A novel approach for the assessment of invertebrate behavior and its use in behavioral ecotoxicology.

Sublethal effects are becoming more relevant in ecotoxicological test methods due to their higher sensitivity compared to lethal endpoints and their preventive nature. Such a promising sublethal endpoint is the movement behavior of invertebrates which is associated with the direct maintenance of various ecosystem processes, hence being of special interest for ecotoxicology. Disturbed movement behavior is often related to neurotoxicity and can affect drift, mate-finding, predator avoidance, and therefore population dynamics. We show the practical implementation of the ToxmateLab, a new device that allows monitoring the movement behavior of up to 48 organisms simultaneously, for behavioral ecotoxicology. We quantified behavioral reactions of Gammarus pulex (Amphipoda, Crustacea) after exposure to two pesticides (dichlorvos and methiocarb) and two pharmaceuticals (diazepam and ibuprofen) at sublethal, environmentally relevant concentrations. We simulated a short-term pulse contamination event that lasted 90 min. Within this short test period, we successfully identified behavioral patterns that were most pronounced upon exposure to the two pesticides: Methiocarb initially triggered hyperactivity, after which baseline behavior was restored. On the other hand, dichlorvos induced hypoactivity starting at a moderate concentration of 5 µg/L - a pattern we also found at the highest concentration of ibuprofen (10 µg/L). An additional acetylcholine esterase inhibition assay revealed no significant impact of the enzyme activity that would explain the altered movement behavior. This suggests that in environmentally realistic scenarios chemicals can induce stress - apart from mode-of-action - that affects non-target organisms' behavior. Overall, our study proves the practical applicability of empirical behavioral ecotoxicological approaches and thus represents a next step towards routine practical use.

Commercial pesticides for urban applications induced population growth and sub-cellular alterations in Raphidocelis subcapitata (Chlorophyceae) at concerning environmental concentrations.

Information regarding the safety and environmental risks of pesticides intended for urban use remains limited. This study aimed to assess the effects of four common pesticides on the microalga Raphidocelis subcapitata: DIAZINON® 25% C. E., Roundup®, URBACIN® 20C. E., and VAPODEL® 20% C. E., which are commercial formulations of diazinon, glyphosate, dichlorvos, and cypermethrin, respectively. According to 96-h inhibition of population growth bioassays, the four pesticide toxicities exemplified the following order: DIAZINON® (diazinon) > Roundup® (glyphosate) > VAPODEL® (dichlorvos) > URBACIN® (cypermethrin). Increasing pesticide concentrations elicited alterations in the specific growth rates (µmax). The macromolecule contents and photosynthetic pigments increased in groups exposed to the highest concentrations of DIAZINON® 25%, Roundup®, and URBACIN® 20 compared to the control group, despite these treatments inducing lower population growth rates. VAPODEL® 20% induced higher growth rates and lower macromolecule content compared to the control. Since active ingredients were not quantified, certain comparisons may prove limiting, but it is important to assess the effects of the whole mixtures in the form that they enter the environment, especially for urban-intended applications or generic formulations with higher additive contents. Finally, this study demonstrated that commercial pesticide formulations designed for urban applications might pose a threat to freshwater microalgae due to their underestimated toxic potential, but further studies are required.

Hepatoprotective and Renoprotective effect of Moringa oleifera Seed Oil on Dichlorvos-induced Toxicity in Male Wistar rats.

