TUGAS

MATA KULIAH : DESAIN PENELITIAN

TEMA : Jenis-Jenis Penelitian
Pengajar : Dyah Maharani, S.P.t, MP., Ph.D., Ir., IPM.

Oleh

Yusni Khairani Tampubolon 449060

Tugas Kuliah Pascasarjana

Diserahkan guna memenuhi nilai tugas yang diperlukan

FAKULTAS PETERNAKAN

UNIVERSITAS GADJAH MADA

2019
 1. a. Contoh Penelitian Dasar
Judul :
Freshwater use in livestock production-to be used for food crops or livestock feed
Sumber referensi :
Ylva Ran,Corina E.,Van Middelaar, Mats Lannerstad, Mario Herrero, Imke J.M. de
Boer. 2017. Freshwater use in livestock production—To be used for food crops
or livestock feed?. Agricultural Systems 155 (2017), 1–8.
b. Contoh Penelitian Terapan

Judul :
Use of naturally sourced feed additives (lactobacillus fermentation products and
enzymes) in growing and finishing steers: Effects on performance, carcass
characteristics and blood metabolite

Sumber referensi :
T.,Rana,B.,W.,M.,S.,Gomaaa,C.,Y.,Z.,Shena,D.,A.M.,Saleema,E.,W.,Z.,Yanga,T.,A.,
McAllistera. 2019. Use of naturally sourced feed additives (lactobacillus
fermentation products and enzymes) in growing and finishing steers: Effects
on performance, carcass characteristics and blood metabolite. Animal Feed
Science and Technology 254 (2019) 114190.

2. a. Judul penelitian : Freshwater use in livestock production-to

be used for food crops or livestock feed

Penelitian ini merupakan penelitian dasar (basic research) karena menggunakan metode baru
dalam penelitiannya yaitu metode untuk menghitung persaingan air bersih yang digunakan antara
tanaman pangan dan makanan ternak. Pada jurnal ditunjukkan dibagian abstrak, “This article presents a
new method to account for the competition for freshwater use between food crops and animal feed, while
assessing freshwater use in livestock production systems”. Dan juga terdapat pada bagian latar belakang
“The method presented in this study, focuses on water resource use in livestock production systems and
the competition between food and feed production. Different from other water assessment studies, water
resources are categorized with regard to land use, taking into account the opportunity costs of land for
food crop production. In this way livestock production is compared to food crop production based on their
contribution to the production of human digestible proteins per unit of water resource used”

Berdasarkan pendekatan karakteristik, penelitian tersebut merupakan penelitian korelasional.

Mengetahui persaingan penggunaan air bersih yang berasal dari air hujan (air hijau) dan air tanah (air biru)
pada tanaman pangan dan makanan ternak yang berhubungan dengan jumlah maksimum protein cerna
manusia yang berasal dari tanaman pangan yang mengkonsumsi air untuk menghasilkan 1 kg sumber
makanan ternak . Adapun tiga jenis pemeliharan yang dilakuakan yaitu pemeliharaan dengan pasture
alami, padang rumput dan pemeliharaan intensif. Pada jurnal ditunjukkan dibagian deskripsi metode
umum, “The developed method is illustrated in a flowchart in Fig. 2. The method calculates CWU during
plant growth of feed crops on cultivated land and grass growth on pastures. Water used for feed
production is either green water, i.e. rainwater on crop or grasslands, or blue water, e.g. groundwater or
surface water used for irrigation of primarily cropland (Fig. 2). Cropland and associated green and blue
water resources can be used directly to cultivate feed crops, food crops or other crops (e.g. fuel or fiber),
whereas water resources used on grassland that is suitable for crop production could provide animal feed
as grass, but could also support crop growth”.

Berdasarkan kedalaman analisisnya, penelitian tersebut merupakan penelitian deskriptif karena

tidak ada uji lanjut atau uji statistik setelah mendapatkan hasil perhitungan dari penggunaan air konsumtif.
Pada jurnal ditunjukkan dalam bagian konsep metode hanya menghitung ratio penggunaan air tanpa ada
analisis lanjutan.

The WUR is calculated according to Eq. (1):

× HDP
∑ n ∑m −3
CWU m y−1
i=1 j=1 ( ij j )
WUR =
HDP of one kg of ASF
3
CWUij is the consumptive water use in m , evapotranspired over land suitable to produce food crops that is required to produce feed
ingredient i (i = 1,n) in country j (j = 1,m) used to produce 1 kg of ASF.

Berdasarkan pendekatan analisis, penelitian ini merupakan penelitian kuantitatif karena

menggunakan angka dalam penelitiannya dan menyajikannya hasil dalam bentuk tabel angka dan diagram
angka yang merupakan hasil dari perhitungan rumus mencari penggunaan air konsumtif. Dalam jurnal
tersebut ditunjukkan pada bagian hasil, “illustrates the CWU to produce 1 k of beef for each of the three
Uruguayan beef production systems, categorized per type of water and land and expressed as liters of
water per kilo of meat. All production systems depend almost entirely on green water, with blue water
resources only representing about 1% of total CWU”.

b. Judul penelitian: Use of naturally sourced feed additives (lactobacillus fermentation

products and enzymes) in growing and finishing steers: Effects on performance,
carcass characteristics and blood metabolite

Penelitian ini merupakan penelitian terapan karena mengimplementasi probiotik komersial

kedalam pakan ternak untuk melihat performance sapi. Hal ini ditunjukkan didalam bagian latar
belakang yaitu “Bio-Lac Plus (BL) and Boviglo (BG) are commercial feed additives that contain lactic
acid bacteria (LAB), lactobacillus fermentation products (LFPs), and enzymes. We hypothesized that
feeding BL to implanted cattle would have additive effects on growth performance, and that BG may act as
an alternative to the current use of implants and in-feed antibiotics in beef cattle. We also hypothesized
that feeding of BL and BG may impact carcass traits, possibly by exogenous enzymes increasing fiber
digestion which could alter fat deposition. The objectives of this study were to examine BL and BG as
alternatives to in-feed antibiotics and to investigate their effects on growth performance, carcass traits,
blood metabolites and antioxidant status of growing”.

Berdasarkan pendekatan karakteristik, penelitian ini merupakan penelitian eksperimental karena

menggunakan lima perlakuan untuk penelitiannya, ditunjukkan pada bagian materi dan metode, “five
treatments:1) control (basal diet); 2) implant (IM; Elanco-Component TE-100 with Tylan, containing 100 mg
of trenbolone acetate USP, 10 mg of estradiol USP and 29 mg of tylosin tartrate; Elanco Canada Limited,
ON, Canada); 3) implant + antibiotics (330 mg/steer/d monensin +110 mg/steer/d chlortetracycline; IMAT);
4) implant + BL (30 g/steer/d BL; IMBL); and 5) BG (5 mL/steer/d)”.
 Berdasarkan kedalaman analisisnya, penelitian tersebut merupakan penelitian inferensial karena data
yang didapatkan dari hasil perlakuan tehadap ternak diuji menggunakan SAS. Di jurnal ditunjukkan pada
bagian analisis statistic, “For the growing phase, data of DMI, BW, ADG, and G:F were analyzed using the
Mixed procedure of SAS (Version 16.0.0, SAS Inst. Inc. Cary, NC) as a completely randomized design with
treatment, period (days on-feed), and their interaction as fixed effects and steers within treatment as a
random effect. For the finishing phase, data of DMI, BW, ADG and G:F were analyzed using the Mixed
procedure of SAS with treatment, period (days on-feed) and their interaction as fixed effects and steers
within treatment as a random effec”.

Berdasarkan pendekatan analisis, penelitian ini merupakan penelitian kuantitatif karena data yang
ditampilkan berbentuk angka dan melakukan analisis statistik pada hasil data yang diperoleh. Pada jurnal,
angka dapat ditemukan dalam semua bagian contohnya pada bagian hasil pertumbuhan performan “In the
growing phase, DMI tended (P < 0.08) to be greater for IM, IMBL and BG steers than control and IMAT
steers from d 29 to 56, but did not differ over the entire growing phase (Table 2). Body weight was higher
(P < 0.02) for IM, IMAT and IMBL, inter-mediate with BG and lowest with control steers at d 84 and 112.
There was a treatment × days on-feed (P < 0.05) interaction for ADG. The IM, IMAT and IMBL steers had
greater (P < 0.05) or tended to have greater (P < 0.10) ADG than control steers on most of the days on-
feed and as a result, the overall ADG was greater (P < 0.01)”.

Agricultural Systems 155 (2017) 1–8

Contents lists available at ScienceDirect

 Agricultural Systems
journal homepage: www.elsevier.com/locate/agsy

Freshwater use in livestock production—To be used for food crops or MARK

livestock feed?
a,b, a c,1 d a
Ylva Ran , Corina E. van Middelaar , Mats Lannerstad , Mario Herrero , Imke J.M. de Boer
a Animal Production Systems Group, Wageningen University, PO Box 338, 6700 AH Wageningen, The Netherlands
bStockholm Environment Institute, Linnégatan 87D, 115 23 Stockholm, Sweden
c International Livestock Research Institute, PO Box 30709, 00100 Nairobi, Kenya
dCommonwealth Scientific and Industrial Research Organization, Bioscience Precinct Carmody RD, St. Lucia, 4067, Queensland, Australia

ABSTRACT

Current approaches to estimate freshwater use in livestock production systems generally fail to consider the competition for
water resources with alternative uses, such as production of food crops food or other ecosystem services. This article presents
a new method to account for the competition for freshwater use between food crops and animal feed, while assessing
freshwater use in livestock production systems. The developed water use ratio (WUR) is defined as the maximum amount of
human digestible protein (HDP) derived from food crops from the consumptive water use (CWU) appropriated to produce 1
kg of animal-source food (ASF) over the amount of HDP in that 1 kg of ASF. The CWU for livestock production is first
categorized according to the land over which it is consumed, based on the suitability of that land to produce food crops. Then,
the method assesses food-feed competition by determining the amount of HDP that could have been produced from food
crops, using the same CWU currently used to produce ASF. The method enables identification of livestock production systems
that contribute to global food supply without competing significantly over water resources with food production, based on
their CWU. Three beef production systems in Uruguay are used to illustrate the method. During the backgrounding and the
finishing stages, which are analyzed in this study, cattle can be kept on natural pasture (NP), seeded pasture (SP) or in feedlots
(FL). The following three systems were analysed: i) NP-NP, ii) SP-SP and iii) SP-FL. Results show that the NP-NP system
uses the largest amount of water per kg of beef output. However, results also show that the SP-SP and SP-FL systems can
potentially produce more HDP by growing food crops than by producing beef. Based on the traditional measure for water
productivity, i.e. the quantity of CWU per kilo of beef produced, we would conclude that the NP-NP system is least e fficient,
whereas based on the WUR the NP-NP system is the only system producing HDP more efficiently than food crops.
Sustainable intensification not only implies improving agriculture and livestock productivity per unit of resource used, but also
improving the number of human beings nourished. Results from this study illustrate the importance of considering
competition and trade-offs with other uses when evaluating water use efficiency of livestock systems to promote sustainable
intensification.

