassland under Different Land Uses in
Qinghai-Tibet Plateau
Tao Li, Lei Ji, Tao Liu, Zhongqi Song, Shujing Yang and Youmin Gan
105
Chemical and Sensory Properties of Olive Oil as Influenced by Different Sources of Irrigation Water
Salam Ayoub, Saleh Al-Shdiefat, Hamzeh Rawashdeh and Ibrahim Bashabsheh
113
Ethical Trading: The Implications of the Human Rights Watch Report on South African Fruit
Exports
Portia Ndou and Ajuruchukwu Obi
126
Physiological and Phytosanitary Potentials of Coriander and Radish Seeds

Jucilayne Fernandes Vieira, Francisco Amaral Villela, Orlando Antonio Lucca Filho and Raifer Simes
Campelo
131
Response of Amaranth to Irrigation and Organic Matter

Jimmy Akinfemi Osunbitan
140
Quantitative Changes in Protein and Cholesterol in Haemolymph of the Red Palm Weevil
Rhynchophorus ferrugineus after Treatment LeucokininII
Mona Mohammed Saleh Al-Dawsary
146
The Role of Cellulase and Pectinase in Apricot Canker Caused by Hendersonula torulidi and
Phiaoacremonium aleophillium
Nidhal Y. M. Al-Morad
151
Innovation and Technical Efficiency in the Smallholder Dairy Production System in Ethiopia
Amlaku Asres, Johann Slkner and Maria Wurzinger
165
Richness and Diversity of Ants and Beetles in Genetically Modified Cotton Field in Brazil
Carla Cristina Dutra, Marcos Gino Fernandes, Josu Raizer and Camila Meotti
Journal of Agricultural Science and Technology A 3 (2013) 83-98

Earlier title: Journal of Agricultural Science and Technology, ISSN 1939-1250
D
DAVID
PUBLISHING
Effect of Biofertilizer on Growth, Productivity, Quality

and Economics of Rainfed Organic Ginger (Zingiber
officinale Rosc.) Bhaisey cv. in North-Eastern Region of
India
Nongmaithem Jyotsna1, Mainak Ghosh2, Dulal Chandra Ghosh3, Wahengbam Ingo Meitei1 and Jagadish Timsina4
1. College of Agriculture, Central Agricultural University Imphal, Manipur 795001, India
2. Department of Agronomy, Bihar Agricultural University, Bihar 813210, India
3. Institute of Agriculture, Visva-Bharati, Sriniketan-731236, West Bengal, India
4. IRRI-Bangladesh Office, Banani DOHS, Dhaka-1206, Bangladesh
Received: November 13, 2012 / Published: February 20, 2013.

Abstract: Ginger yields in the NorthEastern region of India are low because the extremely poor farmers of the region can not afford
to apply any chemical fertilizers and hence apply only the locally-available farmyard manures to ginger fields. Biofertilizers may be a
cheap source of fertilizers for ginger cultivation as they can increase nutrient availability and improve rhizome quality and are
required in small quantity. An investigation was thus undertaken to study the effect of different biofertilizers on growth, productivity,
quality and economics of organic ginger grown under rainfed condition in NorthEastern region of India. Seed treatment with
biofertilizers enhanced growth, increased rhizome yield by 19.0% and resulted in 32.4% higher net profit over control. Among the
seed treatments, Azotobacter 5.0 kg ha-1, Azospirillum 3.75 kg ha-1 and Phosphotica 3.75 kg ha-1 were found optimum in improving
most of the growth attributes, increasing yield components and yield of rhizome by 5.6%-13.5%. They also improved rhizome
quality by increasing specific gravity, oleoresin and dry matter content and by decreasing crude fibre in rhizome. They resulted in
higher net return by 4.0%-12.0% as compared to their other levels. Combined use of Azotobacter 5.0 kg ha-1 along with Phosphotica
3.75 kg ha-1 was found to be the best treatment combination which greatly improved growth and yield attributes of ginger and
ultimately recorded markedly higher productivity (2.0%-23.5%) over other combinations. This treatment combination improved the
quality of the produce and resulted in the highest gross return ($4,905 ha-1), net return ($3,525 ha-1) and return per dollar (3.55)
invested in ginger cultivation. It appears that growing organic ginger by treating the seed rhizome with Azotobacter 5.0 kg ha-1 along
with Phosphotica 3.75 kg ha-1 can result in good growth and high productivity of improved quality rhizome and ultimately result in
maximum net profit and thus can be recommended for the NorthEastern region of India.
Key words: Biofertilizers, growth, productivity, quality, economics, ginger (Zingiber officinale Rosc.).
1. Introduction
Ginger (Zingiber officinale Rosc.) is a tropical
rhizomatous high value spice crop adapted for
cultivation in tropical and subtropical climate. The
NorthEastern region of India with the subtropical
climate, where it is the main cash crop, has
Corresponding author: Jagadish Timsina, Ph.D., research
field: agronomy. E-mail: timsinaj@hotmail.com.
tremendous potential for ginger production and hence

can support the livelihoods and improve the economic
level of many ginger growers. The average yield (5.8 t
ha-1) of ginger in this region, however, is considered
to be low [1]. The soil, climate and other ecological
factors in the NorthEastern region favour the growth
and development of this crop and there is a
tremendous scope to increase its yield and total
84
Effect of Biofertilizer on Growth, Productivity, Quality and Economics of Rainfed Organic Ginger
(Zingiber officinale Rosc.) Bhaisey cv. in North-Eastern Region of India
production. One possible reason for its low yield in

NorthEastern region could be due to the poor nutrient
management practices adopted for this crop. The
ginger production in the NorthEastern region is
organic by default because the farmers of the region
apply only the locally available farmyard manures
(e.g., cow dung manure, pig manure, poultry manure
and rabbit manure) and do not apply any chemical
fertilizers or pesticides [2]. In contrast, ginger being a
nutrients supply through greater mineralization due to

higher microbial activities [12, 15]. However,
information about the use of different biofertilizers in
ginger production is very limited. The present study
was thus carried out to investigate the effect of
biofertilizer on growth, productivity, quality and
economics of rainfed ginger production in
NorthEastern region of India.
2. Materials and Methods
heavy feeder and an exhaustive crop requires large

quantities of manures and fertilizers. Considering the
2.1 Experimental Site
increasing demand for organic products all over the
The field experiment was conducted during 2007

and 2008 at the Horticulture Experimental Farm,
College of Agriculture, Central Agricultural
University, Imphal, Manipur in NorthEastern India.
The place is located at 2445N latitude, 9356E
longitude with an altitude of 790 m above mean sea
level. The experimental soil was clayey in texture
(15.5% sand, 21.2% silt and 61.1% clay), medium in
fertility status (230, 13.3 and 267 kg ha-1 available N,
P and K, respectively), well-drained with gentle slope.
The experimental site comes under warm humid moist
region where monsoon normally starts from April and
extends up to September. Unpredictable pre-monsoon
shower during March is not uncommon in this region.
The crop was grown on rainfed condition but received
1,341 and 1,207 mm rainfall during the growing
period of 2007 and 2008, respectively (Table 1). The
maximum temperature ranged from 24.1 C to 29.6 C
while the minimum temperature varied from 9.6 C to
22.5 C during the cropping seasons. The relative
humidity varied from 58.5% to 84.7% in 2007 and
58.9% to 88.5% in 2008. Both temperature and
relative humidity remained very conducive for growth
and rhizome productivity of ginger.
world, the ginger farmers can receive higher returns

from their produce if grown organically. Organic
farming
has
attracted
increasing
attention
for
environmental protection, improved quality and better

market demands [3]. However, ginger cultivated with
only organic manures will produce low yields and
would require chemical fertilizers for increased
nutrition or biofertizers for increased availability.
Ginger requires a tropical or sub-tropical humid
climate for its commercial production. The crop is
sensitive to water logging, frost and salinity and
tolerant to wind and drought. It thrives on a wide
variety of soils; but for high yield, it prefers light,
loose, friable and well drained soil rich in humus and
slightly acidic (pH 6.0-6.5) in reaction [1].
Biofertilizers have now emerged as a promising
component of nutrient supply [4-6]. The role of
different biofertilizers like Azospirillum and
Azotobacter cultures in fixing atmospheric nitrogen
has been well established by several workers [7-9].
The microorganisms can build up organic matter of
the soil which can increase the availability of other
nutrients [10, 11] and secrete growth promoting
substances [12]. The use of phosphate solubilizing
microorganisms has shown positive responses in
many demonstrations and field trials [13, 14]. Use of
biofertilizers in organic ginger production may further
enhance its growth and productivity by producing
growth promoting substances and enhancing plant
2.2 Experimental Details

The experiment was laid out in a completely
randomized block design with three biofertilizers
each at three levels along with a common control (no
biofertilizer) in three replications in 3.6 m 3.0 m
Table 1 Mean maximum and minimum air temperature,
relative humidity and precipitation and during growing
season 2007 and 2008.
Month
T max
T min
Relative
Precipitation
(C)
(C)
humidity (%) (mm)
2007 2008 2007 2008 2007 2008 2007 2008
Mar.
Apr.
May
Jun.
Jul.
Aug.
Sept.
Oct.
Nov.
25.1
26.2
28.3
28.7
28.1
28.8
27.8
26.9
24.1
24.9
28.8
29.6
28.6
28.1
28.5
29.4
27.6
25.3
10.7
14.3
19.9
21.9
22.2
22.3
20.9
18.3
13.6
12.5
15.6
19.3
21.3
22.2
22.5
21.3
18.2
9.6
58.5
70.7
74.5
81.6
84.7
82.7
79.7
81.5
74.1
70.2
58.9
69.7
80.3
84.0
83.0
84.3
88.5
83.8
17.6
136.8
233.4
201.0
218.6
112.2
191.0
178.0
52.6
39.6
97.8
94.6
260.2
210.2
244.5
151.1
87.6
21.7
plots. The three levels for each of three biofertilizers

are: (a) Azotobacter (a1 = 2.5, a2 = 3.75 and a3 = 5.0
kg ha-1), (b) Azospirillum (b1 = 2.5, b2 = 3.75 and b3 =
Table 2
5.0 kg ha-1) and (c) Phosphotica (c1 = 2.5, c2 = 3.75

and c3 = 5.0 kg ha-1). Thus there were 28 treatment
combinations. Culture solutions of different
biofertilizers were prepared by dissolving 10 g, 15 g
and 20 g of each biofertilizer (for their respective
doses) and their combinations in 500 mL of water
separately for each biofertilizer treatment, with total of
27 biofertilizer culture solutions (Table 2). Each
biofertilizer treatment (culture solution) was mixed
thoroughly with 8 kg ginger sets (required for each
treatment) of variety Bhaisey and dried in shade
before planting. The rhizomes (20 g set) were planted
on March 16, 2007 and March 18, 2008 with a
spacing of 30 cm 30 cm in 3.6 3.0 m plots.
A general dose of 20 t farm yard manure (FYM) ha-1
containing 0.50% N, 0.13% P and 0.55% K was
Treatment details along with composition of 500 mL biofertilizer culture solution.
Treatments
T1
T2
T3
T4
T5
T6
T7
T8
T9
T10
T11
T12
T13
T14
T15
T16
T17
T18
T19
T20
T21
T22
T23
T24
T25
T26
T27
T28
Combinations
a1b1c1
a1b1c2
a1b1c3
a1b2c1
a1b2c2
a1b2c3
a1b3c1
a1b3c2
a1b3c3
a2b1c1
a2b1c2
a2b1c3
a2b2c1
a2b2c2
a2b2c3
a2b3c1
a2b3c2
a2b3c3
a3b1c1
a3b1c2
a3b1c3
a3b2c1
a3b2c2
a3b2c3
a3b3c1
a3b3c2
a3b3c3
a0b0c0
85
Biofertilizer composition in 500 mL culture solution

Azotobacter 10 g + Azospirillum 10 g + Phosphotica 10 g
Control (No use of biofertilizer)
86
applied during land preparation. The crop received

hand weeding twice along with light earthing up on
May 25 and July 14 in 2007 and May 27 and July 16
in 2008. No chemical fertilizer, pesticide and
irrigation water was applied.
2.3 Observations Recorded
The canopy spread was measured in centimeter
with the help of a linear scale from five randomly
selected clumps of each plot at 90, 150 and 210 days
after planting (DAP). The canopy spread was
calculated by multiplying the length with width of
each canopy in each plot and average canopy spread
(cm-2 clump-1) was estimated for each plot. Four
clumps from each plot were collected at 90, 150 and
210 DAP for recording biomass production. The plant
samples were cleaned and washed in water to remove
surface contamination and separated into stem (dry
leaves + stem), green leaves (lamina) and rhizome.
Fresh weight of rhizome was recorded in g m-2. A
piece of rhizome of each plot was taken; its fresh
weight was noted before chopping. All plant parts
including chopped rhizome were kept in separate
paper packets which in turn placed in an oven for
drying at 65-70 C till constant weights were obtained.
The dry weight of leaves, stems and rhizome was then
recorded in g m-2. The sum of dry weights of these
plant parts were taken as the total dry matter
accumulation (DMA). The area of 10 leaves of each
treatment was measured with a leaf area meter (AM
300, USA), and the leaves were put in an oven for
recording dry weight. The area/dry weight of these
leaves was used for determining leaf area index (LAI)
as suggested by Watson [16]. The crop growth rate
(CGR) during the period of 90-150 and 150-210 DAP
were estimated as: CGR = (W2 W1)/(t2 t1),

expressed in g m-2 day-1, where W2 and W1 were the
final and initial dry weights of the crop per unit land
area at times t2 and t1, respectively [16]. The fresh
weight of rhizome was used for determining rhizome
bulking rate (RBR). The RBR during the period of
90-150 and 150-210 DAP were estimated using the
following formula: RBR = (R2 R1)/(t2 t1) expressed

in g m-2 day-1, where, R2 and R1 were the final and
initial fresh weights of rhizome per unit land area at
times t2 and t1 respectively. The crop was harvested on
November 12 in 2007 and November 14 in 2008 when
the leaves turned yellow and started drying up. Fifty
clumps of each plot were lifted carefully with the help
of a spade and the rhizomes were separated and kept
in shade for 2 days. The fresh weights of rhizome
were recorded in t ha-1. Harvest index (HI) was
estimated as: HI = Rhizome dry weight (kg ha-1)/total
dry biomass (kg ha-1) 100, and expressed in
percentage (%).
2.4 Quality Parameters
The quality parameters like specific gravity, dry
matter content, oleoresin content and crude fibre
content in rhizome were estimated in the laboratory.
The specific gravity of rhizome was measured by water
displacement method. After removing the mud, roots
and shoots, rhizomes were weighed in a balance, and
expressed in gram. A 250 mL measuring cylinder was
filled with distilled water and rhizomes were
submerged into the water. The level of water in the
measuring cylinder was noted before and after the
submergence of rhizomes. The displaced water was
measured in milliliter and specific gravity was
determined by the following equation: Specific gravity
= Weight of rhizomes (g)/amount of water displaced
(mL) and expressed in g mL-1. Equal quantity of fresh
ginger from each treatment was oven dried uniformly
after chopping into thin slices and the dry weights were
recorded. The dry matter content in rhizome was
estimated by the equation: Dry matter content = Dry
weight of the sample/fresh weight of the sample 100,
and expressed in percent. The oleoresin content was
determined by using acetone as a solvent. 10 g of the
dried sample was weighed and transferred to a glass
column (18 mm 450 mm) with stopcock, then 50 mL

of acetone was added and allowed to stand overnight
87
for 16 h at 25 2 C. The filtrate extracted through non

absorbent cotton was collected in a pre-weighed 100
mL beaker. Column was washed with 20 mL of acetone.
The extracts were pooled and evaporated to dry at
80 C over a water bath. The amount of oleoresin was
estimated gravimetrically as per the Official
Analytical Methods of the American Spices Trade
Association [17], and expressed in percentage. The
crude fibre was determined following the acid digestion
method [17]. Equal quantity of fresh ginger from each
treatment was oven dried uniformly after chopping into
thin slices. The dried sample was allowed to digest with
distilled sulphuric acid and sodium hydroxide. The
crude fibre was estimated by weighing the organic
matter in the dried residue remaining after digesting the
sample with distilled sulphuric acid and sodium
hydroxide, and expressed in percent.
parameters such as specific gravity, dry matter,

oleoresin and crude fibre content in rhizome were also
studied. The average values of three replications for
all the treatments of two years data were used for this
purpose.
2.5 Economics
Canopy spread
Leaf area index
(cm-2 clump-1)
90
150
210
90
150
210
DAP DAP DAP DAP DAP DAP
Control
504
1,597 1,965 0.29 2.33 2.55
Treatment 700
1,823 2,306 0.41 3.24 3.56
89.8
107.3 0.03 0.15 0.21
S. Em () 40.3
LSD 0.05 121.8 271.3 323.9 0.08 0.46 0.62
Azotobacter level
a1
540
1,626 2,010 0.31 2.41 2.72
a2
585
1,680 2,100 0.34 2.63 2.96
a3
682
1,825 2,298 0.40 3.32 3.49
29.9
35.8
0.01 0.05 0.07
S. Em () 13.4
LSD 0.05 38.0
84.8
101.2 0.02 0.14 0.19
Azospirillum level
b1
561
1,653 2054 0.32 2.53 2.84
b2
658
1,792 2,250 0.39 3.17 3.40
b3
587
1,686 2,103 0.33 2.66 2.94
29.9
35.8
0.01 0.05 0.07
S. Em () 13.4
LSD 0.05 38.0
84.8
101.2 0.02 0.14 0.19
Phosphotica level
c1
571
1,666 2,077 0.33 2.59 2.90
c2
650
1,781 2,234 0.39 3.11 3.33
c3
585
1,684 2,097 0.34 2.66 2.94
29.9
35.8
0.01 0.05 0.07
S. Em () 13.4
LSD 0.05 38.0
84.8
101.2 0.02 0.14 0.19
*a = Azotobacter; b = Azospirillum; c = Phosphotica; doses of
biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively.
The cost of inputs such as FYM, seed rhizome,

biofertilizer and labour and output (rhizomes) were
estimated as per prevailing market price. The gross
return, net return and return per dollar invested in
different biofertilizer treatments were assessed by
computing the cost of the inputs and price of the
produce (output) to study the economics of rainfed
ginger production.
2.6 Data Analysis
All data were analyzed statistically following the
standard procedures as described by Gomez and
Gomez [18]. The data were tested for analysis of
variance and least significant difference (P = 0.05) to
compare the effect of biofertilizer treatments on
growth, productivity, quality and economics of rainfed
ginger production. The interaction effects were
presented wherever they were significant. Multiple
regression analysis was done to examine the
relationships between the rhizome yield and the
growth attributes like canopy spread and LAI at
different stages. The relationships among the quality
3. Results
3.1 Growth Parameters
Use of biofertilizer exhibited significant effect on
growth attributes of rainfed organic ginger. The
canopy spread increased by 14.2% to 38.9%, LAI by
39.1% to 41.4% (Table 3), CGR by 23.9% to 25.2%
and DMA by 21.2% to 27.9% (Table 4) at different
stages due to biofertilizer treatments over control.
Table 3 Effect of biofertilizer on canopy spread and leaf
area index of ginger at different stages (average data of two
years).
Particulars
88
Among the different biofertilizer treatments, seed

treatment with high dose (5.00 kg ha-1) of Azotobacter
(a3) showed significant improvement of all the growth
attributes over their lower levels. Similarly, seed
treatment with medium dose (3.75 kg ha-1) of both
Azospirillum (b2) and Phosphotica (c2) caused marked
increase in all the growth attributes at most of the
growth stages over those of their higher and lower
levels. Application of low level (2.50 kg ha-1) of
biofertilizers became less effective in improving the
growth parameters as compared to their higher levels.
Combined use of Azotobacter and Phosphotica
showed significant interaction effect on influencing
most of the growth attributes of this crop. The highest
values of canopy spread (Fig. 1A), LAI (Fig. 1B),
CGR (Fig. 1C) and DMA (Fig. 1D) at most of the
stages were recorded with the use of high level (5.00
kg ha-1) of Azotobacter in combination with medium
level (3.75 kg ha-1) of Phosphotica (a3c2). These
values were significantly higher than those obtained
with other treatment combinations except the
combinations of high level of Azotobacter with high
or low levels of Phosphotica (a3c1 or a3c3) in most of
the cases. This treatment combination (a3c2) was
found to be optimum in improving the growth
parameters of ginger under the study.
3.2 Yield and Yield Attributes
The seed treatment with biofertilizer showed
positive and significant effect on influencing yield
components like rhizome growth and rhizome bulking
rate at different growth periods that ultimately
influenced
the
rhizome
yield
over
control.
Biofertilizer increased rhizome yield over control by

15.9% in 2007 and 22.4% in 2008. The highest
rhizome growth and rhizome bulking rate were found
with the highest level (5.0 kg ha-1) of Azotobacter.
The medium level (3.75 kg ha-1) of both Azospirillum
and Phosphotica also showed markedly greater yield
components as compared to its high and low levels at
Table 4 Effect of biofertilizer on crop growth rate (CGR)

and dry matter accumulation (DMA) in ginger (average
data of two years).
Crop growth rate Dry matter accumulation
(g m-2 day-1)
(g m-2)
90-150 150-210
150
210
90 DAP
DAP
DAP
DAP
DAP
Control
6.54
1.51
124.2
507.0 614.2
Treatment 8.10
1.89
153.2
648.4 744.4
0.12
8.6
29.7
36.9
S. Em () 0.47
LSD 0.05 1.43
0.38
26.1
89.6
111.3
Azotobacter level
a1
6.76
1.64
132.0
538.0 643.0
a2
7.16
1.70
137.0
565.0 667.0
a3
8.03
1.77
147.0
630.0 728.0
0.04
2.9
9.9
12.3
S. Em () 0.16
LSD 0.05 0.45
0.12
8.2
28.0
34.8
Azospirillum level
b1
6.95
1.67
135.0
551.0 654.0
b2
7.85
1.74
144.0
615.0 716.0
b3
7.17
1.70
137.0
567.0 668.0
0.04
2.9
9.9
12.3
S. Em () 0.16
LSD 0.05 0.45
0.12
8.2
28.0
34.8
Phosphotica level
c1
7.06
1.65
136.0
559.0 661.0
c2
7.74
1.75
143.0
608.0 709.0
c3
7.17
1.70
137.0
566.0 668.0
0.04
2.9
9.9
12.3
S. Em () 0.16
LSD 0.05 0.45
0.12
8.2
28.0
34.8
a = Azotobacter; b = Azospirillum; c = Phosphotica; doses of
biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively.
Particulars
all the growth stages (Table 5). The increased yield

components at high to medium level of biofertilizer
enhanced rhizome yield of rainfed ginger and
accordingly the high level of Azotobacter increased
rhizome yield by 13.5% over its lower level and 9.2%
over its medium level as evidenced by pooled analysis
of two years data (Table 6). Similarly, medium level
of both Azospirillum and Phosphotica enhanced
rhizome yield by 8.9% and 8.5% respectively over
their lower level and 5.6% and 4.6% over their higher
level (Table 6). The lower levels of biofertilizer were
found less effective in improving yield components
and rhizome yield of rainfed ginger. The harvest index,
however, did not vary much among the different
biofertilizer treatments during both years.
89
Fig. 1 Interaction effect of Azotobacter and Phosphotica on growth attributes of ginger. (A) canopy spread, (B) leaf area
index, (C) crop growth rate and (D) dry matter accumulation; vertical lines show the error bars.
a1, a2 and a3 represent Azobactor levels of 2.50 kg ha-1, 3.75 kg ha-1 and 5.00 kg ha-1, respectively; c1, c2 and c3 represent Phosphotica
levels of 2.50 kg ha-1, 3.75 kg ha-1 and 5.00 kg ha-1, respectively.

the yield attributes and yield of this crop. The highest
values of rhizome growth (Fig. 2A) and rhizome
bulking rate (Fig. 2B) were recorded with the use of
high level of Azotobacter in combination with
medium level of Phosphotica (a3c2). This treatment

combination ultimately produced the higher rhizome
yield (Fig. 2C) in both years (22.06 t ha-1 in 2007 and
22.09 t ha-1 in 2008) except the combinations of high
level of Azotobacter with higher and lower levels of
Phosphotica. Use of lower levels of both Azotobacter
90
Table 5 Effect of biofertilizer on rhizome growth and

rhizome bulking rate of ginger (average data of two years).
Table 6
Effect of biofertilizer on productivity and
efficiency of ginger.
Rhizome bulking rate

Rhizome growth
(g m-2 day-1)
(fresh weight in g m-2)
90
150
210
90-150
150-210
DAP DAP
DAP DAP
DAP
Control
388
1,760
2,293 22.39
8.34
Treatment 438
2,061
2,622 27.57
9.90
86
105
1.14
0.45
S. Em () 21
LSD 0.05 62
260
317
3.44
1.37
Azotobacter level
a1
394
1,777
2,332 22.99
8.65
a2
413
1,877
2,422 24.54
9.20
a3
433
2,079
2,620 27.41
9.50
28.7
35.0 0.38
0.15
S. Em () 6.8
LSD 0.05 19.3
81.2
99.0 1.07
0.43
Azospirillum level
b1
402
1,824
2,373 23.68
8.76
b2
428
2,032
2,579 26.73
9.51
b3
408
1,878
2,423 24.53
9.08
28.7
35.0 0.38
0.15
S. Em () 6.8
LSD 0.05 19.3
81.2
99.0 1.07
0.43
Phosphotica level
c1
404
1,853
2,398 24.15
8.77
c2
427
2,003
2,558 26.29
9.47
c3
409
1,878
2,419 24.50
9.11
28.7
35.0 0.38
0.15
S. Em () 6.8
LSD 0.05 19.3
81.2
99.0 1.07
0.43
biofertilizer 1, 2 and 3 are 2.5 kg ha-1, 3.75 kg ha-1 and 5.0 kg
ha-1, respectively.
Rhizome yield (t ha-1)

Harvest index (%)
2007
2008
pooled 2007
2008
Control
18.97
17.88
18.43
53.8
54.5
Treatment 21.98
21.89
21.94
54.4
54.4
0.83
0.76
1.2
1.4
S. Em () 0.80
LSD 0.05
2.43
2.50
2.29
NS
NS
Azotobacter level
a1
19.42
18.74
19.08
54.2
54.6
a2
20.16
19.50
19.83
54.2
54.5
a3
21.87
21.43
21.65
53.9
54.2
0.28
0.25
0.4
0.5
S. Em () 0.27
LSD 0.05
0.76
0.78
0.71
NS
NS
Azospirillum level
b1
19.76
19.06
19.41
54.3
54.6
b2
21.40
20.87
21.14
53.8
54.2
b3
20.29
19.73
20.01
54.2
54.5
0.28
0.25
0.4
0.5
S. Em () 0.27
LSD 0.05
0.76
0.78
0.71
NS
NS
Phosphotica level
c1
19.74
19.06
19.40
54.2
54.5
c2
21.21
20.86
21.04
54.0
54.3
c3
20.50
19.74
20.12
54.1
54.5
0.28
0.25
0.4
0.5
S. Em () 0.27
LSD 0.05
0.76
0.78
0.71
NS
NS
ha-1, respectively.
Particulars
and Phosphotica recorded the lowest values of the

yield parameters as compared to those of the
other treatment combinations at all the growth stages.
Accordingly, seed treatment with lower levels of both
the biofertilizers produced the lowest rhizome yield
(18.12 t ha-1 in 2007 and 17.62 t ha-1 in 2008)
resulting in 21.7% and 25.4% less yield when
compared with the highest yield in respective years.
Seed treatment with Azotobacter 5.0 kg ha-1 along
with Phosphotica 3.75 kg ha-1 (a3c2) was found the
best for improving yield components and producing
high rhizome yield of rainfed organic ginger.
3.3 Rhizome Quality
The seed treatment with different biofertilizers
exerted favourable effect on quality of ginger rhizome.
Particulars
The biofertilizers treated crop increased the dry matter

content, specific gravity and oleoresin content but
decreased the crude fibre content in rhizome and
thereby improved its quality over control (Table 7).
Application of high and medium doses of Azotobacter
increased the dry matter, specific gravity and oleoresin
content over its low level but decreased the crude fibre
content. Use of medium doses of Azospirillum and
Phosphotica increased the specific gravity and
oleoresin content over those of their higher and lower
levels. The dry matter content in rhizome was also
increased by the application of medium level of
Phosphotica over its higher and lower levels. Medium
level of Phosphotica resulted in the highest dry matter
content (22.1%) in rhizome. The crude fibre content in
rhizome, however, did not vary much among the
different levels of Azospirillum and Phosphotica.
91
Fig. 2 Interaction effect of Azotobacter and Phosphotica on (A) rhizome growth, (B) rhizome bulking rate, (C) rhizome yield
and (D) economics of rainfed ginger; vertical lines indicate the show the error bars.
a1, a2 and a3 represent Azobactor levels of 2.50 kg ha-1, 3.75 kg ha-1 and 5.00 kg ha-1, respectively; c1, c2 and c3 represent Phosphotica
levels of 2.50 kg ha-1, 3.75 kg ha-1 and 5.00 kg ha-1, respectively.
3.4 Economics of Rainfed Ginger

The results showed significant effect of biofertilizer
treatments on increasing gross return, net return and
return per dollar invested in ginger cultivation, but it
had no effect on cost of cultivation of ginger. Seed

treatment with biofertilizers considerably increased
the gross return ($4,886 ha-1), net return ($3,499 ha-1)
and return per dollar invested (3.5) as compared to
those of the control plots ($4,086 ha-1, $2,805 ha-1 and
92
Table 7 Effect of biofertilizer on quality attributes of

ginger (average data of two years).
Oleoresin
Specific
Dry matter
Crude fibre
content
gravity
content (%)
content (%)
-1
(%)
(g cc )*
Control
1.19
20.41
5.20
6.33
Treatment
1.30
22.02
6.41
5.65
0.03
0.52
0.16
0.19
S. Em ()
LSD 0.05
0.08
1.59
0.48
0.57
Azotobacter level
a1
1.22
20.69
5.44
6.17
a2
1.26
21.49
5.98
6.06
a3
1.27
21.49
6.01
5.75
0.01
0.17
0.05
0.06
S. Em ()
LSD 0.05
0.02
0.49
0.15
0.18
Azospirillum level
b1
1.24
21.09
5.74
6.08
b2
1.27
21.29
5.93
5.94
b3
1.24
21.29
5.76
5.97
0.01
0.17
0.05
0.06
S. Em ()
LSD 0.05
0.02
NS
0.15
NS
Phosphotica level
c1
1.24
20.69
5.51
5.98
c2
1.27
22.09
6.32
6.05
c3
1.24
20.89
5.60
5.96
0.01
0.17
0.05
0.06
S. Em ()
LSD 0.05
0.02
0.49
0.15
NS
ha-1, respectively.
Particulars
3.2, respectively). Seed treatment with high dose of

