Efficient in vivo and in silico assessments of antiandrogenic potential in zebrafish.

This study aimed to establish zebrafish-based in vivo and in silico assay systems to evaluate the antiandrogenic potential of environmental chemicals. Zebrafish embryos were exposed to 17α-methyltestosterone (TES) alone or coexposed to TES and representative antiandrogens including flutamide, p,p'-DDE, vinclozolin, fenitrothion, and linuron. We assessed the transcript expression of the androgen-responsive gene sulfotransferase family 2, cytosolic sulfotransferase 3 (sult2st3). The expression of sult2st3 was significantly induced by TES in the later stages of embryonic development. However, the TES-induced expression of sult2st3 was inhibited by flutamide in a concentration-dependent manner (IC50: 5.7 µM), suggesting that the androgen receptor (AR) plays a role in sult2st3 induction. Similarly, p,p'-DDE, vinclozolin, and linuron repressed the TES-induced expression of sult2st3 (IC50s: 0.35, 3.9, and 52 µM, respectively). At the highest concentration tested (100 µM), fenitrothion also suppressed sult2st3 expression almost completely. Notably, p,p'-DDE and linuron did not inhibit sult2st3 induction due to higher concentrations of TES; instead, they potentiated TES-induced sult2st3 expression. Fenitrothion and linuron, which had relatively low antiandrogenic potentials in terms of sult2st3 inhibition, induced broader toxicities in zebrafish embryos; thus, the relationship between developmental toxicities and antiandrogenic potency was unclear. Additionally, an in silico docking simulation showed that all five chemicals interact with the zebrafish AR at relatively low interaction energies and with Arg702 as a key amino acid in ligand binding. Our findings suggest that a combination of zebrafish-based in vivo and in silico assessments represents a promising tool to assess the antiandrogenic potentials of environmental chemicals.

Resveratrol alleviates cardiac apoptosis following exposure to fenitrothion by modulating the sirtuin1/c-Jun N-terminal kinases/p53 pathway through pro-oxidant and inflammatory response improvements: In vivo and in silico studies.

Fenitrothion (FNT), a commonly used organophosphate, can cause oxidative damage and apoptosis on various organs. However, the underlying mechanisms for FNT-induced cardiotoxicity did not formally report. Here, we have evaluated the possible ameliorative roles of resveratrol (RSV) against FNT-induced cardiac apoptosis in male rats through the sirtuin1 (SIRT1)/c-Jun N-terminal kinase (c-JNK)/p53 pathway concerning pro-oxidant and inflammatory cytokines. Forty-eight male rats were equally grouped into control, RSV (20 mg/kg), 5-FNT (5 mg/kg), 10-FNT (10 mg/kg), 20-FNT (20 mg/kg), 5-FNT-RSV, 10-FNT-RSV, and 20-FNT-RSV where all doses administrated by gavage for four weeks. The present findings demonstrated that RSV markedly diminished the level of hyperlipidemia and elevation in lactate dehydrogenase (LDH), total creatine kinase (CK-T), and troponin T (TnT) levels following FNT intoxication. Furthermore, RSV significantly reduced FNT-induced cardiac oxidative injury by reducing malondialdehyde (MDA) level and improving the levels of glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), and acetylcholinesterase (AchE). Also, the levels of interleukin-1ß (IL1ß,), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were significantly attenuated in the co-treated groups. Moreover, RSV alleviated the histopathological changes promoted by FNT and repaired the transcript levels of SIRT1, c-JNK, and caspase-9/3 along with p53 immunoreactivity. In silico study revealed that the free binding energies of RSV complexes with protein and DNA sequences of SIRT1 were lower than docked complexes of FNT. Therefore, RSV reserved myocardial injury-induced apoptosis following exposure to FNT by modulating the SIRT1/c-JNK/p53 pathway through cellular redox status and inflammatory response improvements.

Infection dynamics of insecticide-degrading symbionts from soil to insects in response to insecticide spraying.