1. Introduction
A growing world population, estimated to reach nine billion people by
2050, is increasing the pressure on global agricultural production to ensure
food security for all. Between 2005 and 2050 the demand for meat and milk
products is projected to increase by around 70–80% and the demand for crop
protein by 100–120% (Tilman et al., 2011; Alexandratos and Bruinsma,
2012).
Livestock production requires large amounts of natural resources,
including water and land, and the expected rising demand for animal
sourced foods (ASF) can potentially amplify environmental impacts related to
livestock (Delgado et al., 1999; Godfray et al., 2010; Bouwman et al., 2013;
Westhoek et al., 2014; Herrero et al., 2015).
At present, the global livestock sector uses about 75% of all agricultural
land (Foley et al., 2011), and is responsible for about 30% of global
agricultural water requirements, including rain and irrigation water used for
production of feed and withdrawals for animal husbandry (Mekonnen and
Hoekstra, 2012). At current productivity levels, the expected rise in demand
for animal products will result in a doubling of the land and freshwater
requirement, increasing the water

Corresponding author at: Stockholm Environment Institute, Linnégatan 87D, 115 23 Stockholm, Sweden.
E-mail addresses: ylva.ran@sei-international.org (Y. Ran), research@lannerstad.com (M. Lannerstad).
1Present address: Welanders väg, 7, 112 50 Stockholm, Sweden.
http://dx.doi.org/10.1016/j.agsy.2017.03.008
Received 12 August 2016; Received in revised form 1 March 2017; Accepted 11 March 2017
Available online 06 April 2017
0308-521X/ © 2017 Elsevier Ltd. All rights reserved
Y. Ran et al.
Fig. 1. Conceptual flow chart of land and water resource requirement in livestock production. [Single column.]
resource use competition (Rockström and Barron, 2007; Rockström et al.,
2007). An amplified water use for livestock and crop production can, in turn,
locally increase the risk of water stress (Ridoutt and Pfister, 2010). At present,
> 1.2 billion people already suffer conditions of physical water scarcity
(Molden, 2007).
Livestock require water for e.g. drinking and cleaning services, and for
the cultivation of feed crops or for grass growth (Figs. 1 and 2). In this paper,
we focus on consumptive water use (CWU), which refers to water that is
withdrawn from a watershed, and not discharged to the same watershed
because it evaporates, is embodied in plants or animals, or is discharged to a
different watershed (Falkenmark and Lannerstad, 2005). As a general rule, >
98% of the total CWU in livestock production can be attributed to
evapotranspiration from feed crops and pastures. Only 2–8% of livestock
CWU is drinking, servicing and feed-mixing water (Steinfeld et al., 2006;
Mekonnen and Hoekstra, 2012; De Boer et al., 2013).
To acknowledge the importance of both soil moisture and water
withdrawals from water bodies, water resources can be divided into green
water, which refers to soil moisture available to plant growth, and blue water,
which refers to liquid water in water bodies, as rivers, lakes and aquifers
(Falkenmark, 1995). Green and blue water resources, however, are
interchangeable states, and water can shift from one state to the other, and
back. Green water use does not only affect the availability of soil moisture,
but could also affect the availability of blue water, since part of the soil
moisture, if unused, could drain out of the soil and re-charge water bodies as
blue water. Thus, both green and blue water uses may ultimately alter water
availability in the landscape in different ways, impacting local ecosystem
functioning and, should therefore both be considered in water use assessments
(Milà i Canals et al., 2009).
As illustrated in Fig. 1, livestock products, e.g. beef meat, can be
produced in a variety of production systems that use a wide range of
Fig. 2. Water use in livestock production categorized, considering differences by feed composition for different animal type and production systems and possible trade-offs between feed and food
crops. [Single column.]
2
Agricultural Systems 155 (2017) 1–8
different feeds, which in turn can be grown using different natural resources
and management practices. The use of water resources, and primarily green
water, is tightly connected to the land that is used by a particular livestock
production system. Green water is directly linked to a specific area, available
as soil moisture for plant growth, while blue water is linked to water bodies,
thus the ability in the landscape to store liquid water. Since the majority of
water consumption in livestock systems relates to the cultivation of feed,
water resource use and land use should be considered together, rather than
separately (Ran et al., 2016).
Animal feed can be produced on grasslands such as natural pastures
(grazing livestock) and cropland (all livestock). Grasslands, especially natural
pastures, require primarily green water. However some pastures are irrigated,
thus using additional blue water resources, and some are even cultivated and
occupy land suitable as cropland. All animal feed crops require cropland for
growth, however, some feed crops are rainfed, and thus depend entirely on
green water, while others require irrigation water depend on both green and
blue water.
To prevent unsustainable use and management of water resources, there is
a need to describe the linkages between livestock production and freshwater
use. Understanding and quantification of these links is imperative in order to
increase water productivity in livestock produc-tion, and to identify trade-offs
and synergies between livestock production and other competing water uses,
such as food crop produc-tion. The focus on increased feed efficiency for
livestock to improve resource use efficiencies, and changing consumer
preferences towards more pork and poultry products, has led to a larger share
of human edible plant material in animal feed (De Vries and De Boer, 2010;
Eisler et al., 2014). An increased use of high quality croplands to cultivate
animal feed, in preference to food crops, will further proliferate resource use
competition between food and feed production.
Current estimates of both water and land resource use by livestock
Y. Ran et al.
generally fail to consider the competition for resources between the
production of food crops and animal feed (Van Zanten et al., 2016). To
address such knowledge gaps, Van Zanten et al. (2016) developed a method
that accounts for the competition for land resources between food and feed
production. Based on a land use ratio (LUR), the land use efficiency of
livestock systems is defined as the maximum amount of human digestible
protein (HDP) derived from food crops on all land used to cultivate feed
required to produce 1 kg of ASF, over the amount of HDP in that 1 kg of ASF.
Considering that livestock production systems, in addition to land, use
large amounts of water resources we further develop the method presented by
Van Zanten et al. (2016) to investigate food-feed competition over water
resources in livestock production systems. This requires careful consideration
of the complexity of the hydrological cycle, recognising that water is a
dynamic resource with a strong connection to landscape dynamics and
multiple users competing for its availability.
The method presented in this study, focuses on water resource use in
livestock production systems and the competition between food and feed
production. Different from other water assessment studies, water resources
are categorized with regard to land use, taking into account the opportunity
costs of land for food crop production. In this way livestock production is
compared to food crop production based on their contribution to the
production of human digestible proteins per unit of water resource used. The
method enables identification of livestock production systems that contribute
to global food supply without competing significantly over water resources
with food pro-duction. In this paper, the method is first described in generic
terms and subsequently illustrated, using three beef production systems in
Uruguay.
2. Conceptualization of the method
2.1. Generic description of the method
The developed method is illustrated in a flowchart in Fig. 2. The method
calculates CWU during plant growth of feed crops on cultivated land and
grass growth on pastures. Water used for feed production is either green
water, i.e. rainwater on crop or grasslands, or blue water, e.g. groundwater or
surface water used for irrigation of primarily cropland (Fig. 2). Cropland and
associated green and blue water resources can be used directly to cultivate
feed crops, food crops or other crops (e.g. fuel or fiber), whereas water
resources used on grassland that is suitable for crop production could provide
animal feed as grass, but could also support crop growth. Thus, water use for
feed production on cropland competes directly with food crop produc-tion,
whereas water use on grasslands suitable for crop growth illustrates indirect
food-feed competition (Fig. 2). The developed method calculates and
differentiates the CWU between water resources evapotranspired over land
suitable for crop production, and land that is assumed to be unsuitable for
crop production.
The proposed methodology is a four-step process. First, green and blue
CWU during production of animal feed is quantified, for example by using a
hydrological model or from field measurements. The division into green and
blue water highlights to what extent CWU for animal feed constitutes of soil
moisture from naturally infiltrated rainfall, and to what extent it is water
abstracted from water bodies. Second, the green and blue water required for
production of feed is categorized according to the two agricultural land types
over which it is evapo-transpired, i.e. croplands and grasslands (see Fig. 2).
Third, the opportunity cost of land and water resources, with regard to food-
feed competition, is identified by assessing the suitability of the land and
water resources to produce food crops. The green and blue CWU on crop land
could have been used directly to produce food crops on that land and
represent direct food-feed competition over water resources. Indirect
competition refers to the CWU over grasslands that are partly
3
Agricultural Systems 155 (2017) 1–8
or fully suitable to support crop cultivation. The suitability can be assessed by
using statistical data, like the global agro-ecological data base (FAO, 2016;
Van Zanten et al., 2016), or by field observations. Finally, the water use ratio
(WUR) is calculated to provide a measure of how efficient a production
system uses water resources to produce HDP comparing food crops against
livestock products.
2.2. Water use ratio
The WUR is calculated according to Eq. (1):
n m −3
∑ ∑ CWU × HDP m y−1
i=1 (
j=1 ij j )
WUR =
HDP of one kg of ASF (1)
3
CWUij is the consumptive water use in m , evapotranspired over land
suitable to produce food crops that is required to produce feed ingredient i (i =
1,n) in country j (j = 1,m) used to produce 1 kg of ASF. HDPj is the amount of
human digestible protein (HDP) that can be produced in country j, using the
same water resources, by direct cultivation of suitable food crops in country j
per year. The denomi-nator is the amount of HDP of 1 kg of ASF. A ratio
larger than 1 implies that the water resources for that production system can
generate a larger amount of HDP by producing food crops instead of ASF.
Correspondingly, if the ratio is below 1 the production of HDP through
livestock is more efficient than cultivating food crops using the same water
resources.
2.3. Case study description: Uruguayan beef production
Three beef production systems in the Rocha region in the southeast of
Uruguay are used as a case study to illustrate the new method, with data
adapted from previous studies (Modernel et al., 2013; Ran et al., 2013;
Picasso et al., 2014). The region is a good representation of Uruguay, with
78% of the land use dedicated to beef compared to 77% for the entire nation
(MGAP, 2011; Modernel et al., 2013). For each of the three systems, the two
final stages of the production cycle; back-grounding and finishing are
analyzed (Picasso et al., 2014). During backgrounding and finishing, beef
cattle grow from about 150 kg to their final slaughter weight of around 500
kg. The dressing percentage (i.e. carcass weight / live weight × 100%) of beef
produced in Uruguay was assumed to be 52% (FAO, 2003). The protein
content of beef was assumed to be 17.6 g protein per 100 g of meat, whereas
protein digestibility was assumed to be 94% (Young and Pellet, 1994; USDA,
2015). The cow-calf system was not included in this study.
The backgrounding can be based on either natural pasture (NP) or seeded
pasture (SP), and the finishing system can be based on NP, SP or a feedlot
(FL) system (Picasso et al., 2014). The three analyzed systems, each have a
different combination of a backgrounding and a finishing systems, and are
defined as follows; 1) NP-NP, 2) SP-SP, 3) SP-FL (Table 1). The relative area
for each system to produce required animal feed was calculated from animal
nutritional requirements (NRC, 1996; AFRC, 1993) which are based on initial
and final animal weight, daily weight gain, feed composition and nutritional
characteristics of forages and concentrates (first reported in Mieres et al.
(2004)), as described in Modernel et al. (2013) and Picasso et al. (2014).
In Uruguay, most beef cattle are finished on pasture; only about 10% of
the cattle are finished on feedlots. Natural pastures are assumed to be
unsuitable for crop production, because agricultural expansion in Uruguay,
primarily for soybean production, has reduced the grazing area in the country
and pushed grazing animals to marginal lands (Picasso et al., 2014). Seeded
pastures are cultivated with a crop-pasture rotation, where a crop is sown at
least every fourth year (Modernel et al., 2013), and, therefore, this land is
suitable for both crop and grass growth.
In a global comparison, all three Uruguayan beef systems are rather
extensive; largely depending on grass as animal feed (Seré and
Y. Ran et al.
Table 1
Dietary composition and characteristics for the backgrounding (B) and finishing (F) stages of
Uruguayan beef production systems combined as NP-NP, SP-SP and SP-FL, where NP is
natural pasture, SP is seeded pasture and FL is feedlot ( Modernel et al., 2013; Picasso et al.,
2014).
Beef cattle system NP-NP SP-SP
B F B F
Dietary composition (%)
Natural pasture 100.0 100.0 30.0
Seeded pasture 61.5 93.0
Sorghum grain 9.5 6.5
Rice bran 0.5
Residues, vitamins and minerals
System characteristics
−1 −1
Dry matter intake (kg animal d ) 9.9 12.0 7.9 8.4
a
Days to achieve final weight 486 366 285 214
a
350 kg in B and 500 kg in F.
Steinfeld, 1996). In this study, the animal diet constitutes of grass or a
combination of grass, grain of sorghum (Sorghum bicolor (L.) Moench), rice
bran, rice husk and rice hay (Table 1). Uruguayan grasslands were assumed to
be rainfed, since there was no available data indicating the existence of
irrigated pasture. The category “by-products”, i.e. rice and sorghum straw,
does not result in any corresponding water use since they are a rest product of,
e.g. another food or feed production process. The CWU related to these by-
products, therefore, is embedded in the CWU of the main product. In cases
where by-products have a significant economic or functional value, the
relative CWU can be calculated based on e.g. economic allocation or
biophysical allocation using the HDP or the energy value of the different
products.
2.3.1. Water use assessment
The water requirement per feed ingredient was calculated as the total
CWU for a specific feed type. The CWU per type of vegetation used as feed
was computed by the Lund-Potsdam-Jena managed Land (LPJmL) model
(Gerten et al., 2005; Bondeau et al., 2007; Haberl et al., 2007; Rost et al.,
2008; Fader et al., 2010), accounting for area specific crop water requirement
for each feed type, both irrigated and rainfed, vegetation growth and yield per
pixel at a resolution of 0.5°. The CWU for production of crops was based on
CWU during the growing season. The CWU for production on natural and
seeded pasture was assumed to be evenly distributed throughout the year.
The LPJmL model yields results on CWU per type of vegetation, in m
per ton of fresh matter for crops, or per ton of dry matter for grasses. The
CWU per ton of feed crop was multiplied by the amount of feed crops used to
produce 1 kg of HDP from beef for each beef production system. All crops
were assumed to have a dry matter content of 85%. For results to be
comparable, livestock CWU were also calculated as liters/kg of beef, as
presented in the results section in Fig. 3.
2.3.2. Crop suitability index and maximum HDP from food crops
To determine the amount of HDP from food crops, the spatially defined
crop suitability index (CSI) for global agro-ecological zones (GAEZ) (FAO,
2016) was used to define the suitability of both land and associated green
water resources for food crop production. The GAEZ database operates at a
0.5-degree resolution. In this study the CSI for cultivated land was determined
for baseline climate conditions (1961–1990) and a “high input level” situation,
referring to a market oriented farming system with well managed agricultural
production (FAO, 2016) to reflect Uruguayan crop production. The CSI is
based on input data of climate (i.e. frequency of wet days, temperature and
sunshine), crop water requirements, soil conditions (i.e. pH, soil water
holding capacity and total exchangeable nutrients), applied soil man-agement,
slope, elevation, terrain, land cover, protected areas and administrative areas
(FAO and IIASA, 2012). In this study, land with a
Agricultural Systems 155 (2017) 1–8
SP-FL
B F
30.0
70.0
60.5
12.0
27.5
7.9 13.2
285 102
Fig. 3. Consumptive water use (CWU) in liters per kg of beef for the three Uruguayan beef
production systems in categories of green and blue water over crop and grasslands. Cattle in the
systems are fed on different combinations of natural pastures (NP), seeded pasture (SP) and
feedlot (FL). [Single column.] (For interpretation of the references to colour in this figure
legend, the reader is referred to the web version of this article.)
crop suitability of either “good”, “high” or “very high” (i.e. a CSI > 55) for
cultivation of food crops was regarded as suitable.
For the case study of Uruguay, CSI was assessed for the four major food
crops produced in the country: wheat (Triticum spp.), rice (Oryza sativa),
barley (Hordeum vulgare L) and maize (Zea mays) (MGAP, 2012). Crop
suitability was determined for the Rocha region as all feed was assumed to be
produced within that region. The CSI shows that all four major food crops
have a suitability of > 55. For the water and land resources, used for pastoral
biomass growth, that are also suitable for crop production, we account for the
maximum amount of HDP that could be produced. This is calculated by
combining crop yields per hectare of suitable food crops, i.e. wheat, maize,
barley and rice, with protein content and human digestibility (Van Zanten et
al., 2016). Ideally, data on crop suitability should be obtained at the lowest
possible spatial resolution. However, this case study is based on three beef
production system spatially defined to a region of production and all feed is
assumed to be produced within the same region. For this case study, there is
3 no data available on crop suitability below regional level, thus the relative
suitability of the four identified suitable food crops is based on their relative
cultivated area within Uruguay.
The crop- and grasslands used by the beef systems in this analysis were
preliminary rainfed and only used small amounts of irrigation water for feed
production. However, rice cultivation in Uruguay requires some irrigation
water for production. Since the objective of the WUR is to calculate the
maximum amount of HDP that can be produced using the same amount and
type of water currently used for production of beef in Uruguay, the
requirement of additional blue water resources to cultivate rice will impact the
suitability for rice production in this particular case study. A larger amount of
blue water is required to cultivate rice than what is required for beef
production in any of the three production systems. We therefore did not
assume that rice can be produced satisfactory, only using the CWU currently
used in the analyzed beef production systems. Thus, even though rice has a
CSI > 55 based on the GAEZ database, rice was assumed to be unsuitable for
production considering the water resource availability, and accordingly
excluded from further analysis.
The CWU on cropland and grassland used for beef production was
divided into blue and green water resources. All blue and green water
resources consumed on croplands and seeded pastures (i.e. as compared
4
Y. Ran et al. Agricultural Systems 155 (2017) 1–8

