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International Journal of Entomology and Nematology

Vol. 5(2), pp. 135-141, September, 2019. © www.premierpublishers.org. ISSN: 2326-7262

Research Article
Evaluation of Selected Botanical Extracts against Subterranean Termite,
Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under
Laboratory and Semi-Field Conditions
Weldesenbet Beze Kassie
Department of Forest Protection, Central Ethiopia Environment and Forest Research Institute, P.O.Box 33042, Addis
Abeba, Ethiopia
E-mail: weldesenbetbeze@gmail.com

Coptoterme formosanus is an economically important agricultural and structural pest of warm


and humid regions of the world. The main objective of the study was evaluate seed extracts of
Brassica nigra and leaves extracts of Acokantra schimperi, Croton macrostachyus and Rhamnus
prinoides against C. formosanus workers under laboratory and semi-field conditions. Treatments
were consisted of three concentrations levels (5, 10 and 15 weight of botanical powder (g) per 100
ml volume of water) by three replications. Mortality of termite was counted after 24, 48 and 72
hours exposure for both conditions. The results of all botanical extracts at all concentration levels
showed that caused mortality of C. formosanus workers. Complete mortality (100%) of C.
formosanus was observed after treatment with 15 w/v B. nigra extract at three time intervals under
both laboratory and semi-field conditions. Moreover, A. schimperi at 15 w/v concentration also
resulted 100% mortality after 48-72 hours of exposure. Brassica nigra extract showed least LC50
(5.63g/100ml) value than other botanical extracts after 24 hours exposure under laboratory
condition. Based on their toxicity status extracts of B. nigra > A. schmperi > R. prinoides > C.
macrostachyus leaf extracts.

Keywords: Botanical, extract, mortality, pesticide, pest, termite, subterranean.

INTRODUCTION

Termites are eusocial insects that are the most (Damalas and Eleftherohorinos, 2011).
agriculturally and structurally important insects and which
cause for vast economic loss by feeding on many crops, These adverse effects of synthetic pesticides are bearing
plants and wooden strictures in buildings. They have ability attention for development of botanical pesticides to control
to feed various stages of plant growth (Mitchell, 2002). different insect pests. Botanical extracts and powders from
Subterranean termites are the most successful and different bioactive plants used for insecticidal, repellent
destructive pests that comprise under family and anti-feeding properties (Isman, 2006). Plant based
rhinotermitidae, which inhabit in the ground and forage in pesticides are preferred to control insect pests because of
and aboveground (Chris et al., 2006), which, damage their less harmful nature to non-target organisms due to
homes and other structures in short period of time, their innate biodegradability (Prabakar and Jebanesan,
because their colony members are huge and long lived 2004). The uses of these biologically active botanical
(Horwood and Eldridge, 2005). To control termites, extracts are usually safer to humans and the
synthetic pesticides play important roles. However, some environment than conventional pesticides, with minimal
of synthetic insecticides create number of ecological residual effects and also with least development of
problems, development insect resistance and unsafe to resistance against pests. Therefore, the aim of this study
non-target organisms including human being (Ahmed et was to evaluate the efficacy of B. nigra seeds and A.
al., 2005). Using synthetic pesticides repeatedly to schimperi, C. macrostachyus and R. prinoides leaves
manage termites increase environmental influences, pest extracts against C. formosanus workers both under
resistance and pest resurgence of other insect pests laboratory and semi-field conditions.

Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Kassie WB 136

MATERIALS AND METHODS Filtration has been done repeatedly to obtain maximum
amount of the extract. Then, 25ml extract solution was
The study Design taken and applied independently for each treatment.
Chloropyrifos 48% Emuisifiable Concentrate was diluted in
Laboratory and semi-field experiments were arranged as
water based on the recommended field application rate. In
Complete Randomized Design (CRD) and Complete
all experiments, the same volume (25ml) of Chlorpyrifos
Randomized Block Design (CRBD) respectively, with three
48% E.C. and water were used as a standard check and
replications per treatment for each botanical extracts
untreated check controls, respectively. The percentage of
concentration levels. The same batch of C. formosanus
extracts varies, when the amount of material used were
workers for both laboratory and semi-field experiments
transformed, in same water quantity as described (Sahay
was chosen and collected from newly termite infested field
et al., 2014).
(February-May, 2017) by bucket traps. All botanical
extracts effectiveness was evaluated against workers by
Efficacy of selected botanical extracts against
comparing from standard check (Chloropyrifos 48% E. C)
Coptotermes formusanus under laboratory condition
and an untreated check (water).
Laboratory experiment was done repeatedly to check the
Collection of plant materials
accuracy. Bioassays were carried out to determine the
For efficacy test four locally available botanicals (Table 1) mortality of Formosan workers by each botanical
were selected based on cultural used information and termiticides activities. For laboratory test, 42 cups were
preliminary study. They collected from Debre Markos town labeled and arranged in carton box (120cm x 80cm x
and surrounding from January 21st to 25th in 2017. 40cm), which was inspected for maintenance of required
moisture level. The volume of each container was 1 liter.
Plant materials were brought into the laboratory by Then, a piece of moistened Eucalyptus globules wood was
polyethylene bags. Identification of botanical species has added as food source and five gram soil to maintain
been done by expert. Voucher specimens of the botanical moisture level were placed for survival of termites in each
species were deposited in herbarium at Debre Markos prepared cups. Immediately, 20 workers were counted and
University, Ethiopia. introduced in each 42 prepared cups carefully. For
acclimatization of termites, laboratory room temperature
Table 1: List of plant materials tested against C. (20-28oc) was controlled by using 100 watt electric bulb.
formosanus termite workers. Moisture also adjusted by added drops of water in each
Scientific name Family name Common Part cups to create optimum humidity. The box had covered by
name used sake that able to exchange of air for them and stay for five
Brassica nigra Brassicaceae Black seeds days to check their adaptation in the laboratory. During
Mustard application time, from each prepared filtered solution 25ml
Acokanthera Apocynaceae Poison bush Leaves extract was sprayed in each cup to test the efficacy.
schimperi Mortality of termites was recorded carefully and accurately
Croton Euphorbiaceae Broad Leaves after 24, 48 and 72 hours of exposure. Percentage of
macrostachyus levead- mortality has been calculated by Abbott, (1925) Equation.
croton
Rhamnus Rhamnaceae Dogwood Leaves
prinoides
Efficacy of selected botanical extracts against
Preparation of botanical extracts
Coptotermes formusanus under semi-field condition
Collected botanical parts were washed by sterilized water
to avoid dust particles and tiny organisms that are present Semi-field experiment site was selected at Debre Markos
on them. After that, they were dried in well-ventilated room University, which is protected and was not treated by
under shade for ten days until completely dry at room pesticides. Similar to laboratory experiment, totally 42
temperature (18-25ºC). Dried form of plant materials was prepared containers were inserted 25cm into the ground.
grinded into powder by using pestle and mortar to obtain Then, termite preferred wood and 5 gram soil were added
uniform fine powder and stored in separate plastic to each container to serve as food and to keep moisture
containers. Sensitive balance was used to measure 5g, respectively.
10g and 15g portion of each grinded fine powder. Each
grinded botanical powder was soaked in 100 ml of water After that, 20 worker Formosan termites were counted and
to obtain three concentration levels (5, 10 and 15 w/v) introduced in each container. Every cup was covered by
weight of powder/volume of water, then shacked for five piece of sake separately to prevent direct sun light and
minutes to make homogeneous. After stay two days, each other organisms that can enter to the treatment. After 30
solution supernatant was filtered by Whatman (No.1) to minutes, 25ml filtered solution was sprayed in each
remove impurities. experimental cups to test the efficacy. In semi-field

Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Int. J. Entomol. Nematol. 137

experiment, mortality was counted at evening time (after At 5 w/v botanical extracts concentration level, after 48
11pm) to protect termites from direct sunlight. Similar to hours exposure mean mortality of termites was increased
laboratory experiment, the temperature of semi-field was as 66.67, 33.33, 10.53 and 23.33 mean mortality of
recorded at 20-24oC. Mortality of termites was recorded termites were recorded from B. ngra, A. schmperi, C.
after 24, 48 and 72 hours of exposure. macrostachyus and R. prinoides respectively. At 5 w/v
concentration level of botanical extracts, the highest
Data Analysis (78.33) and the least (26.67) mean mortality of termites
under laboratory condition were registered from B. nigra
Data was subjected to one-way of analysis of variance and C. macrostachyus respectively, after 72 hours
(ANOVA) by using SPSS version 20 software to determine exposure time.
statistically significant differences among treatments.
Significant differences mean were separated by using Mean mortality of termite (96.67%) by B. nigra, (76.67)
Tukey’s studentized range test (α =0.05). The lethal mortality by A. schmperi (40.00)% by C. macrostachyus
concentration (LC50) and (LC90) were analyzed by probit and 73.33 by R. prinoides were recorded under laboratory
analysis to determine botanical extracts toxicity. 50% and condition after 72 hours at 10 w/v concentration. There
90% mortality were calculated after 24, 48 and 72 hours was highly significant difference (P<0.0001), among the
both under laboratory and semi-field conditions. different treatments at different time intervals (24, 48 and
72 hours) due to the potency (termicidal effect) of botanical
extracts. There was no significant difference among B.
RESULTS AND DISCUSSION nigra at 5 w/v, A. schmperi and R. prinoides at 10 w/v
Evaluation of the efficacy of botanical extracts under concentration levels after all (24, 48 and 72 hours of
laboratory conditions exposure time as indicated in Table 2.

Effectiveness of botanical extracts on Formosan workers Maximum termite mortality (100%) from Chloropyrifos
under laboratory condition represented in Table 2. As followed by non-significantly difference 10 and 15 w/v
indicated in Table 2, at 5 w/v botanical extracts concentration of B. nigra and 15 w/v concentration of A.
concentration level after 24 hours exposure highest and schmperi after 72 hours exposure under laboratory
lowest mortality were recorded from B. ngra and R. condition. As indicated in Table 2, 61.67 and 88.33 mean
prinoides respectively under laboratory condition. mortality of termites were recorded from C. macrostachyus
Similarly, after 48 hours exposure of time interval and R. prinoides respectively at 15 w/v concentration level
maximum termite mortality (100%) also was recorded from after 72 hours exposure of time.
standard check.

Table 2: Mean mortality ± SE of C. formosanus workers by botanical extracts under laboratory.


Treatments Concentration Mean mortality ± SE over time (hours) exposure
(w/v) 24 hours 48 hours 72 hours
Brassica nigra 5 53.33 ± 1.67d 66.67 ± 1.67d 78.33 ± 1.67c
10 86.67 ± 1.67 b 90.00 ± 0.00 b 96.67 ± 1.67a
15 100.00 ± 0.00 a 100.00 ± 0.00 a 100.00 ± 0.00a
Acokanthera schmperi 5 21.67 ± 1.67g 33.33 ± 2.89f 41.67 ± 1.67e
10 53.33 ± 3.33d 63.33 ± 1.67d 76.67 ± 1.67c
15 91.67± 2.89b 100.00±0.00a 100.00 ± 0.00a
Croton macrostachyus 5 8.33 ± 1.67 h 18.33 ± 2.89 g 26.67 ± 1.67f
10 30.00 ± 2.89 f 36.67± 1.67 f 40.00 ± 0.00e
15 43.67 ± 1.67 e 53.33 ± 3.33 e 61.67 ± 2.89d
Rhamnut prinoides 5 11.67 ± 1.67 h 23.33 ± 1.67 g 31.67 ± 1.67f
10 56.67 ± 1.67 d 65.00 ± 0.00 d 73.33 ± 1.67c
15 78.33 ± 1.67c 83.33 ± 1.67c 88.33 ± 1.67b
Chloropyrifos 48% E.C 25ml 100.00 ± 0.00a 100.00 ± 0.00a 100.00 ± 0.00a
Untreated check (water) 25ml 0.00 ± 0.00i 0.00± 0.00h 0.00± 0.00g
CV (%) 5.30 3.67 3.54
P-value <0.0001 <0.0001 <0.0001
Means within a column followed by the same letter are not significantly different (Tukey Student Test (HSD) at p=0.05).
Key: SE=Systematic Error, w/v= weight of powder/volume of water.

Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Kassie WB 138

Evaluation of the efficacy of botanical extracts under There was a significant difference among the treatments.
semi-field condition As indicated in (Table 3), at 15 w/v concentration level
after 72 hours exposure time, the highest mean termite
The result of experiments conducted to evaluate mortality (100%) recorded from B. nigra and A. schmperi
effectiveness of four locally available botanical insecticides that without significantly difference from Chloropyrifos 48%
on Formosan termite workers at different concentrations E.C.
under semi-field condition as represented in Table 3.
Table 3: Mean mortality ± SE of C. formosanus workers by botanical extracts under Semi-field condition.
Treatments Concentration Mean mortality ± SE over time (hours) exposure
(w/v) 24 hours 48 hours 72 hours
Brassica nigra 5 38.33± 1.67e 48.33 ± 2.89de 61.67± 1.67d
10 73.33 ± 3.33bc 85.00 ± 0.00b 93.33 ± 2.89ab
15 100.00 ± 0.00 a 100.00 ± 0.00 a 100.00 ± 0.00a
Acokanthera schmperi 5 8.67 ± 2.89 g 16.67 ± 1.67 f 26.67 ± 1.67f
10 40.00± 0.00 e 53.33 ± 21.67 d 61.67 ± 1.67d
15 76.67 ± 1.67 b 86.67± 1.67 b 100.00 ± 0.00a
Croton macrostachyus 5 1.67 ± 1.67 g 10.00 ± 0.00 fg 15.00 ± 0.00g
10 25.00 ± 0.00f 31.33 ± 1.67e 36.67 ± 1.67e
15 56.67 ± 1.67d 66.67 ± 1.67c 73.33 ± 2.89c
Rhamnut prinoides 5 8.33 ± 2.89g 15.00 ± 0.00f 23.33 ± 2.89fg
10 36.67±2.897e 46.67 ± 1.67de 55.00 ± 0.00de
15 66.67± 1.67 c 76.67 ± 1.67 bc 88.33 ± 2.89b
Chloropyrifos 48% E.C 25ml 100.00 ± 0.00 a 100.00 ± 0.00 a 100.00 ± 0.00a
Untreated check (water) 25ml 0.00 ± 0.00g 0.00 ± 0.00g 0.00 ± 0.00h
CV (%) 7.25 6.50 4.78
P-value <0.0001 <0.0001 <0.0001
Mean with in a column followed by the same letter are not different (Tukey Student Test (HSD) at
p= 0.05). Key: SE=Systematic Error, w/v= weight of powder/volume of water
Minimum concentration required to kill 50% and 90% of the
In both laboratory and semi-field conditions, the highest tested worker Formosan termites were calculated for each
and least mean mortality of termite was recorded from B. botanicals after 24, 48 and 72 hours exposure of time with
nigra seeds and C. macrostachyus leaf extracts their lower and upper limits. As indicated in (Table 4),
respectively. No termite mortality was recorded from minimum concentration (3.97g/100ml) that used to kill 50%
untreated check for both laboratory and semi-field of tested termites’ recorded from B. nigra after 72 hours.
conditions. Mean mortality of termites in all botanical Maximum concentration (15.31 g/100ml) that used to kill
extracts concentration levels was also increased based on 50% of tested termites’ recorded from C. macrostachyus
exposure time intervals. after 24 hours exposure.

Dose response bioassay under laboratory condition

The results of the LC50 and LC90, is indicated in (Table 4).

