Effects of carbaryl,chlorpyrifos and endosulfan on growth,reproduction and respiration of tropical epigeic earthworm,Perionyx excavatus (Perrier)

Effects of sub-lethal doses of carbaryl (1-Naphthyl-methylcarbamate), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridinyl-phosphorothioate) and endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin-3-oxide), respectively a carbamate, an organophosphate and an organochlorine insecticide on growth, reproduction and respiration of the tropical earthworm, Perionyx excavatus (Perrier) were investigated under laboratory conditions. The results showed significant reduction in biomass, production and hatching of cocoon and production of juveniles of the worms exposed to 0.75 to 3.03 mg/kg soil of carbaryl, 0.91 to 3.65 mg/kg soil of chlorpyrifos and 3.75 to 15.0 μg/kg soil of endosulfan corresponding to 12.5 to 50 % of LC₅₀ value of the respective insecticide for P. excavatus. Endosulfan was found most dangerous among the three insecticides followed by carbaryl and chlorpyrifos. There was no hatching of the worms at endosulfan treatment 5.0 μg/kg soil (25 % LC₅₀) or above while the highest dose of carbaryl and chlorpyrifos (50 % of LC₅₀) rendered respectively 87.13 and 24.84 % reductions in hatching as compared to control. Chlorpyrifos produced no change in respiration of the worms except at the highest dose, while the worms showed an increase in evolution of CO₂ at all doses of carbaryl and endosulfan. Based on the recommended agricultural dose of each insecticide, it was concluded that application of endosulfan and carbaryl was potentially dangerous to earthworms.     

Acute contact toxicity test of insecticides (Cipermetrina 25, Lorsban 48E, Thionex 35) on honeybees in the southwestern zone of Uruguay

Glyphosate-resistant soybean cultivation is expanding rapidly in Uruguay, with its land area having increased by 95 times during the past 10 years. Because of the region's Neotropical conditions, insecticide use is required to ensure adequate soybean productivity. However, in areas shared by soybean crops and beekeepers - such as the southwestern zone of Uruguay (SWZU) - the use of insecticides can increase the risks of honeybee death and honey contamination. Uruguayan commercial and legal guidelines set out practices and field doses designed to prevent acute intoxication with insecticides. However, honeybees in the SWZU are predominantly a polyhybrid subspecies different from that used to set international reference values, and hence they may have a different acute toxicity response, thus rendering such precautions ineffective. The aim of this work was to assess the acute toxicity response of polyhybrid honeybees in the SWZU to cypermethrin (commercial formulation: Cipermetrina 25 Agrin?), chlorpyrifos (commercial formulation: Lorsban 48E?), and endosulfan (commercial formulation: Thionex 35?). Acute toxicity bioassays were conducted to determine the median lethal dose (LD(50)) of each insecticide for the honeybees. The results indicate that, compared with EU reference values, SWZU honeybees have a higher toxicological sensitivity to chlorpyrifos and endosulfan, and a lower toxicological sensitivity to cypermethrin, based on the commercial formulations tested. However, when these results were adjusted according to their field dose equivalents, only chlorpyrifos emerged as a potential problem for beekeeping, as the maximum recommended field dose of Lorsban 48E? for soybean crops in Uruguay is 23 times the corresponding LD(50) for honeybees in the SWZU.     

Assessment of insecticide contamination in runoff and stream water of small agricultural streams in the main soybean area of Argentina