Azotobacter resulted in higher gross return ($4,811 ha-1)
over its medium ($4,407 ha-1) and low ($4,239 ha-1)
levels (Table 8). The net return and return per dollar
invested also followed a similar trend. High dose of
Azotobacter paid maximum profit ($3,449 ha-1) and
return per dollar invested (3.53) as compared to medium
and low doses. The lower dose of Azotobacter paid the
lowest gross return ($4,239 ha-1), net return ($2,928 ha-1)
and return per dollar invested (3.23) indicating its less
efficiency in ginger productivity. Azospirillum and
Phosphotica also exerted significant effect on gross
return, net return and return per dollar invested in ginger
cultivation. Use of medium dose of Azospirillum and
Phosphotica paid higher gross return ($4,696 and 4,674
ha-1) over their high ($4,447 and 4,472 ha-1) and low
($4,314 and 4,311 ha-1) levels. Both the treatments paid
Table 8 Effect of biofertilizer on economics of ginger

cultivation (average data of two years).
Cost
of Gross
Particulars cultivation return
($ ha-1)*
($ ha-1)
Control
1,281.02
4,085.75
Treatment 1,387.02
4,885.78
191.65
S. Em () 65.47
LSD 0.05 NS
578.78
Net return Return per

$ invested
($ ha-1)
2,804.73
3,498.76
149.21
450.62
3.19
3.52
0.10
0.32
4,239.09
4,406.87
4,811.36
63.88
180.79
2,928.07
3,077.84
3,449.34
49.74
140.76
3.23
3.32
3.53
0.03
0.10
4,313.74
4,696.38
4,447.20
63.88
180.79
2,995.25
3,338.30
3,112.70
49.74
140.76
3.27
3.46
3.33
0.03
0.10
Azotobacter level
a1
a2
a3
1,311.02
1,329.03
1,362.02
S. Em () 21.82
LSD 0.05 NS
Azospirillum level
b1
b2
b3
1,318.49
1,358.08
1,334.50
S. Em () 21.82
LSD 0.05 NS
Phosphotica level
c1
c2
c3
1,318.20
4,311.27 2,993.07 3.27
1,355.92
4,674.49 3,318.57 3.45
1,336.93
4,471.56 3,134.63 3.34
63.88
49.74
0.03
S. Em () 21.82
LSD 0.05 NS
180.79
140.76
0.10
ha-1, respectively; Ginger seed rhizome $330/ton, FYM
$5.5/ton, Biofertilizers $5/kg, Labour charges $1.4/man-day
and price of ginger rhizomes $220/ton.
markedly higher net return ($3,338 and 3,319 ha-1) and

return per dollar invested (3.46 and 3.45) in ginger
cultivation over their high and low levels.
The combined use of Azotobacter and Phosphotica
showed significant interaction effect on gross return,
net return and return per dollar invested in ginger
cultivation. The highest gross return ($4,905 ha-1), net
return ($3,525 ha-1) and return per dollar invested
(3.55) in ginger cultivation were recorded with the
application of high level of Azotobacter along with
medium level of Phosphotica. This treatment
combination paid significantly higher gross return, net
return and return per dollar invested than those
obtained with most of the other treatment
combinations (Fig. 2D). The lowest gross return
($3,971 ha-1), net return ($2,688 ha-1) and return per

dollar invested (3.1) were obtained with combined use
of low level of both Azotobacter and Phosphotica.
4. Discussion
4.1 Growth Parameters
Biofertilizers exhibited significant and positive
effect on growth attributes like canopy spread, LAI,
CGR and DMA in organic rainfed ginger over those
of the control plots. Seed treatment with high dose
(5.0 kg ha-1) of Azotobacter (a3) and medium dose
(3.75 kg ha-1) of both Azospirillum (b2) and
Phosphotica (c2) caused marked increase of all of
these growth attributes at most of the growth stages
over those of their higher and lower levels. The higher
canopy spread and greater LAI of biofertilizer treated
plots might be due to better development of shoots by
greater availability of plant nutrients and growth
promoting hormones released by higher microbial
activities [4]. Similar beneficial effect of biofertilizer
inoculation on growth attributes of different crops was
observed by many workers [9, 11, 19].
The leaf area index is a factor closely related to its
productivity because of the fact that the total leaf area
affects the amount of photosynthate available. Increase
in LAI enables the plant to enhance photosynthetic rate
and results in higher growth and yield. The higher
CGR and DMA in biofertilizers inoculated plants
might be correlated with the increased LAI and canopy
spread. The increased LAI and canopy spread
ultimately enhanced CGR and DMA. Such increase
was attributed to effective N fixation by Azotobacter
and Azospirillum, solubilization of soil available and
native P through production of organic acids by
phosphate solubilizing bacteria (PSB) and release of
growth regulators [4, 10].
most of the growth attributes like canopy spread, LAI,
CGR and DMA. Combined application of high level
(5.0 kg ha-1) of Azotobacter with medium level (3.75
93
kg ha-1) of Phosphotica seemed to be optimum on

influencing the above growth parameters in most of
the cases (Fig. 1). The different types of biofertilizers
in association improved the growth of the crop than
their application in isolation. This is because of the
fact that range of organic acid and plant growth
promoting substances produced by combined use of
biofertilizers increased substantially over their single
application [20]. Different types of acid and growth
promoting substances secreted by the microorganisms
in association are additive in action and synergistic in
effect [10, 21]. The increased growth attributes
obtained at combined use of Azotobacter and
Phosphotica in this investigation was due to their
additive effect. However, the low level (2.5 kg ha-1) of
Azotobacter and Phosphotica application recorded
low values of the growth attributes which was
significantly lower than those obtained with higher
level of Azotobacter and Phosphotica at most of the
growth stages.
4.2 Yield and Yield Attributes
The yield of ginger is a function of rhizome growth
and rhizome bulking rate. The development of
rhizome starts after initial period of establishment and
early shoot growth. Seed treatment with biofertilizer
showed positive effect on rhizome growth and
rhizome bulking rate that ultimately increased the
rhizome yield over control. The high level (5.0 kg ha-1)
of Azotobacter exerted greater effect on increasing
rhizome growth, rhizome bulking rate and rhizome
yield as compared to its lower levels. The medium
levels (3.75 kg ha-1) of both Azospirillum and
Phosphotica also showed markedly greater yield
components and greater rhizome yield than their
higher and lower levels. Increased yield by
biofertilizer inoculation was due to the high canopy
spread and LAI enabling production of relatively large
amount of assimilates for high yield. The rhizome
yield showed very strong and positive correlation with
canopy spread (Figs. 3A, 3B and 3C) and LAI (Figs.
94
Fig. 3 Subfigures [A-C] represent the relationships of rhizome yield with canopy spread at [A] 90, [B] 150 and [C] 210 days
after planting; subfigures [D-F] represent the relationships of rhizome yield with leaf area index at [D] 90, [E] 150 and [F]
210 days after planting.
DAP represents days after planting and LAI indicates leaf area index.
3D, 3E and 3F) at all the growth stages. This showed

very vital role of these growth parameters in the yield
improvement of ginger. The above growth attributes
enhanced by the application of biofertilizers might be
owing to higher availability and efficient use of the
nutrients through out the growing period as a result of

greater microbial activities. This in turn increased the
rhizome growth and rhizome bulking rate at different
periods. Increase in yield could be attributed to
increase in growth and yield attributing characters
resulting from higher fixation of atmospheric N,

dissolution of insoluble phosphates in soil to soluble
forms and production of plant growth hormones and
vitamins by microorganisms [22]. High rhizome yield
of ginger was mainly due to high DMA in crop
resulting from high rhizome growth by greater RBR.
High DMA in biofertilizer treated crop during its
growth period was, thus, responsible for determining
high rhizome yield of ginger. This is in conformity
with the findings of Gupta and Awasthi [23], Stoop et
al. [24] and Wijebandara et al. [25].
showed significant interaction effect on yield
attributes and yield of this crop. Use of high level (5.0
kg ha-1) of Azotobacter together with medium level
(3.75 kg ha-1) of Phosphotica seemed to be the best
treatment combination for improving yield and yield
attributes (Fig. 2). This treatment combination
ultimately produced the highest rhizome yield (22.08 t
ha-1). The increased growth parameters were mainly
responsible for enhancing yield attributes that
ultimately produced high rhizome yield [11, 26].
4.3 Rhizome Quality
The biofertilizer treatments improved the rhizome
quality by increasing specific gravity, dry matter
content and oleoresin content in rhizome and
decreasing the crude fibre content in it. Application of
higher levels of Azotobacter (5.0 kg ha-1 and 3.75 kg
ha-1) increased the specific gravity, dry matter content
and oleoresin content in rhizome over its lower level
(2.50 kg ha-1) while decreased the crude fibre content
with increasing its dose. Similarly, use of medium
levels of Azospirillum and Phosphotica (3.75 kg ha-1)
increased specific gravity and oleoresin content in
rhizome over their higher and lower levels.
Phosphotica was also responsible for increasing dry
matter content but both Azospirillum and Phosphotica
did not affect the crude fibre content in rhizome.
Azotobacter and Azospirillum, apart from their ability
to fix N, produce anti-fungal antibiotics that inhibit
95
the growth of several pathogenic fungi in the root

region and hence improving root growth and crop
nutrition that ultimately improves the quality of the
product [27]. They also produce growth regulating
substances like phytohormones, vitamins etc. that help
in improving the quality of the produce by balancing
the nutrition of the crop [28]. Similarly, Phosphotica
helps in solubilization of soil available and native P
through production of organic acids and improves
crop quality by promoting physiological activities
through better P nutrition [29]. It is interesting to note
that these quality parameters of rhizome were strongly
related to each other either positively or negatively.
The dry matter content was positively correlated with
specific gravity (Fig. 4A) and oleoresin content (Fig.
4B) indicating their compatibility in improving
rhizome quality. It was also noticed that crude fibre
content in rhizome was strongly but negatively
correlated with dry matter (Fig. 4C) and oleoresin
contents (Fig. 4D). This has a good impact on quality
control. Increase in dry matter and oleoresin contents
in rhizome decreases the crude fibre content and thus
improves the rhizome quality. The beneficial effect of
biofertilizer treatment on rhizome quality might be
due to better nourishment of the crop by increasing
nutrient availability in the root zone through greater
microbial activities [30].
4.4 Economics of Rainfed Ginger Cultivation
The seed treatment with different biofertilizers
increased the gross return, net return and return per
dollar invested on ginger cultivation. This was mainly
because biofertilizer application improved growth and
increased rhizome productivity and quality through
better crop nutrition and thus resulted in higher profit.
Azotobacter showed marked effect on gross return, net
return and return per dollar invested in ginger
cultivation and application of high level of Azotobacter
resulted in higher gross return ($4,811 ha-1), net profit
($3,449 ha-1) and return per dollar invested (3.54) over
its medium ($4,407 ha-1, $3,078 ha-1 and 3.31,
96
Fig. 4 Relationships among different quality parameters of rhizome: [A] dry matter content (%) with specific gravity (g
cc-1), [B] dry matter content (%) with oleoresin content (%), [C] dry matter content (%) with crude fibre (%), and [D] crude
fibre content (%) with oleoresin content (%).
respectively) and low ($4,239 ha-1, Rs 2,928 ha-1 and

3.23, respectively) levels. Azospirillum and
Phosphotica also favourably influenced the economics
of ginger cultivation and use of medium level of both
the biofertilizers increased gross return, net return and
return per dollar invested over those obtained at their
high and low levels. Greater availability of plant
nutrients together with increased activities of plant
growth promoting substances in biofertilizers treated
crop might be responsible for higher productivity and
greater income [8, 12]. The high economic return was
attributed to high rhizome productivity resulting from
enhanced nutrient supply through greater microbial
activities induced by biofertilizer treatments [28, 29].
The significant interaction effect of seed treatment
with Azotobacter and Phosphotica on economics of
ginger cultivation showed their compatibility in
microbial activities. The highest gross return ($4,905
ha-1), net return ($3,525 ha-1) and return per dollar

invested (3.55) in ginger resulting from the combined
use of high level of Azotobacter along with medium
level of Phosphotica (a3c2) proved to be the best
treatment combination out of all treatment
combinations (Fig. 2D). This treatment combination
was found to be optimum in exerting maximum
additive and synergistic microbial effects on
enhancing growth, productivity and economic return
of rainfed organic ginger in the north-eastern region of
India.
5. Conclusions
The beneficial effect of biofertilizer on influencing
the growth, productivity, quality and economics of
organic ginger grown under rainfed condition has
been established. The seed treatment with Azotobacter
5.0 kg ha-1 (a3), Azospirillum 3.75 kg ha-1 (b2) and
Phosphotica 3.75 kg ha-1 (c2) improved most of the

growth attributes, increased yield components and
yield of rhizome, improved crop quality like specific
gravity, oleoresin, and increased dry matter content in
rhizome. These treatments also resulted in higher
return as compared to their other levels. Combined use
of high level of Azotobacter along with medium level
of Phosphotica (a3c2) was found to be the best
treatment combination which improved growth and
yield attributes of ginger and ultimately resulted in
markedly higher productivity (22.08 t ha-1) of the crop.
This treatment combination greatly improved the
quality of the produce and resulted the highest gross
return ($4,905 ha-1), net return ($3,525 ha-1) and return
per dollar (3.55) invested in ginger cultivation. The
results suggest growing of organic ginger by treating
the seed rhizome with Azotobacter 5.0 kg ha-1 along
with Phosphotica 3.75 kg ha-1 (a3c2) for obtaining
good growth, high yield of good quality rhizome and
maximum profit in the NorthEastern region of India.
[9]
[10]
[11]
[12]
[13]
[14]
References
[15]
[1]
[2]
[3]
[4]
[5]
[6]
[7]
[8]
Anonymous, Basic Statistics of North Eastern Region,

North Eastern Council, Ministry of Home Affairs, Shillong,
2002, pp. 122-125.
S. Govind, R. Chandra, G.S. Karibasappa, Research on
Spices in NEH Region, ICAR Research Complex for NEH
Region, Umiam, 1998, pp. 9-22.
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D
DAVID
PUBLISHING
Research on the Soil Carbon Storage of Alpine

Grassland under Different Land Uses in Qinghai-Tibet
Plateau
Tao Li, Lei Ji, Tao Liu, Zhongqi Song, Shujing Yang and Youmin Gan
Department of Grassland Science, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Yaan
625014, China
Abstract: In this article, we mainly analysis the soil carbon storage of the alpine grassland under different land uses in Qinghai-Tibet
Plateau. The samples of this investigation include six experimental fields which are fenced mowing grassland, artificial grassland,
winter and spring grazing meadowland, summer and autumn mild grazing land, summer and autumn moderate grazing pasture and
summer and autumn severe grazing land and seven soil layers included 0 cm-5 cm, 5 cm-10 cm, 10 cm-20 cm, 20 cm-30 cm, 30
cm-50 cm, 50 cm-70 cm and 70 cm-100 cm. The results show that the soil carbon storage in different soil layers will gradually
reduce and the difference was remarkable (P < 0.05). What is more, the soil carbon storage of alpine grassland under different land
uses has following sequence: winter and spring grazing grassland > summer and autumn mild grazing land > artificial grassland >
summer and autumn moderate grazing meadowland > summer and autumn severe grazing pasture > fenced mowing meadow, and the
significant difference between them is remarkable (P < 0.05).
Key words: Qinghai-Tibet Plateau, alpine grassland, soil, carbon storage.
1. Introduction
The grassland ecosystem is one of the most
important and widely distributed ecosystem types in
terrestrial ecosystems and playing an important role in
the global carbon cycle and climate regulation [1, 2].
The Qinghai-Tibet Plateau, as the highest and largest
Eurasia geomorphology unit, is an essential
component of the grassland ecosystem and the worlds
concentrated distribution area of low-latitude
permafrost. It is not only sensitive to global climate
change, but also playing an important role in Asia
even global climate change [3]. Due to its unique
geographical location, the Qinghai-Tibet Plateau
widely distributed with such typical alpine grassland
vegetation as the alpine meadow, alpine steppe and
Corresponding author: Youmin Gan, professor, research
fields:
grassland
resources
and
ecology.
E-mail:
ganyoumin1954@163.com.
alpine meadow grassland. These grasslands occupy

more than 60% of the plateau area [4]. The consequent
development of alpine meadow soil, sub-alpine
meadow soil, alpine grassland meadow soil and other
alpine soil are rich in organic matter. Soil carbon
density is significantly higher than that of other
regions [5].
In recent years, with the temperature rising of the
Qinghai-Tibet Plateau, studies [5, 6] have shown that
the plateau permafrost has great potential for
emissions of greenhouse gases such as carbon and
nitrogen, because of the remarkable permafrost
thermal sensitivity. Because of this special
geographical and ecological unit of the Qinghai-Tibet
Plateau and its important role in global change, so the
study of carbon storage of grassland ecosystem in the
Qinghai-Tibet
Plateau
and
its
distribution
characteristics for the evaluation of grassland
100
Research on the Soil Carbon Storage of Alpine Grassland under

Different Land Uses in Qinghai-Tibet Plateau
ecosystems, even the carbon cycle response and

feedback effects of the whole terrestrial ecosystems
have important scientific value to the global carbon
cycle and global climate change.
Different use of grassland is one of the main human
factors of the grassland ecosystem carbon storage [7].
Domestic and foreign scholars have undertaken some
researches on the soil carbon storage under different
grassland uses: Staben et al. [8] reported that after
becoming a perennial artificial grassland, the soil
organic carbon storage of cultivated land would
increase; Mensah et al. [9] said pasture could increase
surface soils organic carbon storage, while reclaimed
farmland would result in reduction of soil organic
carbon storage; Andeoon et al. [10] showed that
grassland during reclamation could lead to large losses
of soil organic storage, which accounted for 30% to
50%; Risse et al. [11] demonstrated that overgrazing
could promote soil respiration, accelerating the carbon
release from soil to the atmosphere; Studies about the
typical temperate grassland in the Xilin River basin of
Inner Mongolia [12] have shown that reclamation and
grazing can reduce the soil carbon and nitrogen
content, but fence rotational grazing can increase the
content of the soil carbon and nitrogen; Studies about
the different grassland soil properties and the density
of the soil carbon, nitrogen, phosphorus in Eastern
Qilian Mountains of the Qinghai-Tibet Plateau [13]
have shown that plant communities and characteristics
of root distribution influenced the density of soil
carbon, nitrogen and phosphorus. The difference is
obvious between the plots and the soil layers. The
above studies have shown that different grassland uses
have an influence on carbon cycle of the grassland
ecosystem. At present, the problem of the carbon
source and sink of the grassland ecosystems has not
been determined. Therefore, the study of carbon
storage has more important theoretical value and
practical ecological significance.
In this article, through the analysis of the alpine
grassland soil carbon storage under different land uses
in Science and Technology Park of Sichuan Academy

of Grassland Science, we intend to explore the main
factors, providing data support for accurately
estimating the carbon storage of grassland ecosystem
in Qinghai-Tibet Plateau in China, analyzing its
variation, as well as researching on the function of
grassland carbon source and sink and the carbon cycle
mechanism in the system.

2.1 General Situation of the Study Area
The Science and Technology Park of Sichuan
Academy of Grassland Science located at the
Hongyuan County, Aba autonomous prefecture,
Sichuan province. The region belongs to the
Qinghai-Tibet Plateau, which average elevation is
3,560 m, the topography is high and smooth, the
annual precipitation is 500 mm-800 mm and the
average annual temperature is -2 C-5 C. It has a
large temperature difference between day and night
and the region belongs to the alpine semi-humid and
humid areas. Grassland vegetation and livestock, in
the region, have many species. The type of the soil
belongs to meadow soil and marsh soil. And the soil
has large amount of organic matter, good texture and
non-saline. Grassland vegetation is meadow and
marsh vegetation mainly formed by Gramineae and
Cyperaceae plant [14, 15].
The Science and Technology Park of Sichuan
Academy of Grassland Science have total land area is
328.14 hm2 and the grassland area of which can make
use is 314.76 hm2. The type of the grassland is mainly
alpine meadow. In recent years, to reasonable using
grassland resources, the park has been fenced
construction, which created favorable conditions for
scientific and rational use and management of
grassland. The park has summer and autumn pastures
about 165.71 hm2, winner and spring pastures about
147.64 hm2, fenced mowing meadow 3.54 hm2 and
artificial grassland. In the research, we select six plots
as objects of the research for observation, sampling

and analysis of soil carbon storage. The plots come

from fenced mowing meadow, artificial grassland,
winter and spring grazing grassland, summer and
autumn grazing grassland (according to the degree of
the use, summer and autumn grazing grassland is
divided into summer and autumn mild grazing
grassland, summer and autumn moderate grazing
grassland and summer and autumn severe grazing
grassland).
2.2 Sampling
We randomly selected ten sample plots which size
is one square meters in the selected plots in August.
And then, we investigated the species and their
coverage within each sample and recorded the
elevation, slope and aspect of the plot. In the sample,
we cut the vegetation that grown above the ground,
and next cleared the residue and impurities leaved on
the soil surface. The method of taking sample that we
adopted is the soil auger [16]. We used a five-diameter
soil auger to get seven soil layers that are
0-5-10-20-30-50-70-100 cm. The upper 4-6 drill
mixed and the lower 3-4 drill mixed. We put the
samples in the ziplock bags by layers and then
removed the plant residues over 2 mm sieve and last
put the samples on the brown paper for shade drying
and analyzing; we used cutting ring to collect soil
samples for determination of soil bulk density [17].
2.3 Laboratory Analysis
Soil samples were dried to constant weight at 65 C.
And we used gravimetric method to determine the soil
bulk density [18]. For determining the soil organic
carbon content [19], we used K2Cr2O7-H2SO4
oxidation and volumetric method.
2.4 Statistical Analysis
We adopted software SPSS (version 17.0) for the
mathematical statistics. In the process, we selected the
LSD test method (0.05 significant levels) for the
multiple comparisons. The formula calculated the soil
organic carbon storage:
101
BDsoil Dsoil iAi

Soil carbon storage CSsoil =
In this formula, BDsoil indicates soil bulk density;
Dsoil shows soil organic carbon content; Ai represents
area of each grassland type; i (1, 2) means the
number of grassland types.
3. Results and Discussion
3.1 Changes of the Soil Carbon Storage in Different
Soil Layers
The soil carbon storage under different grassland

used in the region shows the certain regularity in the
vertical direction and the soil carbon storage of
grassland consistently shows a decreasing tendency
from the surface to the bottom with the gradient. The
significant difference between the layers is P < 0.05
(Table 1). Soil carbon storage in 0 cm-5 cm layer is
2.8388-5.3013 kg m-2; Soil carbon storage in 5-10 cm
layer has 2.2796-5.1381 kg m-2; Soil carbon storage in
10-20 cm layer contains 3.9607-9.7223 kg m-2; Soil
carbon storage in 20-30 cm layer is 3.0538-8.2274 kg
m-2; Soil carbon storage in 30-50 cm has 4.274-15.414
kg m-2; Soil carbon storage in 50-70 cm contains
3.1939-13.1211 kg m-2; Soil carbon storage in 70-100
cm is 4.2221-15.3571 kg m-2.
3.2 Changes of the Soil Carbon Storage under the
Different Uses of Grassland
Under the different uses of the grassland, soil
carbon storage in each soil layer and the total soil
carbon storage show the certain regularity in the
horizontal direction (Table 2). And the soil carbon
storage has significant difference (P < 0.05). In 0
cm-5 cm layer with relatively high carbon storage in
soil, the sequence of carbon storage is winter and
spring grazing grassland > summer and autumn severe
grazing grassland > artificial grassland > summer and
autumn moderate grazing grassland > summer and
autumn mild grazing grassland > fenced mowing
meadow; In 5 cm-10 cm soil layer, the situation of
carbon storage is similar to the layer in 0 cm-5 cm; Of
the carbon storage in 10 cm-20 cm soil layer, winter and

102
Table 1
Soil carbon storage of each soil layers of the different use of grassland kg m-2.
Sampling Fenced mowing

depth
meadow
Artificial
grassland
Mean
SD
0-5cm
2.83880.1083bc
5-10cm
2.27960.0671c
10-20cm 3.96070.0907a
20-30cm 3.05380.0457b
30-50cm 4.2740.2409a
50-70cm 3.19390.0398b
70-100cm 4.22210.4632a
Total
3.40330.1709
Mean
SD
4.25630.0236e
4.06670.0917e
7.69160.0987a
5.62190.0835d
7.26430.0842b
3.71700.0481f
6.34220.1291c
5.56570.3327
Summer and
Winter and spring
autumn mild
grazing grassland
grazing grassland
Mean
SD
Mean
SD
5.30130.032e
2.99870.1062c
5.13810.0207e 2.08140.0602c
9.72230.0528c 5.05210.0458ab
8.22740.053d
3.26960.1343c
15.4140.7047a 5.79630.1633a
13.1210.4235b 4.47050.2464b
15.3570.0999a 5.41170.7212a
10.32590.9155 4.25720.2718
Summer and
autumn moderate
grazing grassland
Mean
SD
3.13330.4526d
3.340.0925cd
4.58420.234bc
4.88030.324b
8.11110.49a
7.83680.8098a
6.92340.4132a
5.54420.4525
Summer and
autumn severe
grazing grassland
Mean
SD
5.27890.013d
4.66950.0518e
7.48280.0458c
4.98670.0638de
8.13630.0294b
7.15070.101c
9.49460.5121a
6.74280.3810
Lowercase in the same treatment of each column in the table show significant difference (P < 0.05).
Table 2
Analysis of variance about the soil carbon storage of the different use of grassland.
Grassland types
Fenced mowing
meadow
Artificial grassland
Winter and spring
grazing grassland
Summer and
autumn mild
grazing grassland
Summer and
autumn moderate
grazing grassland
Summer and
autumn severe
grazing grassland
0 cm-5 cm
5 cm-10 cm
2.83880.1083c 2.27960.0671f
10 cm-20 cm
20 cm-30 cm
30 cm-50 cm
3.96070.0907e 3.05380.0457d 4.2740.2409d
50 cm-70 cm
70 cm-100 cm
3.19390.0398d 4.22210.4632e
4.25630.0236b 4.06670.0917c 7.69160.0987b 5.62190.0835b 7.26430.0842b 3.71700.0481cd 6.34220.1291cd

5.30130.032a
5.13810.0207a 9.72230.0528a 8.22740.053a
15.4140.7047a 13.1210.4235a 15.3570.0999a
2.99870.1062c 2.08140.0602e 5.05210.0458c 3.26960.1343d 5.79630.1633c 4.47050.2464c 5.41170.7212de
3.13330.4526c 3.340.0925d
5.27890.013a
4.58420.234d
4.88030.324c
8.11110.49b
7.83680.8098b 6.92340.4132c
4.66950.0518b 7.48280.0458b 4.98670.0638c 8.13630.0294b 7.15070.101b
9.49460.5121b
Lowercase in the same treatment of each column in the table show significant difference (P < 0.05).
spring grazing grassland > artificial grassland >

summer and autumn severe grazing grassland >
summer and autumn mild grazing grassland > summer
and autumn moderate grazing grassland > fenced
mowing meadow; Of the carbon storage in 20 cm-30
cm, winter and spring grazing grassland > artificial
grassland > summer and autumn severe grazing
grassland > summer and autumn moderate grazing
grassland > summer and autumn mild grazing
grassland > fenced mowing meadow; Of the carbon
storage in 30 cm -50 cm, winter and spring grazing
grassland > summer and autumn severe grazing
grassland > artificial grassland > summer and autumn
mild grazing grassland > fenced mowing meadow; Of
the carbon storage in 50 cm-70 cm, winter and spring
grazing grassland > summer and autumn moderate
grazing grassland > summer and autumn severe

grazing grassland > summer and autumn mild grazing
grassland > artificial grassland > fenced mowing
meadow; In 70 cm-100 cm, the situation of carbon
storage is similar to it in 30 cm-50 cm soil. Total soil
carbon storage of grassland under different use
patterns, winter and spring grazing grassland >
summer and autumn severe grazing grassland >
artificial grassland > summer and autumn moderate
grazing grassland > summer and autumn mild grazing
grassland > fenced mowing meadow.
Carbon storage of fenced mowing meadow is the
lowest and the main reason is that the structure of the
vegetation is simple, thus the carbon fixed and
transformed to the soil are relatively small, whereas
the biomass is larger and the intensity of soil
respiration is bigger in growing season. Ploughing and

planting have caused loses of soil organic carbon of

artificial grassland, therefore, the carbon storage gets
lower than mild grazing grassland. The otherness of
the soil carbon storage under different uses of the land
shows in the Table 2. Grasslands of different grazing
degree would demonstrate a certain differences in the
characteristics of its vegetation and the biomass, so it
is inevitable to result in the differences in carbon
storage and efficiency of fixing and transforming
organic carbon. The vegetation cover and richness of
species in wild grazing grassland, as well as the
biomass, is relatively large, so the efficiency of fixing
and transforming carbon to the soil is higher and the
fixed amount of carbon will be relatively large. The
condition in severe grazing grassland is just the
reverse. Therefore, the soil carbon storage will
increase as the grazing decreases.
4. Conclusions
One of the characteristics of the grassland
ecosystem significantly different from the other
terrestrial ecosystems is that its carbon storage most
concentrated in the soil [20]. Grazing decline the
ability on fixed carbon of grassland vegetation,
promoting soil respiration and accelerating carbon
release from the soil to the atmosphere, which reduce
the soil carbon storage [21]. The experimental results
show that carbon storage of grazing grassland is more
than artificial grassland and fenced mowing grassland
during the vegetation growing season in August in
alpine grassland. The research results are similar to
the results may be due to the following reasons: first,

research grassland ranges are different; second,
research methods are different; third, the sampling
process and determination process has error.
Although China now has many scholars study the
characteristics of grassland carbon storage, but for the
research on carbon storage in alpine grassland is not
enough [23, 24]. Grassland carbon stocks affected by
many factors, in the different geographical regions,
climate conditions and in different seasons, grassland
carbon storage may make different changes; in the
same geographical region, climate conditions and in
the same season, different types of vegetation and
grassland under different land uses can also cause
different soil carbon storage. Therefore, the study of
the characteristics about carbon storage of grassland
under different land uses in alpine grassland is
necessary, it provide data reference for study the
carbon storage of grassland in Qinghai-Tibet Plateau
and it has important significance to establish the
reasonable development policies of animal husbandry
in pastoral areas and ensure the sustainable
development of grassland animal husbandry.
Acknowledgments
This study was supported by Strategic and Pilot
Project of Chinese Academy of Science
(XDA05050404-1-2).
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Pratacultural Science 22 (1) (2005) 4-11. (in Chinese)
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on characteristics of alpine meadow communities in the
eastern Qinghai-Tibetan Plateau, Acta Prataculturae
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carbon and total nitrogen in typical steppe, Pratacultural
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of grassland biomass in china, Acta Phytoecologica
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J. Ni, Carbon storage in grasslands of China, Journal of
Arid Environments 5 (2002) 205-218.