The liver and the kidney are one of the vital organs of body. Drug-induced toxicity is one of the most common problems encountered by these organs. The search for an effective medicine to treat this toxicity without any side effects has led to the use of traditional-based medicine. This study evaluated the effect of ethanolic extract of Moringa oleifera seed oil on hepatic and renal markers in dimethyl 2, 2-dichlorovinyl phosphate (DDVP, known as dichlorvos)-exposed Wistar rats. Twenty-one male Wistar rats were randomly divided into three groups of seven animals each. Group A served as the negative control and were not exposed to dichlorvos. Group B served as the positive control and were exposed to dichlorvos for 2 minutes but received no extract. Group C animals were exposed to the dichlorvos and received 300mg/kg of extract (Moringa oleifera seed oil) for 7days before and 21days after exposure. Exposure to DDVP led to a significant increase in hepatic & renal markers, inflammatory markers, decrease in plasma protein and alteration of plasma electrolyte. Moringa oleifera seed oil regulated and significantly enhanced plasma protein, reduced elevated levels of hepatic & renal markers, inflammatory markers in the study sample. In addition, histopathology observation showed that Moringa seed oil was able to regenerate the hepatorenal damage on exposure to dichlorvos. Conclusion: Moringa oleifera seed oil exhibited hepato-protective, nephroprotective properties and could be explored in nutrition and health.

Moringa oleifera seed oil partially abrogates 2,3-dichlorovinyl dimethyl phosphate (Dichlorvos)-induced cardiac injury in rats: evidence for the role of oxidative stress.

OBJECTIVES: Cardiovascular diseases are major causes of non-infectious diseases globally. The use of pesticides has been linked with the high global burden of non-communicable diseases. Despite the indiscriminate exposure to dichlorvos (DDVP) by inhalation, no report exists on its possible cardiotoxic effect. This study investigated the cardiotoxicity of DDVP exposure by inhalation and the possible role of Moringa oleifera seed oil. METHODS: Twenty-one male rats were randomly assigned into 3 groups. Group A (control) received only standard rat diet and water ad' libitum, group B (DDVP) was exposed to DDVP via inhalation for 15 min daily in addition to rat diet and water, and group C (DDVP + M. oleifera seed oil) received treatment as group B as well as 300 mg/kg of M. oleifera seed oil p.o for 28 days. RESULTS: Significant reductions in body weight gain and cardiac weight were observed in DDVP-exposed animals (p<0.05). Similarly, 28 days of exposure to DDVP led to a significant increase in lactate dehydrogenase, creatinine kinase and troponin (p<0.05). DDVP-exposed rats also showed a significant increase in malondialdehyde, and a significant decline in superoxide dismutase and glutathione peroxidase (p<0.05). However, catalase was comparable in DDVP-exposed and control rats. Histopathological observations of the cardiac tissue revealed that DDVP caused marked fat degeneration and necrosis of the myocardial layer. The changes in DDVP-exposed rats were significantly, though not completely, restored by M. oleifera seed oil administration. CONCLUSIONS: This study provides novel mechanistic information on the cardiotoxicity of DDVP inhalation, and the antioxidant potential of M. oleifera seed oil.

Gut bacteria of the cowpea beetle mediate its resistance to dichlorvos and susceptibility to Lippia adoensis essential oil.

Bacteria inhabiting the gut of insects provide many benefits to their hosts, such as aiding in food digestion, reproduction, and immunity, tissue homeostasis, adaptation to environment and resistance to pathogen and pesticides. The cowpea beetle, Callosobruchus maculatus, is a serious cosmopolitan pest of pulses. This beetle has lent itself as a guinea pig for several ecological studies. It harbors a consortium of bacterial communities in its gut, but the evidence for their role in its physiology is fragmentary. In this work, we hypothesized that gut microbiota mediates C. maculatus resistance to dichlorvos (DDVP or O,O-dimethyl O-2,2-dichlorovinylphosphate) and represent the target of Lippia adoensis (Gambian Tea Bush) essential oil (EO). Symbiotic and aposymbiotic beetles were exposed to artificial cowpea seeds earlier treated with DDVP or EO. Adult mortality and changes in gut bacterial community composition and abundance were examined at F1 and F5 generations. The susceptibility of experimental beetles to DDVP was significantly affected by their symbiotic status. The adult mortality decreased across generations in DDVP treatments, and remained significantly higher in aposymbiotic groups. In EO treatments, the mortality was consistent irrespective of symbiotic status and experimental generations. When compared to DDVP and the Control, EO treatments had significantly lower bacterial richness and diversity, as well as lower abundance of Proteobacteria, Firmicutes, and Bacteroidetes. These results support our hypothesis and describe the responses of gut microbial communities to pesticide treatments. This could be of interest for developing new management strategies of this pest.

