Ecotoxicological characterization of a tropical soil after diazinon spraying

The impact of diazinon spraying in an agricultural tropical soil through the evaluation of both the habitat and retention functions of the soil system was never reported. To fill this gap, five times the recommended dose of a commercial diazinon formulation was sprayed in an agricultural area of Costa Rica, and dilution gradients of the sprayed soil were prepared in the laboratory. Avoidance and reproduction tests with soil organisms (Eisenia andrei, Enchytraeus crypticus and Folsomia candida) to evaluate losses in terrestrial habitat function, and growth and reproduction tests with aquatic organisms (Chlorella vulgaris and Daphnia magna, respectively) to evaluate the retention function of soil were performed. Results demonstrated that regarding habitat function, F. candida reproduction was the most sensitive endpoint (EC₅₀?=?0.288?mg?a.i./kg), followed by avoidance behaviour of E. andrei (EC₂₀?=?1.75?mg?a.i./kg). F. candida avoidance and the reproduction of E. andrei and E. crypticus were not affected by diazinon. The toxicity tests with aquatic organisms showed that the soil retention function was insufficient to prevent effects of diazinon either on microalgae growth (EC₅₀?≤?0.742?mg/L and EC₂₀?≤?0.223?mg/L) and on the reproduction of the cladoceran (EC₅₀?≤?0.00771?mg/L and EC₂₀?≤?0.00646?mg/L). Results suggested that diazinon exerted toxic effects even at the dilution corresponding to the recommended dose, fact which makes its misuse an issue of environmental concern. Furthermore, the present study highlighted the importance and complementary nature of the assessment of both habitat and retention functions to an ecological risk assessment in tropical systems.     

Comparative toxicity of imidacloprid and thiacloprid to different species of soil invertebrates

Neonicotinoid insecticides have come under increasing scrutiny for their impact on non-target organisms, especially pollinators. The current scientific literature is mainly focused on the impact of these insecticides on pollinators and some aquatic insects, leaving a knowledge gap concerning soil invertebrates. This study aimed at filling this gap, by determining the toxicity of imidacloprid and thiacloprid to five species of soil invertebrates: earthworms (Eisenia andrei), enchytraeids (Enchytraeus crypticus), Collembola (Folsomia candida), oribatid mites (Oppia nitens) and isopods (Porcellio scaber). Tests focused on survival and reproduction or growth, after 3–5 weeks exposure in natural LUFA 2.2 standard soil. Imidacloprid was more toxic than thiacloprid for all species tested. F. candida and E. andrei were the most sensitive species, with LC₅₀s of 0.20–0.62 and 0.77?mg/kg dry soil for imidacloprid and 2.7–3.9 and 7.1?mg/kg dry soil for thiacloprid. EC₅₀s for effects on the reproduction of F. candida and E. andrei were 0.097–0.30 and 0.39?mg/kg dry soil for imidacloprid and 1.7–2.4 and 0.44?mg/kg dry soil for thiacloprid. The least sensitive species were O. nitens and P. scaber. Enchytraeids were a factor of 5–40 less sensitive than the taxonomically related earthworm, depending on the endpoint considered. Although not all the species showed high sensitivity to the neonicotinoids tested, these results raise awareness about the effects these insecticides can have on non-target soil invertebrates.     

Multigeneration toxicity of imidacloprid and thiacloprid to <Emphasis Type="Italic">Folsomia candida</Emphasis>

In a recent study, we showed that the springtail Folsomia candida was quite sensitive the neonicotinoid insecticides imidacloprid and thiacloprid. This study aimed at determining the toxicity of both compounds to F. candida following exposure over three generations, in natural LUFA 2.2 standard soil. In the first generation, imidacloprid was more toxic than thiacloprid, with LC₅₀s of 0.44 and 9.0?mg/kg dry soil, respectively and EC₅₀s of 0.29 and 1.5?mg/kg dry soil, respectively. The higher LC₅₀/EC₅₀ ratio suggests that thiacloprid has more effects on reproduction, while imidacloprid shows lethal toxicity to the springtails. In the multigeneration tests, using soil spiked at the start of the first generation exposures, imidacloprid had a consistent effect on survival and reproduction in all three generations, with LC₅₀s and EC₅₀s of 0.21–0.44 and 0.12–0.29?mg/kg dry soil, respectively, while thiacloprid-exposed animals showed clear recovery in the second and third generations (LC₅₀ and EC₅₀?>?3.33?mg/kg dry soil). The latter finding is in agreement with the persistence of imidacloprid and the fast degradation of thiacloprid in the test soil.     