D
DAVID
PUBLISHING
Chemical and Sensory Properties of Olive Oil as

Influenced by Different Sources of Irrigation Water
Salam Ayoub1, Saleh Al-Shdiefat1, Hamzeh Rawashdeh2 and Ibrahim Bashabsheh2
1. Olive Research Department, National Center for Agricultural Research and Extension, P.O. Box 639, Baqa 19381, Jordan
2. Water Management and Environment Research Department, National Center for Agricultural Research and Extension, P.O. Box
639, Baqa 19381, Jordan
Received: October 8, 2012 / Published: February 20, 2013.
Abstract: This research was conducted throughout the years 2008-2010 to study the influence of irrigation water quality on oil
quality of Nabali Muhassan olive trees. Reclaimed municipal wastewater and fresh water were used twice a week using drip
irrigation system. Rainfed olive trees of the same farm were taken as control. No significant differences were observed between
rainfed, fresh water and reclaimed wastewater treatments in terms of acidity, peroxide values and UV absorbance of the extracted
olive oil. Heavy metals and microbiological pathogens were not detected in all tested olive oil samples. Oleic acid was significantly
higher in olive oil obtained from rainfed trees than irrigated trees. Linoleic acid content was significantly higher in reclaimed
wastewater irrigated trees than the rainfed trees. Total polyphenol contents were significantly higher in oil obtained from rainfed
olive trees than oil obtained from the irrigated olive trees. Results of organoleptic analysis showed no significant differences in the
fruity attribute within treatments, while the bitter and pungent attributes were higher in olive oil obtained from rainfed trees as
compared to that obtained from irrigated trees. No negative attributes were observed in oil obtained from the irrigated or rainfed trees
and they were all classified as extra virgin grade.
Key words: Olive oil, irrigation, reclaimed wastewater, fresh water, rainfed trees, total phenols.
1. Introduction
The olive (Olea europaea L.) is one of the most
important trees in the Middle-East region. Olive tree
has been traditionally grown under rainfed conditions
and is considered as one of the best adapted species to
the semi-arid environment [1]. Area under olive
cultivation in Jordan is about 129,000 ha with an
annual production of 222,000 tons of olive fruits [2].
Jordan is facing a drastic water shortage problem
and is classified among the poorest countries with
regard to water availability [3]. Therefore, one of the
strategies to be adopted to alleviate the water shortage
problem in the country is to use treated municipal
wastewater for irrigation purposes. Olive is considered
moderately tolerant to salinity [4-6] and therefore
Corresponding author: Salam Ayoub, Ph.D., research
fields: olive tree physiology and oil quality. E-mail:
sayoub@ncare.gov.jo.
treated wastewater may be a useful option [7].

In a series of studies, it has been demonstrated that
introduction of irrigation to rainfed olive orchards
dramatically increases yields [8, 9]. However, fresh
water in these rainfed areas is scarce and only
low-quality sources of water (saline and reclaimed
wastewater) is available for olive irrigation.
It is well established that the quantity of water used
in irrigating of olive trees affects the chemical and
sensory quality of olive oil [10]. The phenolic
compounds of olive oil are mostly influenced by
irrigation, since their levels decreased as the amount
of applied water increased [11]. Sales et al. [12]
reported that the content of monounsaturated and
polyunsaturated fatty acids was higher in the oils of
the dry-farming olive, while the relative contents of
the saturated fatty acids and phenolic compounds were
higher in the irrigated one.
106
It was reported that treated wastewater available for

irrigation is commonly characterized by high salinity,
excess levels of B and significant but non-uniform
concentrations of both potential plant nutrients (N, P,
K
and
micronutrients)
and
environmental
contaminants including heavy metals, COD and BOD5
[13]. Little information is available regarding the
effect of the reclaimed wastewater on olive oil quality.
Palese et al. [14] reported that the irrigation of olive
trees using treated wastewater during six crop seasons
in Southern Italy did not affect significantly the
quality parameters of the obtained olive oil.
A Tunisian study demonstrated that the use of
treated wastewater increased vegetative growth and
olive yield in comparison to non-irrigation regime
[15]. A study carried out in Southern Italy indicated
that applying treated sewage water for olive trees does
not result in significant microbial contamination of
fruit harvested from the wastewater-irrigated trees [7].
Wiesman et al. [16] found no effect of irrigation
with saline water as compared to fresh water on olive
oil basic quality parameters, such as free acidity,
peroxide value and fatty acids composition. However,
saline water increased the level of certain antioxidant
components (polyphenols and Vitamin E).
Furthermore, Cresti et al. [17] found no effect of
irrigation with saline water on peroxide value and
fatty acids profile, while the oleic-linoleic ratio
decreased in olive oil obtained from trees irrigated
with saline water.
The aim of this research was to study the influence
of using secondary treated wastewater for irrigation of
Nabali Muhassan olive trees on the quality of the
produced olive oil in comparison to irrigation with
fresh water and rainfed conditions.

The experiment was conducted for three successive
seasons 2008, 2009 and 2010 at a private olive
orchard located in Ramtha area in the north of Jordan
(altitude 484 m, latitude 3235N and longitude
3559E), with an average annual rainfall of about 275

mm in a silty clay soil texture.
Fourteen-year-old Nabali Muhassan olive trees
spaced 6 m 6 m were selected for the experiment. A
randomized complete block design with five
replications and three treatments was used. Each
experimental plot consisted of nine trees for each
treatment providing one inner tree for monitoring
and for data collection and surrounded by border trees
receiving the same treatment. Experimental treatments
were carried out to compare between rainfed trees (not
irrigated), irrigation with fresh water and finally
irrigation with reclaimed wastewater.
Reclaimed wastewater was supplied from Ramtha
wastewater secondary treatment plant located near the
experiment orchard. It was pumped from the treatment
plant through a main pipeline to the drip irrigation
system at the olive orchard. Underground well water
was brought by tanker vehicle and reserved in tanks at
the experiment location, then it was pumped through
the drip irrigation system. Rainfed olive trees at the
same orchard were taken as the control treatment.
Quantity of irrigation water was determined
according to the maximum crop evapotranspiration
(ETc), using the FAO method [18, 19] based on
reference crop evapotranspiration (ETo) multiplied by
crop coefficient (Kc) and a plant cover factor
measured as percent surface cover by trees in the
orchard and determined by measuring shaded area at
solar noon. Irrigation water (fresh and reclaimed) was
provided two days per week by drip irrigation method
(seven emitters per tree; 8 L/h). Distances between
emitters were 0.4 m and were equally spaced around
the tree 0.5 m from the main trunk. Irrigation
scheduling and monitoring was controlled manually
through opened valves according to a time schedule.
Irrigation was started on April and finished on
October for each growing season.
Total quantity of fresh water and reclaimed
wastewater applied were similar during the irrigation
period. Based on daily calculation of ETc, the total
quantity of applied water for each type of irrigation

water was 382 mm, 308 mm and 295 mm for the 2008,
2009 and 2010 seasons, respectively. Samples of fresh
water and reclaimed wastewater were taken during the
irrigation period and analyzed for physical, chemical
and biological characteristics.
Olive fruits were harvested by hand during
November of each growing season at a maturity index
around four. Each inner tree from the experimental
plot was harvested separately. Oil extraction was
performed within two days using small scale olive
mill (two phase centrifugal system, Model BuonOlio
Campagnola, Italy).
Oil samples were analyzed for free acidity, peroxide
value, UV absorbance (K270, K232, K), heavy metals,
total polyphenol contents and fatty acid composition
using official methodologies of the International
Standard Organization (ISO) and the International
Olive Council standards [20].
reclaimed wastewater were within the Jordanian

standard for treated wastewater used in irrigation of
fruit trees. However, values of EC, Sodium adsorption
ratio (SAR), Cl and Na were higher than the Jordanian
standard limits (Table 1). Although the EC value of
the reclaimed wastewater was relatively high, olive
trees can tolerate irrigation water salinity of up to 5
dS/m with a SAR of 18 and can produce new growth
at leaf Na levels of 0.4%-0.5% dry weight (d.w.) [23,
24].
Results of oil quality analysis showed no significant
differences among the three treatments throughout the
three seasons in terms of acidity, peroxide values, K270,
K232 and K of the extracted olive oil with the
exception of fresh water treatment for the year 2008
which gave significantly lower free acidity value as
Table 1 Characteristics of water used in irrigation of olive
trees as compared to Jordanian standard for treated
wastewater used for irrigation of fruit trees. Each value is
the mean of six samples standard deviation.
Sensory evaluation of the olive oil samples was

performed by eight trained olive oil tasters from the
accredited Jordanian olive oil tasting panel according
to the official methodology of the International Olive
Council [21]. The mean of each tasting session for
each olive oil sample for the three treatments (two
samples/replicate) was calculated.
Data were analyzed using two-way analysis of
variance (ANOVA) using the statistical package
MSTAT-C [22]. Means were compared using
Duncans multiple range tests. Significance was set at
P 0.05. The means for each sensory olive oil
attribute was also calculated using the IOC statistical
computer program and accordingly the classification
of the oil was determined.

Analysis of irrigation water showed higher electric
conductivity (EC) value for reclaimed wastewater (2.6
dS/m) as compared to fresh water (1.22 dS/m). The
mean values of pH, TSS, cations, anions, N, NO3, B,
heavy metals, BOD5, COD and fecal coliform in
107
Type of irrigation water

Parameter
Fresh water
Reclaimed
wastewater
Maximum limit
(Jordanian
standard, No.
893/2006)*
6.0-9.0
< 2.5
150
9
11.5
8.0
10.0
11.0
30
10.0
70
45
1.0
100
0.01
5
500
200
pH
8.20 0.10 7.97 0.31
EC (dS/m)
1.22 0.11 2.60 0.29
TSS (ppm)
19.67 0.58 47.00 19.92
SAR
2.93 0.42 11.05 3.02
Ca (meq/L)
3.81 0.54 3.43 0.25
Mg (meq/L) 3.01 0.42 3.52 0.16
Na (meq/L)
5.38 0.69 20.59 5.35
Cl (meq/L)
5.92 0.80 18.75 3.31
K (meq/L)
0.28 0.04 1.41 0.12
T-PO4 (ppm) 0.00
0.67 0.75
SO4 (meq/L) 4.91 0.66 6.78 3.03
Na (%)
43.15 4.38 70.41 5.34
T-N (ppm)
28.67 11.55 49.67 20.55
NO3 (ppm)
8.27 5.24 16.73 11.45
B (ppm)
0.24 0.13 0.50 0.27
As (ppb)
< 0.002
< 0.002
Cd (ppm)
< 0.002
< 0.002
Pb (ppm)
< 0.01
< 0.01
COD (ppm)
9.00 1.73 36.67 12.34
BOD5 (ppm) 5.33 1.53 14.20 2.31
Fecal coliform
<2
58.00 15.87 1,000
(100 mL-1)
*Jordanian standard for reclaimed waste water, Ministry of
Water and Irrigation, 893/2006.
108
compared to rainfed and reclaimed wastewater

treatments. The free acidity values for the year 2008
were higher than the years 2009 and 2010. These
results may indicate that the enzymatic lipolysis in the
olive fruits or the attack of parasites was greater in
2008 compared to the years 2009 and 2010. The
peroxide value was significantly higher for the rainfed
treatment for the year 2010 as compared to reclaimed
wastewater treatment (Table 2). The low level of
acidity, peroxide values, K270, K232 and K in this
experiment may be due to the handling process since
the olives were hand-picked and processed within 1-2
days. The K270, K232 and K values measure some of
the oxidation products of fatty acids and reflect the
degree of stability of olive oil [25]. These results
indicated that irrigation of olive trees with reclaimed
wastewater characterized by high water salinity gave
high quality olive oil similar to olive oil obtained from
rainfed or fresh water irrigated trees. Our results agree
with the finding of Palese et al. [14] who reported that
irrigation of olive trees with treated wastewater during
six crop seasons in Southern Italy did not affect
significantly the quality parameters of the obtained
olive oil. Furthermore, Wiesman et al. [16] reported
that olive trees of the cv. Barnea irrigated with
moderately saline water (4.2 dS/m EC) produced oils
of relatively high quality parameters in terms of free

fatty acids, peroxide value and fatty acid profile.
There was no detection of heavy metals in all olive
oil tested samples for the three seasons (Table 2). This
finding is in agreement with Al-Shdiefat et al. [26]
who reported no detection of heavy metals in the
extracted olive oil from trees irrigated with treated
wastewater.
Fatty acid composition values were within the
limits of the International Olive Council standard over
the three seasons (Table 3). Oleic acid content was
significantly higher in oil obtained from rainfed trees
as compared to oil obtained from fresh water irrigated
trees for the three seasons. While linoleic acid content
was significantly higher in oil obtained from fresh
water irrigated trees than rainfed trees for 2008 and
2009 seasons. Stearic acid content was significantly
higher in olive oil obtained from trees irrigated with
reclaimed wastewater or fresh water irrigated trees for
2009 season (Table 3). With respect to 2010 season,
no significant differences were observed among
rainfed, fresh water and reclaimed wastewater
treatments for all measured fatty acids except for oleic
acid and linoleic acid. Oleic acid was significantly
higher in oil obtained from rainfed trees than
that obtained from fresh water or reclaimed wastewater
Table 2 Quality characteristics of olive oil extracted from Nabali Muhassan olive fruits grown under rainfed and
irrigated conditions during the three experimental years (2008, 2009 and 2010).
2008
2009
2010
Fresh Reclaimed
Fresh Reclaimed
Fresh Reclaimed Limit
Rainfed
Rainfed
Rainfed
water wastewater
water wastewater
water wastewater value**
Free acidity as oleic acid (%) 0.60 a* 0.47 b 0.67 a
0.39 a 0.29 a 0.35 a
0.28 a 0.30 a 0.30 a
Max 3.3
Peroxide value (meq O2 Kg-1 oil) 7.18 a 7.20 a 6.00 a
5.54 a 4.18 a 4.16 a
6.24 a 5.62 ab 4.18 b
Max 20
K270
0.15 a 0.15 a 0.15 a
0.10 a 0.11 a 0.14 a
0.10 a 0.10 a 0.12 a
Max 0.22
K232
1.81 a 1.78 a 1.80 a
2.12 a 2.06 a 2.14 a
2.07 a 1.90 a 1.81 a
Max 2.5
K
0.001 a 0.001 a 0.001 a
0.001 a 0.001 a 0.002 a
0.002 a 0.001 a 0.001 a
Max 0.01
Iron (Fe) (mg kg-1)
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1 0.5
< 0.1
< 0.1
< 0.1
Max 3.0
Copper (Cu) (mg kg-1)
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1
< 0.1
Max 0.1
Lead (Pb) (mg kg-1)
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1
< 0.1
Max 0.1
Cademium (Cd) (mg kg-1)
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1 < 0.1
< 0.1
< 0.1
< 0.1
Max 0.1
*Means within rows for each parameter and year having the same letters are not significantly different at 5% probability level
according to Duncans multiple-range test;
**Source: IOC, 2008, trade standard applying to olive oils and olive-pomace oils, International Olive Council, COI/T.15/NC
no.3/Rev. 2, Madrid, Spain.
Parameter
109
Table 3 Fatty acid composition (%) of olive oil extracted from Nabali Muhassan olive fruits grown under rainfed and
2008
2009
2010
Limit
Fresh Reclaimed
Fresh
Reclaimed
Fresh
Reclaimed value**
Rainfed
Rainfed
Rainfed
water wastewater
water
wastewater
water
wastewater
Myristic acid C14:0
0.02 a* 0.02 a 0.02 a
0.02 a
0.02 a 0.02 a
0.02 a 0.02 a 0.02 a
Max 0.05
Palmitic acid C16:0
15.88 b 17.78 a 17.28 ab
15.86 a 16.44 a 15.64 a
16.13 a 15.97 a 17.32 a
7.5-20.0
Palmitoleic acid C16:1
1.18 b
1.44 a 1.32 ab
1.03 a
1.14 a 1.08 a
1.12 a 1.29 a 1.29 a
0.3-3.5
Heptadecanoic acid C17:0 0.04 a
0.04 a 0.04 a
0.04 a
0.04 a 0.04 a
0.05 a 0.05 a 0.05 a
Max 0.3
Heptadecenoic acid C17:1 0.06 a
0.06 a 0.06 a
0.06 a
0.06 a 0.06 a
0.06 a 0.06 a 0.07 a
Max 0.3
Stearic acid C18:0
2.14 a
2.10 a 2.03 b
2.27 b
2.26 b 2.46 a
2.50 a 2.69 a 2.32 a
0.5-5.0
Oleic acid C18:1
64.23 a 60.62 b 62.77 ab
65.87 a 62.94 b 64.43 ab
66.20 a 61.79 b 62.16 b
55.0-83.0
Linoleic acid C18:2
15.01 b 16.50 a 15.06 b
13.70 b 15.81 a 14.71 ab
14.17 b 15.14 ab 16.59 a
3.5-21.0
Linolenic acid C18:3
0.74 a
0.74 a 0.73 a
0.74 a
0.72 a 0.75 a
0.75 a 0.72 a 0.72 a
Max 1
Arachidic acid C20:0
0.34 b
0.36 a 0.36 a
0.42 a
0.41 a 0.40 a
0.40 a 0.44 a 0.40 a
Max 0.6
Gadoleic acid C22:1
0.22 b
0.20 a 0.22 b
0.22 a
0.22 a 0.23 a
0.23 a 0.24 a 0.22 a
Max 0.4
Behenic acid C22:0
0.09 b
0.10 a 0.09 b
0.11 a
0.10 a 0.11 a
0.10 a 0.12 a 0.11 a
Max 0.2
Lignoceric acid C24:0
0.04 b
0.06 a 0.06 a
0.05 a
0.06 a 0.06 a
0.06 a 0.06 a 0.06 a
Max 0.2
according to Duncans multiple-range test;
**Source: IOC, 2008, trade standard applying to olive oils and olive-pomace oils, International Olive Council, COI/T.15/NC
no.3/Rev. 2, Madrid, Spain.
Fatty acids
irrigated trees. Linoleic acid content was significantly

higher in oil obtained from reclaimed wastewater
irrigated trees than that from the rainfed trees (Table
3). These finding agree with Al-Ismail et al. [25] who
reported a decrease in oleic acid and an increase in
linoleic acid content in olive oil samples obtained
from olive trees irrigated with fresh or treated
wastewater when compared to rainfed trees. Our
results indicated that the quality of irrigation water
had no significant effect on fatty acid composition
over the three seasons. Olive oil obtained from trees
irrigated with reclaimed wastewater of high salinity
was not different in fatty acid composition from trees
irrigated with fresh water. This finding agrees with
El-Agaimy et al. [27] who reported that irrigation of
olive trees with saline water up to 6,000 ppm had no
adverse effect on fatty acid composition.
Total polyphenol contents for 2008 and 2009
seasons was higher in olive oil obtained from rainfed
trees than olive oil obtained from trees irrigated with
fresh water or reclaimed wastewater, however, the
difference was not significant. The same trend for
polyphenol contents was observed in the 2010 season
but with significant differences between rainfed

treatment and fresh or reclaimed wastewater
treatments (Table 4). Results of our experiment
indicated that total polyphenol contents were not
affected by the quality of irrigation water. These
results agree with the finding of Al-Ismail et al. [25]
who reported that total phenols decreased in olive oil
samples obtained from trees irrigated with fresh water
or treated wastewater when compared to rainfed
samples, but disagree with their finding that total
phenols content were affected by the quality of
irrigation water, being lower in oil obtained from trees
irrigated with treated wastewater characterized by
high salinity compared with samples obtained from
trees irrigated with fresh water. Furthermore, our
results agree with the finding of Patumi et al. [9] who
reported that, as the amount of water applied to the
olive tree increased, the total polyphenol content in
the oil obtained decreased. This might be due to the
change in the activity of the enzymes responsible for
the phenolics synthesis such as L-phenylanalinine
ammonia type-lyase whose activity is greater under
higher water stress conditions [28].
110
Table 4 Total polyphenol content in olive oil extracted from Nabali Muhassan olive trees grown under rainfed and
Parameter
Total polyphenol contents
(mg L-1 caffeic acid)
Rainfed
213.56 a
2008
Fresh
water
Reclaimed
wastewater
Rainfed
174.41 a
185.20 a
224.91 a
2009
2010
Fresh
Reclaimed
Fresh Reclaimed
Rainfed
water
wastewater
water wastewater
178.17 a 155.24 a
255.99 a 182.5 b 175.23 b
Table 5 Means of positive and negative attributes of olive oil from trees grown under rainfed and irrigated conditions
during the three experimental years (2008, 2009 and 2010).
2008
2009
2010
Reclaimed
Fresh
Reclaimed
Reclaimed
Rainfed Fresh water
Rainfed
Rainfed Fresh water
wastewater
water
wastewater
wastewater
Fruity
4.1 a*
3.9 a
3.7 a
3.0 a
2.8 a
2.6 a
3.2 a
2.9 a
2.8 a
Bitter
2.8 a
1.7 b
1.5 b
1.8 a
1.2 b
1.3 b
1.9 a
1.4 b
1.2 b
Pungent
3.1 a
1.9 b
2.1 b
3.2 a
1.9 b
2.3 b
3.4 a
2.2 b
2.5 b
Fusty
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
Musty
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
Winy
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
Metallic
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
Rancid
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
0.0 a
Olive oil
Extra
Extra
Extra
Extra
Extra
Extra
Extra
Extra
Extra
Grade
virgin
virgin
virgin
virgin
virgin
virgin
virgin
virgin
virgin
according to Duncans multiple-range test.
Parameter
Results of sensory analysis of the olive oil obtained

over the three seasons showed no significant
differences in the fruity attribute, however, the bitter
and pungent attributes were significantly higher in
olive oil obtained from rainfed trees as compared to
olive oil obtained from trees irrigated with fresh or
reclaimed wastewater. This finding can be explained
since the quantity of water in irrigated treatments was
higher than rainfed treatment. In addition, rainfed
trees received water during winter months which
expose the trees to stress conditions during summer
months compared to irrigated trees that receive water
during the whole season. This finding agrees with
Vossen et al. [29] who reported that the intensity
levels of bitterness and pungency declined in olive oil
made from trees receiving more water. The lowest
irrigation levels produced oils that were characterized
by excessive bitterness and high pungency. In addition,
the high bitterness and pungency levels of olive oil
obtained from rainfed trees might be due to high
polyphenol contents reported in these trees. Water
stress has been proven to decrease phenol content in

olive fruits [30]. No negative attributes were observed
in olive oil obtained from fresh water, reclaimed
wastewater irrigated trees or rainfed trees over the
three seasons and they were classified as extra virgin
olive oils (Table 5). These results indicated that
irrigation water quality had no effect on sensory
attributes of the obtained olive oil. The achieved
sensory results for the three treatments were almost
similar to those obtained by Patumi et al. [7] and
Gomez-Rico et al. [8] who reported that olive oil
under irrigated and rainfed conditions were classified
as extra virgin and oil from irrigated regime were less
bitter and less pungent than those obtained from
rainfed trees.
4. Conclusions
Olive oil quality parameters (acidity,
values, K270, K232, and K) were not
significantly among the treatments. Oleic
total polyphenol contents were significantly
peroxide
affected
acid and
higher in
olive oil obtained from rainfed trees than irrigated

trees. Olive oil obtained from rainfed trees or from
trees received fresh or reclaimed wastewater over the
three seasons was free from sensory defects and
accordingly classified as extra virgin olive oil.
The achieved results showed that olive oil produced
from trees irrigated with reclaimed wastewater was
not inferior in terms of chemical and sensory
properties to oil produced from rainfed or fresh water
irrigated trees.
[8]
[9]
[10]
[11]
Acknowledgments
This research was supported by the US Agency for
International Development, Middle East Regional
Cooperation
(MERC)
Program,
Grant
No.
TA-MOU-06-M26-062. The authors wish to thank the
Director General of the National Center for
Agricultural Research and Extension for his support
during the implementation of this study.
[12]
[13]
[14]
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D
DAVID
PUBLISHING
Ethical Trading: The Implications of the Human Rights

Watch Report on South African Fruit Exports
Portia Ndou and Ajuruchukwu Obi
Department of Agricultural Economics and Extension, University of Fort Hare, P. Bag X1314, Alice 5700, South Africa
Abstract: This paper presents an ex-ante assessment of the potential implications of the 2011 Human Rights Watch Report on the
South African fruit industry. The report explicitly mentions the lack of compliance with ethical codes within the fruit industry, the
prevalence of exploitative conditions for farm workers and diverse human rights abuses in farms. The report recommends import ban
on culprits as well as engaging third party auditing to ensure compliance. The high vulnerability of the smallholder farmers justified
the special interest in the implications on their reputation and hence their ability to access profitable export markets. Although
large-scale commercial farmers are potentially at risk of reputational damage due to the Human Rights Watch Report, they are better
able to cope owing to their stronger capital base. The opinions of knowledgeable industry insiders were therefore elicited through
qualitative research that used a five-point Likert scale to assess perceptions about market access. On the basis of the results, it is
probable that the report will lead to improved working conditions for farm workers, improved concern for consumers health, and
enforcement of legislation by the government. The negative implications involve increased competition, possibility of retailers
rationalising their supply base and increased evaluation that ends at the farm gate. There is also high probability of increased
marginalisation of the already disadvantaged smallholder suppliers, and possible increase in costs of auditing and accreditation for
the entire fruit industry. Thus, active collaboration among all stakeholders to ensure the competitiveness of the fruit industry is
inevitable.
Key words: Ethical trading, ethical codes, South Africa, South African fruit exports, Human Rights Watch.
1. Introduction
The Human Rights Watch Report about the
negative ethical conditions in the South African fruit
farms
and
wine
industry
made
strong
recommendations to international retail markets. The
recommendations may have very significant
implications and impact on the nations fruit and wine
market shares in international markets. The
implications can add to existing market access
challenges that include food health and safety
regulations as well as trade-related private standards.
An ex-ante establishment of the stakeholder views and
opinions on the implications of the may result in the
establishment of possible preventive actions and
multi-stakeholder engagements to regain a good
Corresponding author: Portia Ndou, Ph.D., research field:
agricultural economics. E-mail: ndoupoh@yahoo.com.
public image in the export market.