Insecticide resistance is a serious concern in modern agriculture, and an understanding of the underlying evolutionary processes is pivotal to prevent the problem. The bean bug Riptortus pedestris, a notorious pest of leguminous crops, acquires a specific Burkholderia symbiont from the environment every generation, and harbors the symbiont in the midgut crypts. The symbiont's natural role is to promote insect development but the insect host can also obtain resistance against the insecticide fenitrothion (MEP) by acquiring MEP-degrading Burkholderia from the environment. To understand the developing process of the symbiont-mediated MEP resistance in response to the application of the insecticide, we investigated here in parallel the soil bacterial dynamics and the infected gut symbionts under different MEP-spraying conditions by culture-dependent and culture-independent analyses, in conjunction with stinkbug rearing experiments. We demonstrate that MEP application did not affect the total bacterial soil population but significantly decreased its diversity while it dramatically increased the proportion of MEP-degrading bacteria, mostly Burkholderia. Moreover, we found that the infection of stinkbug hosts with MEP-degrading Burkholderia is highly specific and efficient, and is established after only a few times of insecticide spraying at least in a field soil with spraying history, suggesting that insecticide resistance could evolve in a pest bug population more quickly than was thought before.

Combined target site (kdr) mutations play a primary role in highly pyrethroid resistant phenotypes of Aedes aegypti from Saudi Arabia.

BACKGROUND: Pyrethroid resistance is a threat to effective vector control of Aedes aegypti, the vector of dengue, Zika and other arboviruses, but there are many major knowledge gaps on the mechanisms of resistance. In Jeddah and Makkah, the principal dengue-endemic areas of Saudi Arabia, pyrethroids are used widely for Ae. aegypti control but information about resistance remains sparse, and the underlying genetic basis is unknown. Findings from an ongoing study in this internationally significant area are reported here. METHODS: Aedes aegypti collected from each city were raised to adults and assayed for resistance to permethrin, deltamethrin (with and without the synergist piperonyl butoxide, PBO), fenitrothion, and bendiocarb. Two fragments of the voltage-gated sodium channel (Vgsc), encompassing four previously identified mutation sites, were sequenced and subsequently genotyped to determine associations with resistance. Expression of five candidate genes (CYP9J10, CYP9J28, CYP9J32, CYP9M6, ABCB4) previously associated with pyrethroid resistance was compared between assay survivors and controls. RESULTS: Jeddah and Makkah populations exhibited resistance to multiple insecticides and a similarly high prevalence of resistance to deltamethrin compared to a resistant Cayman strain, with a significant influence of age and exposure duration on survival. PBO pre-exposure increased pyrethroid mortality significantly in the Jeddah, but not the Makkah strain. Three potentially interacting Vgsc mutations were detected: V1016G and S989P were in perfect linkage disequilibrium in each strain and strongly predicted survival, especially in the Makkah strain, but were in negative linkage disequilibrium with 1534C, though some females with the Vgsc triple mutation were detected. The candidate gene CYP9J28 was significantly over-expressed in Jeddah compared to two susceptible reference strains, but none of the candidate genes was consistently up-regulated to a significant level in the Makkah strain. CONCLUSIONS: Despite their proximity, Makkah and Jeddah exhibit significant differences in pyrethroid resistance phenotypes, with some evidence to suggest a different balance of mechanisms, for example with more impact associated with CYP450s in the Jeddah strain, and the dual kdr mutations 989P and 1016G in the more resistant Makkah strain. The results overall demonstrate a major role for paired target site mutations in pyrethroid resistance and highlight their utility for diagnostic monitoring.

EFFICACY OF THREE INSECTICIDES ON RAT FLEA (XENOPSYLLA CHEOPIS) INFESTING RODENTS IN GIZA GOVERNORATE, EGYPT.

The extensive use of insecticides in public health and agriculture sectors is the main reason for development of resistance in fleas associated in domestic rodents. The present work was planned to investigate the insecticidal efficacy of Lambda-cyhalothrin, Chlorpyrifos and Fenitrothion against rat flea (Xenopsylla cheopis) infesting rodent species in Giza Governorate, Egypt. The lethal concentration Lc50 and Lc90 of population percent were obtained from the established regression log concentrate-response lines. Data indicated that the values of lethal concentration (Lc50) were 0.293, 1.725 & 2.328% for Lambda-cyhalothrin, chlorpyrifos and Fenitrothion, respectively. The values of lethal concentration (Lc90) were 0.467, 2.839 & 5.197% for Lambda-cyhalothrin, chlorpyrifos and Fenitrothion, respectively.

Unraveling the effect of structurally different classes of insecticide on germination and early plant growth of soybean [Glycine max (L.) Merr].

