Integument CYP genes of the largest genome-wide cytochrome P450 expansions in triatomines participate in detoxification in deltamethrin-resistant Triatoma infestans.

Insect resistance to chemical insecticides is attributed to a combination of different mechanisms, such as metabolic resistance, knockdown resistance, and the cuticular resistance or penetration factor. The insect integument offers an efficient barrier against contact insecticides and its role as penetration factor has been previously reported; however, there is no information about its potential function in the metabolic resistance. Cytochrome P450 genes (CYP) are highly expressed in the fat body of several insects and thus play a key role in their metabolic resistance. Here, we describe new members that belong to the highly genome-wide expanded CYP3093A and CYP4EM subfamilies in the Chagas disease vectors Rhodnius prolixus and Triatoma infestans. We modeled the docking of deltamethrin in their active site and detected differences in some amino acids between both species that are critical for a correct interaction with the substrate. We also knocked down the two constitutively most expressed genes in the integument of resistant T. infestans nymphs (CYP3093A11 and CYP4EM10) in order to find clues on their participation in deltamethrin resistance. This is the first report on the role of the insect integument in detoxification events; although these two CYP genes do not fully explain the resistance observed in T. infestans.

Residue of insecticides in foodstuff and dietary exposure assessment of Brazilian citizens.

The goal of this work is to study the presence of 17 different pyrethroids and chlorpyrifos in animal origin food samples, including chicken, beef, fish, eggs and milk. The samples were analyzed by GC/NCI-MS/MS in order to determine their insecticide concentration levels, the relation between the amount of insecticides and the lipid content, as well as their isomeric composition. Bifenthrin, cypermethrin, cyhalothrin, permethrin, deltamethrin and chlorpyrifos have been detected in real samples. The highest levels find were verified for chlorpyrifos (45.7 µg L-1, 17.5 µg kg-1 ww) and cypermethrin (2.75 µg L-1, 14.7 µg kg-1 ww) in milk and beef, respectively. The estimate of daily intake showed that none of these insecticides exceeded the safety limits of the acceptable daily intake values. A strong correlation between the amount of pesticides and lipid content was found for beef, fish and chicken samples. Regarding the isomers, a predominance of cis isomers in samples contaminated with cypermethrin was verified.

Breast milk intake and mother to infant pesticide transfer measured by deuterium oxide dilution in agricultural and urban areas of Mexico.

Vector-borne diseases have increased pesticide use in urban areas (UA) and agricultural areas (AA) in Mexico. Breast milk can be contaminated by pesticide exposure. The objective of the study was to measure breast milk intake by deuterium oxide dilution as well as organochlorine and pyrethroid transfer from mother to infant in AA and UA of Sonora, Mexico. Human milk intake was determined by the 'dose-to-mother' technique using deuterium oxide (D2O) dilution. Mothers' body composition was also assessed by this technique and the intercept method. Pyrethroids (deltamethrin, cypermethrin and cyhalothrin) and organochlorine pesticide residues (p,p'- DDT, p,p'- DDE, p,p'- DDD) in breast milk samples were measured by gas chromatography. Sixty-two lactating women and their infants participated in the study, 32 lived in the UA and 30 lived in the AA. Breast milk intake was approximately 100 mL higher in the AA than in the UA 799 ± 193 and 707 ± 201 mL/day, respectively (p < 0.05). The concentrations of p,p'- DDT and cypermethrin levels in breast milk were higher in the UA than in the AA (p < 0.05 and p = 0.001, respectively). None of the pyrethroids and organochlorine pesticides studied surpassed the Acceptable Daily Intake (ADI) in milk for humans according to EPA and FAO/WHO. In conclusion, breast milk intake was higher in the AA compared to the UA. The p,p'- DDT and cypermethrin levels in breast milk were higher in the UA compared to the AA. Since pesticide levels in human milk did not exceed the ADI, breastfeeding is still a safe practice and should be encouraged.

Toxic heritage: Maternal transfer of pyrethroid insecticides and sunscreen agents in dolphins from Brazil.

