Due to worldwide production, sales and application, neonicotinoids dominate the global use of insecticides. While, neonicotinoids are considered as pinpoint neurotoxicants that impair cholinergic neurotransmission in pest insects, the sublethal effects on nontarget organisms and other neurotransmitters remain poorly understood. Thus, we investigated long-term neurological outcomes in the decomposer nematode Caenorhabditis elegans. In the adult roundworm the neonicotinoid thiacloprid impaired serotonergic and dopaminergic neuromuscular behaviors, while respective exposures to thiamethoxam showed no effects. Thiacloprid caused a concentration-dependent delay of the transition between swimming and crawling locomotion that is controlled by dopaminergic and serotonergic neurotransmission. Age-resolved analyses revealed that impairment of locomotion occurred in young as well as middle-aged worms. Treatment with exogenous serotonin rescued thiacloprid-induced swimming deficits in young worms, whereas additional exposure with silica nanoparticles enhanced the reduction of swimming behavior. Delay of forward locomotion was partly caused by a new paralysis pattern that identified thiacloprid as an agent promoting a specific rigidity of posterior body wall muscle cells and peripheral neuropathy in the nematode (lowest-observed-effect-level 10 ng/ml). On the molecular level exposure with thiacloprid accelerated protein aggregation in body wall muscle cells of polyglutamine disease reporter worms indicating proteotoxic stress. The results from the soil nematode Caenorhabditis elegans show that assessment of neurotoxicity by neonicotinoids requires acknowledgment and deeper research into dopaminergic and serotonergic neurochemistry of nontarget organisms. Likewise, it has to be considered more that different neonicotinoids may promote diverse neural end points.

Pesticides are widely used to alleviate pest pressure in agricultural systems, and atrazine is a typical diffuse pollutant and serves a sensitivity index for environmental characteristics. Based on the physicochemical properties of parent substances, degradation products of pesticides may pose a greater threat to aquatic ecosystems than pesticides. Atrazine and three primary degradation products (deethylatrazine (DEA), deisopropylatrazine (DIA) and didealkylatrazine (DDA)) were investigated in a semienclosed bay of the western Pacific Ocean. Seasonal surface water and suspended particulate sediment (SPS) samples were collected from the estuary and bay in January, April, and August 2019. The level of pesticide contamination was lower in the bay than in the estuary, and the pesticide concentration in the dissolved phase was higher than that in the adsorbed phase. The average concentrations of atrazine and the three degradation products in the three seasons ranged from 2.42 to 328.46 ng/L in water and from 0.07 to 12.75 ng/L in SPS. The proportion of atrazine among the four detected pollutants decreased from 0.7 to 0.1 in surface water and from 0.3 to 0.1 in SPS over the seasons. As the main degradation products, the concentration proportions of DDA and DEA reached as high as 0.6 in August. The ratio of DEA to atrazine (DEA/ATR) increased from January to August, which indicated the progressive degradation process in the bay. Single-factor analysis of variance and principal component analysis indicated that atrazine degradation was sensitive to temperature, dissolved oxygen, and salinity. These three factors accounted for almost 70% of the seasonal variance in atrazine without a quantification assessment of photolysis or bacteria. The spatial distributions of DEA in the three seasons demonstrated that wind and currents also played important roles in pollutant redistribution. The seasonal temporal and spatial correlations between water and SPS demonstrated the degradation patterns of atrazine in marine conditions, supporting the need for future detailed toxicity studies.

Biofumigation is an effective, non-chemical method to control soil-borne pests and diseases and to maximize crop yield. We studied the responses of soil bacterial and fungal communities, the soil’s nutritional state and strawberry yield, when the soil was biofumigated each year for five consecutive years using fresh chicken manure (BioFum). BioFum significantly increased the soil’s NH4+-N, NO3−-N, available P and K and organic matter. Fusarium spp. and Phytophthora spp. which are known to cause plant disease, were significantly decreased after BioFum. In addition, Biofum increased the soil’s temperature, enhanced chlorophyll levels in the leaves of strawberry plants, and the soluble sugar and ascorbic acid content in strawberry fruit. We used high-throughput gene sequencing to monitor changes in the soil’s bacterial and fungal communities. Although BioFum significantly decreased the diversity of these communities, it increased the relative abundance of some biological control agents in the phylum Actinobacteria and the genera Pseudomonas, Bacillus and Chaetomium. An increase in these biological control agents would reduce the incidence of soil-borne pathogens and plant disease. Although strawberry marketable yield using BioFum was higher in the first three years, the decline in the final two years could be due to the accumulation of P and K which may have delayed flowering and fruiting. Methods to overcome yield losses using BioFum need to be developed in the future. Our research, however, showed that BioFum enhanced soil fertility, reduced the presence of soil pathogens, increased the relative abundance of beneficial bacteria and fungi and improved strawberry quality. Unlike chemical soil treatments that can cause pest and disease resistance when used continuously over many years, our multi-year research program on BioFum showed that this treatment provided significant benefits to the soil, plant and strawberry fruit.