Although a considerable number of studies about the effect of different insecticides on plant physiology and metabolism have been carried out, research work about the comparative action of structurally different classes of insecticide on physiological and biochemical properties of soybean seed germination and early growth has not been found. The objective of this study was to investigate the effect of different classes of insecticides on soybean seed germination and early plant growth. Soybean seeds of Bosuk cultivar were soaked for 24h in distilled water or recommended dose (2mLL(-1), 1mLL(-1), 0.5gL(-1), and 0.5gL(-1) water for insecticides Mepthion, Myungtaja, Actara, and Stonate, respectively) of pesticide solutions of four structurally different classes of insecticides - Mepthion (fenitrothion; organophosphate), Myungtaja (etofenprox; pyrethroid), Actara (thiamethoxam; neonicotinoid), and Stonate (lambda-cyhalothrin cum thiamethoxam; pyrethroid cum neonicotinoid) - which are used for controlling stink bugs in soybean crop. Insecticides containing thiamethoxam and lamda-cyhalothrin cum thiamethoxam showed positive effects on seedling biomass and content of polyphenol and flavonoid, however fenitrothion insecticide reduced the seed germination, seed and seedling vigor, and polyphenol and flavonoid contents in soybean. Results of this study reveal that different classes of insecticide have differential influence on physiologic and metabolic actions like germination, early growth, and antioxidant activities of soybean and this implies that yield and nutrient content also might be affected with the application of different types of insecticide.

Evaluation of Sumithion L-40 against Aedes aegypti (L.) and Aedes albopictus Skuse.

Space spraying of chemical insecticides is still an important mean of controlling Aedes mosquitoes and dengue transmission. For this purpose, the bioefficacy of space-sprayed chemical insecticide should be evaluated from time to time. A simulation field trial was conducted outdoor in an open field and indoor in unoccupied flat units in Kuala Lumpur, to evaluate the adulticidal and larvicidal effects of Sumithion L-40, a ULV formulation of fenitrothion. A thermal fogger with a discharge rate of 240 ml/min was used to disperse Sumithion L-40 at 3 different dosages (350 ml/ha, 500 ml/ha, 750 ml/ha) against lab-bred larvae and adult female Aedes aegypti and Aedes albopictus. An average of more than 80% adult mortality was achieved for outdoor space spray, and 100% adult mortality for indoor space spray, in all tested dosages. Outdoor larvicidal effect was noted up to 14 days and 7 days at a dosage of 500 and 750 ml/ha for Ae. aegypti and Ae. albopictus, respectively. Indoor larvicidal effect was up to 21 days (500 ml/ha) and 14 days (750 ml/ha), respectively, after spraying with larval mortality > 50% against Ae. aegypti. This study concluded that the effective dosage of Sumithion L-40 thermally applied against adult Ae. aegypti and Ae. albopictus indoor and outdoor is 500 and 750 ml/ha. Based on these dosages, effective indoor spray volume is 0.4 - 0.6 ml/m³. Additional indoor and outdoor larvicidal effect will be observed at these application dosages, in addition to adult mortality.

Status of organophosphate and carbamate resistance in Anopheles gambiae sensu lato from the south and north Benin, West Africa.

BACKGROUND: With the rapid spread of pyrethroid resistance in the main malaria vectors from Benin and the various resistance mechanisms involved (metabolic resistance and knock-down resistance (kdr), it is important to foresee effective resistance management strategies. Thus, the knowledge of the insensitive acetylcholinesterase (ace-1R) effects on phenotypes of An. gambiae will help us to strengthen basic and operational research on the development of strategies that will use organophosphates or carbamates as alternatives against pyrethroids-resistant malaria vectors in the field. METHODS: Larvae and pupae of Anopheles gambiae s.l. mosquitoes were collected from the breeding sites in Ouemé , Atacora, and Alibori departments. CDC susceptibility tests were conducted on unfed female mosquitoes aged 2-5 days old. CDC bioassays were performed with stock solutions of fenitrothion (50 µg per bottle) and bendiocarb (12.5 µg per bottle). PCR techniques were used to detect species and Ace-1 mutations. RESULTS: Anopheles gambiae Seme and Kandi populations were susceptible to fenitrothion whereas Anopheles gambiae Tanguieta and Malanville populations were resistant. An. gambiae populations from Seme, Kandi and Malanville were fully susceptible to bendiocarb whereas those from Tanguieta have developed a strong resistance to the same insecticide. A slight decrease in mortality rate was observed with 97.91% in populations of mosquitoes from Malanville. PCR revealed that all specimens tested were Anopheles gambiae s.s.. CONCLUSION: This study demonstrated the need to monitor organophosphate (OPs) and Carbamates resistance among populations of the An. gambiae s.l. in Benin, to determine its spread and anticipate vector control failure where these insecticides are used. However, further studies are needed to understand the current distribution of the Ace-1R mutation in other localities in the south-north transect Benin.