Pyrethroids (PYR) and UV filters (UVF) were investigated in tissues of paired mother-fetus dolphins from Brazilian coast in order to investigate the possibility of maternal transfer of these emerging contaminants. Comparison of PYR and UVF concentrations in maternal and fetal blubber revealed Franciscana transferred efficiently both contaminants to fetuses (F/M > 1) and Guiana dolphin transferred efficiently PYR to fetuses (F/M > 1) different than UVF (F/M < 1). PYR and UVF concentrations in fetuses were the highest-ever reported in biota (up to 6640 and 11,530 ng/g lw, respectively). Muscle was the organ with the highest PYR and UVF concentrations (p < 0.001), suggesting that these two classes of emerging contaminants may have more affinity for proteins than for lipids. The high PYR and UVF concentrations found in fetuses demonstrate these compounds are efficiently transferred through placenta. This study is the first to report maternal transfer of pyrethroids and UV filters in marine mammals.

Is cumulated pyrethroid exposure associated with prediabetes? A cross-sectional study.

Pyrethroids are a class of insecticides used widely for vector control programs. Acute pyrethroid poisoning is rare, but well documented, whereas effects of cumulative exposure are insufficiently described, including possible negative effect on glucose regulation. The objective of this study was to investigate an association between exposure to pyrethroids and abnormal glucose regulation (prediabetes or diabetes). A cross-sectional study was performed among 116 pesticide sprayers from public vector control programs in Bolivia and 92 nonexposed controls. Pesticide exposure (duration, intensity, cumulative exposure) was assessed from questionnaire data. Participants were asked about symptoms of diabetes. Blood samples were analyzed for glycosylated hemoglobin (HbA1c), a measure of glucose regulation. No association was found between pyrethroid exposure and diabetes symptoms. The prevalence of abnormal glucose regulation (defined as HbA1c ≥ 5.6%) was 61.1% among sprayers and 7.9% among nonexposed controls, corresponding to an adjusted odds ratio (OR [95% confidence interval]) for all sprayers of 11.8 [4.2-33.2] and 18.5 [5.5-62.5] for pyrethroid-exposed only. Among sprayers who had only used pyrethroids, a significant positive trend was observed between cumulative pesticide exposure (total number of hours sprayed) and adjusted OR of abnormal glucose regulation, with OR 14.7 [0.9-235] in the third exposure quintile. The study found a severely increased prevalence of prediabetes among Bolivian pesticide sprayers compared with a control group, but the relevance of the control group is critical. Within the spraying group, an association between cumulative exposure to pyrethroids and abnormal glucose regulation was seen. Further studies are needed to confirm this association.

[Determination of insecticide-resistance and resistance mechanisms of Blattella germanica (Dictyoptera: Blattellidae)]. / Determinación de la resistencia a insecticidas y mecanismos de resistencia en cepas de Blattella germanica (Dictyoptera: Blattellidae).

In this paper, the level of resistance to four insecticides of 3 Blatella germanica strains collected from various places in the City of Havana province was evaluated. These strains were resistant to two pyrethroids (cypermethrin and lambda-cyalothrine) and to organophosphorate malathion but susceptible to carbamate propoxur. The values of alpha and beta esterases, acetylcholinesterase and gluthatione-S-transferase were estimated in three strains involved in the study. The results of the study showed high esterase activity in all the strains, mainly beta esterases and two of the three strains presented with high gluthation-S-transferase enzyme. No changes in acetylcholinesterase were demonstrated in relation to the reference strain. The association of levels of resistance to insecticides, the possible resistance mechanisms in each strain and the results of the enzymatic activity were also analyzed.

[Determination in vivo of the role of esterase and glutathione transferase enzymes in pyrethroid resistance of Aedes aegypti (Diptera: Culicidae)]. / Determinación in vivo del papel de las enzimas esterasas y glutation transferasa en la resistencia a piretroides en Aedes aegypti (Diptera: Culicidae).

An in vivo study of two synergists, that is, Triphenil phosphate -specific esterase inhibitor- and ethacrynic acid -specific gluthation transferase inhibitor- was performed to determine if these enzymes were responsible for pyrethroid resistance of Aedes aegypti. To this end, two insecticide resistant Aedes aegypti strains were used, one strain selected with temephos by six selection generations (SAN-F6) and the other strain with delmamethrin by 12 selection generations (SAN-F12), being both strains resistant to pyrethroid insecticices. Through the use of TPP and EA synergists, it was proved that esterase and gluthation-s-transferase (GST) enzymes were responsible for pryrethroid resistance of these strains. These results showed the existence of cross-resistance and multidrug resistance, which should be taken into account for insecticide use strategies aimed at vector control.