Sensitivity of <Emphasis Type="Italic">Eisenia andrei</Emphasis> (Annelida,Oligochaeta) to a commercial formulation of abamectin in avoidance tests with artificial substrate and natural soil under tropical conditions

Obtaining ecotoxicological data on pesticides in tropical regions is imperative for performing more realistic risk analysis, and avoidance tests have been proposed as a useful, fast and cost-effective tool. Therefore, the present study aimed to evaluate the avoidance behavior of Eisenia andrei to a formulated product, Vertimec^® 18 EC (a.i abamectin), in tests performed on a reference tropical artificial soil (TAS), to derive ecotoxicological data on tropical conditions, and a natural soil (NS), simulating crop field conditions. In TAS tests an adaptation of the substrate recommended by OECD and ISO protocols was used, with residues of coconut fiber as a source of organic matter. Concentrations of the pesticide on TAS test ranged from 0 to 7 mg abamectin/kg (dry weight—d.w.). In NS tests, earthworms were exposed to samples of soils sprayed in situ with: 0.9 L of Vertimec^® 18 EC/ha (RD); twice as much this dosage (2RD); and distilled water (Control), respectively, and to 2RD: control dilutions (12.5, 25, 50, 75%). All tests were performed under 25 ± 2°C, to simulate tropical conditions, and a 12hL:12hD photoperiod. The organisms avoided contaminated TAS for an EC_50,48h = 3.918 mg/kg soil d.w., LOEC = 1.75 mg/kg soil d.w. and NOEC = 0.85 mg/kg soil d.w. No significant avoidance response occurred for any NS test. Abamectin concentrations in NS were rather lower than EC_{50, 48h} and LOEC determined in TAS tests. The results obtained contribute to overcome a lack of ecotoxicological data on pesticides under tropical conditions, but more tests with different soil invertebrates are needed to improve pesticides risk analysis.     

A comparison of the effects of silver nanoparticles and silver nitrate on a suite of soil dwelling organisms in two field soils

Nanomaterials are increasingly used in a wide range of products, leading to growing concern of their environmental fate. In order to understand the fate and effects of silver nanoparticles in the soil environment, a suite of toxicity tests including: plant growth with Elymus lanceolatus (northern wheatgrass) and Trifolium pratense (red clover); collembolan survival and reproduction (Folsomia candida); and earthworm avoidance, survival and reproduction (Eisenia andrei) was conducted. The effect of silver nanoparticles (AgNP) was compared with the effect of ionic silver (as AgNO₃) in two agricultural field soils (a sandy loam and a silt loam). Lethal (LC50) or sub lethal (IC50) effect levels are presented for all endpoints and demonstrate that in most cases AgNO₃ (i.e. ionic silver) was found to be more toxic than the AgNP across test species. The difference in effects observed between the two forms of silver varied based on test species, endpoint and soil type. In tests that were conducted across different soil types, organisms in the sandier soil had a greater response to the Ag (ionic and nano) than those in soil with a high silt content. Earthworms (avoidance behavior and reproduction) were the most sensitive to both AgNP and AgNO₃, while plant emergence was the least sensitive endpoint to both forms of Ag. The use of a test battery approach using natural field soils demonstrates the need to better quantify the dissolution and transformation products of nanomaterials in order to understand the fate and effects of these materials in the soil environment.     

Copper toxicity in a natural reference soil: ecotoxicological data for the derivation of preliminary soil screening values

The risk assessment of contaminated soils is conventionally done with the support of soil screening values (SSVs). Since SSVs are still unavailable for many European countries, including Portugal, standardized toxicity tests are urgently claimed for their derivation. Hence, this work aimed the generation of toxicity values for copper (Cu) in a natural reference soil (PTRS1) targeting different terrestrial species, endpoints and soil functions, as to derive a preliminary Cu SSV. For this, the Assessment Factor approach was applied, which allowed calculating predicted no effect concentrations (PNEC) for Cu that will be the basis for SSV proposal. In order to increase the reliability of the PNEC, and hence of the SSV, a lab/field factor was applied to correct the toxicity values used for PNEC determination. Cu affected urease, cellulase and nitrogen mineralization activities. The EC₅₀ values calculated for the invertebrates reproduction were 130.9, 165.1 and 191.6 mg Cu Kg^?1soil_dw for Eisenia andrei, Enchytraeus crypticus and Folsomia candida, respectively. Cu inhibited seed germination mainly for Lactuca sativa, whilst it was toxic for the growth of different plant species (EC₅₀s between 89 and 290.5 mg Cu Kg^?1soil_dw). Based on the outcomes gathered, we proposed SSVs for Cu ranging between 26.3 and 31.8 mg Kg^?1 soil_dw, which is above the background values reported and below all the EC₂₀s recorded for the species and endpoints herein analyzed. Overall, this work describes a procedure that could be easily followed by other European countries wishing to derive SSVs adjusted to their soils.     