Ethical trade is concerned with the responsibility
taken by companies and retailers especially in Europe,
to introduce codes of conduct to cover minimum
working conditions among their suppliers (agriculture
value chains in this instance-export oriented) in
developing countries [1]. Ethical trade has its origin,
labour practices and the rights of employees of supplier
companies around the world, many of whom are based
in developing countries where laws designed to protect
workers rights are inadequate or not enforced.
However, it is commonly used to refer to other
business practices such as treating customers and
vendors fairly, providing transparency of financial
practices, environmental [2] and more social
responsibility.
The labour and supply chains are challenging in
114
Ethical Trading: The Implications of the Human Rights Watch Report on South African Fruit Exports
nature. The modern global supply chain makes the

aspects more complex and can not be addressed by
individual companies singlehandedly. The Ethical
Trading
Initiative
(ETI)
brings
together
Non-Governmental Organisations (NGOs), trade
unions, and corporations in a unique alliance to
collectively tackle these issues [2]. ETI base codes
were developed to address the exploitation of workers
through such conducts as physical abuse, extreme
form of intimidation and forced, bonded or
involuntary prison labour and reflects the most
relevant international standards on labour practices.
The ETI promotes, improves implementation and
encourages adoption of corporate codes of practice
covering supply chain working conditions. The labour
practice codes, which include wages that are enough
to meet basic needs, working hours that are not
excessive, health and safety and the right to join free
trade unions, have been increasing since their
introduction in the early 1990s [3]. Other issues
included in ETI are the ban on child labour,
non-discriminating hiring, compensation, access to
training, promotion, termination or retirement based
on race, caste, national origin, religion, age, disability,
gender, marital status, sexual orientation, union
membership or political affiliation and avoidance of
harsh and inhumane treatment. The codes are adopted
by companies committed to ethical trade.
The implementation of the ethical codes has
numerous challenges in addition to the complexity of
the global supply chain. Many companies expend
large amounts of money in the monitoring and
auditing of the codes of labour [3]. Cost of
compliance with ethical codes is borne by the
producers and not passed to consumers while the
retailers reluctantly squeeze their margins [4]. Though
the interdependencies created by globalisation have
the potential to generate greater global solidarity, the
rules of the new global economy are only partially
written and are themselves the subject of contention
[5]. There are discrepancies of code implementation
activities and the need for bridging performance

between different companies [6]. Very few
organisations have the capacity to effectively work
with companies on code implementation. Differences
in approaches to ethical trade owing to a growing
number of codes and code initiatives have created
significant confusion and duplication of effort on the
ground. The global production systems manifest
complexities of the commercial networks and the
wider social and institutional environment in which
the codes operate [3]. Child labour is more common in
the less developed countries (LDCs) as a function of
poverty and the values esteemed by developed
countries (DCs) to protect children may be unrealistic.
This is manifest in the setting especially of the small
scale farmers that uses all members of the family to
perform the work in the fields [7]. The increase in the
number of working hours for minors under the age of
18 in the USA from 44 h to 48 h a week may mean the
acceptance of the use of child labour to help family
earnings. It is now left to the discretion of the child to
determine if they are willing and able to work for that
long [7]. Actually, retrenching children in some
instances may worsen their livelihoods.
Integration of the codes into the complex systems
of supply chain management and other internal
systems that are deeply integrated within business
strategies is a big challenge [4]. While the codes are
supposedly considered as acceptable international
norms, they represent to a larger degree the retailer
markets of the developed countries [4]. Thus, the ETI
is thought to save the transnational corporations and
retailer agendas. Since the emphasis on conditions of
labour is more toward the supplier companies, the
retailers themselves are spared from the responsibility
of creating an environment of non-exploitation. The
challenge of unjust trade power between the rich and
poor nations is ignored by the ethical trade issues [4].
In addition to ethical challenges, access to
international markets is currently faced with many
challenges among which are, food safety and health
regulations, private standards, need for traceability,

packaging, carbon footprints, green marketing and the
changes in the agro-business environment.
Compliance with many recommendations set from the
consumer side attracts additional costs for most
suppliers especially from the developing countries.
The farmers in developing countries and especially the
smallholder producers are continually contending with
the international Human Rights Watchdogs regarding
the conditions and practices in farms which, however,
present additional challenges to already existing
barriers to market entry. Commercial producers are
equally affected by the negative publicity by the
Human Rights Watchdogs but they are able to meet
the challenges. Investing in recommended practices
increases costs which their robust capital base can
handle.
The South African fruit industry especially the table
grape and citrus industries are export oriented. For
instance, about 54% of citrus produced in South
Africa is exported [8]. The major destinations for
South African citrus fruit for the past five seasons
were Central Europe (28% of total exports), Middle
East and the Mediterranean (27%), Far East and Asia
(25%), UK (13%) and the Americas (6%) [8]. Ethical
trade became a challenge for all South African citrus
suppliers from 2008/2009 season in response to
pressures from the UK-based retailers [9]. The initial
cost implications were between R15,000 and R20,000
per audit, but have declined to between R6,000 and
R9,000 per audit. Besides the cost associated with the
audit, the self-assessment questionnaire makes use of
too much time, on average 2-8 h. There is also
over-emphasis on the audit rather than on a
continuous improvement approach [9]. The industry
was challenged to create a capacity within fruit SA to
drive the Ethical Trading (ET) issues for its members.
The ET is presently reported to be better suited to the
South African agricultural context and better
organised to promote Ethical Trade principles
throughout the supply chain [9].
115
There was unlikely to anticipate threat related to

ruinous cost that might be caused by Ethical Trading
[4]. The implementation of Base Codes was, however,
linked to the possibility of exacerbating dynamics that
widen the inequality gap in the South African
agriculture which concentrates power in the hands of
those who are already wealthy, have ready access to
capital resources and lines of credit [4]. This reduces
the governments efforts on integrating land reform
with the development of commercial agriculture.
Tight regulation of access to premium markets and
emphasis on the ability to demonstrate compliance
with the host array of standards is very critical.
In June 2001, the South African Human Rights
Commission (SAHRC) [10] conducted an inquiry into
the Human Rights Violation in farming communities
of South Africa. The inquiry focused mainly on the
general widespread lack of compliance with labour
legislation, the vulnerability of women and seasonal
workers on farms, unacceptable levels of crime and
violence experienced in farming communities, limited
access to services and housing among many others.
SAHRC [11] made a follow-up investigation to
establish the progress since 2003. Its general findings
were that there was lack of awareness of human rights,
lack of mechanisms to enforce rights, lack of access to
farms and skewed power dynamics between dwellers
and owners. SAHRC [11] also found out that there
was an increasing tendency for farm workers to live
off farm and be employed via labour brokers,
widespread poor conditions of employment in the
industry as well as abuse of non nationals who are
illegally employed in farms. Few farm workers were
members of trade unions and though child labour
existed in farms, there was limited information on
how prevalent it was.
In August 2011, the Human Rights Watch came up
with a report on ethical situations in South African
fruit and wine industries. The Human Rights Watch
carried out an industry survey that is more inclined
towards compliance with the ethical trading codes.
116
This report is very new and its contents are not yet
fully disseminated among the scientific community
and its implications have not been assessed with
regard to the extent to which it can add to market
access challenges, consumer decision making and
South African fruit flows into the high value markets.
The impact can only be known several years from
now by means of ex-poste studies. It is thus
worthwhile to analyse the report and assess the
opinions and views of the stakeholders with regard to
the implications of the recommendations made by the
report on the demand and supply of the South African
fruit into the high value markets.
The main objective of this study is to establish the
stakeholder views and opinions on the implications of
the Human Rights Watch Report about the South
African fruit industries. Representatives of labour and
government are obviously important this study is
primarily concerned with the impact on producers.
The other stakeholders involved in regulation and
support have a role and do respond to such opinions
and that is taken in a separate study. Specifically, the
study seeks to establish the possible implications and
impact of the report on the extent to which it can add
to market access challenges, general economic
situation of South Africa, rural development, poverty
alleviation, welfare of farm workers, international
consumer decision making and the South African fruit
and wine flows into the high value markets. The study
also seeks possible precautionary measures and
interventions that the industry and other stakeholders
may take to regain a good public image especially in
the export market.
2. Methods
This article focuses specifically on the implications
of the Human Rights Watch Report on the South
African fruit industry ethical codes, the corporate
strategies for the management of labour codes and the
responsibility of the producers, exporters and
government. It is an investigation of the implications of
the recommendations made by the Human Rights

Watch upon the South African fruit and wine exports.
First, the paper sets the scene with regard to the Human
Rights Watch Report and recommendations to different
stakeholders based on the findings of its investigations
of labour relations on South African farms. Second, the
article closely investigates the implications of the
recommendations on the fruit exports to the most high
value international market. Third, the study considers
possible strategies and responsibilities for the South
African fruit producers, exporters, other value chain
players and the government.
The aims of the research required a methodology
that would uncover the perceptions of exporters and
producers on the probable implications of the
recommendations made by the Human Rights Watch
Report on labour conditions in South African fruit
farms. The methodology adopted was therefore
qualitative, utilising the expertise of key informants
by means of questionnaires. This study takes
cognisance of the element of multiple stakeholders
involved in the export of fruit and also in the human
rights issues including the government, labour
representatives (Trade Unions), ETI, farm labourers,
retailers and consumers. However, as a preliminary
study, the study sought the perceptions of two groups
of stakeholders. The target groups included fruit
producers and exporters. Fruit producers were chosen
because the immediate changes in fruit trade flows in
response to the recommendations made by the Human
Rights Watch are better viewed from the production
side and export side. Also, the exporters are the
recipients of the strong recommendations to cut on
sourcing fruit from South African fruit producing
farms. Exporters provide the link and may not source
fruit that they will not be able to sell. A survey was
designed through structured questionnaires to
determine the views of fruit exporters and farmers on
the implications of ethical codes of conduct as
enforced by the Human Rights Watch. The
questionnaire focused on information on the
challenges and benefits of compliance, and the

probable strategies that will promote competitiveness
of the South African fruit industry in the export
market. Of the 80 questionnaires distributed, 36 were
returned. The participants comprised of fruit exporters
(24%), table grape producers (37%) and citrus
producers (39%). The study sought to establish the
threats that may rise as a result of the implementation
of the Human Rights Watch Report for the South
African fruit and wine industries. It also sought to
establish strategies to curb the likely losses of
consumer faithfulness, perception and trust.
Respondents were asked to evaluate the implications
of the report on predetermined possible benefits,
conditions and circumstances associated with
compliance. Each aspect was measured using a
five-point Likert scale anchored by one for strongly
decreasing to five for strongly increasing. The
closer to 5 the index is, the stronger the impact and
vice versa for the lower index. The respondents also
provided the possible strategies that the stakeholders
can put into action for the sustainability of the South
African fruit and wine exports.

The results begin with a presentation of the results
of the document analysis with respect to the Human
Rights Watch Report. The opinion survey of
knowledgeable insiders is then presented. The
researchers are aware that the actual impact can only
be assessed ex-poste, but this ex-ante assessment
saves as a basis for identifying the key issues to focus
on in more systematic study.
Farm workers in the South African fruit and wine
industries have been found to be subject to
exploitative conditions and human rights abuses
without sufficient protection of their rights and thus
benefit very little from the success of countrys
valuable fruit and wine industries [12]. The Human
Rights Watch Report for ethical conditions in South
African fruit farms highlights very sensitive issues
117
such as the lack of hand washing facilities, toilets and

lack of access to drinking water and descent housing.
Difficulty for seasonal farm workers to demand wages
and benefits owing to lack of contracts that stipulate
work conditions have been cited as prevalent cases.
According to the report, the lack of government to
promote health and farm worker rights in the Western
Cape Province in fact represents better conditions
compared to other provinces. The violation of the
human rights and lack of compliance with ethical
codes has been cited as against the South African
constitution and other international covenants which
South Africa has ratified [12]. These findings have a
strong bearing on any claims by the fruit and wine
industries to deliver. This is particularly important
because the health issues associated with international
trade prompted the need for such issues as ethical
trade, packaging and traceability among others.
The labour inspectors failure to comply with health
and safety regulations contravenes ethical trade as the
best practices in agriculture are not promoted [12].
The report may have far-reaching implications
especially considering the recommendations to
international consumers which include the inquiry into
the human rights and labour rights of conditions on
farms that grow the products they purchase and the
need to push retailers to only purchase from farms
with working conditions that meet international
standards [12]. Another forceful recommendation to
international consumers is the need to ask for ethical
trading initiatives including strong assurance measures,
as well as independent third-party audits down the
supply chain, so that consumers can be confident that
the ethical trade products they purchase are in fact
made without the exploitation of workers. The report
advises consumers to inquire into the human rights
and labour rights conditions on farms that grow the
products they purchase.
The retailers are urged to put pressure on suppliers
to comply with the law and to improve labour, health,
and housing conditions [12]. It is of great interest that
118
powers are given to the retailers who already have an

unfair dominance in the supply chain, overbearing
influence in the sourcing, setting of private standards
that are difficult to harmonise as well as having
powers to extract favourable pricing terms and
switching suppliers [13], and yet these are not touched
by the ETI Base Code. Also, the retailers that adhere
to the ETI Base Code are urged to ensure that the
standards contained therein, including the freedom of
association, are respected on supplying farms. They
also should establish a safe and transparent complaints
mechanism that allows workers to launch queries,
including intimidation or discrimination based on
union membership. The report advises retailers to
ensure that, following any third-party audits, all
recommendations are followed. In fact, the title of the
report itself is forceful to any reader: Ripe with
abuse! Considering the sensitivity of the report
coupled with the power of the consumers and retailers
in todays global trade environment, serious
consequences may befall the industry if stern
measures are not put in place. Serious losses might be
incurred especially considering the share of the
produce destined for the export market.
While it is good to advocate for the welfare of the
farm workers, the publicity of the lack of compliance
for the South African producers may lead to much
scrutiny and challenges by the importers of South
African fruit. There is evidently bound to be an
increased pressure for the South African fruit
exporters to compete with the heavily subsidised
Northern rivals. The report instils a sense of insecurity
among the South African suppliers. The uncertainties
surrounding the extent to which retailers may treat the
South African suppliers have a bearing on the future
efficiencies to meet the Base Codes and also for the
retention of suppliers within the retailers supply base.
The complexity of monitoring standards, the problem
of traceability coupled with ethical trade creates a
complex chain. The responsibility itself is more
complex. The involvement of retailers in the
monitoring process may make the whole process not

only complex but difficult.
The Human Rights Watch Report has extensive
implications for the South African fruit industry,
economy and policy. The report has the potential of
impacting both negative and positive outcomes. On a
short-term basis, three significant supply risks can be
identified, namely, immediate withdrawal by the
importers, extreme cuts on labour force and increased
scrutiny by the international retailers. On the positive
side, the awareness of non-compliance can raise a
possibility for public policy support as well as
government financial intervention to the aid of the
producers. This is possible only through the
realisation of the fact that the producers already are
faced with high transaction costs especially relating to
transport to export markets [14]. This is crucial
considering the fact that the Northern hemisphere
rivals are heavily subsidised by their governments.
3.1 Implications of the Report on the South African
Fruit Industry
Tables 1 and 2 sum up the possible implications of
the Human Rights Watch Report if the
recommendations put across will be implemented by
the relevant authorities.
The compliance with ethical codes of conduct has
the potential to beset obstacles to entry into high value
fruit markets. This will have negative long term
consequences on rural development, economic growth
and the agrarian reform. The export driven
horticulture has had significant contributions towards
poverty alleviation, export earnings, employment and
growth as well as gross domestic product [15].
Compliance with ethical codes of conduct can
especially promote the marginalisation of the
resource-poor farmers. The failure by the
resource-poor producers to demonstrate compliance
with base codes can further widen the inequality gaps
associated with South African agriculture which
has seen wealth distribution, access to credit, capital
Table 1
119
The positive implications of the Human Rights Watch recommendations on South African fruit.
Positive implications
Score
Ethical trade can also help to raise support for public policies
3.9
Increased human resource capacity
3.5
Networking to develop ethical codes of conduct
3.7
Environmental responsibility
4.8
Ethical premium
3.2
Voluntary codes can encourage ministries of labour to carry out their proper role
3.6
Savings through increased efficiency
2.3
Improved market access
3.6
Voluntary codes and monitoring systems can potentially be used to reinforce existing legislation and encourage governments
4.2
to enforce the legislation
Price competitiveness in international markets
1.8
Long-term co-operative and transparent relationships
3.7
Descent wages and fair working conditions
4.3
Improves government services in farms
3.8
Respect for workers rights
4.7
Increased and urgent need for basic competence training for farm workers and for smallholder farmers and their extension
4.8
officers
Concern for health and fairness to consumers
3.9
Better working conditions for employees
4.2
Investing in labour saving technology
2.6
Codes standardise practices and assure consumers of the highest quality, safe and hygienic produce
3.1
Producers gain credibility with the European markets
3.7
Compliance with codes becomes very critical determinant for competitiveness in the export markets
4.5
Voluntary codes and monitoring systems impact on local labour legislation and its enforcement
3.6
1 = strongly decrease; 5 = strongly increase.
resources and access to high value agriculture being

prevalent in the white farming community [4]. The
procedures associated with adherence to the codes and
their implementation by the main export markets may
serve as a barrier to the smallholder market
participation. The problem of limited information
transfer associated with smallholder farmers may
hamper the implementation of the ethical codes in the
smallholder production units. The riskiness of work
and potentially unsustainable livelihoods are some of
the likely consequences of implementing the
recommendations set by the Human Rights Watch
despite the possible positive impact for workers that
this development might bring along.
The enforcement of the labour laws that might have
remained dormant for some time may be ensured
through such interventions and through the
enforcement of ethical codes of conduct. The
economic significance of the fruit industry may

encourage the government to enforce not only the
national labour laws but also promote the compliance
with international labour standards. Standardisation of
practices and the assurance of consumers of the
highest quality, safety and hygienic produce can be
facilitated through codes of conduct. All reasonable
due diligence to protect consumers, workers and the
environment will be adhered to and the ethical codes
will become a very critical aspect to remain
competitive in the export markets. Compliance with
the standards will gain the producers credibility with
the international markets.
Negatively, codes of conduct can be used to
substitute and destabilise collective bargaining. The
producers are forced to strive for compliance and
competitive advantage in the international market.
They strive also for the improvement and maintenance
120
Table 2
The negative implications of the Human Rights Watch recommendations on South African fruit.
Negative implications
Score
Increased impact evaluation which generally stops at the farm gate and does not reach far enough down the supply chain
4.3
Trade flows
2.7
Revenues
2.7
Increased prices to consumers
3.2
Elimination of small producers
3.9
Increased pressure in competing with heavily subsidized northern hemisphere rivals
4.8
Marginalisation of the already disadvantaged producers and marketing chains
3.7
Increased independent monitoring and evaluation
3.6
Increasing costs of auditing and certification for compliance with the code of practice
3.8
Impact on rural livelihoods
3.5
The code of conduct has a potential to result in many suppliers being cut by retailers
3.2
Lack of capacity among the South African exporters to engage at a technical, strategic and governance level
2.5
Limited amount of information transfer in smallholder production units may hamper the implementation of ethical codes of
2.4
conduct
Procedure imposed on exporters by their main markets may serve as a barrier to the inclusion of the smallholders
3.9
Unfair advantage given to retailers to rationalise their supply base in view of the greater efficiencies to be gained through
4.2
monitoring only a small number of larger suppliers
Contracts cancelled due to boycotts
2.3
Voluntary processes used to undermine public regulation
1.7
Increased need for more coordination in order to meet the requirements
3.6
Increased complexity of monitoring standards against the Base Code in the context of the kinds of vast networks of
3.6
subcontracting relationships
Relocation of multinational exporting companies to other countries if improved conditions lead to cost increases locally
2.3
Exporters assume the role of setting up strict supervisory system and assuming responsibility for rigid enforcement of
3.7
standards
Codes used to replace and thereby undermine collective bargaining
4.2
Increased complexity of traceability as the retailers in the export market have to lay down specifications and details of the
3.3
traded product at the point of production
Failure of membership of the ETI and acceptance of Base Code to resolve supplier insecurity
3.2
The demands for ethical trade may strengthen the economic power of multinational corporations thus squeezing out medium
3.1
and small sized enterprises
1 = strongly decrease; 5 =strongly increase.
of their market share, sustained loyalty and good

reputation. Thus, the stringent market-side demands
encourage individual effort and solitary positioning in
the over-supplied global market. There also might be
shifts, changes and relocations by export-linked
companies if the improved conditions bear cost
implications locally.
The socio-economic situations of different nations
present a challenge for the consideration of smooth
operation of generic code of conduct. The
implementation of codes may be situational biased.
For instance, South Africa has undergone rapid
socio-economic and legal changes since 1994 [1].
Applying the codes of conduct uniformly across
borders may fail to take into account the inherent

socio-economic situations that may hinder the
implementation under certain circumstances. The
encouragement of retailers to purchase only from
compliant suppliers may lead to discrimination in
sourcing which in turn impact on the returns for the
affected group of producers. This is imperative
realising that the South African fruit produce is export
oriented. This will also impact on the South African
economy since the fruit industry is a high foreign
currency earner with a significant contribution
towards the GDP.
The South African agricultural industry is dynamic,
composed of a diverse array of producers ranging
from the resource-poor smallholders to the

well-established large-scale producers. The demands
of ethical trade may be the most easily and profitably
met by large-scale producers. Compliance and
implementation of the ethical trade strengthens the
economic power of the well-established commercial
farmers while squeezing out the resource-poor
smallholder producers. This may seriously undermine
the efforts by the government of South Africa to
ensure equity and fair distribution of wealth and land.
The smallholders as well as the medium sized
producers lack capacity to engage at a technical,
strategic and governance levels.
3.2 Strategies for the Competitiveness of the South
African Fruit Industry
Fig. 1 summarises the institutions or stakeholders
that should be actively involved in the implementation
of the ethical codes of conduct as well as ensuring the
compliance with basic codes by the South African
fruit producers. These key stakeholders include the
121
ETI, fruit exporters, trade unions, the South African

government and fruit producers. The responsibilities
of each are further stated in Table 3.
Fruit exporters, the ETI, trade unions, the South
African government, the international retailers and the
fruit producers need to actively collaborate in ensuring
the competitiveness of the South African fruit industry
in the export market without compromising the labour
conditions. External intervention is especially
important because the implementation of ethical codes
has cost implications which the producers can not
manage single-handedly without aid and support in
the face of the pressures of prices. Leaving the
responsibility of complying with ethical codes to the
producers will severely squeeze out their profit
margins in the face of an oversupplied international
market where prices are dictated by forces of demand
and supply. Table 3 lists some of the strategies that
these bodies can engage in to maintain or improve
compliance with ethical codes and thus improve the
competitiveness of the fruit exports.
Fig. 1 Supporting stakeholders to sustain export competitiveness (own recommendations based on results and approach
used for this study).
122
Table 3
Strategies towards a sustainable competitive position.
Authority
Responsibility
Fruit
exporting Provision of information
companies
Assist producers with support in implementing the codes
Capacity building
Consolidation of international codes of conduct
ETI
Provide incentives for those suppliers who comply with codes
Establishing efficient systems of conformity and enforcement assessment
Trade unions
Educating farm workers on ethical codes
Promotion of skills development
Strengthening the legislations on: basic employment conditions, employment insurance, labour relations and
the right to strike action, ensuring adequate health and safety condition on farms
Capacity building
The South African
Financial support e.g. in the form of soft loans
government
Provide incentives for those suppliers who comply with codes
Enforce compliance with ethical codes and labour laws
Enforcing the provision of descent housing to workers
Ensure protection of immigrant workers
Engagement of independent accreditation bodies
Fruit producers
Engage in self-assessment of compliance with codes
Support producer compliance with codes
Improved communication with suppliers
Retailers
Explaining codes to suppliers and workers
Active in monitoring of audits rather than rely on third parties
The government needs to introduce, implement and

adopt legislative measures to ensure compliance with
labour laws and access to adequate and descent
housing which are already entrenched in the South
African Bill of Rights. The government should
provide financial support for industrial level
investments that are prerequisites to effective and
demonstrable implementation of ethical trade codes.
Gaps and variations leading to greater implication
costs should be eliminated through the establishment
of efficient systems of conformity and enforcement
assessment of the ethical codes. Centralised systems
of implementation and assessment and compliance
with codes of conduct can help in the closing of gaps,
overlaps and any inefficiency associated with a
decentralised system.
While the Human Rights Watch Report rules out
in-house audits through the suggestion of engaging
third party audits, this study still recommends the
incorporation of self-assessment. Despite the lack of
the generation of objective results attributed to
in-house audits, it, however, lessens the problem of
social accounting to the organisational complexity
associated with the global supply chain [13]. Third
party audits attract additional costs if these audit costs

are borne by the producers. The engagement of
independent accreditation bodies, however, can
eliminate bias and ensure fairness in assessment.
4. Conclusions
By way of conclusion, the implications of lack of
compliance with the ethical codes of conduct may ruin
the industrys export sector. There is need for urgent
action to address the perceptions of exporters and
consumers, redress the bad image that may emanate
from the report on the South African farm labour
conditions and equipping exporters for compliance.
On the other hand, strategies for a sustainable
competitive advantage, compliance with the ethical
trading codes and government intervention are
urgently needful.
Recognition is not made for any possible options
available to existing suppliers who may be facing an
initial failure to adhere to the ethical codes. Immediate
removal from the supply base seems to be the only
option available to the retailers as recommended by
the Human Rights Watch. The main challenge is that
it is easier to retain a market than to regain it
especially in the over-supplied and highly competitive

global market which is also characterised by supplier
integrity, consistency and loyalty. The report should
have left the decision to the retailers on how they will
implement the codes to their South African suppliers.
This
study,
however,
does
not
condone
non-compliance but weighs the balance of possible
available options, especially to long-established
relationships.
The terms of trade between retailers and suppliers
are themselves not explicitly incorporated into the
code [13]. It is ambiguous to have the report
proposing trading terms on a trading platform that still
gives additional privileges to the powerful over the
weak rather than moderating the situation by
suggesting means that will enable continued
relationships, while working on possibilities of
complying with the minimum standards. The honours
should have been given to the supermarkets to
establish whether to abruptly stop purchases from
South Africa or to continue while helping the exporter
step-up their compliance levels. It is apparently clear
that the lack of compliance with the ETI Base Code
will be borne by the producers and not passed on to
consumers, and such partial recommendation that
ignores the organisational dynamics of the supply
chain relationships in turn poses some significant
organisational challenges for the South African export
producers and retailers. This is contrary to the ETIs
unique alliance to collectively bring together NGOs
and exporters to tackle ethical issues.
While the producers will be faced with difficulties
of offsetting their production costs, the international
retailers will be reluctantly squeezing their margins.
This distorts the whole purpose of fair trade through
the promotion of unjust power relations between rich
consumer countries and poor supplier countries.
Corporate responsibility is evaded while retailers
distance themselves from their own role in creating an
environment for exploitation [4]. Retailer control and
power can dampen competition and at the same time
123
raise barriers to entry through such instances as

resorting to dealing with a limited number of highly
favoured and reliable suppliers, i.e., exclusive supply.
There is bound to be a one-sided determination of
terms of trade. This worsens the traditional
characteristics of the perishable products producers
who are often small, unconcentrated and
uncoordinated relative to the buyers. Their bargaining
power is thus further reduced by the price inelasticity
of their highly perishable products. Viability and
efficiency can be seriously affected, leaving
detrimental effects upon the fruit sector.
The growth in the market-side requirements and the
need for ethical code compliance may act as a means
to indirectly regulate the activity of the exporters in
the global market. Imposing retailer control as a
means to serve the interests of the producers in the
developing countries for the purposes of
implementation and auditing of the codes are contrary
to the ethical trade. In a highly competitive global
market, ethical trade proves to be another compliance
cost and disguised market entry barrier. The benefits
of fair trade are thus not well-distributed among
players when producers are not only faced with
stringent food safety standards but also the retailers
are directed on how to treat the non-complying. The
rise in costs for the producers may negatively affect
the revenues as oversupplied markets may turn out not
to favour price competitiveness. If the audit costs are
borne by the producers, then the retailers will be
enjoying over dominance in the trade. This justifies
[16] attribution of the strategies, codes and auditing
methods to agents in the developed world.
This study makes several recommendations based
on the findings. In addition to ensuring information
transfer especially to the smallholders, there is need
for coordination and setting up of a strict supervisory
system by both the producers, exporters and the
government. Nationwide compliance with the ethical
trade codes should be ensured with the exporters
assuming responsibility for the setting of standards.
124
There is need for education through education on

ethical codes especially for the smallholders. There is
need for an urgent unified industry approach to the
adoption of codes of practice to defend the industry in
the face of negative publicity. Continual
familiarisation with the Code of Practice is needful to
curb any non-compliance with some clauses that may
be subject to updates and revisions.
Watch Report.
References
[1]
[2]
Contracting producers and suppliers by retailers

may help eliminate insecurity and uncertainty. This
may also be an opportunity for the inclusion of
[3]
smallholders in main supply chains. Contracting the

producers may also ensure continued production and
supply to the international market while continually
[4]
working towards adhering to the ethical codes of

conduct. This may in turn secure the employment of
the labourers in these farms which may otherwise be
[5]
lost when the producers get out of business and are

unable to meet salary and wage payouts.
Interventions should ensure other socio-economic
issues such as rural development, poverty alleviation
[6]
and the general housing conditions in the nation as a

whole. Otherwise, the situation may not yield
improved results if not holistically addressed. The
[7]
possibility of resorting to machinery may be an

available option to producers and exporters, which
may worsen the welfare of the dismissed farm workers
and thus adding on to the poverty problem.
Since this study did not exhaust all the key and
important stakeholders involved in the South African
fruit and wine industries, a recommendation is made
for a ex post evaluation of the impact of the report
which will include the opinions of all stakeholders.
The ex-post study is worthwhile for establishing the
impact of the report on real product flows across
borders and the changes in market shares in certain
export markets of choice after several seasons of fruits
exports post the publishing of the report. Thus this
study has set a stage with regards to the most
important issues to evaluate when carrying out an
ex-post study of the implications of the Human Rights
[8]
[9]
[10]
[11]
[12]
[13]
S. Barrientos, S. McCleneghan, L. Orton, Ethical trade

and South African deciduous fruit exportAddressing
gender sensitivity, The European Journal of Development
Research 12 (1) (2000) 140-158.
L.F. Golodner, Understanding the difference: Ethical trade,
fair trade, sustainable consumption, social responsibility,
National Consumers League, ANSI, USA COPOLCO
Workshop, May 23, 2007, Salvador de Bahia, Brazil, 2007,
pp. 13-23.
S. Barrientos, S. Smith, Do workers benefit from ethical
trade? Assessing codes of labour practice in global
production systems, Third World Quarterly 28 (4) (2007)
713-729.
A. Du Toit, Ethical tradingA force for improvement, or
corporate whitewash? Natural Resources perspectives
Number 71, Oct. 2001, ODI Programmes for Land and
Agrarian Studies (PLAAS), 2001, pp. 1-4.
Ethical issues in food and agriculture [Online], Food and
Agriculture Organization of the United Nations Rome,
2001,
http://www.fao.org/DOCREP/003/X9601E/X9601E00.H
TM.
Ethical Trading Initiative, Key challenges in ethical trade.
Report on the ETI Biennial Conference 2003, Ethical
Trading Initiative, Cromwell House, London UK, 2003, p.
10.
A. Rushton, Child labour is the global recession
squeezing childhood [Online], Aug. 8, 2011,
http://www.ethicaltrade.org/news-and-events/blog/abi-rus
hton/chld-labour-is-the-global-recession-squeezing-child
hood%3F.
Citrus
Growers
Association
[Online],
2008,
www.cga.co.za/site/awdep.asp?depnum=4289.
CGA 2009 Annual Report, Citrus Growers Association of
Southern Africa [Online], 2009, www.cga.co.za.
The South African Human Right Commission (SAHRC),
Final Report on the Inquiry into Human Rights Violations
in Farming Communities, SAHRC, 2003, pp. 1-56.
The South African Human Rights Commission (SAHRC),
Progress made in terms of Land Tenure Security, Safety
and Labour relations in the Farming Communities since
2003, SAHRC, 2008, pp. 14-46.
Human Rights Watch, Ripe with abuse: Human rights
conditions in South Africas Fruit Wine Industries,
Human Rights Watch, New York, USA, 2011, pp. 1-103.
A. Hughes, Multi-stakeholder approaches to ethical trade:
Towards a reorganisation of the UK retailers global
supply? Journal of Economic Geography 1 (2001)
421-437.
[14] P. Ndou, A. Obi, The business environment and
international competitiveness of the South African citrus
industry, Paper presented at the International Food and
Agribusiness Association (IFAMA)s 21st World Annual
Forum and Symposium, June 20-23, 2011, Frankfurt,
Germany, 2011, p. 15.
125
[15] K. Ellies, J. Keane, A review of ethical standards and

labels: Is there a gap in the market for a new Good for
Development label, Overseas Development Institute
(ODI), 11 Westiminster Bridge Road, London, UK.
Working paper 297 (2008) 1-35.
[16] M. Blowfiled, Ethical trade: A review of developments
and issues, Third World Quarterly 20 (1999) 753-770.