Epidemiological relationships between pesticide use and male infertility have been suggested for a long time. Etoxazole (ETX), an oxazoline pesticide, has been extensively used for pest eradication. It is considered relatively safe and has low mammalian toxicity because it specifically inhibits chitin synthesis. However, ETX may have toxic effects on the reproductive system. In this study, we examined the effects of ETX on the reproductive system using mouse testis cell lines (TM3 for Leydig cells and TM4 for Sertoli cells) and C57BL/6 male mice. We confirmed that ETX has anti-proliferative effects on the TM3 and TM4 cell lines. Moreover, ETX induced mitochondrial dysfunction and hampers calcium homeostasis. Western blot analysis of MAPK and Akt signaling cascades was performed to demonstrate the mode of action of ETX at a molecular level. Moreover, ETX induced misregulation of genes related to testicular function. Upon oral administration of ETX in C57BL/6 male mice, testis weight was reduced and transcriptional expression related to testis function was altered. These results indicate that ETX induces testicular toxicity by inducing mitochondrial dysfunction and calcium imbalance and regulating gene expression.

Pesticide exposure is regarded as a contributing factor to the high gross loss rates of managed colonies of Apis mellifera. Pesticides enter the hive through contaminated nectar and pollen carried by returning forager honey bees or placed in the hive by beekeepers when managing hive pests. We used an in vitro rearing method to characterize the effects of seven pesticides on developing brood subjected dietary exposure at worse-case environmental concentrations detected in wax and pollen. The pesticides tested included acaricides (amitraz, coumaphos, fluvalinate), insecticides (chlorpyrifos, imidacloprid), one fungicide (chlorothalonil), and one herbicide (glyphosate). The larvae were exposed chronically for six days of mimicking exposure during the entire larval feeding period, which is the worst possible scenario of larval exposure. Survival, duration of immature development, the weight of newly emerged adult, morphologies of the antenna and the hypopharyngeal gland, and gene expression were recorded. Survival of bees exposed to amitraz, coumaphos, fluvalinate, chlorpyrifos, and chlorothalonil was the most sensitive endpoint despite observed changes in many developmental and physiological parameters across the seven pesticides. Our findings suggest that pesticide exposure during larvae development may affect the survival and health of immature honey bees, thus contributing to overall colony stress or loss. Additionally, pesticide exposure altered gene expression of detoxification enzymes. However, the tested exposure scenario is unlikely to be representative of real-world conditions but emphasizes the importance of proper hive management to minimize pesticide contamination of the hive environment or simulates a future scenario of increased contamination.

Common varieties of genetically modified (GM) cotton increasingly display insect-resistant properties via expression of bacterial-derived toxins from Bacillus thuringiensis (Bt). This necessitates a deeper understanding of the possible effects of these crops on non-target insects. The mirid bug Apolygus lucorum is a major pest in cotton production in China, however, the effect of GM cotton on this non-target species is currently virtually unknown. This insect is exposed to these transgenic plants by consuming genetically modified (GM) leaves. In this study, laboratory experiments were conducted to assess the toxicity of CCRI41 and CCRI45, (genetically modified cotton varieties which express the toxins Cry1Ac and CpTI (Cowpea Trypsin Inhibitor)) on nymphs and adults of A. lucorum. There was no detectable increase in mortality after A. lucorum fed on GM cotton leaves for 20 days. While we detected trace amounts of Cry1Ac proteins in both A. lucorum nymphs and adults (<10 ng/g fresh weight), the expression of genes related to detoxification did not detectably differ from those feeding on non-GM cotton. Our binding assays did not show Cry1Ac binding to receptors on the midgut brush border membrane from either A. lucorum nymphs or adults. Our findings collectively indicate that feeding on leaves of the GM cotton varieties CCRI41 and CCRI45 have few toxic effects on A. lucorum.