Influence of temperature and soil moisture on the toxic potential of clothianidin to collembolan Folsomia candida in a tropical field soil

Climate change can alter the toxic effects of pesticides on soil invertebrates. However, the nature and magnitude of the influence of climatic factors on clothianidin impacts in tropical soils are still unknown. The influence of increasing atmospheric temperature and the reduction in soil moisture on the toxicity and risk of clothianidin (seed dressing formulation Inside FS^®) were assessed through chronic toxicity tests with collembolans Folsomia candida in a tropical field soil (Entisol). The risk of clothianidin for collembolans was estimated using the Toxicity-Exposure Ratio (TER) approach. Organisms were exposed to increasing clothianidin concentrations at 20, 25 and 27?°C in combination with two soil moisture conditions (30 and 60% of the maximum water holding capacity—WHC). The effect of temperature and soil moisture content on clothianidin toxicity was verified through the number of F. candida juveniles generated after 28 days of exposure to the spiked soil. The toxicities estimated at 25?°C (EC₅₀__30%WHC?=?0.014?mg?kg^?1; EC_{50_60%WHC}?=?0.010?mg?kg^?1) and 27?°C (EC_{50_30%WHC}?=?0.006?mg?kg^?1; EC_{50_60%WHC}?=?0.007?mg?kg^?1) were 2.9–3.0-fold (25?°C) and 4.3–6.7-fold (27?°C) higher than those found at 20?°C (EC_{50_30%WHC}?=?0.040?mg?kg^?1; EC_{50_60%WHC}?=?0.030?mg?kg^?1), indicating that clothianidin toxicity increases with temperature. No clear influence of soil moisture content on clothianidin toxicity could be observed once the EC₅₀ values estimated at 30% and 60% WHC, within the same temperature, did not significantly differ. A significant risk was detected in all temperatures and soil moisture scenarios studied, and the TER values indicate that the risk can increase with increasing temperatures. Our results revealed that temperature could overlap with soil moisture in regulating clothianidin toxicity and reinforce the importance of including climatic factors in the prospective risk assessment of pesticides.

     

Individual and combined toxic effects of herbicide atrazine and three insecticides on the earthworm, <Emphasis Type="Italic">Eisenia fetida</Emphasis>

In the present study, we evaluated the individual and combined toxic effects of herbicide atrazine and three insecticides (chlorpyrifos, lambda-cyhalothrin and imidacloprid) on the earthworm, Eisenia fetida. Results from 48-h filter paper test indicated that imidacloprid had the highest toxicity to E. fetida with an LC₅₀ of 0.05 (0.041–0.058) μg a.i. cm^?2, followed by lambda-cyhalothrin and atrazine with LC₅₀ values ranging from 4.89 (3.52–6.38) to 4.93 (3.76–6.35) μg a.i. cm^?2, while chlorpyrifos had the least toxicity to the worms with an LC₅₀ of 31.18 (16.22–52.85) μg a.i. cm^?2. Results from 14-days soil toxicity test showed a different pattern of toxicity except that imidacloprid was the most toxic even under the soil toxicity bioassay system. The acute toxicity of atrazine was significantly higher than that of chlorpyrifos. In contrast, lambda-cyhalothrin was the least toxic to the animals under the soil toxicity bioassay system. The binary mixture of atrazine–lambda-cyhalothrin and ternary mixture of atrazine–chlorpyrifos–lambda-cyhalothrin displayed a significant synergistic effect on the earthworms under the soil toxicity bioassay. Our findings would help regulatory authorities understand the complexity of effects from pesticide mixtures on non-target organisms and provide useful information of the interaction of various pesticide classes detected in natural environment.     