The first- and second-order streams, Brown and Horqueta, respectively, which are located in the main area of soybean production in Argentina were examined for insecticide contamination caused by runoff from nearby soybean fields. The insecticides most widely used in Argentina (chlorpyrifos, cypermethrin and endosulfan) were detected in sediments, suspended particles and water. Highest concentrations in suspended particles were 318 microg/kg for endosulfan in the stream Horqueta, while 226 microg/kg chlorpyrifos and 13.2 microg/kg cypermethrin were measured in the stream Brown. In the Horqueta stream 150 and 53 microg/kg chlorpyrifos and cypermethrin were detected in runoff sediments, respectively. Whereas cypermethrin concentrations in the suspended particles were relatively low, levels in the floodwater of Brown reached 0.7 microg/l. The highest chlorpyrifos concentration in floodwater was 0.45 microg/l in Brown. However, endosulfan was not detected in the water phase. In runoff water the highest concentrations measured were 0.3 microg/l for chlorpyrifos in Horqueta and 0.49 microg/l for cypermethrin in the Brown stream. On five sampling dates during the pesticide application period in Brown stream (2002/2003) the concentration of chlorpyrifos and cypermethrin in runoff and/or floodwater exceeded the water quality criteria for freshwater mentioned in this study. In three cases this insecticide concentration was measured in stream water, indicating an acute risk to aquatic life. The acute toxicity-exposure-ratio (TER) for chlorpyrifos and cypermethrin also shows an acute risk for aquatic invertebrates in the Brown stream. In the Horqueta chlorpyrifos concentrations in the runoff exceeded the safety levels three times during the application period (2001/2002), potentially endangering the aquatic fauna. Effects on aquatic macroinvertebrates after insecticide contamination were reported in earlier studies in Horqueta stream.     

Toxicity assessment of 45 pesticides to the epigeic earthworm Eisenia fetida

This study was conducted to investigate comparative toxicity of 45 pesticides, including insecticides, acaricides, fungicides, and herbicides, toward the epigeic earthworm Eisenia fetida. Results from a 48-h filter paper contact test indicated that clothianidin, fenpyroximate, and pyridaben were supertoxic to E. fetida with LC(50) values ranging from 0.28 (0.24-0.35) to 0.72 (0.60-0.94) μg cm(-2), followed by carbaryl, pyridaphenthion, azoxystrobin, cyproconazole, and picoxystrobin with LC(50) values ranging from 2.72 (2.22-0.3.19) to 8.48 (7.38-10.21) μg cm(-2), while the other pesticides ranged from being relatively nontoxic to very toxic to the worms. When tested in artificial soil for 14 d, clothianidin and picoxystrobin showed the highest intrinsic toxicity against E. fetida, and their LC(50) values were 6.06 (5.60-6.77) and 7.22 (5.29-8.68) mg kg(-1), respectively, followed by fenpyroximate with an LC(50) of 75.52 (68.21-86.57) mgkg(-1). However, the herbicides fluoroglycofen, paraquat, and pyraflufen-ethyl exhibited the lowest toxicities with LC(50) values>1000 mg kg(-1). In contrast, the other pesticides exhibited relatively low toxicities with LC(50) values ranging from 133.5 (124.5-150.5) to 895.2 (754.2-1198.0) mg kg(-1). The data presented in this paper provided useful information for evaluating the potential risk of these chemicals to soil invertebrates.     

Susceptibility of epigeic earthworm Eisenia fetida to agricultural application of six insecticides

Ecotoxicological risks of agricultural application of six insecticides to soil organisms were evaluated by acute toxicity tests under laboratory condition following OECD guidelines using the epigeic earthworm Eisenia fetida as the test organism. The organochlorine insecticide endosulfan (LC₅₀ - 0.002 mg kg⁻¹) and the carbamate insecticides aldicarb (LC₅₀ - 9.42 mg kg⁻¹) and carbaryl (LC₅₀ - 14.81 mg kg⁻¹) were found ecologically most dangerous because LC₅₀ values of these insecticides were lower than the respective recommended agricultural dose (RAD). Although E. fetida was found highly susceptible to the pyrethroid insecticide cypermethrin (LC₅₀ - 0.054 mg kg⁻¹), the value was higher than its RAD. The organophosphate insecticides chlorpyrifos (LC₅₀ - 28.58 mg kg⁻¹), and monocrotophos (LC₅₀ - 39.75 mg kg⁻¹) were found less toxic and ecologically safe because the LC₅₀ values were much higher than their respective RAD.     