Toxicity and neurophysiological impacts of plant essential oil components on bed bugs (Cimicidae: Hemiptera).

Bed bugs (Cimex lectularius L.) are globally important human parasites. Integrated pest management (IPM) approaches, which include the use of essential oil-based insecticidal compounds, have been proposed for their control. This study aimed to define insecticidal activity and neurophysiological impacts of plant essential oil constituents. The topical and fumigant toxicity of 15 compounds was evaluated against adult male bed bugs. Neurological effects of the 6 most toxicologically active compounds were also determined. In both topical and fumigant bioassays, carvacrol and thymol were the most active compounds. The potency of bifenthrin (a pyrethroid insecticide) in topical bioassays was 72,000 times higher than carvacrol, while vapors of dichlorvos (an organophosphate insecticide) were 445 times more potent than thymol. Spontaneous electrical activity measurements of the bed bug nervous system demonstrated neuroinhibitory effects of carvacrol, thymol and eugenol, whereas linalool produced an excitatory effect. Although citronellic acid and (±)-camphor increased baseline activity of the nervous system their effects were not statistically significant. Bifenthrin also caused neuroexcitation, which is consistent with its known mode of action. These comparative toxicity and neurological impact findings provide new information for formulating effective essential oil-based insecticides for bed bug IPM and conducting mode-of-action studies on individual essential oil components.

Latrophilin mediates insecticides susceptibility and fecundity through two carboxylesterases, esterase4 and esterase6, in Tribolium castaneum.

Latrophilin (LPH) is known as an adhesion G-protein-coupled receptor which involved in multiple physiological processes in organisms. Previous studies showed that lph not only involved the susceptibility to anticholinesterase insecticides but also affected fecundity in Tribolium castaneum. However, its regulatory mechanisms in these biological processes are still not clear. Here, we identified two potential downstream carboxylesterase (cce) genes of Tclph, esterase4 and esterase6, and further characterized their interactions with Tclph. After treatment of T. castaneum larvae with carbofuran or dichlorvos insecticides, the transcript levels of Tcest4 and Tcest6 were significantly induced from 12 to 72 h. RNAi against Tcest4 or Tcest6 led to the higher mortality compared with the controls after the insecticides treatment, suggesting that these two genes play a vital role in detoxification of insecticides in T. castaneum. Furthermore, with insecticides exposure to Tclph knockdown beetles, the expression of Tcest4 was upregulated but Tcest6 was downregulated, indicating that beetles existed a compensatory response against the insecticides. Additionally, RNAi of Tcest6 resulted in 43% reductions in female egg laying and completely inhibited egg hatching, which showed the similar phenotype as that of Tclph knockdown. These results indicated that Tclph affected fecundity by positively regulating Tcest6 expression. Our findings will provide a new insight into the molecular mechanisms of Tclph involved in physiological functions in T. castaneum.

Dose-response modeling of reactivating potency of oximes K027 and K203 against a direct acetylcholinesterase inhibitor in rat erythrocytes.

Inhibition of acethylcholinesterase (AChE) as a key molecular event induced by organophosphate (OP) pesticides and nerve agents presents a human health concern. In efficacy testing of experimental oximes, potential antidotes in OP poisoning, reactivation of OP-inhibited AChE is used as specific endpoint. However, according to our best knowledge, so far oximes have not been quantitatively evaluated by comprehensive benchmark dose (BMD) approach, that would improve both identification and quantification of the effect and allow more rigorous comparison of efficacies. Thus, we have examined in vivo dose-response relationship for two promising experimental oximes, K203 and K027, concerning reactivation of erythrocyte AChE inhibited by dichlorvos (DDVP). Groups of Wistar rats were treated with six different doses of oximes (i.m) immediately after DDVP challenge (s.c) and AChE was measured 60 min later. Dose-response modeling was done by PROAST software 65.5 (RIVM, The Nederlands). BMD-covariate method resulted in four-parameter model from both exponential and Hill model families as the best estimate of relationship between AChE activity and oxime dose, with potency parameter being oxime-dependent. Oxime K027 was shown to be 1.929-fold more potent considering that 58% increase in AChE activity was achived with the dose BMD58-K027 = 52 µmol/kg in contrast to BMD58-K203 = 100 µmol/kg.