D
DAVID
PUBLISHING
Physiological and Phytosanitary Potentials of Coriander

and Radish Seeds
Jucilayne Fernandes Vieira, Francisco Amaral Villela, Orlando Antonio Lucca Filho and Raifer Simes Campelo
Department of Plant Science-Sector of Science and Seed Technology, Federal University of Pelotas (UFPel), Pelotas, RS 96001-970,
Brazil
Abstract: Information on seed vigor of the vegetable crops is always important and necessary due to the increasing demand of high
quality seeds for sowing and high-value commercial. The cultivation of these species, conducted intensively, should be established
with seed high potential physiological and health for the development of a more productive and sustainable agriculture. The present
study was conducted to evaluate the potential physiological of seed lots of radish and coriander. The experiment was conducted at the
laboratory of seed analysis and greenhouse of the plant science department of the Federal University of Pelotas in South Brazil. Four
radish seed lots, cultivar Saxa, and four coriander seed lots, cultivar Verdo were used. Germination seed test, first count of
germination, accelerated aging test, electrical conductivity and seedling emergence were used to evaluate the physiological quality of
the lots. The experimental design was completely randomized design with five replications. Means were compared by Tukey test.
The accelerated aging test was the most efficient test in assessing the physiological quality for both lots of radish seeds and coriander
and this test provide coherent results with seedlings emergence.
Key words: Radish (Raphanus sativus L.), coriander (Coriandrum sativum L.), vegetable crops, seed quality, seed vigor.
1. Introduction
Vegetable seeds have a high commercial value, thus
a lot of information about seed vigor are always
important. The cultivation of these species, conducted
intensively, should be established with seed of high
physiological potential and seed health [1].
Researches talking about vegetable seeds have shown
significant development in Brazil since the beginning
of the 1990s, but papers are still less frequent than
those conducted with species of other crops. It is true
that many vegetable crops are grown, but each one has
its own peculiarity, and the volume of available
knowledge is not enough to the importance of this
subject [2]. There are increasing demands on high
quality seeds for the establishment of more productive
and sustainable agriculture, and it is growing also the
Corresponding author: Jucilayne Fernandes Vieira, Ph.D.,

research fields: seed science and technology. E-mail:
laynevieira@yahoo.com.br.
importance of monitoring each stage of the seed

production process of the seed industry [3].
The radish crop (Raphanus sativus L.) is an annual
vegetable crop grown mainly on small farming and in
areas of green belts of the country [4]. It has shorter
cycle among vegetables; and because of this, radish
becomes an excellent choice for the small producer [5].
It has considerable economic importance, but it is still
produced and consumed in volumes less significant in
the country, though, the seeds have great commercial
value, which reinforces the importance of assessing
seed physiological quality. Coriander (Coriandrum
sativum L.) is a crop consumed in various regions of
Brazil, especially in the North and Northeast.
Vegetable widely consumed in Brazil as a condiment
featuring great socio-economic importance. However,
problems related to low seed vigor and crop
establishment are reported in this specie [6].
There are only few studies about the use of vigor
test to estimate the seeds physiological potential of
these species [2, 6, 7]. In this context, the objective of

this study was to evaluate the seed physiological
quality of lots of radish and coriander.

2.1 Materials
The work was conducted at the Seed Analysis
Laboratory and in the greenhouse of the Federal
University of Pelotas. Seeds from four lots of radish
seeds cultivar Saxa and from four lots of coriander
seeds cultivar Verdo were subjected to following
tests and assessments described below.
127
mL plastic cups containing 50 mL of deionized water.

The readings were taken at 2 h, 4 h, 6 h, 8 h and 24 h.
The conductivity value provided by the device was
expressed in mS cm-1 g-1.
2.6 Accelerated Aging with NaCl (Sodium chloride)
Solution
A total of 200 seeds (four replicates of 50 seeds) per
seed lot were distributed over an aluminum screen
placed inside a plastic box (gerbox) containing 40 mL
of NaCl solution, maintained at 41 C for 48 h. This
solution was prepared by adding 40 g of NaCl in 100
mL of water, establishing an environment with relative
2.2 Determination of Moisture Content

The experiment was conducted in an oven with
forced air circulation at 105 3 C for 24 h, in
accordance to the Rules for Seed Analysis RAS [8],
using two samples of 4 g seed for each lot. Results were
expressed as mean percentage weight per lot. The
moisture content of the seed was assessed before and
after the accelerated aging test.
humidity of 76%, according to the procedure proposed

by ref. [9]. After this period, the seeds were put to
germinate following the methodology used in the
germination test described above. The percentage of
normal seedlings was assessed on the fourth (radish)
and seventh (coriander) days after sowing.
2.7 Seedling Emergence in Substrate
The sowing of each lot was manual, in beds, in a
2.3 Germination Test

The experiment was conducted at 20 C with 200
seeds per seed lot (four replicates of 50 seeds). The seeds
were distributed on two sheets of paper, previously
moistened with distilled water in an amount equivalent
to 2.5 times the mass of paper. Evaluations were
conducted for radish at 4 and 10 days and for coriander
at 7th day and 21th day after sowing and the results
expressed in percentage of normal seedlings [8].
2.4 First Count of Germination
The experiment was made in conjunction with the
germination test, by computing the percentage of
normal seedlings on the fourth day after sowing for
radish and seventh day for coriander [8].
2.5 Electrical Conductivity
The experiment was assessed with four replications
of 50 seeds per seed lot. The seeds were soaked in 200
greenhouse, with four replicates of 100 seeds in rows

2.0 m long with spacing of 0.20 m at a depth of 0.01
m to coriander seeds. For the radish seeds were sown
four replications of 50 seeds in furrows spaced at 0.20
m and 0.01 m depth. Evaluations were performed at
12 days after sowing for radish seeds and 30 days for
coriander seeds, computing normal seedlings emerged
and the results expressed as a percentage.
2.8 Statistics
The data expressed in percentages were subjected to
normality tests that indicated they did not need any
transformation. The experimental design was
completely randomized layout with five repetitions. In
the statistical procedure, analysis of variance was
performed separately for each test and the means of the
lots were compared by Tukey test at 5% probability.
The conductivity test was analyzed in a factorial of 4
lots 5 seed imbibition periods.
128

The water content of the seeds was similar between
lots, which allow obtaining more consistent results.
The germination rate of all radish seed lots used was
above 95%, which is above the minimum standard
required for commercialization of most seed vegetable
crops (80%). These germination values indicate high
physiological seed quality and seedling emergence in
the greenhouse ranged from 80% to 88%, depending
on the lot (Table 1). Radish seed lots did not differ on
the percentage of germination (Table 1), showing low
sensitivity of germination tests performed at the
laboratory, to identify differences in performance of
the seeds, often highlighted in the literature, justifying
the use of vigor testing (Table 1). Using the laboratory
germination test was also not possible to differentiate
seed lots of lentil [10] and wheat [11], because the
germination test is conducted under favorable
conditions, maximizing the potential of physiological
seed lots.
However, the first count of germination test
classified the seed lots into two different vigor levels
(low and high). The data obtained by the author [12] in
the first count of germination test in tomato seeds
permitted the separation of the lots in two vigor levels,
showing similar results as they were observed in this
work (Table 1).
The accelerated aging test with saturated solution of
NaCl detected vigor differences between radish seeds
lots, and a similar response was also observed in
seedling emergence (Table 1). These results showed
consistency with those obtained in the seedling
emergence. The seed water content after aging period
showed maximum change of 0.4% points among seed
lots, not interfering, a priori in the results.
Among the available tests, accelerated aging is one
of the most studied and recommended for various crops.
Initially developed for the purpose of estimating the
longevity of stored seeds, it has been extensively
studied with a view to its standardization [13] and its
ability to provide information with high consistency
Table 1 Initial moisture content of the lots (IMC),

moisture content after accelerated aging (MCA),
percentages for germination test (G), first count of
germination (FCG), accelerated aging with NaCl-76% (AA),
emergence of seedlings in substrate (ESS) of the different
lots of radish seeds.
Lots
IMC(%) MCA(%) G(%) FCG(%) AA(%) ESS(%)
1
8.8
10.7
95 a 88 b
83 ab 84 ab
2
8.9
10.9
95 a 87 b
86 a
88 a
3
8.6
10.5
97 a 93 a
81 b
80 b
4
8.3
10.7
96 a 94 a
83 ab 86 ab
CV (%) 3.22 5.25
4.93
5.31
Means followed by the same letter in a column do not differ in
level of 5% probability by the Tukey test.
degree [1]. This method was also efficient for

evaluating seed vigor of different vegetable seeds such
as carrot [13], cucumber [14], tomato [15], melon [16]
and rocket [17]. For the electrical conductivity (Table
2), it was found that there was no significant
interaction between the periods of soaking and lots of
radish seeds, although this test detected differences in
vigor levels into four lots, with superiority for lots 1
and 2 and inferiority for lot 3. With increasing
imbibition time was greater leaching of exudates from
seeds.
However, for black oat seeds, the electrical
conductivity test at the different conditions detected
difference based on seed vigor levels. Furthermore,
there were significant effects only on main factors:
lots of seeds, number of seeds and imbibing periods
[17].
Among vegetable seeds, coriander has a national
standard germination of (60%) and it is considered
low for seed commercialization. Lots tested in this
present work, seed lot germination exceeding the
minimum value and there was a variation between lots
from 71% to 81% (Table 3). In the germination test
was observed that lot 2 showed lower seed
germination than other seed Lots (Table 3). This result
as also reported [18], they found that the germination
test was not sensitive enough to distinguish
differences physiological quality of onion seed lots,
being only able to show the inferiority of one of the
tested lots.

Table 2 Electrical conductivity test with soaking 50 seeds
in 50 mL of distilled water for periods of 2, 4, 6, 8 and 24 h
from four radish seed lots.
Periods of imbibing (h)
2
4
6
8
24
Mdia
1
32.90 42.30 46.80 51.40 62.40 47.44 B
2
32.00 41.00 43.70 47.60 58.70 44.60 A
3
36.80 46.10 51.60 57.40 65.40 51.46 C
4
34.50 43.60 47.20 52.10 64.80 48.44 B
Mdia 34.05 a 43.23 b 47.32 c 52.12 d 62.82 e
CV (%) 9.00
Means followed by same letter in lower case in the line and
capital letters on the column do not differ by Tukey test at 5%
probability.
Lots
Table 3 Initial moisture content of the lots (IMC),

moisture content after accelerated aging (MCA),
percentages for germination test (G), first count of
germination (FCG), accelerated aging with NaCl-76% (AA),
emergence of seedlings in substrate (ESS) of the different
lots of coriander seeds.
Lots
IMC(%) MCA(%) G(%) FCG(%) AA(%) ESS(%)
1
8.8
10.7
81 a 72 a
35 a
50 b
2
9.1
11.9
71 b 63 bc
15 c
35 c
3
8.8
10.7
82 a 59 c
26 b
53 b
4
8.9
11.7
77 a 66 b
35 a
61 a
CV(%) 7.74 8.26
10.0
10.3
Means followed by the same letter in a column do not differ in
level of 5% probability by the Tukey test.
Table 4 Electrical conductivity test with soaking 50 seeds in
50 ml of distilled water for periods of 2, 4, 6, 8 and 24 h
from four coriander seed lots.
Lots
1
2
3
4
Average
CV(%)
Periods of imbibing (h)

2
4
6
8
330 362
388
398
476 512
521
535
317 342
358
365
556 600
625
635
420 a 454 b 473 c 483 c
6.04
24
468
615
432
724
560 d
Average
389 B
532 C
363 A
628 D
478
The first count germination test separated the seed

lots in different vigor classes: high vigor (lot 1), low
vigor (lot 3), and intermediate vigor (lots 2 and 4).
Results similar to those obtained in the present study
were reported [19], working with corn, found that test
first countal lowed to classify lots into three vigor
levels (high, low and intermediate).
129
The accelerated aging test, performed for 48 h at

41 C was excessively drastic because of reducing seed
germination by more than 50% points in relation to
non-aging seeds; this situation can compromise the test
result. However, this test found differences between
seed lots and allowed to separate the three seed lots into
vigor levels (Table 3). Similar results were obtained
[20], with cotton seed, because in less severe aging
conditions, 41 C for 48 h, most of the seed lots had
germination decreased by about 50% points compared
to initial values. Despite drastic effects, aging at 41 C
for 48 hallowed to classify lots into four and three vigor
levels.
For all lots, in general, seedling emergence was low,
ranging from 35% to 61%. The physiological seed
quality is routinely assessed by laboratory germination
test; however, it is believed that this test conducted
under appropriate conditions can super estimate the
seed vigor [21]. In this case, if the ambient conditions
after sowing are not optimal, the percentage of
seedling emergence may be less than determined in
laboratory [22]; and this occurrence was found in this
work. Seedling emergence separated the seed lots into
three vigor levels, highlighting lot 4 as greater vigor,
lot 2 as a lower physiological potential and the other
as intermediaries. The worst performance presented
by the seeds of lot 2 in the test seedling emergence
was also observed in germination and accelerated
aging.
In the electrical conductivity test there was a
significant effect on the interaction seed and imbibing
periods (Table 4). However, it was possible to stratify
the lots based on the vigor. Lot 3 showed higher vigor
and lower lot 4, contradicting the vigor levels
separation obtained in accelerated aging tests and
seedling emergence.
The first count test, accelerated aging and seedling
emergence showed efficient differentiation of
physiological quality of seed lots of coriander,
however, the tests do not separated lots exactly the
same way. The accelerated aging test despite being
130
very dramatic was allowed to rank the lots a manner

similar to the results observed in seedling emergence.
[8]
4. Conclusions
The accelerated aging test using saturated saline
solution showed efficiency in assessing the
physiological quality of both lots of radish seeds and
coriander. This test at 41 C for 48 h is recommended
to be used for both species.
[9]
[10]
[11]
Acknowledgments
[12]
The authors would like to thank the supporting

agency, CAPES (Coordenao de Aperfeioamento de
Pessoal de Nvel Superior), for granting a scholarship
[13]
to the first author and CNPq (ConselhoNacional de

Desenvolvimento Cientfico e Tecnolgico) for
financial support that was indispensable for the
[14]
accomplishment of this work.
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plant field performance, Rev. Bras. de Sementes 28 (2006)
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P.C.F. Linares, M.L. Silva, M.F.S. Pereira, A.K.H.
Becerra, A.C.C. Pavia, Amounts and times of
decomposition of rooster tree on agronomic performance
of radish, Rev. Verde 6 (2011) 168-173.
R.S. Pereira, M.F.B. Muniz, W.M. Nascimento, Aspects
related to coriander seed quality, Hort. Bras. 23 (2005)
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N.L. Menezes, C.M.R. Santos, E.P. Nunes, Germination
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[16]
[17]
[18]
[19]
[20]
[21]
[22]
induced by sodium chloride and pH, Informativo Abrates

2 (2001) 189.
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accelerated aging test for small-seeded crops, Seed. Sci.
and Tech. 25 (1997) 123-131.
R.A. Freitas, W.M. Nascimento, Accelerated aging test
on lentil seeds, Rev. Bras. de Sementes 28 (2006) 59-63.
L.M. Mertz, S.R. Segalin, C. Huth, T.D. Rosa, Electrical
conductivity test in evaluating the physiological potential
of wheat seeds, Infor. Abrates 22 (2012) 35-39.
S.B. Torres, A.R. Peixoto, I.M.S. Carvalho, Sanitary and
physiological quality of tomato seeds from the submedio
So Francisco Region, Cincia e Agrotec 23 (1999)
825-829.
A.B. Rodo, M. Panobianco, J. Marcos Filho, Alternative
methodology for the accelerated aging test for carrot
seeds, Scientia. Agric. 57 (2000) 289-292.
M.C. Bhering, D.C.F.S. Dias, J.M. Gomes, D.I. Barros,
Vigor evaluation methods of cucumber seeds, Rev. Bras.
de Sementes 22 (2000) 171-175.
M. Panobianco, J. Marcos Filho, Accelerated aging and
controlled deterioration of tomato seeds, Scientia. Agric.
58 (2001) 525-531.
S.B. Torres, J. Marcos Filho, Accelerated aging of melon
seeds, Scie. Agric. 70 (2003) 77-82.
N.P. Ramos, E.P.O. Flor, E.A.F. Mendona, K. Minami,
Accelerated aging of Eruca sativa L. seeds, Rev. Bras. de
Sementes 26 (2004) 98-103.
N.L. Menezes, D.C. Garcia, C.A. Bahry, N.M. Mattioni,
Electrical conductivity test in black oat seed, Rev. Bras.
de Sementes 29 (2007) 138-142.
Z. Piana, M.A.A. Tillmann, D. Minami, Physiological
quality evaluation of onion seeds (Allium cepa L.) and its
relationship with vigorous seedlings production, Rev.
Bras. de Sementes 12 (1995) 149-153.
M.V.C. Miguel, J. Marcos Filho, Potassium leakage and
maize seed physiological potential, Scie. Agrcola 59
(2002) 315-319.
E.A.F. Mendona, S.C. Azevedo, S.C. Guimares, M.C.F.
Albuquerque, Vigor tests in upland cotton seeds, Rev.
Bras. de Sementes 30 (2008) 1-9.
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of pea (Pisumsativum L.) seed lots as evaluated by
different procedures, Rev. Bras. de Sementes 12 (1990)
52-75.

D
DAVID
PUBLISHING

Jimmy Akinfemi Osunbitan
Department of Agricultural & Environmental Engineering, Obafemi Awolowo University, Ile-Ife, Osun State 220005, Nigeria
Received: September 25, 2012 / Published: February 20, 2013.
Abstract: The study investigated the response of Amaranth to irrigation depth and irrigation interval as well as poultry waste
incorporation. A replicated 4 3 3 factorial arrangement with treatments consisting of percent organic matter incorporation (0%,
2%, 4% and 6% by weight), irrigation interval (1, 3 and 5 days) and irrigation depth (4, 6 and 8 mm) was used for the experiment
which was conducted in pots in a green house. The result of the experiment showed that percent organic matter incorporation,
irrigation interval, and irrigation depth significantly (P < 0.05) affected the dry matter yield of the vegetable. However, only the
percent organic matter incorporation and irrigation interval were significant (P < 0.05) in their effects on the plant height. The highest
plant height of 12.03 cm occurred when 2% organic matter was incorporated into the soil while the lowest plant height of 9.04 cm
occurred with 6% organic matter incorporation. The maximum plant yield (47.44 kg/ha) occurred when 2% poultry litter was
incorporated into the soil while the lowest plant yield (31.57 kg/ha) occurred with the control experiment. Irrigation interval of three
days as well as irrigation depth of 6 mm resulted in the highest plant height and maximum dry matter yield of the vegetable. From the
result of the experiment, the factor combination of 2% organic matter incorporation with 6 mm irrigation depth applied at three days
irrigation interval resulted in the highest plant height of 16.7 cm above ground surface and maximum yield of 67.8 kg/ha.
Key words: Amaranth, irrigation depth, irrigation interval, organic matter, dry matter yield.
1. Introduction
The rainfall distribution in the SouthWestern
Nigeria is bimodal with peaks in June and September.
The dry season extends from November to March.
During the rainy season, the rainfall is high and
relatively adequate for the production of most crops.
However, towards the end of the rainy season, the
rainfall becomes erratic and reduces to about 500 mm.
In the dry season, there is always an increasing
competition for water by agriculture and domestic
uses and irrigation is the only option for providing
water to crops. Soils with low water holding capacities,
combined with high evaporation rates, result in low
water use efficiency [1]. Water use efficiency is useful
because of its role in sustainable development.
Improving the efficiency of resource use is one of the
means of meeting sustainable development goals.
Corresponding author: Jimmy Akinfemi Osunbitan, Ph.D.,
research fields: soil and water conservation, soil and water
management for improved soil conditions and increased crop
yield,
irrigation,
organic
farming.
E-mail:
josunbit@oauife.edu.ng, josunbit@yahoo.com.
Improved water use efficiency will result in lowering

the water needs to achieve a unit of crop production.
The problem of low water use efficiency may be
minimized by the use of drought-resistant crop variety
mulching and proper irrigation scheduling [2, 3].
Amaranth species are a group of highly popular
vegetables, belonging to many different species. They
are the most commonly grown leafy vegetables of the
lowland tropics in Asia and Africa. Amaranths as a
plant are crop with ancient history and have been
grown for centuries for vegetables in different part of
the world [4]. It is consumed as vegetable in Africa,
Caribbean, China, Greece and Southern Pacific islands
[5]. The species were most commonly utilized as
vegetables have short plant with leaves and small
inflorescence [6]. The leaves are comparable to
spinach (Spinacia oleracea L.) in taste [7]. However,
unlike spinach, an ideal season for producing
Amaranth crop in the temperate climate is during the
hot month of the year. It is highly adapted under
lowland condition, grows well at day temperature
132
above 25 C and night temperature not lower than

15 C. They are quantitative short day plants and
consumes high amount of water and prefers fertile,
well drained soil with a loose structure.
Organic matter content (OMC) incorporation into
the soil has been found to improve soil structure,
increase water holding capacity, soil porosity and low
bulk density which in itself indicates that the soil is
well aerated, easy to till and facilitates root growth.
Osunbitan et al. [8] reported that the moisture
retention of three SouthWestern Nigerian soils
increased
with
increase
in
livestock
manure
incorporation into the soil. Adekalu and Osunbitan [9]

also observed that incorporation of goat dung into
soils reduced the dry density and increased the
hydraulic conductivity at different moisture contents.
It was observed by Millar and Turk [10] that
incorporation of organic matter into the soil generally
increases the water retaining power of the soil, and
decreases the water runoff. Osunbitan and Adekalu
[11] reported that the total depth of water for the
growing period of Jute Mallow decreased with
increase in the poultry waste incorporation into the
soil. According to Brady [12], the incorporation of
organic matter (OM) into the soil is one of the most
effective
soil
management
measures
aimed
at
improving soil fertility and structure, hence, organic

matter curbs soil erosion. Likewise, organic matter has
been found to enhance the stability of soil aggregate
[12] and its effect is being generated in soil having
low clay content [13] as well as unstable soils [14]. As
such, most well known soil scientists agreed that
organic matter plays a very important role in tropical
soils. This is even true of production systems using
high external inputs.
Also, the availability of water is a very essential
condition for growth in plants to derive their nutrients
uptake in soluble form from the soil. In other words,
the amount of water available to plant through the soil
provides the nutrients required for germination,
growth and yield of plants. Thus, the amount of water
(transpiration, evaporation from the soil, drainage loss)

required to produce a unit amount of dry weight
material (e.g., 1 kg corn) is a measure of efficiency
[15]. And also, how efficiently water can be used by
plant is a function of the transpiration ratio.
Furthermore, the level of irrigation depth is also an
essential factor that should be considered in crop
production. Irrigation is the artificial application of
water to the soil usually for assisting in growing crops.
In crop production, it is mainly used in dry areas and
in periods of rainfall shortfalls, but also protects plants
against frost [16]. It also helps to suppress weed
growth on fields. Irrigation is very essential for plant
survival before permanent wilting point is reached and
also, it cools the soil and atmosphere, making the
environment favorable for plant growth. Small scale
irrigation system is gaining grounds in the country
because of the cost of owning and management of the
system. It was observed by Adekalu et al. [2] that
majority of vegetable farmers in the country depend
on hand watering during the dry season when crop
value is high to produce vegetable crops because of
the high cost of fuel. There is scarcity of information
on optimum irrigation scheduling or the effects of
poultry litters on the yield of Amaranth vegetable. The
objective of this study was therefore to investigate the
response of Amaranth to irrigation scheduling
(irrigation depth and interval) and organic matter.

2.1 Study Area and Soil Sampling
Pot experiments were conducted in a green house of
the Faculty of Agriculture, Obafemi Awolowo
University, Ile-Ife, Nigeria, on sandy loam soil
classified as Alfisol. Total annual rainfall of the study
area was about 1,350 mm. While the average daily
minimum temperature ranged between 20 C and
22 C and the average maximum temperature between
27 C and 35 C. The soil is classified at series level
as two series [17]. It was derived from granite and
gneiss parent material. The texture of the plough layer
(0-15 cm) is sandy loam having 80% sand, 5% silt and

15% clay with organic matter content of 1.56%. The
soil samples were collected from the top 15 cm layer,
packed inside sacks and transferred to the laboratory
for the determination of its physical and chemical
compositions. Plant residues and gravels in the soil
sample were removed by passing the soil through a 2
mm sieve so as to obtain a more homogeneous soil
sample.
2.2 Experimental Procedure
The experimentation was a 4 3 3 factorial

arrangement with three replicates. The factors are
organic matter incorporation, irrigation interval and
irrigation depth. The organic matter treatment
consisted of four levels of poultry litters applied at the
rate of 0, 16, 32 and 48 t/ha, respectively, in form of
0%, 2%, 4% and 6% by weight of poultry litter
incorporation into the soil. The three levels of
irrigation interval used in the study were 1, 3 and 5
days while the three levels of irrigation depth were 4,
6 and 8 mm of water. The mixture of soil and poultry
litter at different levels of poultry litter incorporation
was compacted to bulk density of 1.5 g/cm3 inside
perforated plastic pots with surface area of 314.2 cm2
and depth of 45 cm. The soil/poultry litters mixture
was then left for 14 days to allow for some
decomposition of the manure during which water was
sprinkled into the pots. Ten seeds of Amaranth plant
were planted on September 29, 2009 inside each of the
pots. Two weeks after planting, on October 12
thinning of germinated plants was done to five stands
per plot after which irrigation water at the above
mentioned depths and intervals were applied using
small watering cans as is done by the majority of
small-scale farmers in the country.
Before every application of irrigation water,
moisture contents of soils in the pots were determined
using calibrated soil moisture meter about 1 h prior to
irrigation and drainage water through the perforation
of the pots was collected by small plastic bowls placed
133
under the soil pots after each irrigation for the

determination of the evapotranspiration by the crop.
Evapotranspiration (E) was calculated by the water
balance method:
E = I + R + SWD + D + RO
where, I is irrigation (mm), R is rainfall (mm), SWD is
soil water depletion from the beginning to the end of
the season, D is drainage below the root zone (mm)
and RO is run-off. Rainfall and runoff were taken to
be zero since the experiment was conducted in pots
inside green house. Plant height and leaf area were
measured once a week from the 5th week after
planting. The leaves of the vegetable are sometimes
plucked from five weeks after planting for
consumption and marketing by some farmers while
some wait till eight weeks after planting before
harvesting. Plant height was measured from the
ground level to the top most leaf while the leaf area
was measured by tracing the leaf on a while paper and
measuring the traced area using a planimeter. The
crop was finally harvested on November 16, 2009.
After the final harvest, the plants were cut at the
ground level and placed in an oven for 70 C for 24 h
to determine dry matter [18]. Analysis of variance [19]
was performed on the data and factor analyses were
run to determine the effects of treatments and their
interactions. Treatment means and significant
differences were evaluated using Duncan Multiple
Range (DMR) test.