Comparative and combined toxicities of toluene and methyl tert-butyl ether to an Asian earthworm Perionyx excavatus

An earthworm assay was used to assess the toxicity of two main gasoline components. Ecotoxicities of toluene and methyl tert-butyl ether (MTBE) were compared for Asian earthworm Perionyx excavatus separately and in combination. Sum of toxic unit (TU) at 50% mortality for the mixture (LC50mix) was estimated from the dose (TU-based)-response relationships by the Trimmed Spearman-Karber method. Toluene was shown to be about five and two times more toxic to P. excavatus than MTBE in filter paper contact test and natural soil tests, respectively. This is primarily due to the adsorption of toluene to soil particles, leading to less bioavailability of toluene to the earthworm in soil. The combined effect of toluene and MTBE was investigated using toxic unit model, and it was found to be almost additive and synergistic responses to P. excavatus in filter paper contact test and soil tests, respectively. The combined effect of toluene and MTBE in filter paper contact test was not consistent with the results in soil toxicity tests. This phenomenon may be associated with the interaction of soil salts with pollutants. Treatment with toluene and MTBE also affected the behavior and morphology of P. excavatus. It indicates that the VOCs induce metabolic and functional damages in earthworms. Combined effects of gasoline components should be taken into account to soil risk assessment.     

Insecticide residues in cotton soils of Burkina Faso and effects of insecticides on fluctuating asymmetry in honey bees (Apis mellifera Linnaeus)

Four insecticides (acetamiprid, cypermethrin, endosulfan and profenofos) are used quarterly in the cotton-growing areas of Burkina Faso, West Africa. These insecticides were investigated in soils collected from traditionally cultivated and new cotton areas. Also, the effects of insecticide exposure on the developmental instability of honey bees, Apis mellifera, were explored. In soil samples collected three months after insecticide treatments, endosulfan and profenofos concentrations varied in the range of 10-30 μg kg(-1) in the traditionally cultivated zones and 10-80 μg kg(-1) in the new cotton zones, indicating a pollution of agricultural lands. However, only profenofos concentrations were significantly higher in the new cotton zone than the traditionally cultivated zones. In addition, the index of fluctuating asymmetry, FA1, in the length of second tarsus (L(HW)) was increased for bees when exposed to pesticide treated cotton fields for 82d, and their FA levels were significantly higher than those in the control colony in an orchard. The other studied traits of bees exposed to insecticides were not significantly different from controls. Our results indicate that FA may be considered as a biomarker reflecting the stress induced by insecticide treatments. However, the relationship between FA and stressors needs further investigations.     

Enrichment and isolation of endosulfan-degrading microorganism from tropical acid soil

Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,3,4-benzo-dioxathiepin-3-oxide) is a cyclodiene organochlorine currently used as an insecticide all over the world and its residues are posing a serious environmental threat. This study reports the enrichment and isolation of a microbial culture capable of degrading endosulfan with minimal production of endosulfan sulfate, the toxic metabolite of endosulfan, from tropical acid soil. Enrichment was achieved by using the insecticide as sole sulfur source. The enriched microbial culture, SKL-1, later identified as Pseudomonas aeruginosa, degraded up to 50.25 and 69.77 % of α and β endosulfan, respectively in 20 days. Percentage of bioformation of endosulfan sulfate to total formation was 2.12% by the 20th day of incubation. Degradation of the insecticide was concomitant with bacterial growth reaching up to an optical density of 600 nm (OD600) 2.34 and aryl sulfatase activity of the broth reaching up to 23.93 μg pNP/mL/hr. The results of this study suggest that this novel strain is a valuable source of potent endosulfan–degrading enzymes for use in enzymatic bioremediation. Further, the increase in aryl sulfatase activity of the broth with the increase in degradation of endosulfan suggests the probable involvement of the enzyme in the transformation of endosulfan to its metabolites.     

Toxic and sublethal effects of endosulfan and carbaryl on growth and egg production of Moina micrura Kurz (Cladocera: Moinidae)

The toxic and sublethal effects of endosulfan and carbaryl on a cladoceran, Moina micrura, were studied. The dosage-mortality regression lines gave LC(50) values for the pesticides. The cumulative growth and egg production during life were inhibited to 31.8 and 20.7% in endosulfan, and 28.2 and 13.7% in carbaryl-treated animals, respectively. The growth coefficient (K value) and the intrinsic rate of natural increase per day (r value) of control M. micrura were 0.144 and 0.377, respectively; these decreased to 0.121 and 0.346 in carbaryl-, and 0.073 and 0.356 in endosulfan-treated animals, respectively.     