Acute and sublethal effects of sequential exposure to the pesticide azinphos-methyl on juvenile earthworms (<Emphasis Type="Italic">Eisenia andrei</Emphasis>)

The use of organophosphate pesticides is an integral part of commercial farming activities and these substances have been implicated as a major source of environmental contamination and may impact on a range of non-target fauna. The extent to which soil dwelling non-target organisms are affected by exposure to the organophosphate azinphos-methyl was investigated through monitoring selected biomarker responses and life cycle effects under laboratory conditions in the earthworm Eisenia andrei. Standard acute toxicity tests were conducted followed by a sequential exposure regime experiment, in order to assess the effects of multiple pesticide applications on biomarker (cholinesterase activity and neutral red retention time), life-cycle (growth and reproduction) and behaviour (avoidance and burrowing activity) responses. The present study indicates that the time between exposure events was a more important variable than concentration and that a longer interval between exposures may mitigate the effects of pesticide exposure provided that the exposure concentration is low. Additionally, it was shown that E. andrei was unable to avoid the presence of azinphos-methyl in soil, even at concentrations as high as 50% of the LC ₅₀ value, indicating that the presence of azinphos-methyl in the soil pose a realistic threat to earthworms and other soil dwelling organisms. The ChE inhibition test showed a high percentage inhibition of the enzyme in all exposure groups that survived and NRR times of exposed organisms were lower than that of the controls. The present study yielded important results that contribute to the understanding of biological impacts of pesticide pollution on the environment. Extrapolating these results can aid in optimising pesticide application regimes to mitigate the environmental effects thereof and thus ensuring sustained soil biodiversity in agricultural areas.     

Boric acid as reference substance: pros,cons and standardization

Boric acid (BA) has been successfully used as reference substance in some standard test guidelines. Due to the fact that previously selected reference substances present a significant risk to human health and/or are banned for environmental reasons, BA is being discussed for broader adoption in OECD or ISO guidelines. To provide input on BA data and contribute to the discussion on its suitability as a reference substance, in the present study BA was tested with two standard soil organisms, Enchytraeus albidus and Folsomia candida, in terms of survival, reproduction and avoidance. Additionally, published data on other organisms was analysed to derive the most sensitive soil dwelling invertebrate (hazard concentration–HC5). Results showed that BA affected the tested organisms, being two times more toxic for collembolans (LC50 = 96; EC50 = 54 mg/kg) than for enchytraeids (LC50 = 325; EC50 = 104 mg/kg). No avoidance behaviour occurred despite the fact that BA affects earthworms. Actually, it is the recommended reference substance for the earthworm avoidance test. Clearly, the suitable performance of BA in one species should not be generalized to other species. Absolute toxicity is not an important criterion for the selection of a reference substance, but it has been proposed that effects should occur within a reasonable range, i.e. <1,000 mg/kg. We could confirm, compiling previous data that for most soil invertebrates, the EC50 is expected to be below 1,000 mg/kg. From these data it could be derived that the most sensitive soil dwelling invertebrate (HC5, 50%) is likely to be affected (EC10) at 28 (8–53) mg H₃BO₃/Kg, equivalent to 4.6 (1.4–8.7) mg boron/kg.     

Eprinomectin antiparasitic affects survival,reproduction and behavior of Folsomia candida biomarker,and its toxicity depends on the type of soil

The objective of this study was to evaluate the toxicity of the antiparasitic agent eprinomectin in two subtropical soils, using ecotoxicological lethality, reproduction and avoidance behavior tests with springtails (Folsomia candida). Eprinomectin concentrations were 0 (control), 0.5, 1, 2, 4, 8, 12, 16 and 20 mg kg⁻¹ of dry soil combined with either Entisol or Oxisol soils. Statistically significant toxic effects of eprinomectin on springtails were observed in both soils. Eprinomectin was lethal starting at 8 mg kg^-1 of dry soil in Entisol, and 20 mg kg⁻¹ of dry soil in Oxisol, with effects less than 50% at lethal concentrations. Reductions in the reproduction rate of the springtails were also observed starting at 8 mg kg⁻¹ of dry soil in Entisol, and 0.5 mg kg⁻¹ of dry soil in Oxisol. EC_repr50 value calculated for Entisol was 4.38 ± 0.62 mg kg^-1 of dry soil; for Oxisol the EC_repr50 was above the highest tested concentration. For avoidance behavior, the effect occurred from 0.5 mg kg^-1 of dry soil for both soils. In Entisol, all concentrations caused avoidance of more than 95%, and in Oxisol the EC_avoi50 value was 1.33 ± 0.83 mg kg^-1 of dry soil. We conclude that eprinomectin affected survival, reproduction and caused avoidance behavior of F. candida in both soils. The toxic effects were greater as the concentration in the soils increased. The effects in Oxisol were less intense than those in Entisol with respect to the affected springtails. This discrepancy may be attributed to the different physicochemical characteristics of the soils that determine the retention capacity for eprinomectin; in particular, there are greater contents of clay, organic matter and cation exchange capacity in Oxisol.     