Swimming velocity, avoidance behavior and biomarkers in Palaemon serratus exposed to fenitrothion.

The aim of this study was to develop two behavioral tests (swimming velocity and avoidance behavior) specific for the common prawn, Palaemon serratus, and to investigate the effects of sublethal concentrations of fenitrothion on behavior and on several biomarkers. In a first bioassay, mortality was investigated in prawns exposed during 96 h to concentrations of fenitrothion ranging from 39 to 40000 ng L(-1). The effects on swimming velocity and biomarkers were determined in prawns exposed to the sub-lethal concentrations of fenitrothion (from 39 to 625 ng L(-1)). A specific newly bioassay was developed to assess the capability of prawns to avoid the toxicant (avoidance test). Finally, in order to determine the effects on biomarkers during the avoidance test, prawns were collected at different times of exposure (30, 60, 90 and 120 min). Results showed that prawns exposed to the sub-lethal concentrations of fenitrothion exhibited a significant inhibition of swimming velocity with a LOEC of 313 ng L(-1). A significant inhibition of both eye AChE (LOEC=78 ng L(-1)) and muscle ChE (LOEC=156 ng L(-1)) activities were also observed. Results from the avoidance test indicated that animals significantly avoided fenitrothion (78 ng L(-1)). However, this capability was not observed in prawns exposed to 156 ng L(-1) fenitrothion. Prawns exposed to fenitrothion showed alterations in enzymes involved in the production of energy (LDH and IDH) possibly in an attempt to cope with additional energetic demands. Impairment of locomotion and avoidance may lead to alterations at the population level. Thus, the present behavioral tests can be useful as ecologically relevant tools for ecological risk assessment.

Monitoring of five different insecticide resistance status in Turkish house fly Musca domestica L. (Diptera: Muscidae) populations and the relationship between resistance and insecticide usage profile.

OBJECTIVE: The objective of this study is to assess the resistance status and yearly changes involved for house fly populations from six cities in Turkey. METHODS: Field strains of house fly (Musca domestica L. Diptera: Muscidae) were collected in 2004-2006 from cow farms (Antalya, Izmir) and garbage dumps (Adana, Ankara, Istanbul, Sanliurfa) in Turkey. The resistance levels of first and two generation offspring were determined against five insecticides (cypermethrin, cyphenothrin, deltamethrin, permethrin and fenitrothion). RESULTS: While the highest resistance level for pyrethroid was determined for Antalya 2005 strain (851.97 Cypermethrin) and lowest resistance level for Sanliurfa 2004 strain (2.06 Permethrin), the highest fenitrothion resistance was found in Sanliurfa 2004 strain (50.37) and lowest fenitrothion resistance was found in Adana 2004 strain (6.45). CONCLUSION: Our results showed that pyrethroid resistance levels were very high and determined a decreasing trend for Antalya and Istanbul strains and an increasing trend for Adana and Sanliurfa strain for all tested pyrethroid insecticides from 2004 to 2006. Although cypermethrin and cyphenothrin resistance showed a decreasing trend, deltamethrin and permethrin showed an increasing trend for the Izmir strain from 2004 to 2006. The same trend was also determined for the Ankara strain except for permethrin. Fenitrothion resistance was determined to be lower than pyrethroids, but these levels were still high. Flies from cow farms were generally more resistant than those from garbage dumps. Our results also revealed the presence of strong selective pressure on the populations.

Symbiont-mediated insecticide resistance.