Dichlorvos Induced Oxidative and Neuronal Responses in Rats: Mitigative Efficacy of Nigella sativa (Black Cumin).

Poisoning from Organophosphates (OPs), especially Dichlorvos (DDVP) has become endemic due to theincreasing use in house hold and agricultural pests control, with most marked effects in the nervous system. However, it isevidenced that natural antioxidants are efficacious against OPs toxicity. Thus, this study investigated the possible antidotalefficacy of Nigella sativa oil (NSO) in Dichlovos (DDVP) induced oxidative and neuronal damages in Wistar rats. DDVPwas administered at sub-chronic daily dosage of 8.8 mg/kg.bw for 7 days and a post-administration of NSO at 1 ml/kg.bwfor the subsequent 7 days. The rats were euthanized on the 15thday, blood sample collected via cardiac puncture, centrifugedand the plasma used for biochemical analysis of total antioxidant capacity (TAC), reduced glutathione (GSH) and totalreactive oxygen species (ROS), while the frontal, occipital and cerebellar cortices and the medulla were removed for histomorphological examinations. The results showed significant (P≤0.05) decrease in plasma TAC and GSH, while a significant(P≤0.05) increase in ROS was recorded, and some vacuolation around the neurons especially in the frontal and cerebellarcortices following DDVP exposure. However, post treatment with NSO was observed to be efficacious in the recovery ofthe oxidative activities and the neuro-architectural integrities. Thus, it can be concluded that the antioxidant capacity of NSOcould be efficacious against OPs induced oxidative damages, especially in dichlorvos accidents.

Omics for understanding the tolerant mechanism of Trichoderma asperellum TJ01 to organophosphorus pesticide dichlorvos.

BACKGROUD: Though it is toxic to humans, dichlorvos is a widely used chemical pesticide and plays an important role in the control of plant pests. The application of a combination of the biocontrol agent Trichoderma with dichlorvos may reduce the need for chemical pesticides. Therefore, revealing the specific molecular mechanism of Trichoderma tolerance to dichlorvos has become particularly important. RESULTS: In this study, using transcriptome and metabolome analyses, changes in primary and secondary metabolisms in Trichoderma asperellum TJ01 were comprehensively studied in the presence of dichlorvos. A novel C2H2 zinc finger protein gene, zinc finger chimera 1 (zfc1), was discovered to be upregulated, along with a large number of oxidoreductase genes and ABC transporter genes under dichlorvos stress. In addition, gas chromatography-mass spectrometry (GC-TOF-MS), and liquid chromatography-mass spectrometry (LC-QQQ-MS) data revealed the global primary and secondary metabolic changes that occur in T. asperellum TJ01 under dichlorvos stress. CONCLUSIONS: The tolerance mechanism of T. asperellum TJ01 to dichlorvos was proposed. In addition, the absorption and residue of dichlorvos were analyzed, laying the foundation for elucidation of the mechanism by which T. asperellum TJ01 degrades pesticide residues.

Inhibition effects of pesticides on glutathione-S-transferase enzyme activity of Van Lake fish liver.