Avoidance behaviour response and esterase inhibition in the earthworm, Lumbricus terrestris, after exposure to chlorpyrifos

The avoidance response of earthworms to polluted soils has been standardised using a simple and low-cost test, which facilitates soil toxicity screening. In this study, the avoidance response of Lumbricus terrestris was quantified in chlorpyrifos-spiked soils, depending on the pesticide concentration and exposure duration. The inhibition of acetylcholinesterase (AChE) and carboxylesterase (CbE) activities was also determined as indirect measures of pesticide bioavailability. The effects of different chlorpyrifos concentrations were examined in a standardised test (two-chamber system) with 0.6, 3 and 15 mg/kg chlorpyrifos. A modification of the test involved a pre-exposure step (24, 48 or 72 h) in soils spiked with 15 mg/kg. In both protocols, earthworms were unable to avoid the contaminated soils. However, the esterase activities showed that all earthworms were exposed to chlorpyrifos. Acetylcholinesterase activity did not change in earthworms in the standardised behavioural test (0.58 ± 0.20 U/mg protein, mean ± SD; n = 72), whereas the CbE activity was significantly inhibited (62–87 % inhibition) in earthworms exposed to 3 and 15 mg/kg. In the modified test, earthworms had greatly inhibited AChE activity (0.088 ± 0.034 U/mg protein, n = 72), which was supported by reactivation of the inhibited enzyme activity in the presence of pralidoxime (2-PAM). Similarly, the CbE activity was significantly inhibited in earthworms with all treatments. This study suggests that the avoidance behaviour test for organophosphorus-contaminated soils could be supported by specific biomarkers to facilitate a better understanding of pesticide exposure and toxicity during this test.     

Evaluation of cytotoxicity and genotoxicity of pesticide mixtures on lymphocytes

The cytotoxicity and genotoxicity of pesticide mixtures viz. endosulfan?+?chlorpyrifos, chlorpyrifos?+?profenofos, and endosulfan?+?profenofos were evaluated on cultured human peripheral blood lymphocytes using assays for cell viability, and genotoxicity using chromosomal aberrations test and comet assay. The LC₅₀ values for cytotoxicity were 3.50?μM, 4.18?μM, and 10.5?μM for profenofos, endosulfan, and chlorpyrifos respectively. When combined in equimolar concentrations, the LC₅₀ values for cytotoxicity were 1.4?μM, 1.8?μM, and 2.0?μM for endosulfan?+?chlorpyrifos, chlorpyrifos?+?profenofos, and endosulfan?+?profenofos, respectively. Higher concentrations of individual pesticides (0.5–4.0?μM) but very low concentrations of pesticide mixtures caused significant DNA damage. Additive index values indicated a synergistic effect of toxicity for endosulfan?+?chlorpyrifos combination (1.12 TTU). The binary mixture of chlorpyrifos?+?profenofos showed an additive toxicity (0.46 TTU) while an antagonistic effect was observed for endosulfan?+?profenofos combination. Synergism could be due to these complementary pesticides simultaneously acting in different ways, magnifying their efficacy, whereas an additive interaction would imply that the chemicals are acting by the same mechanism and at the same target. Analysis of toxicity of pesticide mixtures may serve as important biomarker for occupational and household exposure to pesticides, with different modes of action.     