Table 4: Toxicity analysis of botanical extracts on C. formosanus workers under laboratory condition.
Treatments After 24 hours After 48 hours After 72 hours
LC50 LC90 LC50 LC90 LC50 LC90
B. nigra 5.63 9.71 4.86 8.75 3.97 6.87
(4.12-6.99) (8.15-12.59) (3.48-6.31) (6.53-9.97) (2.08-4.87) (5.12-8.23)
A. schmperi 9.20 14.72 7.65 12.45 6.65 11.14
(7.60-10.91) (12.66-18.57) (6.15-9.17) (10.66-15.77) (5.15-8.09) (9.45-14.24)
C. macrostachyus 15.31 24.64 13.44 23.15 11.91 21.44
(12.51-22.50) (19.16-43.01) (10.90-18.64) (18.16-37.66) (9.56-15.84) (17.04-33.04)
R. prinoides 10.36 16.42 8.65 14.52 7.98 13.98
(8.66-12.78) (14.08-21.05) (6.95-10.36) (12.42-18.34) (6.24-9.69) (11.88-17.77)

Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Int. J. Entomol. Nematol. 139

Table 5: Toxicity analysis of botanical extracts on C. formosanus workers under semi field condition.
Treatments After 24 hours After 48 hours After 72 hours
LC50 LC90 LC50 LC90 LC50 LC90
B. nigra 7.00 11.61 5.94 10.06 5.06 8.54
(5.50-8.47) (9.89-14.78) (4.48-7.31) (8.48-12.99) (3.68-6.33) (7.12-11.23)
A. schmperi 11.43 17.29 9.76 15.45 8.05 12.61
(9.76-13.50) (14.86-22.41) (8.12-11.53) (13.30-19.55) (6.59-9.54) (10.86-15.89)
C. macrostachyus 13.89 19.95 12.21 19.17 11.06 17.94
(12.07-17.17) (16.82-28.59) (10.32-14.96) (16.05-26.38) (9.22-13.40) (15.11-23.99)
R. prinoides 12.35 17.38 10.82 17.38 9.25 15.20
(10.50-15.01) (16.04-26.12) (9.04-13.00) (14.74-22.88) (7.57-11.05) (12.99-19.37)