Glutathione-S-transferases (GSTs) have a function in xenobiotic metabolism. They are a significant multifunctional family with a wide variety of catalytic activities. In the current study, we determined in vitro inhibition effects of 2,4-dichlorophenoxyacetic acid dimethylamine salt (2,4-D DMA), haloxyfop-P-methyl, glyphosate isopropylamine, dichlorvos, and λ-cyhalothrin on purified GST. For this purpose, GST were purified from Van Lake fish (Chalcalburnus tarichii Pallas) liver with 29.25 EU mg-1 specific activity and 10.76% yield using GSH-agarose affinity chromatographic method. The pesticides were tested at various concentrations on in vitro GST activity. Ki constants were calculated as 0.17 ± 0.01, 0.25 ± 0.05, 3.72 ± 0.32, 0.42 ± 0.06, and 0.025 ± 0.004 mM, for 2,4-D DMA, haloxyfop-P-methyl, glyphosate isopropylamine, dichlorvos, and λ-cyhalothrin, respectively. λ-Cyhalothrin showed a better inhibitory effect compared to the other pesticides. The inhibition mechanisms of λ-cyhalothrin were competitive, while the other pesticides were noncompetitive.

Genetically engineered zebrafish liver (ZF-L) cells as an in vitro source for zebrafish acetylcholinesterase (zfAChE) for the use in AChE inhibition assays.

Zebrafish acetylcholinesterase (zfAChE) preparations employed for the evaluation of acetylcholinesterase inhibition are usually extracted from animal tissues, a procedure suffering from both technical and ethical limitations, which may be alleviated using an in vitro expression system for enzyme generation. For this end, a protocol for stable transfection and selection of zebrafish liver (ZF-L) cells using an adapted expression plasmid "ZF-L Exp" was developed. After insertion of zfAChE cDNA, the enzyme was efficiently expressed in transgenic ZF-L cell lines, which were then used as a high yield source of zfAChE activity for acetylcholinesterase (AChE) inhibition assays. An adapted assay protocol was used to demonstrate the effects of carbaryl, dichlorvos and caffeine as model AChE inhibitors towards zfAChE. Dimethyl sulfoxide (DMSO) was also strongly inhibitory towards zfAChE. Finally, we provide data on the stability of zfAChE enzyme preparations. The novel test system provides a promising in vitro test system for the assessment of zfAChE inhibition.

Cytotoxicity testing of aqueous extract of bitter leaf (Vernonia amygdalina Del) and sniper 1000EC (2,3 dichlorovinyl dimethyl phosphate) using the Alium cepa test.

BACKGROUND: The unrefined nature of the herbal preparations from Vernonia amygdalina (VA) and toxicity potentials of Sniper may both have severe consequences on the biochemical and genetic systems. OBJECTIVES: To assess the microscopic and macroscopic effects of these substances. METHODS: VA leafs and Sniper were prepared and dissolved in distilled water to give different concentrations. Series of baseline tests were carried out to establish concentration range for root growth. Series of twelve onion bulbs of three per series was prepared, with a series of three onion bulbs serving as control. Chromosomal aberrations were statistically analysed using chi- squared test. Root bundle mean length was obtained after 96 hours and EC50 values at 95% confidence interval was determined from a plot of root length against sample concentrations using Microsoft Excel software. RESULTS: Total cytotoxic effect was induced by 2% sniper and 70% VA. EC50 for VA and sniper were 33.07 and 0.346 respectively. The two substances induced chromosomal aberrations and the effect was concentration dependent. CONCLUSION: There are risks of these widely used substances for therapeutic and environmental purposes.

Deltamethrin-Incorporated Nets as an Integrated Pest Management Tool for the Invasive Halyomorpha halys (Hemiptera: Pentatomidae).

Long-lasting insecticide nets (LLINs), which have insecticide incorporated within the fibers, have been widely used for control of malaria and other insect-vectored diseases. Only recently have researchers begun exploring their use for control of agricultural pests. In this study, we evaluated the toxicity of a deltamethrin-incorporated LLIN, ZeroFly (Vestergaard-Frandsen, Washington, DC) for control of the brown marmorated stink bug, Halyomorpha halys (Stål). In the lab, exposure to the ZeroFly net for 10 s resulted in >90% mortality of H. halys nymphs and >40% mortality of H. halys adults. Longer exposure to the net resulted in higher mortality. In another experiment, a 15-cm2 sheet of ZeroFly net placed inside of the stink bug trap provided long-lasting kill of H. halys adults equal to or better than standard dichlorvos kill strip. Potential for the use of ZeroFly nets for H. halys IPM is discussed.