Role of enhanced detoxication in a deltamethrin-resistant population of Triatoma infestans (Hemiptera, Reduviidae) from Argentina.

Deltamethrin and other pyrethroids have been extensively used in Argentina since 1980, for the chemical control of Triatoma infestans Klug (Hemiptera: Reduviidae). Recently, resistance to deltamethrin was detected in field populations by the survival of bugs exposed by topical application to the diagnostic dose estimated on the CIPEIN susceptible strain. Results of the current study showed low resistant ratios (RRs) to deltamethrin for the resistant populations (RR ranged from 2.0 for San Luis colony to 7.9 for Salta colony). Biochemical studies were made on the most resistant colony (Salta) and the susceptible strain (CIPEIN), in order to establish the importance of degradative mechanisms as a cause of the detected resistance. Esterase activity was measured on 3 days old first instars through phenylthioacetate and a-naphtyl acetate activities. The results showed a significant difference in no cholinesterase esterase activity from susceptible (7.6 +/- 0,7 micro M S./i.min.) and Salta resistant colony (9.5 +/- 0.8 microM S./i.min.). Cytochrome p450 mono-oxygenase (p450) activity was measured on individual insects through ethoxycoumarine deethylase (ECOD) activity using a fluorescence microplate reader. The dependence of ECOD activity on age and body region of the nymphs, and pH and time of incubation were studied in order to optimize the measurement. As a result, comparative studies were performed on abdomens of 2 days old first instars at pH 7.2 and 4 h incubation time. ECOD activity of first nymphs was significantly lower in the susceptible colony (61.3 +/- 9.08 pg ECOD/ insect) than in the resistant one (108.1+/- 5.7 pg ECOD/ insect). These results suggest that degradative esterases (no-cholinesterase) and mono-oxygenases cytochrome p450, play an important role in the resistance to deltamethrin in Salta colony from Argentina.

Vitamin C protects against in vitro cytotoxicity of cypermethrin in rat hepatocytes.

The objective of this study was to investigate the effect of ascorbic acid (AA) on the in vitro cytotoxicity of cypermethrin (CM), and on glutathione (GSH) metabolism in rat hepatocytes. In vitro cell viability, lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) leakage were measured, as indicators of hepatic damage, at 1, 15 and 30 min of exposure to CM. Glutathione and the activities of glutathione-S-transferase (GST) and gamma glutamyl transpeptidase (gamma-GT) were also measured. CM hepatotoxicity increased in a time and dose-dependent manner. In the presence of 30 microM CM, ALT and AST also increased 49 and 130% (P < 0.05), respectively, indicating metabolic hepatocyte damage. AA (1 mM) was capable to preserve 100% of cell integrity and modulated ALT and AST. Furthermore, CM induced a 27% reduction in the endogenous antioxidant GSH, and increased 203% GST and 283% gamma-GT (P < 0.05), indicating an oxidative insult. The presence of AA showed chemopreventive capacity against CM, recovering 60% of GSH and a 54% decrease in gamma-GT activity. These results suggest that AA in a 1:33 (CM:AA) ratio can modulate up to 90% of the damage caused to the cells by CM. It also demonstrates that AA can act as a primary antioxidant and hepatoprotector in rat hepatocytes.

Pyrethroid residues in milk and blood of dairy cows following single topical applications.

The presence of the pyrethroid insecticides flumethrin, deltamethrin, cypermethrin and cyhalothrin in milk and blood of 10 cows was determined after single dermal applications of recommended doses. Milk and blood samples were collected every 7 days over a 35-d period and analyzed by high-performance liquid chromatography. The highest residues in milk were found on day 28 for flumethrin and day 1 for deltamethrin, cypermethrin and cyhalothrin, while in blood the highest concentrations were present on day 28 for flumethrin and deltamethrin, the first day for cypermethrin, and day 14 for cyhalothrin.