Organic matter, irrigation interval, and irrigation
depth affected dry matter yield, depth of drainage and
total water used by the plant (Table 1). Only the
organic matter and irrigation interval were significant
for plant height. Leaf area was not significantly
affected by the three factors considered in this study.
3.1 Total Water Used
Depth of water used by Amaranths at different
irrigation depths in response to irrigation interval for
134
Table 1
Analysis of variance for growth and yield of Amaranth grown under different organic waste incorporation.
Source of variation
Organic matter (OM)
Irrigation interval (II)
Depth (ID)
OM II
OM ID
II ID
OM II ID
DF
3
2
2
6
6
4
12
F value
Leaf area (cm2)
NS
0.70
2.15 NS
0.14 NS
0.06 NS
0.02 NS
0.21 NS
0.07 NS
Plant height (cm) Dry matter (Kg/ha) Total water use (cm)
**
5.45
13.96**
0.23 NS
0.49 NS
0.25 NS
0.19 NS
0.24 NS
> 5,000
155**
4,077**
114 **
990**
7.23**
21.91**
**
**
26.48
759.84**
38.96**
11.55**
1.47NS
12.35**
12.22**
Drainage (cm)
6.74**
> 5,000**
3,696**
2.33NS
1.47NS
958**
1.28NS
NS, ** represent not significant and significant at P < 0.05 respectively.
(a)
(b)
(c)
Fig. 1 Total depth of water used at different irrigation depths, (a) 4 mm, (b) 6 mm, (c) 8 mm irrigation depths and irrigation
interval in response to organic matter.
the four levels of organic matter incorporation is

shown in Fig. 1. Total water used by the crop varied
significantly (P < 0.05) with the three factors
considered in the study. Total depth of water used by
the crop increased with decrease in the irrigation
interval. On the average, the lowest depth of 29.02
mm of water was used by the vegetable when the
irrigation interval was five days while the highest
depth of 44.06 mm of water was used by the plant
when irrigation water was applied daily. According to
Duncan Multiple Range (DMR) test, the three levels
of irrigation interval were significantly different in
their effects on the total water used by the plant.
However, the total depth of water used by the plant
did not vary linearly with the remaining two factors
considered in this study. On the average, the highest
water used by the vegetable occurred when the
irrigation depth was 8 mm while the lowest average
water used occurred when the irrigation depth was 6
mm. The three levels of irrigation depth were also
significantly different (DMR test) in their effects on
the total water used by the plant. Moreover, when
organic matter was considered as a factor, the highest
water used by the plant (37.11 mm) was recorded
when 4% of poultry litter was incorporated into the
soil while the lowest water used by the plant (33.59
mm) was recorded when the percent incorporation of
organic matter was 6%. The control experiment (0%
organic matter incorporation) and 4% organic matter
incorporation were not significantly different in their
effects on total water used. However, these two levels
of poultry litter incorporation were significantly
different from the remaining two levels of organic
matter incorporation in their effects on total water
used.
3.2 Plant Height
The effect of the organic matter incorporation into
the soil, depth of irrigation and irrigation interval on
the plant height is shown in Fig. 2. Plant height varied
significantly (P < 0.05) with organic matter and
135
irrigation interval. The highest plant height of 16.7 cm

was recorded when 2% organic matter was
incorporated and 6 mm of water applied every three
days. This is similar to what was observed by Adekalu
et al. [2] that growth and yield were higher when
irrigation was scheduled at 30% soil water depletion.
On the average, the lowest plant height of 9.19 cm
was observed when the irrigation interval was five
days while the highest plant height of 12.30 cm was
recorded when the irrigation interval was three days.
According to Duncan Multiple Range (DMR) test, the
irrigation intervals of one day and five days were not
significantly different in their effects on the plant
height while three days irrigation interval was
significantly different from the remaining two levels
of irrigation intervals. Moreover, 6 mm irrigation
depth resulted in the highest (10.51 cm) plant height
on the average while 8 mm irrigation depth gave the
lowest (10.06 cm) plant height. However, the three
levels of irrigation depth considered in this study were
not significantly (DMR test) different in their effects
on the plant height of the vegetable. Organic matter as
a factor significantly affected the plant height. The
highest plant height (12.03 cm) occurred when 2%
organic matter was incorporated into the soil while the
lowest plant height (9.04 cm) occurred with 6%
organic matter incorporation. Manure exerts favorable
effects upon granulation and aeration in the soil for
good moisture status to ensure free flow of nutrients
and improves plant growth [9]. According to DMR
test, the 2% organic matter incorporation was
significantly different in its effect on the plant height
from the remaining three levels of organic matter
levels. The remaining three levels were not
significantly different in their effects on the plant
height.
3.3 Dry Mater Yield
The effect of the organic matter incorporation into
the soil, depth of irrigation and irrigation interval on
the plant yield is shown in Fig. 3. Plant yield varied
136
(a)
(b)
(c)
Fig. 2 Plant height as affected by the different irrigation depths, (a) 4 mm, (b) 6 mm, (c) 8 mm irrigation depths and
irrigation interval in response to organic matter incorporation.
significantly (P < 0.05) with organic matter, depth of

irrigation and irrigation interval. On the overall, when
all the factors were considered together, the maximum
dry matter yield of 67.8 kg/ha occurred with 2%
organic matter incorporation when 6 mm irrigation
depth was applied every three days while the
minimum dry matter yield of 30.0 kg/ha was observed

when no poultry litter was added with 4 mm of
irrigation water applied daily. However, on the
average, the minimum plant yield of 36.88 kg/ha was
observed when the plant was irrigated everyday while
the maximum yield of 38.89 kg/ha was recorded when
137
(a)
(b)
(c)
Fig. 3 Plant yield as affected by the different irrigation depths, (a) 4 mm, (b) 6 mm, (c) 8 mm irrigation depths and
irrigation interval in response to organic matter incorporation.
the irrigation interval was three days. Efficient

irrigation interval decreased deep percolation and
evaporation [20] in other crops. According to Duncan
Multiple Range (DMR) test, all the irrigation intervals
were significantly different in their effects on the dry
matter yield. Likewise, the three levels of irrigation

depths considered in this study were significantly
different in their effects on the dry matter yield of the
plant. On the average, irrigation depth of 6 mm
resulted in the maximum (43.78 kg/ha) dry matter
138
Table 2
Relationship between percent organic matter content (OM) and dry matter yield (DMY).
Irrigation depth (mm)

4
8
*
Irrigation interval (day)

1
3
5
1
3
5
1
3
5
Equation*
DMY = 0.43 OM3 5.55 OM2 +19.11 OM + 16.3
DMY = 2.55 OM3 21.15 OM2 +52.6 OM 1.4
DMY = 1.65 OM3 14.65 OM2 +39.4 OM + 4.3
DMY = 7.65 OM3 64.05 OM2 +165.2 OM 76.7
DMY = 14.88 OM3 120.20 OM2 +291.30 OM 153
DMY = 10.15 OM3 83.65 OM2 +208 OM + 103
DMY = 2.7 OM3 24.15 OM2 +64.35 OM 12.4
DMY = 5.27 OM3 43.10 OM2 +103.60 OM 31.3
DMY = 4.10 OM3 34.70 OM2 +87.20 OM 24.9
Equations are significant at 0.05 level of significance; DMY and OM are dry matter yield and organic matter content respectively.
yield followed by 8 mm irrigation depth (35.41 kg/ha)

while the minimum yield (34.15 kg/ha) occurred when
the irrigation depth was 4 mm. Moreover, the four
levels of organic matter incorporation were
significantly different (P < 0.05) in their effects on the
dry matter yield. The maximum plant yield (47.44
kg/ha) occurred when 2% poultry litter was
incorporated into the soil while the lowest plant yield
(31.57 kg/ha) occurred with the control experiment, i.e.,
when no organic matter was incorporation. Yield of
crops is related to water use efficiency, and this
probably explains the higher yield obtained at 2%
organic matter incorporation with higher water use
efficiency [11]. The remaining two levels of organic
matter incorporation, 4% and 6%, resulted in dry
matter yield of 37.41 kg/ha and 34.63 kg/ha,
respectively. Table 2 gives the relationship between
the percent organic matter incorporation (OM) and
observed dry matter yield (DMY). For all the irrigation
depths and intervals, the relationships between DMY
and OM were polynomial of the third order. There was
also high correlation between DMY and OM with R2
values of close to 1.00 for the correlation of DMY
against OM for the three irrigation depths and
irrigation intervals. Using these relationships, the
optimum level of poultry waste for the highest yield is
between 1.8% and 2.2%. This is in agreement with the
suggested value by researchers [11, 21, 22].
incorporation into loamy sand soil on the yield of

Amaranth. Total depths of water used by the crop
increased with decrease in the irrigation interval. Plant
height varied significantly (P < 0.05) with organic
matter and irrigation interval. The highest plant height
occurred with 2% organic matter incorporation while
the lowest plant height occurred with 6% organic
matter incorporation. Plant yield varied significantly
with organic matter, depth of irrigation and irrigation
interval. The maximum dry matter yield occurred with
2% organic matter incorporation when 6 mm
irrigation depth was applied every three days while the
minimum dry matter yield was observed when no
poultry litter was added with 4 mm of irrigation water
applied daily. It is therefore imperative that the
nutrient uptake from the different levels of organic
matter incorporated into the soil should be estimated.
References
[1]
[2]
[3]
[4]
4. Conclusions
[5]
The study investigated the effects of irrigation

depth and irrigation interval as well as poultry waste
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and tillage system on yield and water use efficiency of
cowpea, Communication in Soil Sci. and Plant Analysis
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Afolayan, Response of Okra to irrigation and mulching,
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A.A. Al-Masoum, I.M. El-Gharib, Plastic mulches for
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(1995) 61-67.

D
DAVID
PUBLISHING
Quantitative Changes in Protein and Cholesterol in

Haemolymph of the Red Palm Weevil Rhynchophorus
ferrugineus after Treatment LeucokininII
Mona Mohammed Saleh Al-Dawsary
College Science and Humanities, Salman Bin Abdul Aziz University, Al-Kharj- Kingdom Saudi Arabia

Abstract: We estimated quantitative changes to the content of protein and cholesterol in the Haemolymph of adult Red Palm Weevil
after being fed on sugar cane treatment with different concentrations of LeucokininII. In males, 0.05% has recorded significant
increase in total protein, then 0.25% concentration compeer control, while the maximum high of females 4.846 mg at 0.05% compeer
control. The effect of leucokininII on content of haemolymph cholesterol has shown result that 0.05% concentration and have a clear
impact on cholesterol concentration for both sexes with an average reduction of 37.989 mg in males compeer with 120.123 for
control, and 57.263 in females compeer with 96.087 mg for control.
Key words: LeucokininII, red palm weevil, protein, cholesterol, reduction.
1. Introduction
The red palm weevil (RPW) Rhychophorus
ferrugineus (Olivier) (Coleoptera: Curculionidae) is
considered as one of the most damaging invasive insect
species [1, 2]. The geographic origin of RPW was
claimed to be South East Asia and Melanesia [3-6].
Multiple introductions of RPW to the Middle East, the
Mediterranean Basin and US have occurred since mid
1980s through movement of infested offshoots [2].
Recently, the RPW is being considered as a major pest
of date palms (Phoenix dactylifera) and different
ornamental palm species in the Middle East and the
Mediterranean basin, Caribbean (Island of Curacao,
Netherland Antilles), Lebanon and USA [2, 6-9].
Insect neuropeptides are involved in almost all
physiological aspects in insects, such as diereses,
ecdysis, pheromone biosynthesis and control of
muscle activity. Thus, these peptide hormones and
Corresponding author: Mona Mohammed Saleh
Al-Dawsary, Ph.D., research fields: insects physiology, sense
organs and pharmacology. E-mail: wisdom1425@yahoo.com.
their receptors are promising targets for a novel

generation of selective and non-neurotoxic
insecticides. However, due to poor bioavailability,
pharmacokinetics and short half-life, the peptides
themselves can not be used as insect control agents.
The past two decades have witnessed an increase in
research into the discovery of nonpeptide small
molecules that function as mimics for neuropeptides.
As early as the Polish scientist Kopec [10] proposed
that substances in the brain control of the moulting
and metamorphosis processes. Today, we know that
numerous physiological processes in insects are
controlled by small, bioactive peptides. Because these
regulatory peptides are synthesized in modified
neurons, they are called neuropeptides. Neuropeptides
are chemical messengers which are released from the
neurons into the Haemolymph of the insect to reach
their distant target organs. Neuropeptides are
ubiquitos in the nervous system of insects and they are
by far the most diverse signaling substances, both
structurally and functionally [11]. Neurohormone
Quantitative Changes in Protein and Cholesterol in Haemolymph of the Red Palm

Weevil Rhynchophorus ferrugineus after Treatment LeucokininII
regulated processes, can, however, be grouped into

four major functional categories: 1) growth and
development, 2) behavior and reproduction, 3)
metabolism and homeostasis, and 4) muscle
movement [12]. The leucokinins are considered
cephalomyotropic peptides, to indicate their isolation
from the head of the cockroach Leucophaea maderae
and their ability to increase muscular contractions in
the cockroach hindgut [13]. Curious agents which
stimulate hindgut contractions might also enhance
excretory activity in Malpighian tubules upstream, we
have discovered that leucokinin has diuretic effects in
Malpighian tubules of the yellow fever mosquito
Aedes aegypti [14]. Given the diverse roles of insect
leucokinins, elucidation of the mode of action of these
peptides via their cognate G protein-coupled receptors
(GPCRs) are of importance. Furthermore, as
leucokinins have only found in invertebrates, it is
likely that careful design of leucokinin antagonist or
agonist analogues will avoid interactions with
mammalian species. While identification of
leucokinins and their cognate receptors have been
successfully undertaken in some insects [15-17],
including the genetically tractable Dipteran,
Drosophila melanogaster [18, 19], less progress has
been made in studies of leucokinin signaling in
biomedical relevant insects. As importance of
physiological peptides and their roles on insects
metabolism, have focused this study on quantitative
changes of total protein and lipids in haemolymph
adults red palm weevil after treatment leucokininII,
which may support the vital role of peptide.

2.1 Weevils
Adult insects, which were free from previous
pesticide exposure, were obtained from infected date
palms with the help of the Ministry of Agriculture and
Water located in Al-Kharj, Kingdom of Saudi Arabia.
Breeding of larvae was conducted using different
varieties in accordance with modern established
141
methods [20, 21] in laboratory College Science and

Humanities, Salman Bin Abdul Aziz University,
Kingdom of Saudi Arabia.
2.2 LeucokininII
LeucokininII was obtained from American peptide
company, INC. 777E. Evelyn Ave. Synnyvale, CA
94086 USA. The peptide was dissolved in 3 mL of
acetone
solution
and
preparation
different
concentrations: 0.05%, 0.25%, 0.4% respectively for

treatment of newly adults from cocoon in accordance
with the author [22].
2.3 Treatments
2.3.1 Treatment of Sugar Cane
Pieces of sugar cane were treated high with the
weight of the different known concentrations of
LeucokininII, and distributed each concentration of
each LeucokininII alone to pieces sugar cane using a
thin needle to make sure the distribution of solution
over the entire piece. We have left pieces of sugar
cane in the laboratory atmosphere for a period of time
to make sure the smell of the solvent evaporate
completely.
Sections of sugar cane were treated with various
concentrations of the LeucokininII and carefully
distributed using a thin needle. The sections of sugar
cane were then allowed to remain in the laboratory
atmosphere to permit the solvent to completely
evaporate.
2.3.2 Treatment of Insects
After the treatment of sugar cane pieces of various
concentrations of LeucokininII, they were transferred
to the boxes for rearing. A packet was placed inside a
piece treatment of sugar cane and a pair of insects. We
have conducted 15 replicates for each concentration
separately, as well as a control group treated only by
solvent. Adults were fed for one week on sugar cane
treatment, then, they were transferred into a diet of
untreated sugar cane. We have collected haemolymph
from insects in preparation for physiological studies.
142

2.3.3 Samples for Biochemical Studies

After treatment of insects with different
concentrations leucokininII collected haemolymph
from males and females severally, with these samples
accordance the requirements of the procedures for
each test.
2.3.3.1 Total Proteins Determination
Total proteins determination in haemolymph of
insects treatment and untreatment via proteins kit from
Jermany Human Company, it depended on biuret
method where estimation is as follow:
-Prepare samples for determination take 20 L from
haemolymph (haemolymph collected from fore leg via
capillary tube, added in eppendorf tube contain phenyl
thiourea crystals) in clear tube and added for it 1,000
L from color reagent are ready;
-Prepare standard solution added 20 L from
standard solution are ready in kit to 1,000 L from
color reagent the same samples;
-Use 1,000 L from color reagent blank for
spectrophotometer uv-120-02 shimadzu at 546 nm;
-Mix, incubate for 10 min at 20 C to 25 C.
Measure the absorbance of the sample and standard
against the reagent blank within 30 min;
-Reading is recorded for each sample and calculate
8 A sample
A standard
quantitative protein according to the equation:
C=
(1)
2.3.3.2 Cholesterol Determination

Total cholesterol determination in haemolymph of
insects treatment and untreatment via Cholesterol-kit
from Jermany Human Company where estimation is
as follow:
-Prepare samples for determination take 10 L from
haemolymph as previously in clear test tube and added
for it 1,000 L from color reagent are ready for use;
-Prepare standard solution added 10 L from
standard solution are ready for use to 1,000 L from
color reagent the same samples;
-Use 1,000 L from color reagent blank for
spectrophotometer uv-120-02 shimadzu at 500 nm;
-Mix, incubate for 10 min at 20 C to 25C.

Measure the absorbance of the sample and standard
against the reagent blank within 60 min;
-Reading is recorded for each sample and calculate
quantitative protein according to the equation:
C=
200 sample
A standard
(2)

Results were analyzed by F test in one way using
the program MSTAT [23].
3. Results
3.1 Effect of LeucokininII on Total Proteins and
Cholesterol Concentration in Haemolymph of R.
ferrugineus
3.1.1 Total Proteins
Table 1 shows significant difference between total
protein concentration in Haemolymph when treating
males and females with different concentrations of
leucokininII. These treatments have significant effects
on protein content in Haemolymph for adult females
compared with control, where concentration recorded
significant increase, 0.05% in the protein content
followed by 0.4% compared control. Also, the
leucokininII have significant increase when treating
females with 0.25% that decreased recorded in protein
content 0.110 g mg-1. These results show
leucokininII have an active effect in creating protein
in heamolymph. It is clear that the changes in protein
content in insects tissues fall under hormonal system
control of insect treatment which is the juvenile
hormone and ecdysteroid hormone. With exception to
the females treated with 0.25% concentration which
recorded a decrease in the protein content 0.110 g
mg-1, it indicates suppression in creating protein
[24-26].
3.1.2 Cholesterol
Table 2 shows effect of leucokininII on cholesterol
concentration in haemolymph for adults of red palm
weevil. That results showed a significant decrease in

Table 1 Effect of treatment of LeucokininII on total
protein concentration in haemolymph of adult red palm
weevil R. ferrugineus.
Concentration
0.05%
0.25%
0.4%
Cont.
Males
Mean g mg-1 SD
3.872* 1.00
0.500* 0.18
1.679* 0.04
0.232* 0.00
Females
Mean g mg-1 SD
4.846 0.13
0.110 0.00
1.296 0.00
0.134* 0.00
Significant of 0.001.
Table 2 Effect treatment of LeucokininII on cholesterol
concentration in haemolymph adult red palm weevil R.
ferrugineus.
Concentration
0.05%
0.25%
0.4%
Cont.
Males
Mean g mg-1 SD
37.989* 1.12
114.243* 0.29
82.960* 0.56
120.123* 22.91
Females
Mean g mg-1 SD
57.263* 0.48
63.967* 0.00
82.681* 2.79
96.087* 5.58
Significant of 0.001.
cholesterol concentration in haemolymph for both

sexes compared to the control group. Decreased rate
in cholesterol concentration of males treatment 37.989
g mg-1, 82.970 g mg-1, 114.24 g mg-1 at
concentrations 0.05%, 0.4%, 0.25%, while in females
57.263 g mg-1, 63.967 g mg-1, 82.681 g mg-1 at
concentrations 0.05%, 0.25%, 0.4% respectively. Also,
these results show that 0.05% concentration recorded
very low cholesterol concentration in both sexes
compared with the control group. This low rate
resulted of the leucokininII effect on secretion
juvenile hormone from CA and defected metabolism
of lipids [27].
143
larvae [22]. In this study, we have focused on the

effect of leucokininII on total proteins and cholesterol
concentration in haemolymph of adults R. ferrugineus.
These results showed in Table 1 that defect happened
with leucokininII on total protein in haemolymph
because these concentration worked high of protein
content in haemolymph both sexes, so leucokininII
effected on neurosecretory glands hormone that
controlling proteins metabolism, or effect leucokininII
attributable to effected enzymes of neuropeptides on
metabolism for neuropeptides in insects [28] that
enzymes of neuropeptides are keys of indicators
systems. It was the changes in the protein content of
the tissue insects under control of the hormonal
system of the insect [10, 24, 25]. This defect in the
protein content of decreased or increased due to
irregular of these hormones for the control group.
Also, these results coincided with indicated by
previous studies of link metabolism proteins with
hormonal regulation for secretory nerve glands
specially CA, CC and Prothoracicgland. Also, this
study showed results in Table 2 significant reduction
of cholesterol concentrations in haemolymph in both
sexes after treatment with different concentrations
from leucokininII. These reductions are attributable to
effected leucokininII on secretion juvenile hormone
from CA and defected metabolism for lipids according
to references [27, 29, 30]. And, these effected for
lipids may be leucokininII effect on Adipokinetic
hormone that regulated lipids metabolism which
secretion from CC.
4. Discussion
Neuropeptides are chemical messengers which are
released from neurons into the haemolymph of the
insect to reach their distal target organs. Insect
neuropeptides are involved in almost all physiological
process in insects, such as dieresis, pheromone
biosynthesis, and control of muscle activity. In
previous study, we have studied leucokininII effect on
some biological aspects of red palm weevil, and
observed leucokininII, and some deformation on
5. Conclusions
LeucokininII effected significantly on total proteins
and cholesterol concentration in haemolymph for Red
Palm Weevil, it has effect on nerve gland secretion
hormones that regulated metabolism of these
compounds.
Acknowledgments
The authors extend their sincere thanks to the
144

deanship of Scientific Research, the University for its

support to this research and is part of the project No.
37/H/1432H, which was supported by the Deanship.
[13]
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D
DAVID
PUBLISHING
The Role of Cellulase and Pectinase in Apricot Canker

Caused by Hendersonula toruloidea and
Phiaoacremonium aleophillium
Nidhal Y. M. Al-Morad
Department of Plant Protection, College of Agriculture and Forestry, University of Mosul, Iraq
Abstract: Significant increasing in cellulase and pectinase activity were observed in response to infection with Hendersonula
toruloidea and Phiaoacremonium aleophillium in apricot trees comparing with uninfected trees. Pectinase highest activity was
recorded at the first week after inoculation with H. toruloidea (1.76 unit/g f.w.), while cellulase highest activity was recorded after
two weeks of inoculation with P. aleophillium (6.49 unit/g f.w.). Peroxidase and polypolyphenol oxidase activity were significantly
increased after inoculation with H. toruloidea and P. aleophillium, Peroxidase highest activity was recorded after 48 h of inoculation
with H. toruloidea (1.77 unit/g f.w.), while polypolyphenol oxidase highest activity was recorded after two weeks of inoculation with
P. aleophillium (3.33 unit/g f.w.). The result also showed that total phenol contents was significantly increased as a result to
inoculation with H. toruloidea and P. aleophillium, highest total phenol contents was recorded after 48 h of inoculation with H.
toruloidea (1.61 mg/g f.w.).
Key words: Cellulase, pectinase, peroxidase, polypolyphenol oxidase, apricot, Hendersonula toruloidea, Phiaoacremonium
aleophillium.
1. Introduction
Phytopathogenic fungi that attempt to colonize higher
plants must contend with physical barriers of the host as
surface waxes and cell wall [1], phytopathogenic fungi
produce several pectolytic enzymes, which are capable
to degrade plant cell wall components during plant
pathogenesis. These enzymes are called pathogenic
enzymes mostly cellulolytic and pectinolytic enzymes
by the activity of these enzymes, which may facilitate
fungal growth and provide the fungus with nutrients [2,
3]. In a number of systems, a strong correlation has been
found between the presence of pectinolytic enzymes and
disease symptoms and disease virulence [4, 5]. Many
phytopathogenic fungi possess the enzymes destroying
cellulose, hemicelluloses and pectin. Wood-destructive
fungi possess active cellulases, while the most active

enzymes in grassy plant parasites are pectinases [6-8].
Apricot is one of the most popular fruit in Iraq, apricot is
greatly hindered due to a number of diseases. Majority
of the diseases is fungal origin, among of the fungal
diseases branch wilt and decline diseases caused by
Phiaoacremonium aleophillium and Hendersonula
toruloidea, which attack branches forming typical
necrotic canker of brown to black color [9-11]. Hence,
this work deals with the enzymatic profile of isolates of
P. aleophillium and H. toruloidea and its activity in the
infected apricot plants addition to activity of some
defense related enzymes oxidative enzymes peroxidase
(PO) polypolyphenol oxidase (PPO).

2.1 Plant Material, Pathogen Fungi
Corresponding author: Nidhal Y. M. Al-Morad, assistant

professor, research field: plant pathology. E-mail:
nidalalmurad@yahoo.com.
Apricot trees were used as plant material. P.
The role of cellulase and pectinase in apricot canker caused by Hendersonula

toruloidea and Phiaoacremonium aleophillium
aleophillium and H. toruloidea isolated from naturally

infected apricot trees were maintained on PDA at 25
2 C.
2.2 Preparation of Enzyme Extract from the Pathogen
The isolates were cultivated in Czapeck Dox (CD)
medium by replacing the indigenous source of the
medium and were harvested at 15th day. In order to
elicit the secretion of specific enzymes, related
substrates like 5% of cellulose and pectin were
replaced for carbon source in the (CD) medium for
inducing cellulase and pectinase. The mycelium was
harvested twice using Whahnam No. 1 filtration, the
supernatant was then dried and weighted, the filtered
extract was then used as the respective enzyme source.
2.3 Effect of pH on Enzyme Activity
The effect of pH on enzyme activity was
determined by performing the enzyme assay with
reaction mixtures of which the pH ranged 4-6-7.
2.4 Preparation of Enzyme Extract from the Healthy
and Infected Plants
Samples from each apricot treatment, healthy or
infected, were collected 0, 24, 48, 72, one week, two
weeks after inoculation with P. aleophillium and H.
toruloidea.
2.5 Detection of Pectinase Enzyme Activity
Pectinase activity was expressed as changes in
147
Enzyme extract was obtained by grinding plant

tissues in 0.1 M sodium phosphate buffer at pH (7.1)
(2 mL/ tissues) in a porcelain mortar. The extracted
tissues were strained through four layers of
cheesecloth. Filtrates were centrifuged at 3,000 rpm
for 20 min. PO activity was expressed as changes in
absorbance/min at 420 nm according to the method
[14]. PPO activity was expressed as changes in
absorbance/minat 420 nm according to the method
[15].
2.8 Determination of Total Phenol
Total phenol was obtained from healthy and infected
tissues by homogenizing the tissues (1 g) with 80% (v/v)
methanol and agitated for 15 min at 70 C, 1 mL of the
methanolic extract was added to 5 mL of distilled water
and 250 L of Folin-Ciocalteau reagent (1N), the
solution was kept at 25 C. The abortion of developed
blue colure was measured at 725 nm catechol was used
as the standard as g catechol mg-1 FW [16].
All the grouped data were statistically evaluated
using SAS 9. Hypothesis testing methods included are
one way analysis of variance (ANO VA) followed by
Duncan with P < 0.05 were considered to be
statistically significant. All the results were expressed
as the mean of six replicated values.
absorbance/min at 235 nm according to the method

[12].
2.6 Detection of Cellulases Enzyme Activity
3.1 Effect of pH on Pectinase and Cellulase Activity

The optimum pH for pectinase and cellulase
The reaction mixture contained 0.1 g cellulose

powder; 1 mL 0.05 M citrate buffer (pH 4.8) and 0.5
mL enzyme source was incubated for 1 h at 50 C.
One unit of cellulose is defined as the amount of
enzyme which liberates 1 mg reducing sugar per 1
h/0.1 g fresh weight [13].
2.7 Determination of PO and PPO Enzymes Activity
Fig. 1
Effect of pH on pectinase and cellulase activity.
148

activity was 4 and 6, respectively (Fig. 1).

3.2 Hydrolytic Enzymes
The results presented in Table 1 show that the
relative activities of pectinase and cellulase were
significantly increased in response to infection with H.
toruloidea, P. aleophillium compared to healthy plants
(control). The highest activity of pectinase in infected
apricot trees was recorded at the first week after the
inoculation with H. toruloidea, P. aleophillium and
with both fungi (1.76, 1.88, 1.98 unit/g f.w.)
respectively. While the highest activity of cellulase in
infected apricot trees was recorded at the first week
after the inoculation with H. toruloidea (5.80 unit/g
f.w.) and recorded at the second week after the
inoculation with P. aleophillium and with both fungi
H. toruloidea and P. aleophillium (6.49 and 7.27
unit/g f.w.), respectively.
3.3 Oxidative Enzymes
In the present study, the induction of PO activity in
infected apricot trees over their uninfected controls
has been revealed (Table. 2). The highest induction
was recorded at 48 h after inoculation with H.
toruloidea (1.77 unit/g f.w.) and 24 h after inoculation
with P. aleophillium (1.74 unit/g f.w.) and with both
fungi (1.69 unit/g f.w.) where approximately 2-fold
increase over uninfected counterparts. The highest
induction activity of PPO in infected apricot trees was
recorded at second week after inoculation with H.
toruloidea (1.02 unit/g f.w.) and at first week after
inoculation with P. aleophillium (3.33 unit/g f.w.) and
second week after inoculation with both fungi (1.15
unit/g f.w.).
3.4 Total Phenol
As can be seen from the results presented in Table 2,
infection of apricot trees with P. aleophillium or H.
toruloidea induced significant increases in total
phenol contents as compared to the uninfected
(control). The highest increases in total phenol
Table 1 Hydrolytic enzyme activity in Apricot trees

infested with H. toruloidea and P. aleophillium under
greenhouse condition.
Enzymes activity
(unit/g fresh weight)
Fungi
Period
Pectinase
Cellulase
24 h
0.92fg
3.9 gi
48 h
1.40de
5.23ef
1.52cd
5.56cd
H. toruloidea 72 h
One week 1.76ac
5.80cd
Two weeks 1.72a-d
5.76ce
24 h
1.04f
4.44gf
48 h
1.52cd
5.86ce
1.64a-d
6.23ad
P. aleophillium 72 h
One week 1.88ab
6.45ac
Two weeks 1.62cd
6.49ac
24 h
1.18ef
4.97ef
1.62cd
6.56ac
1.82 ac
6.98ab
and
H. 72 h
toruloidea
One week 1.98a
7.23a
Two weeks 1.66ad
7.27a
24 h
0.44h
2.33j
Control
48 h
0.92g
2.56j
72 h
0.64gh
2.82j
One week
0.64gh
3.10j
Two weeks 0.6h

3.41hj
Mean within a column followed by the same letter are not
significantly different according to Duncan test P < 0.05.
Table 2 Oxidative enzymes activity and phenolic contents
in Apricot trees infested with H. toruloidea and P.
aleophillium on under greenhouse condition.
Enzyme activity Total
(unit/g fresh weight) phenol
Fungi
Period
mg/g f.w.
PO
PPO
24 h
1.22d
0.58c
0.70f
48 h
1.77a
0.62c
1.61a
H. toruloidea
72 h
1.39bc
0.71c
1.56b
One week 1.61a
0.53c
1.30c
Two weeks 1.24cd
1.02c
1.21d
24 h
1.74a
1.18c
0.99e
48 h
1.47b
2.01b
1.44bc
1.12d
1.87b
1.25d
One week 1.71a
3.33a
1.34c
Two weeks 0.89d
1.15c
0.93e
24 h
1.69a
0.58c
0.84f
48
h
1.41bc
0.71c
1.56b
P.aleophillium
1.1d
0.53c
0.90e
and
H. 72 h
toruloidea
One week 1.31 cd
1.02c
1.25d
Two weeks 0.90d
1.15c
1.33c
24 h
0.75e
0.38d
0.44g
48 h
0.81e
0.31d
0.74f
72 h
0.85d
0.28d
0.27g
Control
One week 0.71e
0.32d
0.39g
Two weeks 0.69e
0.35d
0.49g
Mean within a column followed by the same letter are not
significantly different according to Duncan test P = 0.05.

contents were recorded at first week after inoculation

with H. toruloidea and P. aleophillium (1.30 and 1.34
mg/g f.w.) respectively and second week after
inoculation with both fungi (1.33 mg/g f.w.).
Significant correlation was well demonstrated earlier
between pathogenicity causing severe tissue
maceration and the extracellular pectinase and
cellulases activities. Pectic enzymes that have the
ability to split the 1,4 bonds between the galacturonic
acid moieties in the pectic fraction of the cell wall
remain the only enzyme confirmed to cause plant
tissue maceration [17-19].
Our results indicate that hydrolytic enzymes
pectinase and cellulases induction oxidative enzymes in
apricot trees. We show that combinations of pectic
enzymes and cellulases cooperate in the induction
oxidative enzymes in apricot trees. Time-course activity
of peroxidase in apricot tree infected with P.
aleophillium and H. toruloidea. The induction of PO
activity has been repeatedly reported in several plant
species in response to pathogen infection. The
peroxidase activity, in general increases under different
stress conditions like wounds, fungi infections, salinity,
water stress and nutritional disorders, inducing also the
lignin increment and production of ethylene and induce
the increase of the production of phenols oxidized at the
cell wall [20]. This activity, suggests a cell effort for the
establishment of a physiochemical barrier, able to
isolate the infected area [21]. Similar to the present
report on oxidative enzymes, previous works also
indicated that these oxidative enzymes are involved in
the defense role [22]. With resistance in many plants
[23], an immediate response of plants to injury, in most
cases, is the accelerated accumulation of oxidative
enzymes required for scavenging toxic radicals [24].
Plants were reported to possess efficient antioxidant
defense system due to the presence of pathway for
catalase and peroxidase [25].
4. Conclusions
The present results indicate that pectinase and
149
cellulases
have
been
inducted
peroxidase,
polypolyphenol oxidase in apricot trees. We also find
that combinations of pectic and cellulases enzymes
cooperate in the induction oxidative enzymes in
apricot trees.
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D
DAVID
PUBLISHING
Innovation and Technical Efficiency in the Smallholder