Comparative acute toxicities of selected pesticides to Anguilla anguilla

Abstract

The acute toxicities (24, 48, 72 and 96 hr) of eight pesticides to Anguilla anguilla were determined. The organochlorine pesticide, endosulfan was the most toxic, with LC50 values in the range of 0.042 to 0.041 mg/L Endosulfan was followed in order of decreasing toxicity by diazinon, fenitrothion, chlorpyrifos, lmdane, methidathion, trichlorfon and methylparathion. When fishes were exposed to the pesticides tested they exhibited signs of restlessness, erratic swimming, convulsions and difficulty in respiration. This response was more persistent in fishes exposed to organophosphorus pesticides.     

Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats

Two experiments were conducted in male SD rats (225-250 g) to determine changes in the activities of endogenous antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and concentrations of glutathione (GSH) in tissues after exposure to low doses of endosulfan and chlorpyrifos using a whole body exposure technique. In both experiments, 6 rats/group were exposed 3 hr/day, 5 days/week for 30 days to: 0 (control), 5, 10, 20, 40 and 60% of LD50 of either pesticide in 50% ethanol; actual concentrations were: endosulfan = 0, 0.5, 1.0, 2.0, 4.0, 6.0 mg/250 g body weight; chlorpyrifos = 0, 1.9, 3.8, 7.6, 15.2, and 22.8 mg/250 g body weight. Endosulfan decreased erythrocyte SOD by 21% in all groups and chlorpyrifos increased SOD by 18% in groups 40 and 60. Liver SOD was 12%-20% lower after endosulfan exposure; lung SOD was altered: endosulfan decreased activity by 21% and 51% and chlorpyrifos by 58 and 75% in the 40 and 60 groups, respectively (P < or = 0.05). Both pesticides increased plasma GPX activity at lower levels and reduced it by 26% and 19% in groups 40 and 60, respectively (P < or = 0.05). Liver GPX increased in the 60 group and lung GPX declined between 20% and 38% after endosulfan exposure. GSH in the liver and lung: endosulfan reduced GSH by about 30% at lower levels and increased by 41% or 70% at higher levels; chlorpyrifos decreased GSH by 28-40% in 20 and 60 groups, respectively (P < or = 0.05). Exposure to low, increasing levels of endosulfan and chlorpyrifos can differentially modify endogenous antioxidants SOD, GPX and GSH, which may lead to the development of oxidative stress in some tissues.     

Response of thermal stressed bobwhite to organophosphorus exposure

Constant and intermittent cold stressed juvenile northern bobwhite (Colinus virginianus) were exposed to chlorpyrifos, an organophosphorus insecticide, following avian dietary LC(50) test procedures. Chlorpyrifos concentrations ranged from 100 to 1000 ppm; ambient temperatures (T(a)) included 35.0 (control), 32.5, 30.0 and 27.5 degrees C. Survival decreased significantly under constant cold conditions. Non-survivors averaged 33% weight loss, but survivor weight dropped with increasing dose and decreasing T(a). Control quail at all temperatures maintained a constant body temperature (T(b)); however, T(b) of dosed quail fell with increasing dose and decreasing T(a). Non-surviving cold stressed quail had higher cholinesterase (ChE) activity that non-survivors at 35 degrees C; in addition, dead quail failed to average the 50% inhibition of ChE used to implicate a ChE depressant in death. Daily exposure of quail to 12 h of warmth increased survival. Survivors of intermittent cold stress experienced less weight loss than constant cold survivors due to reduced metabolic costs. Synergistic effects were evident in juvenile northern bobwhite exposed simultaneously to chlorpyrifos and cold stress; however, mortality and certain sublethal effects were mitigated when cold exposure was intermittent.     

Persistence of chlorpyrifos in a mineral and an organic soil

Chlorpyrifos (Lorsban emulsifiable concentrate) was applied at 3.4 kg AI/ha and incorporated into sand and muck soil contained in small field plots. Soil samples were taken at intervals over 2 yr. Radishes and carrots, seeded yearly, served as indicator crops for absorption of insecticide residues. Samples were extracted and analyzed, by gas-liquid chromatography, for chlorpyrifos, oxychlorpyrifos, and 3,5,6-trichloro-2-pyridinol. Chlorpyrifos residues declined rapidly, with 50% of the initial application remaining after 2 and 8 wk in sand and muck, respectively, and 4 and 9% after 1 yr. Pyridinol residues increased to 13 and 39% of the initial chlorpyrifos application in sand and muck after 1 and 8 wk, respectively, and declined thereafter. Oxychlorpyrifos was detected in the 2 soils at very low levels only in immediate posttreatment samples. In the first year of the study low levels (less than 0.1 ppm) of chlorpyrifos and the pyridinol were detected in radishes and carrots.     