to natural pastures) were assumed to be suitable for crop production. The

amount of HDP that can be produced from food crops was determined by
dividing the CWU suitable for crop production with spatially explicit crop
water requirements for the suitable food crops, which were assessed with the
LPJmL model. Crop yields were subse-quently multiplied by protein content
and digestibility to determine HDP yield. National production and yield data
for food crops were used since regional data was not available. Production
and yield data were derived from the Uruguayan ministry of livestock,
agriculture and fisheries (MGAP, 2012). Protein content and digestibility for
selected crops were obtained from literature (Young and Pellet, 1994).

3. Results
Fig. 4. Water use ratio (WUR), as kg HDP from food crops/kg HDP in ASF, compared with
Fig. 3 illustrates the CWU to produce 1 k of beef for each of the three land use (LUR) in HDP from food crops/HDP in ASF and consumptive water use (CWU) in
Uruguayan beef production systems, categorized per type of water and land 10,000 l per kilo of beef calculated for three Uruguayan beef production systems. The NP-NP
and expressed as liters of water per kilo of meat. All production systems system does not appear in the WUR and LUR results, because they are equal to 0. Single
depend almost entirely on green water, with blue water resources only column.
representing about 1% of total CWU. The NP-NP system requires the largest
amount of water; 28,000 l of green water per kilo of beef and no blue water. SP system. In the SP-FL system, seeded pastures only constitutes 30% and
The SP-FL system requires 13,800 l of green water and 430 l of blue water green water on cropland for production of sorghum, instead constitutes a
per kilo of beef and the SP-SP system requires the least water; 13,500 l of major part of the total CWU for the system.
green water and 20 l of blue water per kilo of beef. The WUR was calculated to account also for indirect competition over
water resources, i.e. competition where water is currently consumed over
Direct competition over water between food and feed crops is illustrated grasslands that could potentially support crop growth (Fig. 4). Results show
in Fig. 3 by the categories green and blue water from cropland. The NP-NP that the NP-NP system has a WUR of 0. This implies that the CWU to
system does not include feed from croplands so there is no direct competition produce 1 k of beef yields no HDP from food crops, which is logical because
over water in this system. In case of the SP-SP system, however, croplands the NP-NP system does not use any cropland or grassland suitable for crop
constitute about 16% of the total CWU, in comparison to 53% for the SP-FL cultivation. This livestock system, therefore, produces more HDP per liter of
system. Direct competition with production of human food crops, therefore, is CWU than a crop system could have done. The SP-SP system had a WUR of
highest in the SP-FL system. 2.4, whereas the SP-FL systems had a WUR of 2.7, implying that the water
required to produce 1 kg of HDP from beef could yield 2.4 kg of HDP from
In Table 2 the CWU bars in Fig. 3 are further disaggregated and present food crops in case of the SP-SP and 2.7 k of HDP in case of the SP-FL
the green and blue resources behind each feed type used in the three beef system.
production systems. Results in Table 2 indicate that the relative distribution of This depends on that these two systems, as can be seen in Table 2,
CWU between feed composition and production system vary greatly. For although still using mostly green water resources, to a large extent use green
example, while the CWU for the NP-NP system entirely comes from green water evapotranspired over crop and grasslands that are suitable to support
water on natural pastures, green water on seeded pastures corresponds to crop growth, and thus can be used for HDP production directly. Based on the
almost 70% of the total CWU in the SP- traditional measure for water productivity, i.e. liters of CWU per kilo beef
produced, we would conclude that the NP-NP systems is less efficient than
Table 2 the other systems, whereas based on our new WUR, the opposite conclusion
Consumptive water use for three Uruguayan beef production systems combined as NP-NP, SP- can be drawn (Fig. 4).
SP and SP-FL, where NP is natural pasture, SP is seeded pasture and FL is feedlot, categorized Our WUR results show a comparable pattern to results based on the land
according to dietary composition of each system and summarized for green and blue water on
use ratio (LUR; Fig. 4) (Van Zanten et al., 2016). Both the WUR and LUR
crop and grassland.
results indicate that it is more efficient to produce HDP from food crops than
Beef cattle NP-NP SP-SP SP-FL Water from livestock for the SP-SP and SP-FL systems, and that livestock
a
system productivity production is the most efficient way to produce HDP in the NP-NP system.
CWU (l/kg Dietary l/kg beef l/kg beef l/kg beef l/kg However, for the two more intensive production systems, the LUR results (5.7
beef) composition crop/grass
for SP-SP and 6.2 for SP-FL) are significantly higher than the WUR results
Green water Natural 28 014 2056 2056 533 (2.4 for SP-SP and 2.7 for SP-FL) (Fig. 4). A higher LUR in relation to WUR
pasture indicates that the system can yield a higher amount of HDP based on its land
Seeded 9269 4180 533 use relative to the amount of food crops it can yield based on its water use.
pasture
Rice 31 642 786
WUR and LUR results also indicate that the SP-SP system use both water and
Sorghum 2182 6947 1195 land resource more efficiently than the SP-FL system, when considering food-
Blue water Rice 20 429 469 feed competition.
Sorghum <1 <1 <1
Green water 28 014 11 325 6236 Since natural grasslands are assumed not suitable for crop produc-tion, the
on
grassland
NP-NP system generates a WUR and LUR of 0. The results from this study
Green water 2213 7589 indicate that the NP-NP beef production system could be important from a
on food security perspective, since it does not compete, directly or indirectly,
cropland with human food production. This finding is not identified using traditional
Blue water on 20 429
CWU assessment methods where, as in this case study, the NP-NP system
cropland
Total 28 014 13 558 14254 seems to be the least efficient beef production system in terms of water use.

a Crop and grass water productivities has been inverted to liters of water per kg of output to It should be noted that this is because the NP-NP system relies
enable easier comparison with CWU estimates.