Toxicity of profenofos to the springtail, <Emphasis Type="Italic">Folsomia</Emphasis><Emphasis Type="Italic">candida</Emphasis>, and ammonia-oxidizers in two agricultural soils

Extensive use of organophosphorus insecticide profenofos (PFF) for agricultural and house-hold purposes has led to serious environmental pollution, with potential risk to organisms in the ecosystem. This study examined the toxicity of PFF to the soil springtail Folsomia candida and ammonia-oxidizers through a series of toxicity tests conducted on two agricultural soils. It was found that the survival, reproduction, hsp70 gene expression of F. candida and the soil potential nitrification rate (PNR) were sensitive to the PFF, whereas no apparent change was observed in the abundance of ammonia-oxidizers. The reproduction of F. candida was the most sensitive endpoint (mean 0.10 mg/kg of EC₅₀ value) for PFF, although the test was more time-consuming. The results of the acute toxicity tests suggested that the survival of F. candida could be considered as the most suitable bioindicator for fast screening of PFF toxicity because of its fast and easy test procedure. In addition, the hsp70 gene expression in F. candida and the PNR could be used as important parameters for assessment of PFF toxicity. The threshold concentration based on the obtained endpoints differed in the two soils, and consequently the soil property should be considered in toxicity assessments of contaminated soils.     

Bioaccumulation and toxicity of silver nanoparticles and silver nitrate to the soil arthropod Folsomia candida

The growing use of silver nanoparticles (Ag-NP) triggered an increasing interest in their environmental fate and possible ecotoxicological impacts. To investigate the potential risk of Ag-NP to soil organisms, the springtail Folsomia candida was exposed to Ag-NP (reported diameter size 3–8 nm) and AgNO₃ in Lufa 2.2 natural soil for 28 days to determine effects on survival and reproduction. Also, the kinetics of uptake and elimination of Ag were studied for F. candida exposed in Lufa 2.2 soil to Ag-NP (at 168 mg Ag/kg dry soil) and AgNO₃ (at 30 and 60 mg Ag/kg dry soil). AgNO₃ was toxic with an LC50 was 284 mg Ag/kg dry soil for effects on survival and EC10 and EC50 values of 47.6 and 99.5 mg Ag/kg dry soil, respectively for the effect on reproduction. These values did correspond with porewater concentrations of 0.801, 0.042 and 0.082 mg Ag/l, respectively. No effects on survival and reproduction of Ag-NP were observed up to 673 mg Ag/kg dry soil, although porewater concentration was similar to the EC50 for AgNO₃. Exposure to both Ag forms caused a fast uptake of Ag, but the Ag elimination rate was significantly higher for Ag-NP than for AgNO₃. Bioaccumulation factor was higher for AgNO₃ (on average 5.64) than for Ag-NP (1.12). These findings indicate that silver ions are more toxic than Ag-NP and have a higher potential to accumulate in F. candida.     

Cadmium toxicity for terrestrial invertebrates: taking soil parameters affecting bioavailability into account

Acute and chronic ecotoxicity tests with cadmium were conducted with the earthworm Eisenia fetida, the potworm Enchytraeus albidus and the springtail Folsomia candida. To assess the influence of the soil type on cadmium bioavailability, these tests were carried out in a standard artificial soil, in a sandy and a loamy field soil. It was not possible to evaluate the influence of soil parameters on the bioavailability on the basis of the experiments that were conducted in only three different soil types, therefore, literature data were also included. However, even in the same standard artificial soils, toxicity data in the literature for Eisenia fetida and Folsomia candida varied considerably. Consequently, no models could be developed that allow a normalization of the ecotoxicity of cadmium to parameters controlling bioavailability. In contrast to zinc, effect concentrations of cadmium for terrestrial invertebrates were always much higher than background concentrations. As the effect of aging on the bioavailability of cadmium was never taken into account, because toxicity experiments were always carried out in freshly spiked soils, these effect concentrations may even be regarded as conservative. Furthermore, the zinc–cadmium ratio in soils is usually so high that the risk of zinc ecotoxicity for terrestrial invertebrates will usually be much greater in comparison to cadmium ecotoxicity.     

Resistance to conventional insecticides in Pakistani populations of Musca domestica L. (Diptera: Muscidae): a potential ectoparasite of dairy animals

The house fly, Musca domestica L., is an important hygienic pest of humans and dairy animals with the potential to develop resistance to most chemical classes of insecticides. Six adult house fly strains from dairy farms in Punjab, Pakistan were evaluated for resistance to selected insecticides from organochlorine, organophosphate, carbamate and pyrethroid classes. For a chlorocyclodiene and two organophosphates tested, the resistance ratios (RR) at LC₅₀ were in the range of 5.60–22.02 fold for endosulfan, 7.66–23.24 fold for profenofos and 2.47–7.44 fold for chlorpyrifos. For two pyrethroids and one carbamate, the RR values at LC₅₀ were 30.22–70.02 for cypermethrin, 5.73–18.31 for deltamethrin, and 4.39–15.50 for methomyl. This is the first report of resistance to different classes of insecticides in Pakistani dairy populations of house flies. Regular insecticide resistance monitoring programs on dairy farms are needed to prevent field control failures. Moreover, integrated approaches including the judicious use of insecticides are needed to delay the development of insecticide resistance in house flies.     