Development of insecticide resistance has been a serious concern worldwide, whose mechanisms have been attributed to evolutionary changes in pest insect genomes such as alteration of drug target sites, up-regulation of degrading enzymes, and enhancement of drug excretion. Here, we report a previously unknown mechanism of insecticide resistance: Infection with an insecticide-degrading bacterial symbiont immediately establishes insecticide resistance in pest insects. The bean bug Riptortus pedestris and allied stinkbugs harbor mutualistic gut symbiotic bacteria of the genus Burkholderia, which are acquired by nymphal insects from environmental soil every generation. In agricultural fields, fenitrothion-degrading Burkolderia strains are present at very low densities. We demonstrated that the fenitrothion-degrading Burkholderia strains establish a specific and beneficial symbiosis with the stinkbugs and confer a resistance of the host insects against fenitrothion. Experimental applications of fenitrothion to field soils drastically enriched fenitrothion-degrading bacteria from undetectable levels to >80% of total culturable bacterial counts in the field soils, and >90% of stinkbugs reared with the enriched soil established symbiosis with fenitrothion-degrading Burkholderia. In a Japanese island where fenitrothion has been constantly applied to sugarcane fields, we identified a stinkbug population wherein the insects live on sugarcane and ≈8% of them host fenitrothion-degrading Burkholderia. Our finding suggests the possibility that the symbiont-mediated insecticide resistance may develop even in the absence of pest insects, quickly establish within a single insect generation, and potentially move around horizontally between different pest insects and other organisms.

Irritability of malaria vector, Anopheles sacharovi to different insecticides in a malaria-prone area.

OBJECTIVE: To determine the susceptibility and irritability level of malaria vector Anopheles sacharovi (An. sachrovi) to different insecticides in a malaria-prone area. METHODS: Susceptibility and irritability levels of field collected strain of An. sacharovi to WHO standard papers of DDT 4%, dieldrin 0.4%, malathion 5%, fenitrothion 1%, permethrin 0.75%, and deltamethrin 0.05% were determined in East Azerbaijan of Iran during reemerging of malaria as described by WHO. RESULTS: Results showed that at the diagnostic dose of insecticides this species exhibited resistance to DDT, tolerant to dieldrin and but somehow susceptible to fenitrothion, malathion, permethrin and deltamethrin. The results of irritability of this species to DDT, lambdacyhalothrin, permethrin cyfluthrin and deltamethrin revealed that DDT had had the most and deltamethrin the least irritancy effect. The average number of take offs/fly/minutes for DDT was 0.8±0.2. The order of irritability for permethrin, lambdacyhalothrin, cyfluthrin and deltamethrin were 0.7±0.2, 0.5±0.2, 0.5±0.3, and 0.2±0.1, respectively. CONCLUSIONS: Results of this study reveals the responsiveness of the main malaria vector to different insecticides. This phenomenon is depending on several factors such as type and background of insecticide used previously, insecticide properties, and physiology of the species. Careful monitoring of insecticide resistance and irritability level of species could provide a clue for appropriate selection of insecticide for malaria control.

Surface contact toxicity and synergism of several insecticides against different stages of the tropical bed bug, Cimex hemipterus (Hemiptera: Cimicidae).

BACKGROUND: Five formulated insecticides (lambda-cyhalothrin at 10 mg m⁻², bifenthrin at 50 mg m⁻², fipronil at 10 mg m⁻², fenitrothion at 50 mg m⁻², imidacloprid at 5 mg m⁻²) and one active ingredient (DDT at 500 mg m⁻²) were evaluated using a surface contact method against early and late instars and adults of two strains of the tropical bed bug, Cimex hemipterus (F.). Synergism of lambda-cyhalothrin and fipronil using piperonyl butoxide (PBO) was also assessed. RESULTS: The order of susceptibility of different stages of bed bugs was as follows: early stage-lambda-cyhalothrin > bifenthrin = imidacloprid > fipronil > fenitrothion > DDT; late stage-lambda-cyhalothrin > bifenthrin > fenitrothion > imidacloprid > fipronil > DDT; adult-lambda-cyhalothrin > imidacloprid > bifenthrin > fenitrothion > fipronil > DDT. The late instars exhibited significantly higher LT50 among the life stages. The addition of PBO to fipronil increased the susceptibility of the insects. CONCLUSIONS: Lambda-cyhalothrin, bifenthrin, fenitrothion and fipronil at the recommended application rates were effective against C. hemipterus. Although imidacloprid demonstrated good initial response against C. hemipterus, the insects showed substantial recovery 72 h post-treatment. The late instars (fourth and fifth instars) should be used as the model for toxicological evaluation.