5
Y. Ran et al.
entirely on natural grassland with a crop suitability index well below the
minimum level of “good” that was used to determine if land was suitable or
unsuitable for crop production in this study. In reality, all land has some
suitability for producing crops and would thus have a LUR/WUR above zero.
Extensive systems that use concentrates and cultivated roughage, although to
a very small extent, would also generate a ratio above zero.
4. General discussion
This study aimed to investigate water resource use in livestock production
and the competition over water resources for production of food crops. In the
past, water use assessments primarily focused on withdrawals from water
bodies and groundwater, for agriculture, industry, municipal or domestic uses
(Shiklomanov, 2000). These assessments did not account for the large
amounts of green water, i.e. naturally infiltrated rainfall in the soil. For
livestock production, green water resources constitute 90% of the total CWU
on a global average, looking at grazing, mixed and industrial livestock
production systems (Mekonnen and Hoekstra, 2012). Today, the concepts of
green and blue water are widely used to describe and assess water use in
agriculture, including livestock production (e.g. Mekonnen and Hoekstra
(2012); Ran et al. (2013, 2016)).
4.1. Capturing the complexity of green water use
Most of the variation in results from water use assessment studies of
livestock relates to whether or not green water is included, partially included
or excluded from assessments (Ran et al., 2016). Some studies argue that all,
or most of the green water use should be excluded (e.g. Ridoutt et al. (2012);
Ridoutt and Pfister (2013)), because a large amount of the consumptive green
water is used for production of human non-edible biomass, such as grass,
produced on non-cultivated fodder land and grasslands. The water
evapotranspiring over such land areas would be consumed for biomass
growth regardless if the biomass was used as animal feed, or not (Deutch et
al., 2011; Ran et al., 2013). Others highlight the importance of looking at both
green and blue water resources to identify areas of improvement (e.g.
Hoekstra and Mekonnen (2012); Mekonnen and Hoekstra (2012)). They argue
that the location of where the blue and green water is consumed does not
matter significantly, since the focus for policy-relevant water use studies
should be on decreasing the total water use for food production globally
(Hoekstra, 2014).
Recent studies also indicate that we should account for the local and
environmental impacts associated with water use (De Boer et al., 2013;
Schyns et al., 2015), for example by using water stress-related indexes (e.g.
Ridoutt et al. (2012); Ridoutt and Pfister (2013); Zonderland-Thomassen et al.
(2014)) and water scarcity assessments (Mekonnen and Hoekstra, 2016).
These measures, however, only focus on the scarcity of blue water resources,
with the aim to assess environmental impacts (i.e. direct impacts on river flow
or aquifer levels), and not green and blue resource use efficiency caused by
abstractions of blue water.
This perspective does not include the role of surplus soil moisture
contributing to blue water recharge, and the opportunity costs of both blue
and green water usage (Deutch et al., 2011). Recent reviews of freshwater use
in agriculture argue that there is a need to further develop methods that deal
with the efficiency of green water use, as well as the scarcity of green water
resources and indicators to measure that scarcity (Kounina et al., 2013;
Schyns et al., 2015).
In this study we, therefore, seek to capture the green water efficiency and
complexity by developing and applying the WUR. This is a measure that
addresses different debated aspects, in particular for green water resources,
related to water productivity and green water use, such as water use efficiency
of feed production, the ability to convert human non-edible feed products into
food, and the opportunity
6
Agricultural Systems 155 (2017) 1–8
cost of resource use for agricultural production (Van Zanten, 2016). The CWU
estimations for livestock production in this study are well
within the range of previous estimates, ranging from 13,000 l to 30,000 l of
water per kg of beef (e.g. Molden et al. (2007); Mekonnen and Hoekstra
(2012); Ran et al. (2013)). The hydrological model and crop suitability data
set both operates at a 0.5 spatial degree resolution. Ideally, for a regional
analysis, water modelling as well as crop suitability data should be collected
for a lower spatial resolution to deliver more precise national/sub-national
results on crop and grass water requirements and spatial variability within the
region.
Another future improvement would be to include herd dynamics of the
analyzed livestock production systems. Due to insufficient data sources, the
cow-calf system was not considered in this study. Since the cow-calf phase is
similar for all three systems and constitute only a small fraction of total CWU
for feed, an inclusion would not largely impact the comparison of CWU for
different systems. However, it may impact the WUR of each system and
should therefore be included in further analyses.
4.2. Considering multiple resources and competitive uses
The results shown by the WUR method are similar to the results generated
when using the LUR approach to assess the food-feed competition related to
the use of land resources. A comparison of the WUR and LUR results in the
studied Uruguayan beef production systems show that the potential
contribution to HDP by producing food crops is higher when based on land
resources, than when based on water resources. This indicates that land, rather
than water, is the limiting resource in the compared systems under prevailing
conditions. These differences highlight that the food-feed ratio depends on the
natural resource under study. However, it is the natural resource that limits
production under prevailing conditions that will determine the actual food-
feed competition of that particular system and time. This will differ from
system to system, and will change dependent on management practices and
resource availability at the point of analysis. Therefore, resource use
assessments should preferably consider multi-ple resources, since efficient
use of one resource is not necessarily efficient use of another resource.
The opportunity cost of resource use can also change with altered
parameters in a production system. Access to irrigation water and nutrients
can transform currently unproductive land to suitable farming land for food
crops. Such change could also be captured in WUR/LUR calculations to
enable comparison between different points in time and identifying
opportunities to increase the number of human beings that can be nourished
per unit of input, e.g. water resources (Van Zanten et al., 2016). This cannot
be achieved only by increasing production efficiency. Optimizing resource
allocation by identifying alternative uses, multiple users and multiple benefits
can be crucial.
The methodology presented in this paper can help identify oppor-tunities
to feed the world sustainably in several ways. Primarily the method can be
used to identify livestock production systems that use natural resources with
low opportunity costs for other uses. This will make it possible to identify and
value production systems that use crop residues, food waste or grass produced
on marginal lands, in compar-ison to systems that increase their efficiency by
using nutritious feed crops that can be directly consumed by humans.
The methodology can also help to identify unsustainable uses of e.g. blue
water resources. Through alterations in feed composition, such use can be
shifted to a more sustainable use of green water resources. Lastly, the method
can highlight potential situations where livestock production systems could
benefit from an additional use of blue water resources to increase the water
use efficiency in the system.
Studies of environmental impact of beef production do not success-fully
capture all ecosystem benefits (Eshel et al., 2014). Thus, the method presented
in this study should be developed to also capture other competitive uses, such
as fuel and fiber production, and poten-
Y. Ran et al.
tially competition with ecosystem functions, e.g. by including competi-tion
with regulating and cultural ecosystem services, moving the concept of
sustainable intensification to also include eco-efficiency, that is to produce
more value with less impact (Tittonell, 2014). For example, overgrazing by
cattle may impose a threat to the ecosystem in terms of land degradation,
which may also cause large water losses in the long term (Saxton and Rawls,
2006; Bossio et al., 2007). Such an extended method could also be used, for
example, to assess potential benefits of integrated crop-livestock systems in
comparison with intensive agriculture (Lemaire et al., 2014).
4.3. Production of food crops or livestock feed
The methodology presented in this paper adds two new features to the
concept of quantifying CWU of livestock products: 1) a categorisa-tion of
water resources in classes, defined by land use, which enable identification of
how much of the total CWU could have been used for human food production
directly, 2) identification of indirect competi-tion over resources, by
calculation of a WUR, based on the potential of the water and land resources
that are currently used by the livestock system, to be used for another, more
beneficiary way to produce HDP.
The developed method enables identification of livestock systems that use
large amounts of green water with a low opportunity cost for the production
of food crops, and thus appear to be efficient in comparison to systems that
use water resources with higher opportunity costs. As livestock is increasingly
fed on human edible products such as grains, and productive cropland is
dedicated to animal feed production, it is important to demonstrate the
efficiency of water use for livestock in terms of food supply, e.g. by showing
how much of the total CWU could have been used to produce human food
crops more productively. This is not properly captured if water resources are
just quantified, even if they are categorized into blue and green water.
A study by Cassidy et al. (2013) indicate that the global calorie
availability could be increased by up to 70% if crops are directly consumed
by humans rather than used as animal feed and biofuel production. This study
also suggests that shifting meat consumption away from beef towards more
poultry and pig meat could potentially nourish more people per ha. The
results presented in our study, however, indicate that ruminants can play an
important role in future food security as they convert human non-edible
biomass to nutritious food. In terms of maximizing HDP per unit of water or
land, such production systems can be regarded as resource use efficient.
The WUR results presented in this paper, however, also show that, in
regard to water resources, it would be more efficient to produce HDP from
food crops rather than livestock for the two more intensive beef cattle
systems, SP-SP and SP-FL. The results are mostly dependent on the large
amount of sorghum that is fed to cattle in the feedlot systems and the use of
water for grass production on seeded pasture lands that are also suitable for
crop production. Only the livestock system relying entirely on natural pasture,
produce HDP more efficiently than food crops could. Results indicates that
alterations in feed composition may change the resource competition
significantly, which can be of great local importance and contribute to more
sustainable resource use in agriculture.
5. Conclusion
This study argues that water resource use for livestock should be analyzed
and considered based on three criteria. First, water resources use should be
distinguished as blue and green water resource use. Second, they should be
categorized according to the land over which they are evapotranspired. Third,
the competition over resources should be included in assessments to bring
significance to the large use of green water in livestock systems.
To tackle these issues we developed a water use ratio that enables
comparison of livestock production and plant production systems for
7
Agricultural Systems 155 (2017) 1–8
best use of water to produce human edible proteins. Results from this study
show that based on the traditional measure for water productiv-ity, i.e. liters
of CWU per kg beef produced, we would conclude that the most extensive
Uruguayan beef production systems use water resources less efficient than the
more intensive systems, whereas based on our new water use ratio, the
opposite conclusion would be drawn.
This study shows that livestock, and livestock production systems that
produce HDP from human non-edible biomass appropriating CWU from land
with none, or very low suitability for crop cultivation, can play an important
role in food security. It also indicates that some livestock production systems
use resources that may be more suitable for competing purposes, and that
multiple resources should be con-sidered, in order to contribute to the
identification of trade-offs and opportunities for improvement and sustainable
intensification.
Acknowledgements
This work was supported by the Commonwealth Scientific and Industrial
Research Organization [grant number R-04205 (Food Systems and Global
Change)], the CRP Livestock and Fish (L & F) and the CRP Water, Land and
Ecosystems (WLE). We would also like to thank Jens Heinke and Pablo
Modernel for support with data and modelling.
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 Animal Feed Science and Technology 254 (2019) 114190