Cell cycle activation in p21 dependent pathway: An alternative mechanism of organophosphate induced dopaminergic neurodegeneration.

In the previous study, we demonstrated that dichlorvos induces oxidative stress in dopaminergic neuronal cells and subsequent caspase activation mediates apoptosis. In the present study, we evaluated the effect and mechanism of dichlorvos induced oxidative stress on cell cycle activation in NGF-differentiated PC12 cells. Dichlorvos exposure resulted in oxidative DNA damage along with activation of cell cycle machinery in differentiated PC12 cells. Dichlorvos exposed cells exhibited an increased expression of p53, cyclin-D1, pRb and decreased expression of p21suggesting a re-entry of differentiated cells into the cell cycle. Cell cycle analysis of dichlorvos exposed cells revealed a reduction of cells in the G0/G1 phase of the cell cycle (25%), and a concomitant increase of cells in S phase (30%) and G2/M phase (43.3%) compared to control PC12 cells. Further, immunoblotting of cytochrome c, Bax, Bcl-2 and cleaved caspase-3 revealed that dichlorvos induces a caspase-dependent cell death in PC12 cells. These results suggest that Dichlorvos exposure has the potential to generate oxidative stress which evokes activation of cell cycle machinery leading to apoptotic cell death via cytochrome c release from mitochondria and subsequent caspase-3 activation in differentiated PC12 cells.

In vitro test systems supporting the development of improved pest control methods: a case study with chemical mixtures and bivalve biofoulers.

Using the bivalve macrofouler Corbicula fluminea, the suitability of in vitro testing as a stepping stone towards the improvement of control methods based on chemical mixtures was addressed in this study. In vitro cholinesterase (ChE) activity inhibition following single exposure of C. fluminea tissue to four model chemicals (the organophosphates dimethoate and dichlorvos, copper and sodium dodecyl phosphate [SDS]) was first assessed. Consequently, mixtures of dimethoate with copper and dichlorvos with SDS were tested and modelled; mixtures with ChE revealed synergistic interactions for both chemical pairs. These synergic combinations were subsequently validated in vivo and the increased control potential of these selected combinations was verified, with gains of up to 50% in C. fluminea mortality relative to corresponding single chemical treatments. Such consistency supports the suitability of using time- and cost-effective surrogate testing platforms to assist the development of biofouling control strategies incorporating mixtures.

Behavior of field-applied triadimefon, malathion, dichlorvos, and their main metabolites during barley storage and beer processing.

The behavior and fate of 3 pesticides (triadimefon, malathion, and dichlorvos) and the main metabolites (triadimenol and malaoxon) during barley storage or beer processing were assessed using a pilot-plant equipment. The residues of all products were determined using liquid chromatography coupled with tandem mass spectrometry. Field investigation of the dissipation rate kinetics for triadimefon and malathion during storage indicated that their half-life was twice as high when 5 times the recommended dosage was used. Milling had little effect on the removal of dichlorvos and malathion residues, whereas these were substantially removed when the spent grains were filtered after mashing. The calculated processing factors were all <1, indicating the residual ratios of dichlorvos and malathion were reduced during the entire process. In conclusion, storage and processing considerably reduced pesticide residue levels in barley and beer; however, greater focus needs to be paid to the toxicity of their metabolites in commercial by-products.

To Catch a Fly: Landing and Capture of Ceratitis capitata in a Jackson Trap with and without an Insecticide.