Microplastic pollution is a major global environmental problem in both aquatic and terrestrial environments. Pesticides are frequently applied to agricultural soil to reduce the effects of pests on crops, but may also affect the degradation of plastics. In this study, we generated microplastics from polyethylene (PE) film and biodegradable poly(butylene adipate-co-terephthalate) (PBAT) film and determined (1) the effect of prothioconazole on degradation of the microplastics, and (2) the adsorption and release characteristics of heavy metals (Cr, Cu, As, Pb, Ba, and Sn) by the microplastics during degradation process. Changes of surface functional groups and morphologies were measured by FTIR and SEM, while metal concentrations were determined by ICPMS. Prothioconazole was found to promote plastic degradation. PBAT degraded faster and adsorbed more heavy metals from the soil than PE. Whether the microplastics adsorb or release heavy metals depended on the metal and their concentrations. Prothioconazole inhibited the adsorption of Cr, As, Pb and Ba by microplastics, promoted the adsorption of Cu, and had no significant effect for Sn. These results can help to assess the ecological risk of microplastic pollution from plastic mulch when combined with heavy metals.

Leaf-cutting ants are important pests of forests and agricultural crops in the Neotropical region. Atta sexdens colonies can be composed of thousands of individuals, which form a highly complex society with a single reproductive queen. Successful control of this species is achieved only if the queen is affected. Few data are available on the lethal or sublethal effects of toxic compounds on leaf-cutting ant queens. Azadirachtin has been claimed as an effective biopesticide for insect control, but its action on leaf-cutting ants has been little explored. This study shows that azadirachtin affects oviposition in A. sexdens queens, impairing egg development by decreasing protein reserves. Azadirachtin inhibits the synthesis of vitellogenin, the major yolk protein precursor. The negative effects of azadirachtin on the reproduction of leaf-cutting ant queens suggest a potential use for the control of these insects.

Pyrethroid-resistant Hyalella azteca with voltage-gated sodium channel mutations have been identified at multiple locations throughout California. In December 2013, H. azteca were collected from Mosher Slough in Stockton, CA, USA, a site with reported pyrethroid (primarily bifenthrin and cyfluthrin) sediment concentrations approximately twice the 10-d LC50 for laboratory-cultured H. azteca. These H. azteca were shipped to Southern Illinois University Carbondale and have been maintained in pyrethroid-free culture since collection. Even after 22 months in culture, resistant animals had approximately 53 times higher tolerance to permethrin than non-resistant laboratory-cultured H. azteca. Resistant animals held in culture also lacked the wild-type allele at the L925 locus, and had non-synonymous substitutions that resulted in either a leucine-isoleucine or leucine-valine substitution. Additionally, animals collected from the same site nearly three years later were again resistant to the pyrethroid permethrin. When resistant animals were compared to non-resistant animals, they showed lower reproductive capacity, lower upper thermal tolerance, and the data suggested greater sensitivity to, 4, 4′-dichlorodiphenyltrichloroethane (DDT), copper (II) sulfate, and sodium chloride. Further testing of the greater heat and sodium chloride sensitivity of the resistant animals showed these effects to be unrelated to clade association. Fitness costs associated with resistance to pyrethroids are well documented in pest species (including mosquitoes, peach-potato aphids, and codling moths) and we believe that H. azteca collected from Mosher Slough also have fitness costs associated with the developed resistance.

The widespread cultivation of transgenic Bt cotton makes assessing the potential effects of this recombinant crop on non-target organisms a priority. However, the effect of Bt cotton on many insects is currently virtually unknown. The plant bug Adelphocoris suturalis is now a major pest of cotton in southern China and the beetle Haptoncus luteolus is one of the most ancient cotton pollinators. We conducted laboratory experiments to evaluate the toxicity of the Bt cotton varieties ZMSJ, which expresses the toxins Cry1Ac and Cry2Ab, and ZMKCKC, which expresses Cry1Ac and EPSPS, on adult A. suturalis and H. luteolus. No significant increase in the mortality of either species was detected after feeding on Bt cotton leaves or pollen for 7 days. Trace amounts of Cry1Ac and Cry2Ab proteins could be detected in both species but in vitro binding experiments found no evidence of Cry1Ac and Cry2Ab binding proteins. These results demonstrate that feeding on the leaves or pollen of these two Bt cotton varieties has no toxic effects on adult A. suturalis or H. luteolus.