Dairy Production System in Ethiopia
Amlaku Asres, Johann Slkner and Maria Wurzinger
Department of Sustainable Agricultural Systems, University of Natural Resources and Life Sciences (BOKU), Gregor-Mendel-Str. 33,
1180 Vienna, Austria
Abstract: This study provides estimates of smallholder households production efficiency and its determinants, and separately
analyses the technical efficiency of dairy technology adopting and non-adopting farmers using data from Ethiopia. Cobb-Douglas
stochastic frontier production function was modeled in the context of local level agricultural innovation systems framework and
estimated using 2011 milk production data on 304 dairy farmers. Results show that the mean level of technical efficiency among the
sampled farmers was about 26%. This result suggests that there is room for significant increases of production through reallocation of
existing resources. Despite significant variation among farmers, these results also indicate only 19% of farmers have mean efficiency
scores ( 50%), implying a need to focus on creating innovation capacity that pushes the production frontier outward in the dairy
production system. It is also revealed that individual farm households efficiency varied widely across dairy technology adoption
status, gender and districts. The significant gamma () statistic, of 0.9985 in the analysis indicates that about 99.85% variation in the
output of milk production would be attributed to technical inefficiency effects (those under farmers control) while only 0.0015%
would be due to random effects, i.e., beyond the farmers control and hence calling for a focus on efficiency enhancing investments.
Education, farm size, extension visit and off-farm income opportunity were found to be efficiency enhancing. The study recommends
that different components of an agricultural innovation system have to interact to improve the innovation capacity of different actors
and thereby improve the estimated technical inefficiencies.
Key words: Agricultural innovation systems, dairy, Ethiopia, stochastic frontier analysis, technical inefficiency.
1. Introduction
Developing-country agriculture is frequently
characterized by low innovation capacity [1], low
productivity [2, 3], demographic pressures [4],
small-scale subsistence farming, and low levels of
market integration and value addition [3]. However,
there is significant variation across developing
countries [2]. This suggests a need for a better
understanding of the factors that influence
productivity and variations in productivity among
countries, development sectors and farm enterprises.
Ethiopia is one of the most populous countries in the
developing world and agriculture is central in its
Corresponding author: Amlaku Asres, M.Sc., research
field: innovation capacity in the livestock systems. E-mail:
asresamlaku@yahoo.com.
economy. The agriculture sector is a major source of

livelihood for 80% of the population in the country [5].
The livestock sector in particular is an indispensable
component to sustain the agricultural system, accounts
for about 45% of the agricultural GDP [6] and directly
supports the livelihoods of a large proportion of rural
as well as urban and peri-urban households in Ethiopia.
Nonetheless, there is a great concern that
productivity of livestock, especially the dairy sector,
in Ethiopia is still very low compared to other
neighboring countries. For example, milk productivity
is among the lowest in East Africa. It is estimated to
be 270 L per cow per lactation versus 498 L and 480 L
in neighbouring countries like Kenya and Sudan,
respectively [7]. A wide gap exists between actual
dairy farm production and potential productivity
152
identified in research stations, pointing to potential

technical inefficiency in their current dairy production
practices. There is, however, lack of adequate
empirical evidence regarding the production
efficiency of farmers in Ethiopia.
A number of studies have examined the potential of
the Ethiopian dairy sector to meet the expected growth
in demand as well as to improve the incomes of the
farmers [8-10]. Many of those studies, however, focus
on technological constraints of the sector including poor
genotype of local breeds, animal diseases, availability of
feed, input and output markets, and related policies. The
studies ignore an important source of growth-improving
the technical efficiency of farmers.
There are considerable inefficiency challenges that
have greatly retarded the productivity of the livestock
sector in Ethiopia. Livestock agriculture lacked the
policy level attention it deserves [11]. For example, the
Ethiopian public agricultural research staff allocated to
crop research accounts for 56.8% whereas only 14.2%
researchers focused on livestock [12]. Slow innovation
and technology transfer are observed such as shortage
of genetic material, insufficient supply of forage crop
seeds and feed concentrates. Complementary services
such as extension, credit, breeding, veterinary service,
and input-output marketing are poor [13]. All these
constraints will be considered to evaluate where further
efficiency gains are possible.
This study addresses this issue by performing a
production function analysis within a comprehensive
local level innovation systems approach to dairy
production. The innovation systems approach
examines sets of heterogeneous actors who interact in
the generation, exchange, and use of dairy-related
knowledge in processes of social or economic
relevance, as well as the institutional factors that
condition their actions and interactions [14].
Using variables that characterize a given dairy farm,
we utilize a stochastic frontier production function
analysis to estimate the production possibility frontier
under a given innovation system and a given level of
input use to determine where each farm stands in

relation to this frontier. The objective of this paper,
therefore, is to examine the extent and determinants of
technical efficiencies in the context of smallholder
dairy farmers. In addition to the overall technical
efficiency estimates, we estimate a stochastic frontier
production function to analyze differences in technical
efficiency between dairy technology adopters and
non-adopters, men and women smallholder farmers
and across districts in the study area. Our general
hypothesis in this study is that the different
components of the local level agricultural innovation
system will significantly affect the technical efficiency
of dairy production.
The rest of the paper is organized as follows.
Section two discusses the theoretical and analytical
framework that contributes to technical efficiency
analysis (productivity growth) in the context of
local-level agricultural innovation system. Section
three presents data and the empirical model used in
the econometric estimation. It is a two-step
Cobb-Douglas
stochastic
production
function
estimation model presented to analyze the technical
efficiency of dairy farmers in an innovation systems
perspective. Section four focuses on results and
discussion. Section five concludes the paper and
presented associated recommendations.
2. Theoretical and Analytical Framework

Technical efficiency is a measure of a farms
productive performance [15-17]. In the context of this
study, it can be defined as the ability of a smallholder
dairy farmer to obtain maximal output from a given set
of inputs. Technical inefficiency on the other hand is
the deviation of an individual smallholder farms
production from the best practice frontier. The level of
technical efficiency of a particular farm is based upon
deviations of observed output from the efficient
production frontier [18]. If the actual production point
lies on the frontier, it is perfectly efficient. If it lies
below the frontier, then it is technically inefficient. The
distance between the actual to the achievable optimum

production from given inputs, indicates the level of
production inefficiency of the individual firm [18, 19].
We consider local level (district) agricultural
innovation system perspective as a theoretical
construct that contributes to productivity growth
through four main components: knowledge and
education, business and enterprise, bridging
institutions, and the enabling environment, based
broadly on a construct developed by Arnold and Bell
[20] and adapted to the sphere of agriculture and
agricultural development by World Bank [21]. In this
study, the key elements which proxy the local level
agricultural innovation system are described as
follows. The knowledge and education domain
captures the contribution of education to technological
change and proxies by education of the household
head. The business and enterprise domain captures the
set of input-output market actors and activities that
leverage dairy production inputs to farmers and milk
outputs for markets. Within this component, group
membership of the household head assumed to proxy
the marketing role is included. Bridging institutions
represent the domain in which individuals and
organizations that leverage public extension services
in the innovation process. This component
incorporates two variables: extension visited by
extension agent and technical training given to the
farmer on dairying. Circumscribing these domains are
the enabling or frame conditions that foster or impede
innovation, including public policies on innovation
and dairy enterprise, for example, credit availability,
off-farm income opportunity and land availability.
Generally, two approaches are used to obtain
estimates of farm household efficiency: parametric
and non-parametric. The non-parametric approach is
implemented using data envelopment analysis (DEA)
while the parametric approach uses econometric
techniques. These two methods have a range of
strengths and weaknesses which may influence the
choice of methods in a particular application and the
153
constraints, advantages and disadvantages of each

approach have been discussed by Coelli [22] and
Coelli and Perelman [23]. However, it is well
documented that the DEA approach works under the
assumption of absence of random shocks in the data
set. Since farmers always operate under uncertainty,
the present study employs a stochastic production
frontier approach introduced by Aigner et al. [16];
Meeusen and Broeck [17] and refined by Battese and
Coelli [24]. Following their specification, the
stochastic production frontier can be written as:
yi f xi ; i 1,2,........N
th
(1)
farm,
xi is the i th input used by the j th farm and is

a vector of unknown parameters and is a
where yi is the output of milk for the j
i vi ui
composed error term which can be written as:

where
vi
(2)
is a systematic random error which
represents random variations outside the control of the

farmer such as disease, weather condition, natural
disaster, luck, fires, and other exogenous random
factors [25] and assumed independently and identically
distributed with zero mean and constant variance
N 0 , v2 . The error term
non-negative term ui 0 representing the

deviations from the frontier production function, which
is attributed to controllable factors (technical
inefficiency). This one sided error term can assume
various distributions such as truncated-normal,
half-normal, exponential, or gamma [16, 17]. However,
in this paper, it is assumed ui to be distributed
2
identically and independently half-normal N 0, u
ui is one sided
as typically done in the applied stochastic frontier

literature. Furthermore, the two components vi and
ui are also assumed to be independent of each other.
For a detailed review of the literature on stochastic
production function for developing country agriculture,
see Bravo-Ureta and Pinheiro [26], Coelli [27].
Stochastic production frontier functions have been
widely used to assess the economic efficiency of
agricultural production in recent years [28-38].
154
3. Data and the Empirical Model

3.1 Data and Variable Definition
The data used for our empirical analysis are drawn
from the household survey of 304 rural households in
four districts of Northwest Ethiopia (Amhara region).
Three villages were randomly selected from each of
the four districts and therefore the study has been
conducted in a total of 12 villages. The study focused
on innovations that were introduced by the
Ethio-Austrian Integrated Livestock Development
Project (ILDP) that has been implemented for 10 years
(1998-2008) in three phases at 14 districts. ILDP is an
Austrian government financed program implemented
by the Regional Bureau of Agriculture. The project
supports the governments endeavor to improve
livestock productivity and income so as to contribute
to the food security conditions of the farmers in
Northwest Ethiopia.
The surveys were conducted using structured
interviews with multistage stratified sampling
technique to collect quantitative household level
Table 1
Description of output, input and technical inefficiency variables.
Variables
Ln output (Y)
Inputs
Ln roughage
Ln concentrate
Ln labor
Ln health
Ln breeding cost
Ln hay
Inefficiency variables
Age
Age2
Family size
Education
Farm size
Credit availability
Extension visit
Training
Off-farm income
Group membership
a
information. Within each district, a two-stage

selection process had been followed, selecting first
two villages purposively on the bases of their relative
importance in having more project beneficiaries and
one non-beneficiary village, and finally randomly
selecting households (HHs) within each of the
selected villages. The data collection followed
two-pronged approach. First, sample households were
randomly selected from a list of 922 farmers who
participated in dairy technology interventions
(adopters, n = 80) from the ILDP project beneficiaries.
Second, a wider sub-sample of smallholders, that had
no direct intervention with the project was randomly
selected (non-adopters, n = 224) from the same
districts.
The data collected are cross-sectional data obtained
through the above mentioned procedure. The
questionnaires were administered to dairy farmers and
were designed to elicit information on the
socio-economic characteristics of the respondents and
also on the operational systems adopted. Details of all
variables are presented in Table 1.
Description
Natural log. of household total milk output in Birra
Expected sign
Animal feed intake in kg (produced on the farm)

Animal feed intake in kg (purchased industrial by-product)
Number of adults working on the farm (ages between 15-64)
Veterinary service expense in Birr
Breeding service expense in Birr
Animal feed expenditure in Birr (purchased feed)
+
+
+
+
+
+
Age of the household head in years

A proxy for years of farming experience of the household head
+/Number of family members in the household
Years of formal schooling of the household head
Total land owned by the household in hectares
1 if the farmer gets credit in the production year; 0 otherwise
Number of times the household visited for advice by the extension agents during the
production year
1 if the household attended any dairy production training sessions; 0 otherwise
1 if the household gets income from sources other than farming; 0 otherwise
1 if the household is a member of any kind of farmers group; 0 otherwise
-
Negative sign in the inefficiency variables indicates a positive effect on efficiency impact.
A priori expectations about the relationship of the

variables used in determining the factors influencing
technical efficiency are based on the analytical
framework and from previous empirical results. The
data are described as follows:
In the production function models, the dependent
variable, output (Y), is the natural logarithm of the
annual milk produced per farm measured in the value
of total milk production in Ethiopian Birr. Average
market price of the districts is used to estimate output
values.
The various inputs for milk production are: (1)
produced on farm dairy feed (roughage) measured in
terms of the quantity (kg) of total fed to milking cows
on the farm in a given year; (2) purchased dairy
concentrate feed measured in terms of the quantity (kg)
of total fed to cows on the farm in a given year; (3)
family labor measured in number of persons working
in the farm; (4) animal expenses consisting of
veterinary medicine, breeding services and
supplementary feed cost (hay). The estimated
coefficients on all inputs are hypothesized as positive.
Although accurate data on such milk production
inputs are not easily obtainable in the Ethiopian
traditional agricultural sector in general because of
measurement problems, an endeavor was made to
reduce the error of margin. We first collect the data
(for example, number of heap of roughage produced
on farm) subjectively from the holder and then
recorded after correcting it with the agreed conversion
rate into kilogram with the field experts in each
village.
The inefficiency variables that were used to explain
the character and performance of a given dairy farmer
in the study area were classified into two groups: (1)
household characteristics and (2) factors that can
reflect (proxy) the capacity of the local level
agricultural innovation system (AIS).
They are described as follows: of the household
characteristics, age of the household head proxies for
experience and predisposes farmer to better farming
155
techniques and is assumed to increase the productivity

of the farm and the higher the farmers experience, the
greater the technical efficiency was assumed, such that
the estimated parameter is predicted as positive for
both age and age squared term. Family size was
hypothesized to have positive effect to the technical
efficiency, because bigger household size could mean
a more secure labor source for livestock production.
Of the variables that proxy local level agricultural
innovation system, education was considered as the
number of years of formal schooling and was
supposed to have positive relationship with level of
efficiency. Availability of credit in time would
facilitate farmers to procure inputs timely thereby
increasing productivity and decreasing inefficiency.
Regular visits of an extension agent would spur
farmers to increase the efficiency. Access to technical
trainings was hypothesized to reduce the inefficiency.
It was assumed that the farmer with off-farm income
augments its access to a financial capital to purchase
inputs, which lead to higher efficiency. Group
membership was considered as a mechanism for
information sharing and makes members more
efficient than the non-members. The larger the farm
size (land) was hypothesized to have positive effect to
the technical efficiency because farmers may have
more fodder production to feed their animals and
maintain the productivity of the farm. The estimated
coefficients on all inefficiency variables are, therefore,
hypothesized as positive.
3.2 Characteristics of Sample Farmers
Table 2 presents the descriptive statistics of
dependant and independent variables used in the
stochastic frontier production analysis and on the
determinants for the dairy farm household efficiency
analysis. The data set contains information of 304
sample households (281 men and 23 women) and the
average age of a household head is 48 years. The
average experience of a farmer in dairy farming is 23
years, but farmers have experiences ranging from 5 to
156
Table 2 Summary statistics of the variables for dairy farmers in northwest Ethiopia in 2010/11.
Variables
Milk output (Birr)
1. Inputs
Roughage (kg)
Concentrate (kg)
Labor (man days)
Health expense(Birr)
Breeding expense(Birr)
Hay purchase(Birr)
2. Inefficiency factors
2.1 Household characteristics
Age
Experience in farming (years)
Family size (no.)
2.2 Proxy factors to AIS
Education (years)
Farm size (ha)
Credit availability (1/0)
Extension visit (no.)
Training (1/0)
Off-farm income (1/0)
Group membership (1/0)
Dairy tech. adopters

Mean
Range
4,354 (5,003)
0-30,000
Dairy tech. non-adopters

Mean
Range
1,595 (2,940)
0-32,000
All respondents
Mean Range
2,321 0-32,000
2,759 (2,050)
500 (1,044)
4 (2)
120 (105)
14 (36)
1,433 (1,506)
0-10,000
0-7,200
2-8
0-500
0-300
0-7,000
2,041 (3,104)
195 (370)
4 (2)
108 (106)
7 (14)
806 (1,587)
0-40,000
0-2,400
1-10
0-720
0-74
0-13,500
2,230
275
4
111
9
971
0-40,000
0-7,200
1-10
0-720
0-300
0-13,500
47.2 (9.5)
23.17 (9.6)
7.36 (1.75)
28-75
8-56
4-11
48.4 (11.9)
24.85 (12.2)
6.76 (1.8)
22-83
5-60
2-12
48.11
24.41
6.92
22-83
5-60
2-12
4.54 (2.27)
1.64 (0.78)
0.4 (0.49)
2.93 (1.3)
0.66 (0.48)
0.28 (0.45)
0.96 (0.19)
0-11
0-3
0-1
1-5
0-1
0-1
0-1
3.55 (2.66)
1.46 (0.82)
0.43 (0.5)
2.69 (1.16)
0.62 (0.49)
0.2 (0.4)
0.85 (0.36)
0-12
0-3.75
0-1
1-5
0-1
0-1
0-1
3.81
1.51
0.42
2.75
0.63
0.22
0.88
0-12
0-3.75
0-1
1-5
0-1
0-1
0-1
Standard deviations are given in parenthesis.
60 years. On average, dairy farmers have five

household members, four years of formal schooling,
and own 1.51 hectare of land. Of the 304 farmers,
63% have attended technical training on dairying,
42% and 22% have had access to credit and off-farm
income, respectively, and 88% are members at least in
one farmers group. On average, mean milk output
value in the study area is 2,321 Birr, with a very high
variation among farms. Farmers used on average
2,230 kg of roughage, 275 kg of concentrate, and hay
at a value of 971 Birr per farm, but the variation is
quite large.
The descriptive statistics of variables for adopters
and non-adopters show that there is mean output and
input use difference. This is because adopters of dairy
technology have a higher use of land and intermediate
inputs to increase productivity. The mean output of
milk for farmers within the adopters group was about
nearly three-fold than non-adopter farmers. The
roughage, concentrate, health, breeding and hay used

by the farmers within the adopters were about 35%,
156%, 11%, 100%, 78% greater than that used by
non-adopters.
Characteristics of household related variables (age,
farmers experience in dairying and household size)
and local level agricultural innovation system
indicators such as education, farm size, number of
extension visit, those participated in technical training,
off-farm income availability and membership in a
farmers group vary slightly across groupings, and
adopters exhibit the highest. More non-adopters (43%)
than adopters (40%) reported having credit access.
3.3 Empirical Model
As in Battese and Coelli [24], this paper follows a
two step estimation model. The first step involves the
specification and estimation of the stochastic frontier
production function and the prediction of the technical
inefficiency effects, under the assumption that these

inefficiency effects are identically distributed. The
second step involves the specification of a regression
model for the predicted technical inefficiency effects.
Step 1 Estimating the Stochastic Frontier
We introduce here the Cobb-Douglas form of a
standard stochastic frontier production function model.
The Cobb-Douglas functional form is chosen because
it provides an adequate representation of the
production process, since we are interested in an
efficiency measurement and not an analysis of the
production structure [39]. In addition, the
Cobb-Douglas (CD) functional form (in spite of its
restrictive properties) is used because its coefficients
directly represent the elasticity of production. It is also
widely applied in farm efficiency analysis for both
developing and developed countries [26, 40-42].
The following model is estimated using Frontier
4.1c program [22]:
ln Yi o k ln X ik Vi U i
6
k 1
(3)
where Yi is the value of milk output for observation

1
i in Ethiopian Birr ; X 1 quantity of roughage fed
to cows in kg, X 2 quantity of concentrate fed to
cows in kg, X 3 family labor man days, X 4
veterinary expense in Birr, X 5 breeding cost in Birr,
X 6 value of hay purchased to fed the cows in Birr;
o is intercept and k is an 1xk vector of
parameters to be estimated; Vi is iidN 0 , v2

random stochastic disturbance term, independently
distributed of the U i ; and U i is a non-negative
random variable associated with the technical

inefficiency of production which is assumed to be
independently distributed.
Step 2 Identifying Sources of Technical Inefficiency
for explaining technical inefficiency 1 TEi for

Cobb-Douglas stochastic frontier production function:
The following multiple regression model was fitted
U i 0 i X i j
(4)
where, U i is technical inefficiency; X i represents

1
Ethiopian Birr = US$0.06.
157
explanatory variables including: age of the household

head (years), farmers experiences in farming (years),
family size (no.), education, i.e., years of schooling of
the household head, farm size (ha), availability of credit
(binary), access to extension services (categorical),
availability of technical training opportunity on
dairying (binary), off-farm income (binary), affiliation
to farmers group (binary); 0 is the intercept; i is
the unknown parameters to be estimated; and j the
unobservable random disturbance term.
The maximum-likelihood estimates for all the
parameters of the stochastic frontier and inefficiency
model, defined by Eqs. 3 and 4, are simultaneously
obtained by using the program, Frontier Version 4.1c
[22], which estimates the variance parameters in terms
of the parameterization:
and
s2 v2 u2 ;
2
u
2
s
and 0 1
(5)
(6)
where the parameter must lie between 0 and 1.

The technical efficiency of production of the
th
i farmer in the appropriate data set, given the level
of his inputs, is defined in terms of the observed

output by Yi * to the corresponding frontier output
Yi , that is
TEi Yi / Yi * exp xi Vi U i / exp xi Vi exp U i (7)
Therefore, the technical efficiency of a farmer is

between 0 and 1 0 TE 1 and is inversely
related to the level of the technical inefficiency effect
[32]. The technical efficiencies can be predicted using
the Frontier program which calculates the
maximum-likelihood estimator of the predictor for Eq.
7 that is based on its conditional expectation [43].

The maximum likelihood (ML) estimates of the
parameters of the Cobb-Douglas stochastic frontier
production function and the inefficiency model are
presented in this section. The results are presented for:
(1) pooled data; (2) dairy technology adopters and
non-adopters; (3) men and women dairy farmers; and
158
(4) variation across districts. First, we present the

coefficients for the stochastic production function and
then present the technical inefficiency coefficients and
its determinants.
4.1 Pooled Data
4.1.1 The Stochastic Production Frontier Estimation
of Smallholder Dairy Farmers
The results of the maximum likelihood estimates of
the stochastic frontier production functions for dairy
milk production are presented in Table 3. Findings
reveal that coefficients of concentrate, labor and
breeding are found to be positively significant to the
dependant variable dairy milk output. The positive
coefficient of concentrate, labor and breeding with

respect to milk production implies that the higher the
use of these inputs, the higher the total level of milk
production.
From the nature of the Cobb-Douglas production
function fitted, since the model is a log linear model,
the coefficients represent elasticity of output with
respect to the respective inputs. Production elasticities
indicate the percentage change in output relative to a
percentage change in input if other factors are held
constant. Accordingly, the elasticity of milk output
with respect to concentrate is 0.1078 meaning that
10% change in the total concentrate use will bring
about 1.08% change in the output of milk production
Table 3 ML estimates of the Cobb-Douglas production frontier and inefficiency models.

Variable
Estimate
Standard error
Stochastic frontier production
Constant
7.7526
0.4670
Ln roughage
-0.0445
0.0425
Ln concentrate
0.1078
0.0321
Ln labor
0.4599
0.2141
Ln health
0.0694
0.0546
Ln breeding
0.1826
0.0646
Ln hay
-0.0041
0.0242
Inefficiency model
Constant
-19.9794
42.9463
Household characteristics
Age
-0.7184
1.4639
Age2
0.9751
1.4815
Family size
1.3106
1.2822
Knowledge and education domain
Education
-1.6999
1.6947
Business and enterprise domain
Group membership
1.9282
7.8305
Bridging institutions domain
Extension
-0.4203
1.7345
Training
0.3434
4.6725
Enabling environment domain
Credit availability
1.1133
3.3495
Off-farm income
-5.1824
7.4664
Farm size
-4.7861
5.1519
Variance parameters
2
136.0373
125.1240
0.9985
0.0014
Log likelihood function
-701.7271
LR test (one-sided error)
153.0634***
*** significance at the 1% level; ** significance at the 5% level.
t-statistics
16.6016***
-1.0477
3.3558***
2.1483**
1.2707
2.8274***
-0.1682
-0.4652
-0.4908
0.6582
1.0221
-1.0031
0.2462
-0.2423
0.0735
0.3324
-0.6941
-0.9290
1.0872
736.9893***
if other factors are held constant. Labor has an

elasticity of 0.4599 meaning that for 10% change in
labor input; output of milk will change by 4.6%. The
same goes for breeding input with an elasticity of
0.1826 meaning that a 10% change in the expenditure
on breeding will bring about a 1.83% change in the
output of milk production in the study area.
4.1.2 Inefficiency Model
The inefficiency effects described above were
estimated against the components of the household
characteristics and the local level innovation systems
approach (Table 3). The result shows that all the
selected variables in the model produced
non-significant coefficients to the inefficiency model.
The inefficiency model, although statistically not
significant, age, education, farm size, extension visit
and off-farm income opportunity are having negative
sign as expected, indicating that these factors led to
decrease in technical inefficiency or are important
factors to increase production efficiency in the dairy
production system. The coefficients in the inefficiency
function are inefficiency effects and therefore a
positive coefficient implies a negative effect on
performance while a negative sign indicates a positive
impact on efficiency.
Among the variables representing efficiency effects
from the AIS framework in Table 3, measures of
education in the knowledge and education domain
shows the expected effects in reducing inefficiency.
The role of education in technology adoption has been
extensively documented. Schooling has been shown to
provide substantial externality benefits by increasing
farm output and shifting the production frontier
outwards [32, 44, 45]. In the business and enterprise
domain, group membership of the household head is
found to be non-significant and negatively related
with dairy production efficiency despite our
expectation that it increases dairy production
efficiency by facilitating input-output marketing. This
could partly be due to innovation is supply-driven by
extension rather than market-driven by product
159
demand and also may be due to the disaggregated

structure of dairy input-output marketing systems in
Ethiopia. Extension from the bridging institutions
domain is not significant, but it had the expected sign
which indicates that the involvement of extension
agents to visit dairy farmers tends to reduce the
technical inefficiency of milk production. In the
enabling institutions domain, off-farm income and
farm size are not significant, but they had the expected
signs which indicate both variables had positive
contribution to dairy production efficiency.
The significant gamma () statistic, which is a

measure of the overall, of 0.9985 indicates that about
99.85% variation in the output of milk production
would be attributed to technical inefficiency effects
(those under farmers control) alone while only
0.0015% would be due to random effects, i.e., beyond
the farmers control (Table 3). The high value of
parameter highlights the importance of inefficiency
effects in explaining the total variance in the model.
The level of technical efficiency is predicted
simultaneously with the estimated production function
and it was found that the mean technical efficiency is
about 26% (Table 4). Thus in the short run, there is a
scope for increasing dairy milk production by about
74% using the same level of inputs, but improved
management and resource reallocation. One of such
measures is addressing, the issue of negative elasticity
of dairy feed (roughage and hay), other non-significant
input (veterinary service) and improving the innovation
Table 4 TE frequency distribution and deciles range of
dairy farmers.
Efficiency level
0.80-0.89
0.70-0.79
0.60-0.69
0.50-0.59
< 50
Total
Mean efficiency
Minimum
Maximum
Frequency
4
21
13
20
%
1.3
6.9
4.6
6.6
246
80.6
304
0.2617
0.02
0.85
100
160
capacity of the local level agricultural innovation

system actors. The cumulative and frequency
distribution of dairy farmers efficiency scores are
presented in Table 4. Above average (> 50%)
efficiency for 19.4% of the farmers in dairy
production could be the result of ILDP implemented
in the study area to reduce poverty and support the
enhancement of service delivery. Those activities
could have a long-term impact that could have
spillover effects to other non-project farmers.
4.2 Technical Efficiency of Dairy Technology Adopters
and Non-adopter Farmers
4.2.1 The Stochastic Production Frontier Estimation
The stochastic production frontier model estimates
and those for the technical inefficiency model for
adopters and non-adopter farm households are
presented in Table 5.
In the model, a coefficient of breeding is found to
be positively significant to the dependant variable
dairy milk output to both groups. The coefficient of
health and breeding are found to be significantly
positive in case of adopters. Similarly, the coefficients
of concentrate, labor and breeding are significantly
positive in case of non-adopter farmers. The
negatively non-significant coefficient of roughage and
positive but non-significant coefficient of hay inputs
in both groups imply no effect to the output.
The output elasticities of inputs for both adopters
and non-adopter farmers are variable. For the adopters
group, output elasticity of inputs was highest for
breeding (0.1798), followed by health input (0.1105).
In the non-adopters group, output elasticity of inputs
was highest for labor (0.5706), followed by breeding
Table 5 Maximum-likelihood estimates for parameters of the Cobb-Douglas stochastic frontier production functions for
dairy technology adopter and non-adopter farmers.
Variable
Adopters
Non-adopters
Stochastic frontier
Coefficient
Std.dev
Coefficient
Std.dev
Constant
8.4665
0.0696
7.0735
0.6692
Ln roughage (kg)
-0.0296
0.0402
-0.0012
0.0598
Ln concentrate (kg)
0.0470
0.0613
0.1186***
0.0457
Ln labor (man days)
0.2094
0.3088
0.5706***
0.3046
Ln health expense(Birr)
0.1105***
0.0437
0.0316
0.0843
Ln breeding expense(Birr
0.1798***
0.1741
0.1790**
0.1013
Ln hay purchase(Birr)
0.0072
0.0509
0.0054
0.0400
Inefficiency model
Constant
0.1528
0.9992
-20.5590
29.29
Age
0.1548
0.2274
-0.5387
0.9774
Age2
-0.0996
0.2354
0.6632
0.9295
Family size (number)
-0.7839
0.7971
1.8860*
1.2468
Education (years)
-1.2337***
0.4255
-0.2010
0.4045
Farm size (ha)
1.1279
0.9694
-3.4067
2.7478
Credit availability (1/0)
-2.1958**
1.1189
2.7738
3.2894
Extension (number )
0.3813
0.8949
0.1217
1.3739
Training (1/0)
-0.9522
1.0179
3.0090
3.3153
Off-farm income (1/0)
- 3.3906***
1.2709
1.0874
4.2103
Group membership (1/0)
1.2093
1.0353
3.8079
5.2752
Variance parameters
2
17.2752***
0.9451
78.1963*
53.3134
0.9999***
0.0000
0.9958***
0.0031
Log likelihood function
-146.93
-531.73
LR test (one-sided error)
85.87***
88.96***
The coefficients in the inefficiency function are inefficiency effects and therefore a positive coefficient implies a negative effect on
performance while a negative sign indicates a positive impact on efficiency. Significant at * 10%, ** 5%, *** 1%.
(0.1790), and concentrate (0.1186). The overall results

indicate that four inputs, concentrate, labor, health and
breeding have a major influence on milk output of
both adopters and non-adopter farmers.
4.2.2 Inefficiency Model
In the inefficiency model (Table 5), although
statistically not significant, age, farming experience
(age2), family size, education, farm size, credit,
technical training, and off-farm income are negative
indicating important factors to increase production
efficiency in one or the other groups.
The bridging institutions domain (extension visit)
and
business
and
enterprise
domain
(group
membership) proxies do not seem to be important

factors affecting farm household efficiency in both
groups. The explanation to this could be, extension in
Ethiopia has a supply driven nature and its quality is
very low and group memberships have no direct effect
and are less likely contribute to technology adoption
decisions [45].
Knowledge and education domain (education) and
among the enabling environment domain (credit
access and off-farm income opportunity) variables are
positive and significant factors affecting farm
household efficiency in the adopters group. The role
of education in technology adoption has been
extensively documented. Credit and off-farm access
also contributes to farmer adoption of new
technologies and practices by easing farmers liquidity
constraints.
Unlike the adopters group, the positive but
statistically significant coefficient for family size
variable in the non-adopters group indicating possible
excessive use of man power which is a problem of
allocative efficiency. The negative sign on age shows
that younger head of households are productive in the
non-adopters group and hence important factor to
increase production efficiency.
The parameter 2 and results show
significant at 1% level for both groups (except for
non-adopters group 2 that was significant at 10%).
161
The significance value of the 2 shows the presence

of inefficiency effects in dairy milk production in the
study area while the significant of 0.9999 and
0.9958 indicates that about 99.99% and 99.58%
variation in the output of the dairy milk production
would be attributed to random effects, for adopters
and non-adopter farmers, respectively.
The mean predicted technical efficiency for farmers
within the adopters group was estimated to be 0.40
with 0.25 standard deviations, while for non-adopter
farmers, the mean technical efficiency was 0.21
(Table 6). The fact that both groups have technical
efficiency levels below 50%, suggests a relatively
very low level of innovation capacity in the dairy
production system which resulted substantial technical
inefficiency in dairy production operations in both
adopters and non-adopter farmers given the available
technologies.
4.3 Technical Efficiency of Men and Women Dairy
Farmers
Disaggregating the dairy production data by gender
shows that women headed households were more
technically efficient than male headed households in
the study area. This could be due to the fact that
women spend substantial amounts of time doing
livestock activity [46]. This is denoted in Table 7 with
an efficiency score for the women farmers of 43.89%
compared to 23.54% for men farmers.
Table 6 Technical efficiency frequency distribution and
deciles range of dairy farmers by adoption status.
Deciles range Adopters (N = 80)
of TE
Frequency %
0.80-0.89
1.2
0.70-0.79
17.5
0.60-0.69
7.5
0.50-0.59
13.8
60
50
Total
100
Mean TE
0.40
Std.dev.
0.25
Minimum
0.03
Maximum
0.85
Non-adopters (N = 224)
Frequency
%
0.9
3.1
3.1
5.0
87.9
100
0.21
0.20
0.02
0.85
162
Table 7
Summary of farm efficiency descriptive statistics.
Variable
Efficiency combined
Efficiency adopters
Efficiency non-adopters
Efficiency men
Efficiency women
Efficiency by District Gonderzuria
Efficiency by Lay armachiho
Efficiency by Wogera
Efficiency by Debark
Mean
0.2617
0.4000
0.2124
0.2354
0.4389
0.2190
0.2904
0.3092
0.2061
StD
0.2280
0.2481
0.1987
0.2274
0.3757
Min
0.02
0.03
0.02
0.02
0.03
0.03
0.02
0.02
0.02
Max
0.85
0.85
0.85
0.85
0.85
0.85
0.85
0.85
0.77
4.4 Technical Efficiency of Dairy Farmers across