Bendiocarb, a potential alternative against pyrethroid resistant Anopheles gambiae in Benin, West Africa.

BACKGROUND: Anopheles gambiae, the main malaria vector in Benin has developed high level of resistance to pyrethroid insecticides, which is a serious concern to the future use of long-lasting insecticidal nets (LLIN) and indoor residual spraying (IRS). In this context, one of the pathways available for malaria vector control would be to investigate alternative classes of insecticides with different mode of action than that of pyrethroids. The goal of this study was to evaluate under field conditions the efficacy of a carbamate (bendiocarb) and an organophosphate (fenitrothion) against pyrethroid-resistant An. gambiae s.s. METHODS: Wild populations and females from laboratory colonies of five days old An. gambiae were bio-assayed during this study. Two pyrethroids (deltamethrin and alphacypermethrin), an organophosphate (fenitrothion), a carbamate (bendiocarb) and a mixture of an organophosphate (chlorpyriphos + a pyrethroid deltamethrin) were compared in experimental huts as IRS treatments. Insecticides were applied in the huts using a hand-operated compression sprayer. The deterrency, exophily, blood feeding rate and mortality induced by these insecticides against An. gambiae were compared to the untreated control huts. RESULTS: Deltamethrin, alphacypermethrin and bendiocarb treatment significantly reduced mosquito entry into the huts (p < 0.05) compared to untreated huts. Blood feeding rates in huts treated with fenitrothion and the mixture chlorpyriphos/deltamethrin were reduced from 10.95% respectively to 3.7% and 4.47% three months after treatment and from 10.20% to 4.4% and 2.04% four months after treatment. Exophily rates in huts with deltamethrin, alphacypermethrin and the mixture chlorpyriphos/deltamethrin were significantly higher than in the huts with fenitrothion. Deltamethrin and alphacypermethrin had the lowest mortality rate while fenitrothion killed 100% of An. gambiae (in the first month) and 77.8% (in the fourth month). Bendiocarb and the mixture chlorpyriphos/deltamethrin mortality rates ranged from 97.9 to 100% the first month and 77.7-88% the third month respectively. CONCLUSION: After four months, fenitrothion, bendiocarb and the mixture chlorpyriphos/deltamethrin performed effectively against pyrethroid-resistant Anopheles. These results showed that bendiocarb could be recommended as an effective insecticide for use in IRS operations in Benin, particularly as the mixture chlorpyriphos/deltamethrin does not have WHOPES authorization and complaints were mentioned by the sleepers about the safety and smell of fenitrothion.

Insecticide resistance in Anopheles gambiae: data from the first year of a multi-country study highlight the extent of the problem.

BACKGROUND: Insecticide resistance in malaria vectors is a growing concern in many countries which requires immediate attention because of the limited chemical arsenal available for vector control. The current extent and distribution of this resistance in many parts of the continent is unknown and yet such information is essential for the planning of effective malaria control interventions. METHODS: In 2008, a network was established, with financial support from WHO/TDR, to investigate the extent of insecticide resistance in malaria vectors in five African countries. Here, the results of bioassays on Anopheles gambiae sensu lato from two rounds of monitoring from 12 sentinel sites in three of the partner countries are reported. RESULTS: Resistance is very heterogeneous even over relatively small distances. Furthermore, in some sites, large differences in mortality rates were observed during the course of the malaria transmission season. Using WHO diagnostic doses, all populations from Burkina Faso and Chad and two of the four populations from Sudan were classified as resistant to permethrin and/or deltamethrin. Very high frequencies of DDT resistance were found in urban areas in Burkina Faso and Sudan and in a cotton-growing district in Chad. In areas where both An. gambiae s.s. and Anopheles arabiensis were present, resistance was found in both species, although generally at a higher frequency in An gambiae s.s. Anopheles gambiae s.l. remains largely susceptible to the organophosphate fenitrothion and the carbamate bendiocarb in the majority of the sentinel sites with the exception of two sites in Burkina Faso. In the cotton-growing region of Soumousso in Burkina Faso, the vector population is resistant to all four classes of insecticide available for malaria control. CONCLUSIONS: Possible factors influencing the frequency of resistant individuals observed in the sentinel sites are discussed. The results of this study highlight the importance of standardized longitudinal insecticide resistance monitoring and the urgent need for studies to monitor the impact of this resistance on malaria vector control activities.