Contents lists available at ScienceDirect

Animal Feed Science and Technology

journal homepage: www.elsevier.com/locate/anifeedsci

Use of naturally sourced feed additives (lactobacillus fermentation products

T
and enzymes) in growing and finishing steers: Effects on performance,
carcass characteristics and blood metabolites
a,b a,c a,d a,e a,
e Ran , W.M.S. Gomaa , Y.Z. Shen , A.M. Saleem , W.Z. Yang , T.A.
a
McAllister
iii) Lethbridge Research and Development Centre, Lethbridge, AB, T1J 4B1, Canada
iv) Institute of Subtropical Agriculture, the Chinese Academy of Sciences, Changsha, 410125, China
v) Department of Animal Nutrition and Clinical Nutrition, Assiut University, Assiut, Egypt
vi) College of Animal Science and Technology, Hebei Agricultural University, Baoding, 071001, China
vii) Department of Animal and Poultry Production, South Valley University, Qena, 83523, Egypt

ARTI CLE INF O AB STRACT

Keywords: An experiment was conducted to evaluate the use of naturally sourced feed additives that con-sisted of
Antibiotics lactobacillus fermentation products (LFPs), plant based enzymes and prebiotics as al-ternatives to antibiotics
Carcass characteristics in the diets of growing and finishing beef steers. The feed additives were two commercial products: Bio-Lac
Enzymes Plus (BL) and Boviglo (BG). During growing, 75 crossbred steers (initial body weight, 279 ± 4.9 kg) were
Growth performance blocked by weight and randomly allocated into five treatments: control; implant (IM; Elanco-Component
Probiotics
TE-100 with Tylan); implant + anti-biotics (330 mg monensin +110 mg chlortetracycline/steer/d; IMAT);
Steers
implant + BL (30 g/steer/ d BL; IMBL); and BG (5 mL/steer/d). Steers were fed a basal diet consisting of
600 g/kg corn silage, 350 g/kg dry-rolled barley grain, and 50 g/kg protein, vitamin and mineral supplement
(dry matter [DM] basis). When the 112 d growing period finished, steers were maintained on the same
treatments and gradually adapted to finishing diet within one month. Steers were not implanted during the
112 d finishing period. The finishing diet consisted of 100 g/kg corn silage, 870 g/kg dry-rolled barley grain,
and 30 g/kg vitamin and mineral supplement (DM basis). Steers were housed in individual pens with feed
intake measured individually over the 224 d experi-ment. During the growing period, no treatment effect on
DM intake was observed; however, final body weight, ADG and G:F were highest (P < 0.05) with IM, IMAT
and IMBL, intermediate with BG and lowest with control. A treatment × day on feed interaction (P < 0.05)
occurred for ADG, which was higher (P < 0.01) with IMBL than control or other treatments during first 14
days on-feed. Feeding BG improved (P < 0.01) ADG and G:F versus control animals. In addition, the need
to use therapeutic antimicrobials was lower (P < 0.01) for all treatments, in particularly with BL versus
control. During finishing, s neither BL or BG effected growth performance or carcass

Abbreviations: ADF, acid detergent fiber; ADG, average daily gain; BFT, back fat thickness; BG, Boviglo; BL, Bio-Lac Plus; BW, body weight; CAT, catalase; CP,
crude protein; DM, dry matter; DMI, dry matter intake; G:F, gain to feed ratio; GSH-Px, glutathione peroxidase; HCW, hot carcass weight; IM, implant; IMAT,
implant + antibiotics; IMBL, implant + Bio-Lac Plus; LAB, lactic acid bacteria; LFPs, lactobacillus fermentation pro-ducts; LM, longissimus muscle; TMR, total
mixed ration; NEFA, non-esterified fatty acids; aNDF, neutral detergent fiber analyzed with heat-stable α-amylase; OM, organic matter; T-AOC, total antioxidant
capacity
This research was funded by AAFC GF2 - Alternatives to Beef Antimicrobials, Bio-Ag Consultants and Distributors Inc. (Wellesley, ON) and Natures Wave
(Milverton, ON).
Corresponding author.
E-mail address: wenzhu.yang@canada.ca (W.Z. Yang).

https://doi.org/10.1016/j.anifeedsci.2019.05.013
Received 19 January 2019; Received in revised form 16 April 2019; Accepted 23 May 2019
0377-8401/ © 2019 Elsevier B.V. All rights reserved.
T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

characteristics. Greater (P < 0.05) antioxidant capacities were observed in steers fed IMAT than steers fed
IMBL and BG. These results indicate that supplementation of BL or BG improved ADG and feed efficiency
during the stressful, early portion of the growing phase, but had no effect during finishing. The results
suggest that both BL and BG have the potential to be fed as alter-natives to antimicrobial growth promoters
in growing steers.

1. Introduction

Hormonal anabolic implants and ionophore antibiotics have been widely used as growth promoters in North American beef production
(Johnson and Beckett, 2014; Ashima et al., 2016). There is increasing public concern over the use of these growth enhancing technologies, even
though there is no scientific evidence that they pose a serious food safety or public health risk. The European Union and some other countries
have banned the use of hormones and antibiotics for growth promotion (Herago and Agonafir, 2017). Consequently, there has been increased
interest in the use of feed enzymes, probiotics and prebiotics in ruminant production (Chaucheyras-Durand and Durand, 2010; Gaggia et al.,
2010). Addition of some probiotics to cattle diets can benefit growth performance and health, through the prevention of ruminal acidosis (Seo et
al., 2010). Moreover, some probiotics have been shown to inhibit foodborne pathogens such as E. coli O157:H7 (Wisener et al., 2015).

Meanwhile, the use of exogenous enzymes in animal production has attracted growing attention (McAllister et al., 2000; Beauchemin and
Holtshausen, 2011). Cellulases, hemicellulases, proteases and ferulic acid esterase activity are of primary interest in ruminants as they have the
potential to improve fiber digestion (Beauchemin and Holtshausen, 2011). Yang et al. (1999) reported promising responses to exogenous
fibrolytic enzymes in lactating dairy cows. However, compared to swine and poultry, the com-mercial use of exogenous feed enzymes in
ruminants is limited and the results are inconsistent (McAllister et al., 2000; Beauchemin and Holtshausen, 2011). Recently, mixtures of
exogenous enzymes and probiotics have become commercially available. However, reports about the effects of feeding mixtures of exogenous
enzymes and probiotics on the growth performance and carcass char-acteristics of beef steers are scarce. Bio-Lac Plus (BL) and Boviglo (BG)
are commercial feed additives that contain lactic acid bacteria (LAB), lactobacillus fermentation products (LFPs), and enzymes. We
hypothesized that feeding BL to implanted cattle would have additive effects on growth performance, and that BG may act as an alternative to
the current use of implants and in-feed antibiotics in beef cattle. We also hypothesized that feeding of BL and BG may impact carcass traits,
possibly by exogenous enzymes increasing fiber digestion which could alter fat deposition. The objectives of this study were to examine BL and
BG as alternatives to in-feed antibiotics and to investigate their effects on growth performance, carcass traits, blood metabolites and antioxidant
status of growing

Table 1
Ingredients and chemical composition of growing and finishing diets.

Item Diet

Growing Finishing

Ingredients, g/kg DM
1
Corn silage 600 100
2
Barley grain, dry-rolled 350 870
Barley, ground 8.4 16.4
Canola meal 18.9 2.9
Calcium carbonate 17.3 7.3
3
Feedlot premix 0.3 0.3
Molasses 0.7 0.7
Salt 1.5 1.5
Urea 3 0.6
Vitamin E (500,000 IU/kg) 0.02 0.2
Canola oil 0.3 0.3
Chemical composition, g/kg DM
DM 548 843
OM 934 969
NDF 363 187
ADF 167 56
Starch 341 546
CP 123 125

2 Composition (g/kg, DM basis): 312 DM, 453 NDF, 220 ADF, 281 starch, 95 CP, measured based on 5
samples composited by period.
3 Composition (g/kg, DM basis): Growing phase, 900 DM, 978 OM, 170 NDF, 46 ADF, 577 starch and 139
CP; finishing phase, 902 DM, 978 OM, 176 NDF, 42 ADF, 583 starch and 130 CP, measured based on 5 samples
composited by period.
4 Supplied (g/kg, DM basis): 15 mg Cu, 65 mg Zn, 28 mg Mn, 0.7 mg I, 0.2 mg Co, 0.3 mg Se, 6000 IU
vitamin A, 600 IU vitamin D, and 47 IU vitamin E.

2
T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

and finishing cattle.

2. Materials and methods

The study was reviewed and received approval of the Institutional Animal Care and Use Committee at the Lethbridge Research and
Development Centre and was conducted in accordance with the guidelines of the Canadian Council on Animal Care (2009).

2.1. Additive descriptions

Two commercial products, BL and BG developed, respectively, by Bio-Ag Consultants Ltd. (Wellesley, ON, Canada) and Natures Wave
(Milverton, ON, Canada), were examined during the experiment. The BL was a naturally sourced feed additive that contains LFPs, prebiotics
and plant based enzymes (cellulase, protease and amylase). The BG was in liquid form and consisted of a wide range of vitamins and trace
minerals, fermented lactobacillus product, naturally occurring plant enzymes, selenium and whey concentrates. Detailed information of the LFPs
and enzyme activity of the BL and BG was not available due to commercial confidentiality.

2.2. Animals and experimental design

Seventy-five Charolais and Angus cross steers (aged 8 ± 0.5 months) were purchased from a local auction market, housed in individual pens
(4.9 × 1.8 m) bedded with wood chips at the Agriculture and Agri-Food Canada, Lethbridge Research and Development Centre feedlot unit.
After arrival, steers were ear tagged and vaccinated against IBR, PI3, Haemophilus somnus (Resvac 2/Somubac, Pfizer Animal Health) and
Clostridium spp. (Tasvax 8, Schering-Plough Animal Health, Upper Hutt, NZ).
The experiment was a randomized complete design, and consisted of growing and finishing phases, with 112 d for each phase. At the start of
the experiment, steers (initial body weight [BW] 279 ± 4.9 kg) were weighed on two consecutive days, blocked by weight and randomly
allocated to one of five treatments:1) control (basal diet); 2) implant (IM; Elanco-Component TE-100 with Tylan, containing 100 mg of
trenbolone acetate USP, 10 mg of estradiol USP and 29 mg of tylosin tartrate; Elanco Canada Limited, ON, Canada); 3) implant + antibiotics
(330 mg/steer/d monensin +110 mg/steer/d chlortetracycline; IMAT); 4) implant + BL (30 g/steer/d BL; IMBL); and 5) BG (5 mL/steer/d). Bothe
IM or IMAT are standard industry practices in North American feedlots, thus, IMBL was designed to determine the additional benefit of adding
BL, but BG was designed to assess whether BG can be fed as an alternative to implant or implant plus antibiotics. Steers were fed a total mixed
ration (TMR) once daily containing 600 g/kg corn silage, 350 g/kg dry-rolled barley grain and 50 g/kg protein, vitamin and mineral supplement
(Table 1; dry matter [DM] basis). The diet was formulated to meet the nutrient requirements of beef cattle (National Academies of Sciences and
Engineering, and Medicine (NASEM, 2016) for a targeted gain of 1.2 kg/d during the growing phase. Antibiotics were mixed with 100 g carrier
(90 g dry-rolled barley and 10 g molasses) and top-dressed during feeding. The same amount of carrier was also provided to steers on the other
treatments. The BL was administered by mixing it into the TMR during formulation just prior to feeding, whereas BG (5 mL/steer/d) was diluted
in 50 mL of tap water and sprayed onto the ration at mixing. When the growing phase finished, steers were maintained on the same treatments
during the finishing phase, but were not re-implanted. Steers were adapted to the final finishing diets that consisted of 100 g/kg corn silage, 870
g/kg dry-rolled barley grain, and 30 g/kg protein, vitamin and mineral supplement (Table 1; DM basis) over a 4-wk period by gradually replacing
corn silage with dry-rolled barley grain. Steers had free access to clean water over the 224 d experiment.