Insecticide residues in the blood serum and domestic water source of cacao farmers in Southwestern Nigeria

The blood serum of cacao farmers and their domestic water sources were analyzed for insecticide residues in selected cacao growing communities of Southwestern Nigeria. The farmers were grouped into five exposure periods based on their years of involvement in insecticide application, viz, <5 years, 5-9 years, 10-14 years, 15-19 years and >20 years. The residue analyses revealed that 42 out of the 76 farmers had residues of diazinon, endosulfan, propoxur and lindane in their blood; and 47.6% out of these farmers belonged in the >20 years exposure duration period. About 34% of the farmers had diazinon with a mean concentration of 0.067 mg kg(-1), 29% endosulfan (mean=0.033 mg kg(-1)), 23% propoxur (mean=0.095 mg kg(-1)), and 17% lindane (mean=0.080 mg kg(-1)) in their blood. The residues of lindane, endosulfan and propoxur in all the exposure duration categories were found to be far below the no observable adverse effect level (NOAEL) while diazinon residues detected in the blood serum of the farmers in all the exposure duration categories exceeded the NOAEL of 0.02 mg kg(-1) for the insecticide. The study also revealed that the sources of drinking water had been contaminated with dazinon and propoxur in some of the farmers' localities; and the concentrations of the insecticides exceeded the acceptable daily intake (ADI). It is concluded that cacao farmers in Southwestern Nigeria may have been occupationally exposed due to insecticide application for mirid control in their cacao plantations; and the exposure at times is of such magnitude as to be hazardous to the farmers and their respective communities.     

Effect of some technical and formulated insecticides on microbial activities in soil

Abstract

The influence of 11 formulated and technical insecticides at 10 μg/g soil on growth and activities of microorganisms was determined. The populations of bacteria and fungi initially decreased with some pesticide treatments but recovered rapidly to levels similar to or greater than those of controls after three weeks. Both formulated and technical chlordane, chlorpyrifos and cypermethrin stimulated fungal growth. No inhibition on nitrification after two wks and sulfur oxidation after three wks was observed in treatments with either grade of insecticide. The effect of different treatments on respiration was equal to or greater than that of control sample. Less effect was observed with technical insecticides than the formulated ones on microbial populations and activities in the soil.     

Pesticide effects on crabs: How environmental concentrations of endosulfan and chlorpyrifos affect embryos

The objectives of this work were to evaluate the effects of environmentally relevant chlorpyrifos and endosulfan, concentrations in the incubation period, effective hatching and survival of embryos and neonates of the freshwater burrowing crab, Zilchiopsis collastinensis (Decapoda, Trichodactylidae). Both pesticides were prepared from commercial and technical grade products. The exposure to about 100, 200, and 400 ng endosulfan L^?1, and 48, 240, and 1,200 ng chlorpyrifos L^?1 did not cause differences in the incubation period or in effective hatching but decreased survival of neonates, especially in the concentrations prepared from the technical grade product. Even if these concentrations are below the median lethal concentration (LC₅₀) values for embryos, these caused a significant decrease in the survival of neonates, i.e. when crabs are outside the egg and not protected by chorion. The decrease in the neonate population caused by these concentrations, which could be found in the environment, might impact aquatic communities.     