Attractant-based traps are a cornerstone of detection, delimitation and eradication programs for pests such as tephritid fruit flies. The ideal trap and lure combination has high attraction (it brings insects to the trap from a distance) and high capture efficiency (it has a high probability of capturing the insect once it arrives at the trap). We examined the effect of an insecticide (DDVP) in combination with a pheromone lure (trimedlure) on capture of Ceratitis capitata using 1) digital images of surfaces of a Jackson trap analyzed via computer vision, and 2) counts of the number of flies caught in the trap and in the area under the trap. Our results indicate no significant difference in trap capture without or with insecticide (means ± SD = 324 ±135 and 356 ±108, respectively). However, significantly more dead flies were found around the trap with insecticide (92 ±53 with insecticide compared with 35 ±22 without), suggesting a possible decrease in trap efficiency due to mortality before insects enter the trap. Indeed, the average number of flies detected on all surfaces of the traps with insecticide was lower than that for lure-only (4.15±0.39 vs 8.30±1.18), and both were higher than control (no lure: 0.76 ±0.08). We found that the majority of fly sightings, 71% of the total, occurred on the inside panels of the lure-only traps, suggesting that increased efficiency of the Jackson trap may be obtained by adding a contact insecticide to those surfaces.

Weathering and Chemical Degradation of Methyl Eugenol and Raspberry Ketone Solid Dispensers for Detection, Monitoring, and Male Annihilation of Bactrocera dorsalis and Bactrocera cucurbitae (Diptera: Tephritidae) in Hawaii.

Solid male lure dispensers containing methyl eugenol (ME) and raspberry ketone (RK), or mixtures of the lures (ME + RK), and dimethyl dichloro-vinyl phosphate (DDVP) were evaluated in area-wide pest management bucket or Jackson traps in commercial papaya (Carica papaya L.) orchards where both oriental fruit fly, Bactrocera dorsalis (Hendel), and melon fly, Bactrocera cucurbitae (Coquillett), are pests. Captures of B. dorsalis with fresh wafers in Jackson and bucket traps were significantly higher on the basis of ME concentration (Mallet ME [56%] > Mallet MR [31.2%] > Mallet MC [23.1%]). Captures of B. cucurbitae with fresh wafers in Jackson and bucket traps were not different regardless of concentration of RK (Mallet BR [20.1%] = Mallet MR [18.3%] = Mallet MC [15.9%]). Captures of B. dorsalis with fresh wafers, compared with weathered wafers, were significantly different after week 12; captures of B. cucurbitae were not significantly different after 16 wk. Chemical analyses revealed presence of RK in dispensers in constant amounts throughout the 16-wk trial. Degradation of both ME and DDVP over time was predicted with a high level of confidence by nonlinear asymptotic exponential decay curves. Results provide supportive data to deploy solid ME and RK wafers (with DDVP) in fruit fly traps for detection programs, as is the current practice with solid TML dispensers placed in Jackson traps. Wafers with ME and RK might be used in place of two separate traps for detection of both ME and RK responding fruit flies and could potentially reduce cost of materials and labor by 50%.

Hex1-related transcriptome of Trichoderma atroviride reveals expression patterns of ABC transporters associated with tolerance to dichlorvos.

OBJECTIVES: The tolerance to organophosphate pesticide, dichlorvos, is essential for the application of Trichoderma in bioremediation and integrated pest management, although the molecular events associated with the tolerance process have not yet been elucidated. RESULTS: RNA-seq analysis of wild-type Trichoderma atroviride T23 and the hex1-deleted mutant under dichlorvos stress was designed to search for genes involved in the tolerance process. A total of 5382 differentially expressed genes were identified, highlighting the complex transcriptional changes of T. atroviride in response to dichlorvos stress. 137 genes were regulated by dichlorvos and hex1, encoding major facilitator superfamilies, cytochrome P450, glutathione-S-transferase, flavoprotein, Hsp70, Hsp90, etc. Pathway and expression analysis indicated that ABC transporters were affected by the disruption of hex1 gene and might play a vital role in the tolerance process. Expression patterns of seven selected ABC transporter genes were confirmed by qRT-PCR after exposure to dichlorvos for 2, 6 and 24 h. CONCLUSIONS: The present study provides insights into the genetic basis of dichlorvos tolerance in Trichoderma that may be exploited for further development of bioremediation or biocontrol agents.