RNAi validation of resistance genes and their interactions in the highly DDT-resistant 91-R strain of Drosophila melanogaster.

4,4'-dichlorodiphenyltrichloroethane (DDT) has been re-recommended by the World Health Organization for malaria mosquito control. Previous DDT use has resulted in resistance, and with continued use resistance will increase in terms of level and extent. Drosophila melanogaster is a model dipteran that has many available genetic tools, numerous studies done on insecticide resistance mechanisms, and is related to malaria mosquitoes allowing for extrapolation. The 91-R strain of D. melanogaster is highly resistant to DDT (>1500-fold), however, there is no mechanistic scheme that accounts for this level of resistance. Recently, reduced penetration, increased detoxification, and direct excretion have been identified as resistance mechanisms in the 91-R strain. Their interactions, however, remain unclear. Use of UAS-RNAi transgenic lines of D. melanogaster allowed for the targeted knockdown of genes putatively involved in DDT resistance and has validated the role of several cuticular proteins (Cyp4g1 and Lcp1), cytochrome P450 monooxygenases (Cyp6g1 and Cyp12d1), and ATP binding cassette transporters (Mdr50, Mdr65, and Mrp1) involved in DDT resistance. Further, increased sensitivity to DDT in the 91-R strain after intra-abdominal dsRNA injection for Mdr50, Mdr65, and Mrp1 was determined by a DDT contact bioassay, directly implicating these genes in DDT efflux and resistance.

Changes in pesticide occurrence in suburban surface waters in Massachusetts, USA, 1999-2010.

Pesticide occurrence was determined in two suburban surface waters in eastern Massachusetts, USA during 2009 and 2010. Out of 118 collected samples, 45 samples showed detections of one or more target pesticides. Among the herbicides, 2,4-D was the most frequently detected and imidacloprid was the most frequently detected insecticide. Comparison with data from 1999 to 2000 indicated fewer pesticide detections and lower pesticide toxicity index values in the current study. Regulatory phaseout of chlorpyrifos and diazinon from residential use by 2004 was reflected in the results by the absence of chlorpyrifos detections and lower detection frequencies of diazinon. Detected pesticide levels were below aquatic life benchmarks.

Maternal preconception PFOS exposure of Drosophila melanogaster alters reproductive capacity, development, morphology and nutrient regulation.

Perfluorooctanesulfonic acid (PFOS) is a persistent synthetic surfactant widely detected in the environment. Developmental PFOS exposures are associated with low birth weight and chronic exposures increase risk for obesity and type 2 diabetes. As an obesogen, PFOS poses a major public health exposure risk and much remains to be understood about the critical windows of exposure and mechanisms impacted, especially during preconception. Here, we leverage evolutionarily conserved pathways and processes in the fruit fly Drosophila melanogaster (wild-type Canton-S and megalin-UAS RNAi transgenic fly lines) to investigate the window of maternal preconception exposure to PFOS on reproductive and developmental toxicity, and examine receptor (megalin)-mediated endocytosis of nutrients and PFOS into the oocyte as a potential mechanism. Preconception exposure to 2 ng PFOS/female resulted in an internal concentration of 0.081 ng/fly over two days post exposure, no mortality and reduced megalin transcription. The number of eggs laid 1-3 days post exposure was reduced and contained 0.018 ng PFOS/egg. Following heat shock, PFOS was significantly reduced in eggs from megalin-knockdown transgenic females. Cholesterol and triglycerides were increased in eggs laid immediately following PFOS exposure by non-heat shocked transgenic females whereas decreased cholesterol and increased protein levels were found in eggs laid by heat shocked transgenic females. Preconception exposure likewise increased cholesterol in early emerging wildtype F1 adults and also resulted in progeny with a substantial developmental delay, a reduction in adult weights, and altered transcription of Drosophila insulin-like peptide genes. These findings support an interaction between PFOS and megalin that interferes with normal nutrient transport during oocyte maturation and embryogenesis, which may be associated with later in life developmental delay and reduced weight.

Perfluorooctanesulfonic acid (PFOS) and perfluorobutanesulfonic acid (PFBS) impaired reproduction and altered offspring physiological functions in Caenorhabditis elegans.

Perfluorobutanesulfonic acid (PFBS), a shorter chain Per- and polyfluoroalkyl substances (PFASs) cognate of perfluorooctanesulfonic acid (PFOS), has been used as replacement for the toxic surfactant PFOS. However, emerging evidences suggest safety concerns for PFBS and its effect on reproductive health is still understudied. Therefore, the current work aimed to investigate the effect of PFBS, in comparison to PFOS, on reproductive health using Caenorhabditis elegans as an in vivo animal model. PFOS (≥10 µM) and PFBS (≥1000 µM) significantly impaired the reproduction capacity of C. elegans, represented as reduced brood size (total egg number) and progeny number (hatched offspring number), without affecting the hatchability. Additionally, the preconception exposure of PFOS and PFBS significantly altered the embryonic nutrient loading and composition, which further led to abnormalities in growth rate, body size and locomotive activity in F1 offspring. Though the effective exposure concentration of PFBS was approximately 100 times higher than PFOS, the internal concentration of PFBS was lower than that of PFOS to produce the similar effects of PFOS. In conclusion, PFOS and PFBS significantly impaired the reproductive capacities in C. elegans and the preconception exposure of these two compounds can lead to offspring physiological dysfunctions.

Golfer exposure to chlorpyrifos and carbaryl following application to turfgrass.

Exposure of golfers to pesticides following their application to turfgrass is of concern to regulators, turfgrass professionals, and consumers. Multipathway exposures were evaluated for golfers on turfgrass treated with chlorpyrifos and carbaryl. Air concentrations and transferable foliar residues (TFRs) were measured to assess potential respiratory and dermal exposures, respectively. At the same time, exposure to individuals simulating the play of golf was determined by dosimetry and urinary biomonitoring. Individual golfer exposure was determined in 76 rounds of golf following eight applications of chlorpyrifos and two applications of carbaryl. Estimated exposures to golfers following full course and full rate applications of chlorpyrifos and carbaryl were 19-68 times below current U.S. EPA acute reference dose (Rfd) values, indicating safe exposures under U.S. EPA hazard quotient criteria. Dermal exposure was determined to be the dominant exposure pathway to golfers, accounting for approximately 60% of the chlorpyrifos absorbed dose and 100% of the carbaryl absorbed dose. This study also provides a set of transfer factors (TFs) that may be used to determine dermal exposure of golfers to pesticides using transferable residue data.