Effect of triclosan on reproduction,DNA damage and heat shock protein gene expression of the earthworm Eisenia fetida

Triclosan (TCS) is released into the terrestrial environment via the application of sewage sludge and reclaimed water to agricultural land. More attention has been paid to its effect on non-target soil organisms. In the present study, chronic toxic effects of TCS on earthworms at a wide range of concentrations were investigated. The reproduction, DNA damage, and expression levels of heat shock protein (Hsp70) gene of earthworms were studied as toxicity endpoints. The results showed that the reproduction of earthworms were significantly reduced (p < 0.05) after exposure to the concentrations ranges from 50 to 300 mg kg^?1, with a half-maximal effective concentration (EC₅₀) of 142.11 mg kg^?1. DNA damage, detected by the comet assay, was observed and there was a clear significant (R² = 0.941) relationship between TCS concentrations and DNA damage, with the EC₅₀ value of 8.85 mg kg^?1. The expression levels of Hsp70 gene of earthworms were found to be up-regulated under the experimental conditions. The expression level of hsp70 gene increased, up to about 2.28 folds that in the control at 50 mg kg^?1. The EC₅₀ value based on the Hsp70 biomarker was 1.79 mg kg^?1. Thus, among the three toxicity endpoints, the Hsp70 gene was more sensitive to TCS in soil.     

Isomers and their metabolites of endosulfan induced cytotoxicity and oxidative damage in SH‐SY5Y cells

As an organochlorine insecticide, endosulfan has been widely banned or restricted, but it is still largely used in many developing countries. Previous studies have shown multiple adverse health effects of endosulfan. However, the neurotoxicity of endosulfan has not been fully elucidated. In this study, endosulfan isomers (α‐/β‐endosulfan) and their major metabolites (endosulfan sulfate, endosulfan diol, and endosulfan lactone) were, respectively, exposed to human neuroblastoma SH‐SY5Y cells. Results showed that both α‐endosulfan and β‐endosulfan caused decrease of cell viability and morphological damages in a dose‐dependent manner. Their median effective concentrations (EC_50s) were respectively 79.6 μM (α‐endosulfan) and 50.37 μM (β‐endosulfan) for 72 h exposure. EC_50s of α/β‐endosulfan mixture were lower than that of the single isomer. However, EC_50s of its metabolites were higher than that of technical endosulfan. Endosulfan and its metabolites caused increases of reactive oxygen species and the lipid peroxidation, but decrease of superoxide dismutase in a dose‐dependent manner. These results indicate that α‐endosulfan exhibits higher neurotoxicity than β‐endosulfan. Mixture of endosulfan isomers shows stronger cytotoxicity than the single isomer. After endosulfan is degraded, cytotoxicity of its metabolites decreases gradually. The neurotoxicity of endosulfan and its metabolites is closely related to oxidative damage and antioxidative deficit. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 496–504, 2016.     

Acute and sublethal effects of two insecticides on earthworms (Lumbricus terrestris L.) under laboratory conditions

Earthworms (Lumbricus terrestris L.) were exposed to commercial formulations of endosulfan and aldicarb for 2, 7, and 15 days, and the LC(10), LC(25), and LC(50) were determined. Worms were then exposed to LC(10), LC(25), and LC(50) concentrations of endosulfan and LC(10) and LC(25) concentrations of aldicarb. The growth rate and total protein content were determined and related to endosulfan and aldicarb residues in soil and earthworms. Aldicarb was more toxic than endosulfan under the experimental conditions. The residues of endosulfan and aldicarb caused a significant reduction in the growth rate and total protein content of earthworms. The residues of endosulfan and aldicarb were monitored in soil and earthworms after 2, 7, and 15 days of exposure. The residues remaining in the soil after the experiments ranged between 37.75% and 68.54% of the applied concentration for endosulfan and between 10.13% and 67.71% of the applied concentration for aldicarb. Small amounts of both insecticides were detected in worms, and accumulation was more important for endosulfan. This study proposes the use of growth rate and total protein content as biomarkers for contamination by endosulfan and aldicarb.