2.3. Feed sampling and chemical analyses

Feed offered and refused were recorded daily at 0800 for each steer, and daily intake was calculated. Samples of diets and refusals were
collected weekly and subsampled for DM determination. Weekly samples (TMR and refusals) were pooled over 4-wks, mixed, subsampled and
dried in an oven at 55 °C for 48 h (AOAC, 2005; method 930.15). The samples were then ground through a 1 mm screen using a Wiley mill
(standard model 4; Arthur Thomas Co., Philadelphia, PA, USA), and analyzed for DM, organic matter (OM), neutral detergent fiber (aNDF) with
heat-stable α-amylase and sodium sulfite used, acid detergent fiber (ADF), starch and crude protein (CP) as describe by Yang et al. (2010). Ash
content was determined by combustion at 550 °C for 5 h, and OM content was calculated as 100 minus the proportion of ash ( AOAC, 2005;
method 942.05).

2.4. Growth performance

Steers were weighed individually on two consecutive days at the start and the end of each study phase, and every subsequent 28-d. In the
growing phase, an additional weighing was conducted on d 14 to investigate treatment e ffects when stress was highest at the beginning of the
trial. Individual DM intake (DMI) was calculated as the DM offered minus the DM refused. Averaged daily gain was calculated by dividing the
BW gained (final BW – initial BW) by the number of days on-feed. Feed efficiency (G:F) was estimated by dividing ADG by DMI. Net energy
for maintenance (NEm) content of each diet were estimated from animal weights, DMI and ADG as described by Zinn et al. (2002) using the
0.75
retained energy formula for large-framed yearlings (0.0437 × BW × ADG1.097; NRC, 1984). The NEm was converted to NEg according to
Zinn et al. (2002; NEg = NEm × 0.877 – 0.41).

3
T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

2.5. Fecal pH, total generic E. coli and IgA

Fresh feces (approximately 400 g wet) were randomly collected before the morning feeding from 10 individual steers of each treatment on d
®
56 and 112 of the finishing phase. After collection, sample was immediately measured for pH using a portable pH meter (pHep , Hanna
Instruments, Calgary, AB, Canada), and then subdivided into two portions for analyzing generic E. coli and fecal IgA, respectively. For total
enumeration of generic E. coli, 1 g of fecal material was weighed into a Hungate culture tube containing 9 mL of 1 g/L peptone dilution water,
−6 -4 -5 -6
vortexed vigorously and serial dilutions to 10 were prepared in peptone water. One milliliter of 10 , 10 and 10 dilution was dispensed onto
an E. coli Petrifilm plate (3 M Canada, London, ON, Canada) and incubated at 37 °C for 24 h. Plates that fell within the range of 15 to 150
7
colonies were counted. All blue colonies were reported as E. coli (10 cells/g feces). For determining IgA, 1 g of feces was weighed and placed
immediately in ultra-purified water at a concentration of 1 g/mL by adding 9 mL of distilled water in a 15 mL centrifuge tube. The tubes were
vortexed vigorously and incubated overnight before centrifuging at 2000 × g for 15 min at 4 °C. The supernatants were then collected and
analyzed for total IgA using a commercial ELISA kit (Bovine IgA ELISA Quantitation Set, Bethyl Laboratories, Montgomery, TX, USA).

Table 2
Effects of implant, antibiotics or lactobacillus fermentation products and enzyme-based additives on growth performance of growing and finishing steers.

1
Item Treatments SEM P<

Control IM IMAT IMBL BG

Number of steers 15 15 15 15 15
Growing
DMI, kg/d
d0-14 5.33 5.12 5.27 5.43 5.6 0.214 0.61
d15-28 5.79 6.20 6.48 6.82 6.28 0.235 0.45
d29-56 7.85 8.38 7.52 8.44 8.35 0.272 0.08
d57-84 9.05 8.89 8.82 9.36 8.86 0.266 0.57
d85-112 9.38 9.51 9.37 9.6 9.39 0.293 0.97
Overall 7.95 8.11 7.90 8.38 8.13 0.225 0.57
BW, kg
Initial 279 279 279 279 279 4.9 1.0
d14 287 291 290 293 289 5.4 0.94
d28 302 310 313 318 305 6.2 0.43
d56 329 347 343 350 332 6.3 0.08
b a a a ab
d84 359 386 380 387 370 6.8 0.02
b a a a ab
d112 387 420 414 417 400 7.6 0.02
ADG, kg/d
b ab ab a ab
d0-14 0.54 0.81 0.77 0.99 0.76 0.071 0.05
b ab a a b
d15-28 1.10 1.38 1.69 1.78 1.11 0.074 0.05
b a b ab b
d29-56 0.98 1.33 1.03 1.14 0.96 0.048 0.05
b a a a a
d57-84 1.05 1.39 1.35 1.31 1.35 0.041 0.05
b ab a ab ab
d85-112 0.99 1.18 1.22 1.06 1.06 0.033 0.05
c a a a b
Overall 0.99 1.28 1.23 1.25 1.11 0.052 0.01
G:F, g/kg
c b b a bc
d0-14 101 158 146 182 136 7.6 0.01
bc b a a c
d15-28 190 223 261 261 177 8.2 0.01
bc a ab bc c
d29-56 131 165 159 141 120 7.7 0.01
b a a a a
d57-84 116 157 154 139 153 7.1 0.01
d85-112 111 125 128 112 115 7.0 0.29
c a a a b
Overall 124 157 155 150 136 3.8 0.01
Drug treatments
#steers (/15) 8 2 3 0 2 … …
a b b c b
% 53.3 13.3 20.0 0 13.3 … 0.05
Finishing
b a ab a b
Initial BW, kg 428 459 449 457 433 8.5 0.02
Final BW, kg 606 634 633 630 617 10.5 0.27
ADG, kg/d 1.58 1.56 1.65 1.54 1.65 0.075 0.43
DMI, kg/d 10.6 10.6 10.5 10.7 10.9 0.39 0.94
G:F, g/kg 149 145 156 145 150 4.8 0.35
2
NEg , Mcal/kg 1.29 1.32 1.41 1.30 1.30 0.033 0.09
2
NEm , Mcal/kg 1.94 1.98 2.07 1.95 1.95 0.037 0.09

a,b,c
Means within a row with different superscripts differ (P < 0.05).
b IM = Implant; IMAT = Implant + Antibiotics (330 mg/steer/d monensin and110 mg/steer/d chlortetracycline); IMBL = Implant + BioLac Plus (30 g/steer/d); BG
= Boviglo (5 mL/steer/d). Treatment × feeding period were significant at P < 0.05 for ADG and G:F.

4
T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

2.6. Blood metabolites and antioxidant activity

The same 10 steers used for fecal sampling were used to collect blood samples before the morning feeding on d 56 and 112 of the finishing
phase. One 10-mL vacuum tube containing Na heparin, and one 10-mL vacuum tube without additive (Vacutainer, Becton Dickinson, Franklin
Lakes, NJ, USA) were collected from the jugular vein. Plasma samples were centrifuged at 3000 × g for 20 min at 4 °C, and serum samples were
centrifuged at 2000 × g for 15 min at 4 °C. Collected plasma and serum were frozen at −20 °C until analyzed. Blood glucose, urea N, and non-
esterified fatty acids (NEFA) were determined as described by Yang et al. (2010). Anti-oxidant status in blood was characterized by analyzing
antioxidant enzymes including glutathione peroxidase (GSH-Px), total an-tioxidant capacity (T-AOC), and catalase (CAT) in plasma (He et al.,
2012).

2.7. Carcass characteristics

At the end of the experiment, steers were shipped to a commercial abattoir for slaughter. Carcass characteristics including hot carcass weight
th
(HCW, with kidneys removed), dressing percentage, 12 -rib back fat thickness (BFT), longissimus muscle (LM) area, marbling score, quality
grade, saleable meat yield and abscessed livers (including severely abscessed) were recorded for each carcass. Dressing percentage was
calculated individually as HCW divided by final live BW × 100. Marbling scores were estimated according to pictorial standards from 1 (devoid)
to 10 (abundant marbling; USDA, 1989). Quality grade was carried out as Canada AAA standards: Canada grade A is approximately equivalent
to USDA Standard; AA is approximately equivalent to USDA Select; and AAA is approximately equivalent to USDA Choice. Saleable meat
yield was estimated with consideration for the length, width, and fat cover of the LM between the 11th and 12th rib, as estimated lean yield =
57.96 – 0.027 HCW + 0.202 LM – 0.703 BFT. Liver abscess scores were determined based on the ranking scale used by the Canadian Beef
Grading Agency. Severe liver abscesses were defined as liver with at least four small abscesses or at least one abscess with diameter larger than
2.5 cm.

2.8. Statistical analysis

For the growing phase, data of DMI, BW, ADG, and G:F were analyzed using the Mixed procedure of SAS (Version 16.0.0, SAS Inst. Inc.
Cary, NC) as a completely randomized design with treatment, period (days on-feed), and their interaction as fixed effects and steers within
treatment as a random effect. For the finishing phase, data of DMI, BW, ADG and G:F were analyzed using the Mixed procedure of SAS with
treatment, period (days on-feed) and their interaction as fixed effects and steers within treatment as a random effect. Days on feed were removed
from the model because there was no interaction between days on feed and treatment. Carcass data were analyzed using the Mixed procedure of
SAS with treatment as a fixed effect and steers within treatment as a random effect. Meat quality grade, liver abscesses and drug treatment data
were analyzed using the GLIMMIX procedure. Data for blood metabolites, antioxidants and fecal measurements were analyzed using the Mixed
procedure of SAS with treatment as a fixed effect and steers within treatment as a random effect, and with day as a repeated measure. For
repeated measures, various covariance structures were tested and AR(1) was selected based on the lowest value for Akaike’s information criteria.
Least square means were compared using the Tukey correction for multiple comparisons, and treatment e ffects were declared significant at P ≤
0.05 and trend at 0.05 < P ≤ 0.10.