Insecticide residues in bats along a land use-gradient dominated by cotton cultivation in northern Benin,West Africa

Many regions in Africa are currently being converted from subsistence to cash crop farming such as cotton. Agricultural intensification is usually accompanied by increased use of pesticides, which can have an impact on non-target organisms. Bats are particularly sensitive to insecticide loads while providing substantial ecosystem services as predators of herbivorous insects. In this study, pesticide residues in bats in a landscape in northern Benin were investigated, which spanned a land use gradient from an agricultural zone dominated by cotton farms, through a buffer zone, and into a national park. Insecticides used in cotton cultivation, such as endosulfan, chlorpyrifos, flubendiamide, and spirotetramat, as well as persistent insecticides such as bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT), lindane, and aldrine, were analysed. Insecticide residues detected in bats comprised DDT, endosulfan, and their corresponding transformation products. Maximum concentrations in the sampled bats were 11.2 mg/kg lipid of p,p′-DDE (median: 0.0136 mg/kg lipid) and 0.797 mg/kg lipid of β-endosulfan (median: below detection limit [DL]). While insecticide concentrations were below lethal concentrations our data suggest that DDT had probably been recently used in the study region, and larger scale use would pose an increased risk for bat populations due to the high biomagnification of DDT.     

Temporal variability in particle-associated pesticide exposure in a temporarily open estuary, Western Cape, South Africa

This study assesses the risk of current-use pesticides in a temporarily open estuary in South Africa by developing probabilistic risk estimates. Particle-associated pesticides (chlorpyrifos, prothiofos, cypermethrin, fenvalerate, endosulfan and p,p-DDE) and physicochemical parameters (salinity, temperature, flow, and total organic content (TOC)) were measured in the Lourens River estuary (Western Cape) twice a month over a period of two years and equilibrium partitioning theory was applied to calculate concentrations of pesticides in the water. The 90th percentile concentrations of pesticides associated to suspended particles and the calculated concentrations in water were 34.0microg kg(-1) and 0.15microg l(-1) for prothiofos, 19.6microg kg(-1) and 0.45microg l(-1) for chlorpyrifos and 18.6microg kg(-1) and 0.16microg l(-1) for endosulfan. Highest average concentrations were found around the summer season due to higher application rates and as a result of the low flow during this season. Species sensitivity distribution indicated a 1.5-2.8 times higher toxicity (hazardous concentration HC5) for marine organisms compared to freshwater organisms. The calculated concentrations in the water exceeded all threshold values suggested by international water guidelines. Chlorpyrifos and endosulfan posed the highest acute risk to the Lourens River estuary. No sufficient toxicity data and threshold values were found for prothiofos.     

Toxicity of the mosquito control insecticide phenothrin to three life stages of the grass shrimp (Palaemonetes pugio)

Phenothrin is a synthetic pyrethroid used as a contact insecticide in mosquito control programs. This study compared the toxicity of phenothrin to adult, larval and embryonic grass shrimp (Palaemonetes pugio) and examined oxidative stress responses in adult and larval grass shrimp. The adult 24-h LC50 was 0.341 μg/L (95 % confidence intervals 0.282-0.412) and the 96-h LC50 was 0.161 μg/L (95 % CI 0.128-0.203 μg/L). The larval 24-h LC50 was 0.50 μg/L (95 % CI 0.441-0.568) and the 96-h LC50 was 0.154 μg/L (95 % CI 0.139-0.170 μg/L). In the presence of sediment, the 24-h LC50 was 6.30 μg/L (95 % CI 5.00-7.44 μg/L) for adults and 0.771 μg/L (95 % CI 0.630-0.944) for larvae. The sublethal biomarkers glutathione and lipid peroxidase (LPx) were examined after 96-h phenothrin exposure at five concentrations, and there were no statistically significant differences in these levels in adults or larvae compared to controls. There was a significant downward trend in larval LPx levels. This research confirms that phenothrin is highly toxic to grass shrimp and suggests that both adult and larval grass shrimp are appropriate life stages for risk assessments.     

Seasonal variation in amylase, invertase, cellulase activity and carbon dioxide evolution in a tropical protected grassland of Orissa, India, sprayed with carbaryl insecticide

A distinct seasonal variation in the enzyme activities and carbon dioxide evolution in soil, with a maximum in summer, was observed in soil treated with carbaryl and in control soil. There was no significant difference in the rate of enzyme activity between 0 and 10 cm and 10 and 20 cm depth of soil. Carbaryl insecticide applied at a normal agricultural dose did not have any inhibitory effect on soil enzyme activity, or on the rate of CO(2) evolution. However, the cellulase activity was greater in the surface soil of the control plot than in the treated plot.