Dose response bioassay under Semi-field condition study, B. nigra aqueous seeds extract (LC50= 5.63 g/100ml
water and LC90=9.71 g/100ml) displayed highest lethal
As indicated in Table 5, B. nigra seed extract showed least effect against C. formusanus workers under laboratory
LC50 (5.06) and LC90 (8.54) values than other botanicals condition after 24 hours exposure.
after all 24, 48 and 72 exposure of time. Inversely, the
highest LC50 value (13.89) and LC90 value (19.95) were In others investigation, 70% ethanol B. nigra seeds extract,
recorded from C. macrostachyus extract after 24 hours possessed good microbial activities (Rajesh and Vikas,
exposure of time. Based on the LC50 and LC90 values the 2014). Whereas water extracts of elder flowers of
most toxic extract was B. nigra and the least toxic was C. Sambucas nigra can repel granary weevils (Sitophilus
macrostachyus under both laboratory and semi-field granarius) at 5, 10 and 15 w/v concentrations in 200 ml
conditions. The order of toxicity was B. nigra, A. schmperi, water (Ignatowicz and Wesolowska, 2002). In the present
R. prinoides and C. macrostachyus in both Table 4 and 5. study, C. formosanus termite mortality was recorded in
seed extract of B. nigra and leaf extracts of A. schmperi,
In the present study, tested botanical extracts showed C. macrostachyus and R. prinoides after 24 hours
mortality on the C. formusanus workers. As indicated in exposure under laboratory condition (LC50=5.63, 9.20,
Table 2 and 3, there was highly significant difference 15.31 and 10.36g/100ml) and (LC90= 9.71, 14.72, 24.64
(P<0.0001) between different treatments at different time and 16.42g/100 ml water) respectively.
of exposure (24, 48 and 72 hours) due to the potency of
botanical extracts at all concentration levels. There was no In present experiment, C. macrostachyus leaf extracts at
mortality of termites observed in the untreated check over 5, 10 and 15 w/v concentration levels after all time of
the entire exposure (72 hours) of the experiment. exposure showed less toxic effect among all botanical
Maximum mean mortality of termites (100%) was extracts. This result was in lined with the finding of Tadele
registered from Chloropyrifos 48% E. C, which highly Shiberu et al., (2014) who reported that seed extracts of C.
significantly differences from botanicals treatments that macrostachys at 25% concentration level was also less
cause mortality against C. formosanus workers. This is in toxic effect on Macroterms Spp. (Isoptera: Termitidae),
agreement with Tadele Shibru, et al., (2014), reported that even after 5 days under laboratory conditions.
Chloropyrifos 48% E. C gives 100% mortality on
Macroterms Spp. under laboratory condition. Croton macrostachyus leaf extract of the present study
gave 26.67% and 15.00% mortality on C. formosanus
The present experiment showed that, mean mortality of workers after 72 hours exposure at 5 w/v concentrations
termites was increased, when the concentration level of under laboratory and semi-field conditions respectively.
botanicals was increased and time of exposure increased While in the other findings, aqueous leaf extracts of C.
(Table 2 and 3). Based on the present results, maximum macrostachyus at 5 w/v concentration level achieved
termite mortality in both laboratory and semi-field 100% mortality on African bollworm, Helicover pagrmiyera
conditions were observed at higher concentrations (15 Hubner after 72 hours (Ngussie Lulie and Nagappa, 2012).
w/v) after 72 hours exposure. This result is in agreement
with Upadhyay, (2013) and Sattar et al., (2014), who
reported that mortality of termites, were directly proportion CONCLUSION
to botanical extract concentrations and exposure time of
treatments. The present study findings had important implications in
the practical control of Formosan subterranean termite by
In the present study, mean mortality (100%) on C. using botanical extracts, which are easy to prepare. From
formosanus workers by positive control was not the present study results, it can be concluded that, the
significantly different from 15 w/v B. nigra both under efficacy of botanicals depending on the amount of
laboratory and semi-field conditions after all time of concentration and time of application (acting in short,
exposure and A. schmperi under laboratory condition after medium and long period of time after treated). The result
48 and 72 hours exposure time (Table 2). In the current showed that, B. nigra and A. schmperi were more effective

Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Kassie WB 140

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RECOMMENDATIONS Nigussie Lulie and Nagappan, R. (2012). Evaluation of
Certain Botanical Preparations against African
Aqueous extract of tested botanicals has promising in the Bollworm, Helicover pagrmiyera Hubner (Lepidopera:
laboratory and under semi field conditions. Therefore, wide noctuidae) and Non Target Organisms in Ckickpea,
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ACKNOWLEDGMENTS agent against seed microorganisms. International
Journal of Pharmaceutical 3:117-122.
I would like to express my deepest gratitude to Dr. Sahay, N. Prajapati, C. and Singh, P. (2014). Anti-termite
Sundarsan Reddey for his consistent comments from potential of plants selected from the SRISTI database
starting to the completion of my work and valuable of Grassroots Innovations. Journal of Biological Pest
assistance in SPSS programs and for identification of Control 7:164-169.
Formosans termite species by using different identification Sakasegawa, M., Hori, K. and Yatagi, M. (2003).
methods. Composition and anti-termite activities of essential oils
and Melaleuca species. Journal of Wood Science
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Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions
Int. J. Entomol. Nematol. 141

Accepted 26 July 2019

Citation: Kassie WB (2019). Evaluation of Selected


Botanical Extracts against Subterranean Termite,
Coptotermes formosanus Shiraki (Isoptera:
Rhinotermitidae), Under Laboratory and Semi-Field
Conditions. International Journal of Entomology and
Nematology, 5(2): 135-141.

Copyright: © 2019 Kassie WB. This is an open-access


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Evaluation of Selected Botanical Extracts against Subterranean Termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), Under Laboratory and
Semi-Field Conditions

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