3. Results

3.1. Growth performance

In the growing phase, DMI tended (P < 0.08) to be greater for IM, IMBL and BG steers than control and IMAT steers from d 29 to 56, but
did not differ over the entire growing phase (Table 2). Body weight was higher (P < 0.02) for IM, IMAT and IMBL, inter-mediate with BG and
lowest with control steers at d 84 and 112. There was a treatment × days on-feed (P < 0.05) interaction for ADG. The IM, IMAT and IMBL steers
had greater (P < 0.05) or tended to have greater (P < 0.10) ADG than control steers on most of the days on-feed and as a result, the overall ADG
was greater (P < 0.01). Steers fed BG versus control tended (P < 0.08) to have greater ADG at d 14 and greater (P < 0.05) ADG at d 84 and
overall, but had a lower (P < 0.05) ADG than IM and IMAT steers. Similar to ADG, a signi ficant (P < 0.05) treatment effect and treatment × day
on-feed interaction on G:F was detected. The G:F was greater for steers with IM, IMAT and IMBL on d 14 and 28 than control steers, and
greater G:F with all four treatment groups than control group after 84 d on feed. As a result, overall G:F was greatest (P < 0.01) for steers
receiving IM, IMAT and IMBL treatments, intermediate with BG and lowest with control steers. During the first two months of the growing trial,
® ® ®
some steers often required additional antimicrobials including Resflor , Micotil or Nuflor for treating undifferentiated bovine respiratory
disease. The numbers of steers that required drug treatments was lower (P < 0.05) with treatments as compared to control.

In the finishing phase, the initial BW was greater (P < 0.02) for steers receiving IM and IMBL than for control and BG steers which was due
to the treatment effects in the growing phase (Table 2). However, final BW did not differ among treatments, and no treatment effects on DMI,
ADG and G:F were observed. Steers receiving IMAT tended (P < 0.09) to have greater NEg and NEm than the other treatments.

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3.2. Carcass characteristics

Carcass characteristics including HCW, dressing percentage, BFT, LM, and saleable meat yield were not affected by treatments except that
marbling score tended (P < 0.07) to be greater for IM steers than IMAT and IMBL steers (Table 3). Moreover, the IMAT steers had lower (P <
0.01) meat quality grade than steers on the other four treatments. Proportion of total abscessed livers did not di ffer among treatments; however,
the proportion of severely abscessed livers was greater (P < 0.05) with BL and BG than control, but did not differ among the other treatments.

3.3. Blood metabolites and antioxidant activity

Blood urea N and NEFA concentrations did not differ among treatments (Table 4). Blood glucose concentration was greater (P < 0.03) for
BG steers than IM and IMAT steers. Blood antioxidant was greatest (P < 0.01) with IMAT, followed by IM, IMBL and BG treatments and
control steers, respectively. The IMAT steers had greater (P < 0.01) CAT than IMBL, BG and control steers. The IMBL and BG steers had less (P
< 0.01) GSH-Px than IM, IMAT and control steers.

3.4. Fecal pH, E. coli counting and IgA

Fecal pH and total E. coli counts in feces were not affected by treatment (Table 4). However, fecal IgA contents were greater (P < 0.04) with
control and BG steers than with IMAT and IMBL steers.

4. Discussion

Both BL and BG are LFPs and plant sourced enzyme-based feed additives, and commercially available for use in North American feedlots.
However, there is no published data on the use of these products in feedlot cattle. Furthermore, both products are a mixture that might have
multiple mechanisms to benefit the host and may have beneficial effects on a wider portion of the cattle population, making it very difficult to
attribute responses to any one component in the mixture. Therefore, direct comparisons of the present results with other experiments are not
evident.

4.1. Growth performance

Overall the increased final BW and ADG and improved G:F with steers receiving IM, IMAT or IMBL treatments confirmed the
improvements in beef cattle performance through use of anabolic implants. With the present diet, supplementing monensin and chlortetracycline
in implanted steers did not show additional benefits for ADG over the implant alone. Implants and monensin promote feed efficiency in cattle via
different mechanisms (Bretschneider et al., 2008). Anabolic implants improve growth perfor-mance by increasing levels of somatotropin and
IGF-1, which increase synthesis of muscle tissue and reduce fat deposition (Duckett and Andrae, 2001). Feeding monensin increases feed
efficiency by improving animal health and reducing feed intake and by

Table 3
Effects of implant, antibiotics or lactobacillus fermentation products and enzyme-based feed additives on carcass traits of finishing steers.
1
Item Treatments SEM P<

Control IM IMAT IMBL BG

Number of steers 15 15 15 15 15
HCW, kg 385 403 397 400 391 7.1 0.43
Dressing, g/kg 587 589 588 588 584 0.5 0.66
Back fat, mm 24.5 22.3 24.4 26.0 26.8 1.33 0.16
2
LM area, cm 86.5 87.8 90.1 87.7 86.3 1.96 0.68
2
Marbling score 5.96 6.33 5.69 5.87 6.22 0.174 0.07
3
Saleable meat yield , g/kg 478 492 482 466 460 1.1 0.27
4 a a b a a
Quality grade AAA , % 100 100 86.7 100 100 … 0.01
Liver abscess score
5
Abscessed livers , % 53.3 53.3 33.3 40.0 53.3 … 0.76
6 c b b b a
Severely abscessed , % 0 6.7 6.7 13.3 33.3 … 0.05

a,b,c
Means within a row with different superscripts differ (P < 0.05).
1 IM = Implant; IMAT = Implant + Antibiotics (330 mg/steer/d monensin and110 mg/steer/d chlortetracycline); IMBL = Implant + BioLac Plus (30 g/steer/d); BG
= Boviglo (5 mL/steer/d).
2 According to pictorial standards from 1 (devoid) to 10 (abundant marbling) (USDA, 1989).
3 Estimated lean yield = 57.96 - 0.027 HCW + 0.202 LMA - 0.703 BFT.
4 1 = Canada grade A (approximately equivalent to USDA Standard); 2 = AA approximately equivalent to USDA Select); and 3 = AAA (ap-proximately equivalent
to USDA Choice).
5 The percentage of liver with at least 1 abscess.
6 The percentage of liver with at least 4 small abscesses or at least 1 abscess with a diameter greater than 2.5 cm.

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T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

Table 4
Effects of implant, antibiotics or lactobacillus fermentation products and enzyme-based feed additives supplementation on blood metabolites and antioxidants of
finishing steers.
1
Item Treatments SEM P<

Control IM IMAT IMBL BG

Number of steers 10 10 10 10 10
Blood metabolites
Blood UN, mg/dL 12.80 13.99 13.61 13.75 13.48 5.421 0.61
ab b b ab a
Glucose, mg/dL 81.80 70.91 72.14 76.56 89.33 0.045 0.03
NEFA, mmol/L 0.20 0.19 0.18 0.24 0.21 0.018 0.18
Blood antioxidants
c b a b b
Antioxidant, mM 0.09 0.11 0.14 0.11 0.11 0.006 0.01
2 bc ab a bc c
CAT , nmol/min/mL 17.5 19.9 23.5 16.8 14.0 1.46 0.01
2 a a a b b
GSH-Px , nmol/min/mL 179.6 184.8 173.9 157.2 153.5 4.88 0.01
Fecal
pH 6.27 6.31 6.36 6.15 6.07 0.095 0.095
a ab b b a
IgA, μg/g 2.46 1.92 0.78 1.02 2.13 0.433 0.433
7
E. coli, × 10 cfu 10.97 9.95 15.02 7.74 8.33 2.891 2.891

a,b,c
Means within a row with different superscripts differ (P < 0.05).
1 IM=Implant; IMAT=Implant + Antibiotics (330 mg/steer/d monensin and110mg/steer/d chlortetracycline); IMBL = Implant+BioLac Plus (30g/steer/d);
BG=Boviglo (5 mL/steer/d).
2 Catalase = CAT; GSH-Px = Glutathione Peroxidase.

stabilizing rumen fermentation and altering fermentation patterns so as to produce greater molar proportion of propionate at the expenses of
acetate (Duffield et al., 2012). Bretschneider et al. (2008) suggested an additive response to monensin and implant with 17-β-estradiol was
expected based on their distinct site and mode of action. However, in agreement with the present findings, Utley et al. (1976) reported no
synergistic effect between monensin and implant (zeranol or testosterone-estradiol) during growing and finishing. Bretschneider et al. (2008)
concluded in a review that there was no evidence of combined benefits as the result of the use of both monensin and implants because of the
reduced effect of monensin on ADG of cattle fed high-quality forages. In the current study, steers were fed high quality silage, which might
partially explain the similar results between IM and IMAT.
The additive effect of feeding LFPs to implanted cattle on growth performance was not apparent. Although ADG of IMBL steers was
improved as compared to control steers, the lack of a difference between IM and IMBL suggested no additive effects as a result of this additive.
Lactic acid bacteria have been shown to inhibit pathogens, enhance host immune responses and favorably alter ruminal fermentation (Krehbiel et
al., 2003). However, compared to IM, it is noteworthy that IMBL steers tended to exhibit greater ADG and improved feed efficiency during the
first 28 d of the growing phase. Furthermore, no IMBL steers required treatment with anti-microbials as compared to the IM and control steers.
These results suggest that supplementation with BL may have potential growth and health benefits for steers during the early feeding period
where stress is often highest. It is widely accepted that LAB can maintain the intestinal microbial balance and prevent the establishment of
opportunistic pathogenic bacterial populations (Dicks and Botes, 2010). The LAB enhance energy efficiency by altering ruminal fermentation
and decreasing the use of energy for tissue turnover in the gastrointestinal tract (Elam et al., 2003). This mode of action is consistent with the
improved G:F.
Although BG steers had lower BW, ADG and G:F than IM, IMAT and IMBL steers, overall ADG and G:F of these steers was higher than
control steers. Similar results were reported by Aydin et al. (2009), where supplementation of diets with 20 g of a mixture of probiotic
(Lactobacillus acidophilus, L. casei, L. plantarum, Bacillus subtilis) and enzyme (amylase, protease, cellulose, lipase, pectinase) improved G:F
of young bulls, without affecting DMI. Greater overall ADG was also observed by Ponce et al. (2011) in steers receiving BeefPro, a product
containing LAB and digestive enzymes (amylase, proteases, hemi-cellulases, cellulose). Kocyigit et al. (2016) recently suggested feeding
combinations of probiotics and exogenous enzymes to calves had a beneficial effect on growth perfor-mance and feed efficiency of young
ruminants. The steers supplemented with BG also required less therapeutic antimicrobials as compared to the control steers. Fermented
lactobacillus products may have prebiotic effects, plant enzymes may have positive effects on fiber digestion, while whey concentrates may help
improve gastrointestinal immune status (Perez-Cano et al., 2007). Those researchers reported that whey protein concentrate modulates
maturation of immune system in suckling rats (Perez-Cano et al., 2007). Finally, BG contained a wide range of highly-available vitamins and
trace minerals, that may not only meet vitamin and mineral requirements, but also have a functional role in improving immune responses.