Changes in bacterial diversity and community structure following pesticides addition to soil estimated by cultivation technique.

An experiment was conducted under laboratory conditions to investigate the effect of increasing concentrations of fenitrothion (2, 10 and 200 mg a.i./kg soil), diuron (1.5, 7.5 and 150 mg a.i./kg soil) and thiram (3.5, 17.5 and 350 mg a.i./kg soil) on soil respiration, bacterial counts and changes in culturable fraction of soil bacteria. To ascertain these changes, the community structure, bacterial biodiversity and process of colony formation, based on the r/K strategy concept, EP- and CD-indices and the FOR model, respectively, were determined. The results showed that the measured parameters were generally unaffected by the lowest dosages of pesticides, corresponding to the recommended field rates. The highest dosages of fenitrothion and thiram suppressed the peak SIR by 15-70% and 20-80%, respectively, while diuron increased respiration rate by 17-25% during the 28-day experiment. Also, the total numbers of bacteria increased in pesticide-treated soils. However, the reverse effect on day 1 and, in addition, in case of the highest dosages of insecticide on days 14 and 28, was observed. Analysis of the community structure revealed that in all soil treatments bacterial communities were generally dominated by K-strategists. Moreover, differences in the distribution of individual bacteria classes and the gradual domination of bacteria populations belonging to r-strategists during the experiment, as compared to control, was observed. However, on day 1, at the highest pesticide dosages, fast growing bacteria constituted only 1-10% of the total colonies number during 48 h of plate incubation, whereas in remaining samples they reached from 20 to 40% of total cfu. This effect, in case of fenitrothion, lasted till the end of the experiment. At the highest dosages of fenitrothion, diuron and at all dosages of thiram the decrease of biodiversity, as indicated by EP- and CD-indices on day 1, was found. At the next sampling time, no significant retarding or stimulating effect was detected. However, in case of CD values the higher differences were observed. The significant impact of pesticides on the physiological state of soil bacteria was not found. They were generally in dormant state (lambda < 0.5), but immediately after pesticides application, the additional reduction of frequency of bacterial cell proliferation (max. decrease of lambda value to 0.15 for thiram on day 14) and prolonged retardation time of colony appearance (max. increase of t(r) value to 1.39 for fenitrothion on day 1) on agar plates were found.

[Metabolic resistance to organophosphate insecticides in Anopheles aquasalis Curry 1932, Libertador municipality, Sucre State, Venezuela]. / Resistencia metabólica a insecticidas organofosforados en Anopheles aquasalis Curry 1932, municipio Libertador,estado Sucre, Venezuela.

INTRODUCTION: A study of insecticide resistance was undertaken at focal level in the localities Catuaro, Guayana, Platanito and Rio de Agua, Libertador County, Sucre State, Venezuela, a region with malaria transmission, where Anopheles aquasalis is the main vector. OBJECTIVE: Insecticide resistance was assessed in the organophosphate insecticides fenitrothion and pirimiphos methyl, both of which are used in the control of Anopheles aquasalis. MATERIALS AND METHODS: In adult mosquitoes, biological tests were performed and identification of resistance mechanisms in vitro by biochemical tests. RESULTS: Elevated levels of alpha and beta esterases were detected, as well as altered acetylcholinesterase activity. Multifunction oxidase enzymes in populations of Anopheles aquasalis in three of the locations evaluated were also altered; therefore, both enzyme systems may be involved in the expression of resistance to organophosphate insecticides in the study populations. The enzyme activity of glutathione-S-transferase was noted only in Rio de Agua. CONCLUSIONS: A better understanding of the resistance to insecticides was obtained in this species of medical importance. These findings will assist the implementation the practice of insecticide rotation as a strategy within an integrated management program.

[Toxic effect of beta-cipermethrin, deltamethrin and fenitrothion in colonies of Triatoma dimidiata (Latreille, 1811) and Triatoma maculata (Erichson, 1848) (Hemiptera, Reduviidae)]. / Efecto tóxico de beta-cipermetrina, deltametrina y fenitrotión en cepas de Triatoma dimidiata (Latreille, 1811) y Triatoma maculata (Erichson, 1848) (Hemiptera, Reduviidae).