Districts
It is also possible to infer something about the
general status of TE in each district. For example,
Table 7 shows the predicted TE values. Values vary
from 20.6% in Debark to 30.9% in Wogera district.
Districts relatively with higher technical efficiencies
above the overall average are those progressing with
breeding, education, access to off-farm income
opportunity and group membership.
varied widely across dairy technology adoption status,

gender and districts. Accordingly, dairy technology
adopting farmers have relatively high mean efficiency
scores (40%) compared to non-adopters (21%).
Similarly, women farmers have relatively high mean
efficiency scores (44%) compared to men farmers
(24%) and hence in both cases calling for a focus on
efficiency enhancing investments. Education, farm
size, extension visit and off-farm income opportunity
were found to be efficiency enhancing.
We examined separately technical efficiencies of
dairy
technology
households.
The
adopting
results
and
non-adopting
indicated
significant
difference in productivity changes and technical

inefficiencies between adopters and non-adopter
farmers, which could be explained by various farm
specific and household characteristic variables. The
results show veterinary and breeding services are
important inputs and are strongly associated with total
output. The mean technical efficiencies calculated for
adopters (40%) and non-adopter farmers (21%),
5. Conclusions
The paper uses a Cobb-Douglas stochastic frontier
analysis of production functions to estimate the level
of technical efficiency of dairy farmers in four
districts during 2011 production year. The stochastic
production function was modeled in such a way that
local level agricultural innovation systems framework
and indicators of its different domains (the knowledge
and education domain, the business and enterprise
domain, the bridging institutions domain and the
enabling environment domain) serve as an
environment that determines the level of technical
inefficiency. The production function and the
inefficiency effects were estimated simultaneously.
The results showed that the overall mean efficiency
score among sampled dairy farmers is about 26% and
there is room for significant increase of production by
reallocation of the existing resources. Despite
significant variation from the frontier, it is also
revealed that individual farm households efficiency
indicating both groups working under inefficient

condition, with high potential for reducing inputs or
increasing outputs in the range of 60%-79%. The
obtained
significant
measures
of
differences
efficiency
in
indicate
productivity
that
changes
between adopter and non-adopter households are

attributed to previous project support by ILDP.
The results suggest that the government might have
to address the technological and institutional
constraints of its extension services. Additionally,
since farmers are yet far from reaching the production
frontier, government may consider reforming the
current agricultural extension system by involving a
large number of service providers, including allowing
more private service delivery actors. This is supported
by the finding that in the adopters and non-adopters,
access to extension and group membership (a proxy to
marketing) was not at all responding to higher
household farm efficiency. We find that quality of the
agricultural extension and marketing services are
important if farmers in the dairy sector are to

significantly increase productivity. However, this
should be supported by reforming the current
agricultural extension system to address institutional
and policy issues that constrain effective agricultural
innovation system.
We note that the present study is based on data
from a single production period and specific locality, a
follow-up is recommended to examine national level
technical efficiency in the Ethiopian dairy system.
This would help policy makers in strengthening the
capacity of and investing more in extension and
marketing services.
[8]
[9]
[10]
[11]
Acknowledgments
The authors greatly acknowledge OEAD-Gmbh
(Austrian Agency for International Cooperation in
Education and Research), Sustainable Resource
Management Program (SRMP) of North Gondar Zone,
and Sustainable Water Harvesting & Institutional
Strengthening Project (SWHISA), both in Amhara
region of Ethiopia, for financing this study and
facilitating logistics support.
[12]
[13]
[14]
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D
DAVID
PUBLISHING
Richness and Diversity of Ants and Beetles in

Genetically Modified Cotton Field in Brazil
Carla Cristina Dutra, Marcos Gino Fernandes, Josu Raizer and Camila Meotti
Department of Entomology, Biological Science Faculty, Federal University of Grande Dourados, Rodovia Dourados-Itahum, Km 12,
Cidade Universitria, PO-Box 533, CEP 79804-970, Dourados, MS, Brazil
Abstract: Genetic engineering has created many genetically modified (GM) crop varieties that express the cry toxin from the
bacterium Bacillus thuringiensis (Bt). The cry toxin, synthesized during plant growth, has insecticidal properties, and can be
expressed anywhere in the plant. This study aimed to ascertain the richness and species diversity of edaphic Formicidae and
Coleoptera in GM cotton fields compared with the conventional non-transformed cotton crop. We analyzed data from commercial
cotton fields located in the municipality of Maracaju, Mato Grosso do Sul, Brazil. The experiment was conducted during the
reproductive period of cotton, employed two treatments: Bt cotton and non-Bt cotton. Samples were collected with pitfall traps.
Formicidae species richness in the Bt area was lower than in the non-Bt area, but species composition did not differ between the two
treatments. Species composition of Coleoptera communities also differed between the treatments because some species were more
abundant in the Bt cotton area. On the other hand, the species richness of this group was similar in both areas.
Key words: Bacillus thuringiensis, bollgard cotton, Bt crops, Coleoptera, Formicidae.
1. Introduction
Cotton, Gossypium hirsutum (Malvaceae), has been
genetically modified (GM) by the insertion of the cry
genes, which code for insecticidal proteins cry (type
-endotoxin) from a common soil bacterium, Bacillus
thuringiensis (Bt). The cry proteins kill insects that feed
on Bt modified crops by causing the osmotic rupture of
the epithelial layer of the digestive tract, thus acting as
an efficient population control agent [1-3].
Bt cotton is cultivated in many parts of the world,
including Argentina, Australia, China, India, South
Africa and United States [3]. Nowadays, Brazil is the
second largest producer of GM crops, with production
area of 30.3 million hectares, trailing only the United
States (69 million ha) [4]. In Brazil high scale
production are soybeans, maize and GM cotton.
Cotton is the transgenic plant most extensively
cultivated in the mid-west of Brazil, this region is
Corresponding author: Carla Cristina Dutra, Ph.D., research
fields: integrated pest management and risk assessment of
transgenic plants. E-mail: carlacristina.dutra@gmail.com.
responsible for 80% of the cotton produced in the

country [5]. The GM cotton Bollgard (MON531),
legalized in Brazil in 2005 [6], expresses the Cry1Ac
protein, effective against caterpillars such as Heliothis
virescens (Lepidoptera: Noctuidae), Alabama
argillacea (Lepidoptera: Noctuidae) and Pectinophora
gossypiella (Lepidoptera: Gelechiidae).
The little available data on the effects of GM plants
on the biota indicates that transgenic crops have an
impact on the microorganisms that consume the
metabolic wastes they leak into the environment [7].
The main risk associated with Bt cotton, which results
from the accumulation of toxins in the soil, seems to
be associated with the decrease of local biodiversity
and the simplification of the functional dynamics of
soil organisms [8]. It is widely acknowledged that Bt
cotton plants produce the cry protein in the leaves and
flower buds [9], and exude them through the roots
[10]. Several factors modulate the accumulation of
transgenic toxins in the soil, for instance the amount
of toxins in the plant tissues, toxin resistance to
166
degradation, as well as the physical and chemical

properties of the soil [11, 12]. Toxin accumulation in
the soil may have an effect on the organisms
responsible for recycling organic matter, i.e.,
decomposers, thus reducing or even preventing
fragmentation
of
composts
like
cellulose,
hemicelluloses and lignin, with negative consequences
to plant productivity [10].
The soil encompasses more than 90% of the
biodiversity in agroecosystems [13], and the edaphic
fauna impacts the soil in various ways, as for instance
through excavation, ingestion and transport of organic
matter
[14].
Two
different
groups
of
insects
representing the edaphic fauna are ants (Formicidae)

and beetles (Coleoptera). Ants, one of the dominant
edaphic groups [15], play an important role not only in
the redistribution of particles, nutrients and organic
matter in the soil, but also in the improvement of soil
permeability to water and air by the construction of
pores and underground ducts [16]. Besides their role in
soil maintenance, some generalists ants help protect
crops [17], for example by preying upon cotton pest
insects [18]. In a manner similar to ants, beetles, the
most diverse group of insects, also modify the physical
structure of the soil, prey upon different groups of
invertebrates [19], and have a role in the decomposition
of organic matter. Thus, ants and beetles, as other
organisms that are not targeted by the Bt toxins, can be
beneficial to agricultural settings [20].
Some field studies show no effect of Bt toxin from
plants on the diversity and abundance of non-target
insects [21-23]. However, on the field, there are
continuous exposures of non-target organisms to cry
protein from Bt plants. Know the diversity of
arthropods associated with crops is essential for
ecological studies and integrated pest management
(IPM).
This research aimed to describe the richness and
diversity of species of ants and soil beetles in two
contiguous cotton areas, one cultivated with GM
cotton crop resistant to insects (Bt cotton) and the
other area cultivated with conventional cotton crop

(non-Bt cotton).

2.1 Description of the Experimental Area
Sampling was conducted in a commercial cotton
field located in the municipality of Maracaju, Mato
Grosso do Sul, Brazil (2136'52''S, 5510'06"W,
elevation 384 m). Cotton sowing took place in
November, 2007. Two contiguous areas (20 m 100
m each) were delimited: one of them in a field
containing the Bt cotton (NuOpal, Bollgard,

MON531) cultivated in 50 hectares, and the other
cultivated with conventional cotton (DeltaOpal) in
13 hectares. Two clearings were made manually and
insecticides and herbicides were not applied during
the developmental period of the plants.
2.2 Samplings
Beetles and ants were collected by pitfall traps
distributed between on the two treatment areas. Traps
were made from pet plastic bottles (2 L volume) cut in
half, with the upper part turned upside-down and
inserted on the lower half, making a funnel. The entire
trap was then inserted into the soil, maintaining the
upper border at ground level. The inside of the trap
was filled to 2/3 with sodium hypochlorite (0.1%), a
preservative, and a few drops of detergent (to break
the water tension and allow the insects to submerge).
Each trap was protected from the rain and other
impurities with a piece of wood (15 cm 15 cm)

secured 2.5 cm above the trap with metal rods. The
liquid inside the traps was replaced every three days
between January 8 and February 14, 2008
(corresponding to the reproductive period of the
cotton). There were 20 pitfall traps for each sampling
area, and the distance between the traps was random.
The insects collected were put in 70% ethanol for
posterior triage and initial identification at the
Entomology Laboratory at the Universidade Federal
da Grande Dourados (UFGD).
2.3 Species Identification

Species identification was conducted by taxonomists
at the Universidade Federal do Paran (UFPR).
Morphospecies were assigned to the entities that could
not be identified to species. Voucher specimens from
167
Table 1 Frequency of species of Formicidae registered in

pitfall traps on a field of Bt and another non-Bt cotton,
during the reproductive season, Maracaju-MS, Brazil,
2007/2008 crop season.
Taxon
Bt cotton
Fields
Non-Bt cotton
3. Results
Formicinae
Brachymyrmex sp.1
66.66%
Camponotus crassus
11.11%
Camponotus ruphipenis
11.11%
Camponotus sp.1
22.22%
Dolichoderinae
Dorymyrmex sp.1
100%
Dorymyrmex sp.2
11.11%
Ecitoninae
Labidus sp.1
0
Myrmicinae
Pheidole oxyopsis
22.22%
Pheidole sp.1
100%
Pheidole sp.2
22.22%
Pheidole sp.3
0
Solenopis invicta
11.11%
Solenopsis sp.1
100%
Solenopsis sp.2
11.11%
Ponerinae
Hypoponera sp.1
11.11%
Hypoponera sp.2
0
Pachycondyla striata
11.11%
Pseudomyrmecinae
Pseudomyrmex sp.1
11.11%
The species used in the statistical analysis,
three traps, are indicated in grey.
With respect to ants, a total of 18 species in six

subfamilies were collected during the reproductive
period of cotton (Table 1). The nine species present in
at last three traps, used in the statistical analysis, are as
follows: Brachymyrmex sp.1, Camponotus sp.1,
Dorymyrmex sp.1, Labidus sp.1, Pheidole sp.1,
Pheidole sp.2, Pheidole oxyops, Solenopsis sp.1 and
Solenopsis invicta. Labidus sp.1 was found
exclusively in non-Bt cotton and Pheidole sp.1 and
Solenopsis sp.1 were found in all samples from Bt and
non-Bt areas.
Ant species richness was significantly higher in the
non-Bt treatment area (t = 3.05; gl = 8; P = 0.016),
with the exception of one sampling day, when the
reverse trend was observed. On that day, overall species
richness was the highest (Fig. 1). Most species

occurred with similar frequencies in both areas. In
only two samples, we registered a species that
occurred exclusively in one treatment (Fig. 2).
Therefore, the species composition of ants was very
similar in both treatments.
Regarding beetles, a total of 707 individuals in 54
species and 15 families (Table 2) were collected. Of
these, 27 species were present in at least three samples.
Four species were prevalent in the non-Bt area,
Calosoma granulatum, Tetracha sp. 1, Eriopis
connexa and Canthidium sp.1, and four other species
were ubiquitous in Bt cotton samples (Table 2):
Phloeonemus sp.1, Scirtes sp.1, Conoderus malleatus
and Heteroderes sp.1.
this study were deposited in the Entomology Museum

of the UFGD and in the entomology collection Pe.
Jesus Santiago Moure, UFPR.
2.4 Statistics
In order to ascertain the main patterns of variation in
community structure, we only took into consideration
species that were presented in at least three traps. To
compare the means of both treatments, we used a
paired t-test per collecting day. Seeking to obtain
gradients of Formicidae and Coleoptera species
composition, we considered five samples from the
cotton genetically modified area (Bt), and five samples
from the conventional cotton area. Each sample
corresponded to four traps taken randomly from the
total of 20 pitfalls available for each area (drawing
without replacement). The frequency of occurrence on
the sample corresponds to the number of traps where
each species occurred (range from 0 to 4).
88.88%
0
0
11.11%
88.88%
11.11%
33.33%
44.44%
100%
33.33%
22.22%
66.66%
100%
11.11%
0
22.22%
0
0
present in at least
168
with similar abundance in both areas. However, some

species occurred with higher abundances in samples
of Bt cotton field, while other species were more
abundant in non-Bt cotton field (Fig. 4).
4. Discussion
Fig. 1 Ant species richness in a field of genetically

modified cotton (Bt) and a conventional cotton field (nBt).
Each point pairs are corresponding to nine days during the
cotton reproductive season, Maracaju-MS, Brazil,
Beetle species richness did not significantly differ

between Bt and non-Bt treatment areas (t = 0.496; gl =
8; P = 0.633) (Fig. 3). Most of beetle species occurred
The following ant genera, regularly encountered in

cotton crops of the Northeast and Midwest Brazil [24,
25], were most frequent in our samples: Solenopsis,
Pheidole, Camponotus, Labidus, Brachymyrmex and
Dorymyrmex. The species richness differences
between Bt and non-Bt area in this research are not in
agreement with the results of a similar study
conducted in the Cauca Valley, Colombia, where ant
species richness was higher in Bt cotton fields [26].
Other similar studies, have found little or no
differences in community composition of arthropods
[27, 28].
The ant genus Labidus is a generalist predator that
was found exclusively in samples from the non-Bt
cotton field, possibly due to the greater abundance of
prey in the conventional cotton field, which had not
Fig. 2 Species composition of ants in 10 samples (occurrences in four pitfalls by samples), five in a Bt cotton field and five in
a non-Bt cotton field, Maracaju-MS, Brazil, 2007/2008 crop season.
169
Table 2 Number of specimens of Coleoptera registered in pitfall traps on a field of Bt and another non-Bt cotton, during the
reproductive season, Maracaju-MS, Brazil, 2007/2008 crop season.
Taxon
Field
Bt Cotton
Non-Bt Cotton
Taxon
Anthicidae
Erotylidae
Formicilla sp.1
5
2
Pselaphacus sp.
Carabidae
Histeridae
Calosoma granulatum
3
29
Phelister sp.1
Galerita sp.1
1
2
Nitidulidae
Scarites sp.1
1
3
Carpophilus sp.
Tetracha sp.1
1
13
Stelidota sp.
sp.4
5
1
Scarabaeidae
sp.5
1
2
Ataenius sp.1
sp.6
0
1
Leucothyreus kirbyanus
sp.7
0
1
Canthidium sp.1
sp.8
1
0
Canthon chalybaeus
Chrysomelidae
Canthon histrio
Brasilaphthona sp.
5
8
Canthon sp.1
Systena sp.1
1
0
Coprophanaeus cyanecens
Phaedon consimilis
0
1
Coprophanaeus horus
Colaspis joliveti
5
1
Pseudocanthon sp.
Myochrous sp.
0
1
Eutrichillum sp.
sp.9
3
0
Scirtidae
Coccinelidae
Scirtes sp.1
Cycloneda sanguinea
2
2
Silvanidae
Eriopis connexa
4
14
Ahasverus s.p
Hippodamia convergens
6
4
Staphylinidae
Hyperaspis festiva
3
4
Pinophilus sp.
Scymnus (Scymnus) sp.
4
11
sp.12
Colydiidae
sp.13
Phloeonemus sp.1
7
0
sp.10
Curculionidae
sp.11
Conotrachelus sp.
0
1
Tenebrionidae
Rhyssomatus sp.
0
1
Ctesia hirta
Sternechus subsignatus
1
0
Lagria villosa
Elateridae
Blapstinus sp.1
Aeolus sp.1
2
2
sp.14
Aeolus sp.2
1
1
Conoderus malleatus
11
4
Heteroderes sp.1
4
0
Total of individuals
The species used in the statistic analysis, present in at least three traps, are indicated in grey.
been treated with insecticides. This result is consistent

with other studies that have found a gradient in the
abundance of predator species in non-Bt cotton not
treated with insecticides (lower abundance), when
compared with Bt cotton or non-Bt cotton treated with
insecticides (highest abundance) [29, 30]. However,
our results indicate that the exclusive occurrences of
ant species are sporadic in both fields.
Field
Bt Cotton
Non-Bt Cotton
141
13
143
10
2
1
19
2
1
2
1
0
1
0
4
0
81
2
0
6
0
3
1
10
36
14
1
1
0
1
2
0
0
1
1
7
1
0
1
1
1
1
1
0
310
397
The species P. oxyops was recorded in abundance

in both treatments. It is justifiable because it is a
species reported as the most abundant in several
agroecosystems [31].
The predominance of Pheidole and Solenopsis
during the reproductive period of cotton had been
previously noted in Brazil [25]. The two genera are
amongst the most diverse in the neotropic [32] and
170
Fig. 3 Beetle species richness in a field of genetically

modified cotton (Bt) and a conventional cotton field (nBt).
Each point pairs are corresponding to nine days during the
cotton reproductive season, Maracaju-MS, Brazil,
their component species are not only highly tolerant to

the physical conditions of the environment [33], but
also are very efficient in colonizing habitats with low
structural complexity, for example, environments have
suffered high anthropic disturbance [34]. Furthermore,
species of these genera play an important role in
agroecosystems, as they prey on larvae of A.
argillacea (Lepidoptera), an important pest of cotton
[35], as well prey other pests, non-target, such as
Helicoverpa armigera (Lepidoptera) [36] and
Anthonomus grandis (Coleoptera) [37].
Assessments of the presence of the endotoxin
Cry1Ab from Bt cotton in seven species of Coleoptera
(Carabidae), namely Agonum placidum, Bembidion
rupicola, Clivina impressefrons, Cyclotrachelus
iowensis, Harpalus pensylvanicus, Poecilus chalcites
and Poecilus lucublandus have shown that all
individuals collected in the field presented signals of
the cry protein, demonstrating that non-target species
can accumulate this compound. The implications of
this contamination are however not yet clear [38].
Most coleopterans collected from the non-Bt
cotton field are predatory species (e.g., C.
granulatum, Tetracha sp.1 and E. connexa). It is

possible that the strong presence of predators on the
conventional cotton field is due to the greater
abundance of prey that can not thrive as well in the
adjacent field because of the deleterious effects of
the cry protein to the target pests. In contrast, the
most species of Coleoptera collected in the Bt field
are herbivores (Phloeonemus sp.1, C. malleatus and
Heteroderes sp.1). This pattern may be determined
by the absence of some predators (as recorded in this
study) in the Bt field. Also in Brazil was conducted
field study that indicate there are no effects of Bt
maize on predators, for example, earwig (Dermaptera:
Forficulidae)
and
ladybird
(Coleoptera:
Coccinellidae) [39].
When attempting to understand the richness of the
fauna on the Bt cotton, it is important to understand
how different organisms are affected by the
cultivation of these plants. Experiments using
organism that are vulnerable to several different toxins
in the soil, as for example earthworms (Annelida),
have prevailed in studies trying to understand how
transgenic plants affect edaphic, non-target species.
The Cry1Ab was not found to affect the total number
of nematodes and protozoan cultures in the digestive
tract of earthworms in soils containing B.
thuringiensis [40]. In addition, another study using
earthworms reared in Bt cotton fields concluded that
this treatment does not affect the growth or
development of Aporrectodea caliginosa (Oligochaeta)
[41]. Experiments using edaphic mite also arrived to
the conclusion that the Bt toxin does not alter the
survival and developmental rates of Scheloribates
praeincisus (Acari: Oribatida: Scheloribatidae) [42].
In regards to predators, the Bt toxin was also not
found to affect the survival or development of the
neuropteran Chrysoperla carnea (Chrysopidae) [20].
In conclusion, similarly to earthworms, Acari, and
neuropterans, ants and beetles are not targets of the Bt
protein, but it is possible that some characteristics of
their populations may be affected by it.
171
Fig. 4 Beetle species composition, in 10 samples (occurrences in four pitfalls trap by sample), five a Bt cotton field and five
in a non-Bt cotton field, Maracaju-MS, Brazil, 2007/2008 crop season. We only present the species with more than five beetles,
this facilitates the knowledge of the species turnover pattern between the areas. The gradient generated from the association
these plots of relative abundance, recovered a pattern where species that occurred with highest abundances in samples of the
Bt cotton field are represented in the figure base. Species with highest abundances in samples of the non-Bt cotton field are
the other extremity in the figure top.
5. Conclusions
Through this study, we verified the species that
occurred in reproductive period of cotton in one
commercial field cotton (Bt and non-Bt plants) farm
in the Midwest of Brazil. Therefore, additional studies
are necessary through the years and in different
locations, thereby, to make clear enough about the
richness and diversity of ants and beetles. These
insects are very important in maintaining soil quality,
Geovan Henrique Corra, Angelico F. Asenjo Flores

and Fernando Leivas for the identification of
specimens of Coleoptera; Stela de Almeida Soares for
the identification of the ants; Vitor Lemos Landeiro
(Instituto Nacional de Pesquisas da Amaznia) for the
help in the statistical analysis and Fundao de
Apoio ao Desenvolvimento do Ensino, Cincia e
Tecnologia do Estado de Mato Grosso do Sul
(FUNDECT) for the Masters scholarship to the
senior author.
protecting plants from pests among other ecological

interactions.
Acknowledgments
The authors thank the UFPR team members: Dr.
Germano H. Rosado Neto, Paschoal Coelho Grossi,
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Effect of Biofertilizer On Growth, Productivity, Quality and Economics of Rainfed Organic Ginger (Zingiber Officinale Rosc.) Bhaisey Cv. in North-Eastern Region of India

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Effect of Biofertilizer On Growth, Productivity, Quality and Economics of Rainfed Organic Ginger (Zingiber Officinale Rosc.) Bhaisey Cv. in North-Eastern Region of India

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Editorial Board Members (in alphabetical order):

Chuah Tse Seng (Malaysia)

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David Publishing Company

Effect of Biofertilizer on Growth, Productivity, Quality and Economics of Rainfed Organic

Physiological and Phytosanitary Potentials of Coriander and Radish Seeds

Response of Amaranth to Irrigation and Organic Matter

Journal of Agricultural Science and Technology A 3 (2013) 83-98

Effect of Biofertilizer on Growth, Productivity, Quality

Received: November 13, 2012 / Published: February 20, 2013.

tremendous potential for ginger production and hence

production. One possible reason for its low yield in

nutrients supply through greater mineralization due to

2. Materials and Methods

heavy feeder and an exhaustive crop requires large

2.1 Experimental Site

increasing demand for organic products all over the

The field experiment was conducted during 2007

world, the ginger farmers can receive higher returns

environmental protection, improved quality and better

2.2 Experimental Details

plots. The three levels for each of three biofertilizers

5.0 kg ha-1) and (c) Phosphotica (c1 = 2.5, c2 = 3.75

Treatment details along with composition of 500 mL biofertilizer culture solution.

Biofertilizer composition in 500 mL culture solution

applied during land preparation. The crop received

were estimated as: CGR = (W2 W1)/(t2 t1),

90-150 and 150-210 DAP were estimated using the

following formula: RBR = (R2 R1)/(t2 t1) expressed

column (18 mm 450 mm) with stopcock, then 50 mL

for 16 h at 25 2 C. The filtrate extracted through non

parameters such as specific gravity, dry matter,

The cost of inputs such as FYM, seed rhizome,

Among the different biofertilizer treatments, seed

Biofertilizer increased rhizome yield over control by

Table 4 Effect of biofertilizer on crop growth rate (CGR)

all the growth stages (Table 5). The increased yield

Combined use of Azotobacter and Phosphotica

medium level of Phosphotica (a3c2). This treatment

Table 5 Effect of biofertilizer on rhizome growth and

Rhizome bulking rate

Rhizome yield (t ha-1)

and Phosphotica recorded the lowest values of the

The biofertilizers treated crop increased the dry matter

3.4 Economics of Rainfed Ginger

had no effect on cost of cultivation of ginger. Seed

Table 7 Effect of biofertilizer on quality attributes of

3.2, respectively). Seed treatment with high dose of

Table 8 Effect of biofertilizer on economics of ginger

Net return Return per

markedly higher net return ($3,338 and 3,319 ha-1) and

($3,971 ha-1), net return ($2,688 ha-1) and return per

kg ha-1) of Phosphotica seemed to be optimum on

3D, 3E and 3F) at all the growth stages. This showed

nutrients through out the growing period as a result of

resulting from higher fixation of atmospheric N,

the growth of several pathogenic fungi in the root

respectively) and low ($4,239 ha-1, Rs 2,928 ha-1 and

ha-1), net return ($3,525 ha-1) and return per dollar

Phosphotica 3.75 kg ha-1 (c2) improved most of the