Usually, less responses were observed with longer feeding duration, with more positive responses in the growing than finishing period. In the
current study, final BW at the finishing phase had the same classification to the end of growing phase; the changes of BW of treatments vs.
control were similar between the two growing phases (growing vs. finishing; 33 vs. 28, 27 vs. 27, 30 vs. 24, and 13 vs. 11, respectively, for IM,
IMAT, IMBL and BG vs. control). The results suggest that the compensation of the lower BW during the finishing feeding was limited. The lack
of statistical significance in final BW was partly due to increased variation among individuals (SEM; 7.6 vs. 10.5), suggesting a greater
individual variation in finishing than the growing phase. Steers were not re-implanted during finishing phase, thus the absence of difference in
growth performance between the previous implanted steers and control is consistent with the observation of no additive e ffect of adding
antibiotics or BL in growing phase, and suggests that responses observed during growing phase were mostly a result of the implants. The present
results agreed with study of Aydin et al. (2009) who

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reported that feeding LAB plus enzymes improved weight gain and G:F for growing, but not finishing bulls. When the LAB and enzyme were
fed alone, the effects of LAB and exogenous enzymes (cellulase, protease and amylase) on growth performance of finishing beef steers varied
among studies. As for LAB, Seo et al. (2010) indicated that the discrepancies among studies may be due to the differences in LAB strains used,
the amount of LAB fed, type of diets, type of animals or their growth stage. No treatment effects of feeding LAB on finishing performance were
reported under controlled experimental conditions (Elam et al., 2003) or in a larger scale commercial feedlot trial (Greenquist et al., 2004).
Interestingly, Swinney-Floyd et al. (1999) reported increased ADG by calves supplemented with a LAB in the early stages of finishing, but the
response diminished over the entire 120-d feeding period. This is because steers faced a challenge of dietary change from high forage to high
concentrate diet during the early finishing period, and LAB may have helped maintain the intestinal microbial balance and improve immunity.
However, with increasing days on-feed, steers adapted to the diet and experimental conditions, thus the e ffect of LAB on intestinal function
could have been diminished. This effect is expected as we observed in the growing phase, where adding IMBL demonstrated beneficial effects
compared to IM and IMAT only in the first 28 days. Although the original concept of feeding LAB to livestock is based primarily on possibly
intestinal effects, their beneficial effects in the rumen has been reported, particular with regard to preventing ruminal acidosis (Krehbiel et al.,
2003). For example, lactate-producing bacteria may help prevent ruminal acidosis by promoting the activity of lactate-utilizing ruminal
microorganisms in the rumen (Ghorbani et al., 2002). Whereas, rumen lactate concentration is often low in cattle adapted to high-grain diets
(Beauchemin et al., 2003), subclinical ruminal acidosis is primarily caused by high VFA concentrations as a result of the rapid fermentation of
starch. Jiao et al. (2017) reported greater in vitro rumen DM digestibility and ratio of A:P when LAB were included in high grain diet, suggesting
an LAB mediated improvement of fiber digestion. The beneficial effect of LAB on fibre digestion was likely due to alterations in fermentation so
as to promote the activity of fibrolytic bacteria (Weinberg et al., 2007). However, improvements in fiber digestibility of a high-grain diet may
have limited effect on growth performance, particularly for cattle fed a diet that was formulated to meet or exceed nutrient requirements
(National Academies of Sciences and Engineering, and Medicine (NASEM, 2016). This may account for the lack of an effect of BL and BG
during the finishing phase.

Response to exogenous enzyme supplementation by finishing steers varied substantially, depending on feed types, enzyme sources, dosages
and application methods (McAllister et al., 2000). Commercially developed feed enzymes for ruminants primarily focussed on fibrolytic
enzymes because fiber digestion is a major limitation to improved feed digestion. However, with the barley-based high grain diet used in this
study, forage fiber accounted for a small proportion of the diet as its primary role is to stimulate chewing and maintain rumen physical and
dynamic function. Furthermore, the enzymes in BL or BG were not specified for their activities. Therefore, any potential improvement in fiber
digestibility due to exogenous enzymes would be expected to be minor. The beneficial effect of adding BG on growth performance in growing
cattle was not apparent during finishing. Additionally, the finishing diet was formulated to meet or exceed tnutrient requirements (National
Academies of Sciences and Engineering, and Medicine (NASEM, 2016) and other than dietary energy, other nutrients are less critical including
the vitamins and minerals contained in BG. Consequently, addition of BL or BG in finishing diet did not benefit the performance of finishing
cattle.

4.2. Carcass characteristics

The carcass traits including HCW, dressing percentage, BFT, LM area, and saleable meat yield did not differ among treatments. The lack of
treatment effects on carcass characteristics is consistent with the similar growth performance among treatments. Similarly, Ponce et al. (2011)
reported that major carcass characteristics were not affected except the HCW tended to be greater for cattle fed LAB and digestive enzymes than
control steers. Several studies reported no alteration in carcass characteristics when LAB were added to the diets of finishing beef steers (Elam et
al., 2003; Greenquist et al., 2004). The carcass weights and traits of feedlot cattle were also not affected by exogenous fibrolytic enzymes
(McAllister et al., 1999). However, the greater incidence of severely abscessed liver in steers fed BL or BG than control steers was unexpected,
and we have no clear explanation as to the cause of this observation. The results need to be confirmed using more animals or in commercial
feedlots. Inconsistent with the present result, Ponce et al. (2011) reported no difference in the proportion of abscessed livers between control and
steers supplemented with LAB and digestive enzymes. Present control of liver abscess in feedlot cattle largely relies on the use of antibiotics and
chlortetracycline is one of five antibiotics approved for prevention of liver abscesses in North America. These antibiotics vary in their
effectiveness in preventing liver abscesses (Nagaraja and Chengappa, 1998). Brown et al. (1975) reported that chlortetracycline was not as
effective as tylosin at preventing liver abscesses. In the present study, although the prevalence of abscessed livers was numerically lower with
IMAT than control (33.3 vs. 53.3%), its would still be considered high and even more severely abscessed livers occurred with IMAT than
control. Amachawadi and Nagaraja (2016) indicated that the incidence of liver abscesses in tylosin-fed cattle may be due to the development of
resistance in Fusobacterium necrophorum or abscesses are caused by other bacteria that are insensitive to tylosin. Nevertheless, these results
should be interpreted with caution due to the relatively small number of animals used.

4.3. Blood metabolites and antioxidant activity

Blood urea N concentrations can be used as an indicator of the efficiency of utilization of dietary protein within a group of animals (Kohn et
al., 2005). In the current study, the blood urea N concentration did not differ among treatments, a result that is consistent with other studies
where LAB have been added to the diet of calves (Quintero-Gonzalez et al., 2003; Bayatkouhsar et al., 2013). Monensin is also known to inhibit
ruminal protein degradation and decrease flow of microbial protein to the intestine (Ruiz et al., 2001), but increase the flow of dietary AA to the
abomasum (Yang and Russell, 1993). The lack of effect of monensin on protein metabolism in the present study is in agreement with its limited
effect on growth performance of finishing cattle. Blood glucose and

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T. Ran, et al. Animal Feed Science and Technology 254 (2019) 114190

NEFA concentrations are important indicators of energy status. Typically, serum NEFA concentrations increase if fat stores are mobilized to
provide energy for the physiological function of tissues. The lack of a difference in blood NEFA among treatments suggests that none of the
additives had a significant impact on the energy balance of finishing steers. Blood glucose concentration did not differ among control, IMBL and
BG treatments, which is in agreement with previous studies that feeding LAB probiotics had no e ffect on blood glucose (Ghorbani et al., 2002;
Beauchemin et at., 2003). Taken together, supplementing of BL or BG did not affect energy status or dietary protein utilization of finishing
steers.
Blood total antioxidant capacity is a marker of a non-enzymatic antioxidant defense system. In antioxidant defense systems, superoxide
dismutase (SOD) is the antioxidant enzyme that catalyzes the dismutation of the highly reactive superoxide anion to less reactive hydrogen
peroxide. Peroxides can be inactivated by CAT and GSH-P×. Increasing antioxidant enzymes like CAT and GSH-Px is a compensatory
mechanism due to increased oxidative stress. In the current study, steers supplemented with BL or BG had greater total blood antioxidant
capacity than control steers, but blood GSH-Px was reduced by supplementing with either BL or BG and CAT tended to be reduced by BG.
These results suggest that steers supplemented with BL or BG had lower oxidative stress compared to control, IM and IMAT steers. Cattle are
often exposed to stressful conditions resulting from nutrition (e.g. high-grain), environment and management (weaning, transportation). Stressful
events have been implicated in promoting oxidative stress through the pro-duction of excessive levels of reactive oxygen species (McGuffey et
al., 2001). Overproduction of reactive oxygen species may result in oxidative stress ( Valko et al., 2007) and changes in the antioxidant defense
systems, including SOD, CAT and GSH-Px (Han et al., 2011). Supplementation of natural antioxidants, such as vitamin E, vitamin C, tea
polyphenols, probiotics can counteract the negative impact of these reactive species on stress in livestock husbandry (West et al., 2008; Deng et
al., 2010). Therefore, feeding of BL or BG that contain probiotics (LAB), prebiotics, vitamin and minerals, has potential to reduce cattle stress,
and thus improve growth performance and animal health.

4.4. Intestinal immune status and fecal microflora

Modulation of host immunity is another mechanism by which probiotics promote intestinal health and overall well-being of the host (Isolauri
et al., 2001). A key strategy of intestinal immune protection is the production of IgA, which is largely produced in mucosal lymphoid tissues and
plays an important role in mucosal immunity (Maeda et al., 2013). In the current study, the similar fecal IgA concentrations among control, IM
and BG steers suggests that feeding BG did not stimulate immunity. However, the lowered fecal IgA in steers fed IMAT or IMBL is somewhat
unexpected as fecal IgA concentration is often used as indicator of mucosal immunity (Suzuki et al., 2004). Shen et al. (2018) reported increased
fecal IgA with monensin because it decreased fecal LPS; while LPS-induced endotoxemia inhibited humoral immune function of the digestive
tract and decreased the number of IgA + B cells.
In the current study, although only generic E. coli were measured, feeding either BL or BG numerically (P < 0.15) reduced the number of
total E. coli. It suggests that feeding BL or BG may potentially reduce faecal E. coli shedding. This result is consistent with previous reports that
feeding LAB is effective in reducing E. coli fecal shedding in feedlot cattle (Younts-Dahl et al., 2004; Peterson et al., 2007). Thus, we speculate
that the decreased fecal IgA concentration may have resulted from reduction in the need for immune responses through direct inhibition of
pathogens. However, the precise mechanisms underlying the beneficial effects of LAB have not been clearly delineated. Sherman et al. (2005)
reported that LAB can benefit the hosts via excluding pathogens from the host epithelium. The reason for the numerically greater fecal E. coli
with monensin is not clear. It has been hypothesised that monensin and tylosin might have activity against gram-positive bacteria, and their
effects on the intestinal environment may promote pro-liferation of gram-negative bacteria such as E. coli, but McAllister et al. (2006)
demonstrated that dietary inclusion of monensin or tylosin, alone or in combination, did not increase the fecal shedding of E. coli O157:H7 or its
persistence in the environment.

5. Conclusion

In growing phase, feeding antibiotics (monensin and chlortetracycline) or BL to implanted steers did not show additional bene fits over the
implant. However, feeding BL tended to have an additive effect over IM alone on growth performance at first 28 days on feed and particularly
reduced the drug treatment compared with implanted steers. These results suggest that BL benefits cattle perfor-mance during the early feeding
period when stress is high. Feeding BG clearly demonstrated beneficial effects to improved ADG and G:F compared with control animals, but
not to the same extent as implants. In the finishing phase, feeding antibiotics, BL or BG had limited effects on finishing performance and carcass
characteristics. Additionally, the effects of BL and BG on fecal IgA, fecal total E. coli and blood antioxidant activities are also of interest and
warrant further evaluation. These results suggest that feeding BL or BG was more beneficial to growing than finishing cattle.

Conflict of interest statement

The authors confirm that there are no known conflicts of interest associated with this publication.

Acknowledgements

We acknowledge the Lethbridge Research and Development Center Individual Feeding Barn sta ff for their care and management of the
animals, and Alastair Furtado, Karen Andrews, and Darrell Vedres for their assistance with the laboratory analyses.

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