INTRODUCTION: The susceptibility to insecticides of triatomine species must be evaluated because of their involvement in the transmission of the Chagas disease. In each region with Chagas endemicity, evaluation of insecticide response is necessary to predict the success of the control campaigns. OBJECTIVE: The baseline susceptibility was determined for the active principles deltamethrin, beta-cypermethrin and fenitrothion in nymphs of first and fifth instar of Triatoma dimidiata and nymphs of first instar of Triatoma maculata. MATERIALS AND METHODS: The insecticide activity in triatomines was evaluated by the technique of topical application. RESULTS: The values of the LD50 in nymphs of first instar for T. maculata, expressed in nanograms per insect (ng/i), were 0.07, 0.05 and 4.12 for deltamethrin, beta-cypermethrin and fenitrothion respectively. The corresponding LD99 values were 1.08, 0.37 and 17.89 ng/i. In T. dimidiata, the LD50 values were 0.44, 0.46 and 16.45 ng/i; the LD99 values were 2.22, 1.97 and 36.07 ng/i. In nymphs of fifth instar T. dimidiata, the LD50 values were 510.7, 1623.6 and 838.9 ng/i; the LD99 values were 9607.5, 11,717.9 and 1525.0 ng/i, respectively. CONCLUSION: In first instar nymphs of T. dimidiata and T. maculata, the pyrethroid insecticides were more effective; in fifth instar nymphs of T. dimidiata, the effectiveness of the pyrethroids and the organophosphate differed in the LD50 comparison--the nymphs required much higher doses compared with the other triatomines and suggested a low susceptibility. The LD99 for the organophosphate (fenitrothion) was significantly lower and may indicate its greater effectiveness in field. Studies of synergistic effects amonst insecticides are important to clarify the role of biochemical mechanisms that determine tolerance to the pyrethroids. Insecticide tolerance represents a new challenge for control campaigns in the Andean and Central American countries where Chagas disease is endemic.

Inhibition and recovery of maternal and foetal cholinesterase enzymes following fenitrothion administration in CD rats.

The purpose of this study was to characterize tissue esterase activity and blood fenitrothion concentrations in the rat dam and foetus following in-utero exposure to the organophosphate insecticide fenitrothion. Time-mated, 8-week-old rats were gavaged on gestation day 19 with 0, 5, or 25 mg fenitrothion kg-1. Fenitrothion was absorbed rapidly from the gastrointestinal tract, with peak maternal and foetal blood levels observed 0.5-1.0 h after dosing. Fenitrothion concentrations in maternal and foetal blood were virtually identical and demonstrated a non-linear dose-response relationship. Acetylcholinesterase and carboxylesterase activities in maternal liver and blood and in foetal liver and brain decreased within 30-60 min of fenitrothion exposure. Esterase inhibition occurred at a fenitrothion dose (5 mg kg-1) that has not been previously associated with reproductive toxicity, suggesting that esterase inhibition should be considered as the critical effect in risk assessments for this pesticide.

Molluscicidal effect of fenitrothion and anilofos on Lymnaea natalensis and Biomphalaria alexandrina snails and on the free larval stages of Schistosoma mansoni.

Psticides; fenitrothion and anilofos (aniloguard) were testd as molluscicides against Lymnaea natalensis and Biomplhalaria alexandrina. The LC10 & LC90 of fenitrothion was 0.12 & 0.21 ppm for L. nalalensis and 0.17 & .26 ppm for B. alexandrina, respectively. The LC50 & LC90 anilofos was 2.61 & 6.47 ppm for Lymnaea and 3.07 & 8.6 ppm for Biomphalaria. The effect of sublethal concentrations (LC0, LC5 & C10) of Feni-rothion on B. alexandrina growth rate, eggs hatchability and on free larval stages of Schistosonma mansoni (miracidia & cerca-riae) were studied. The results obtained showed that sublethal concentrations of fenitrothion caused reduction in growth rate of B. alexandrina and reduction in the hatchibility of snails eggs. The mortality rates of miracidia and cercariae were elevated by increasing both the concentrations of fenitrothion and the time of exposure. The results showed that fenitrothion was more toxic to the free larval stages of S. mansoni than to their snails. The results showed a significant reduction in total protein of treated snails when compared with controls in haemolymph while there was an increase of protein contents of the tissue. The AlkP enzyme activity was slightly increased in the haemolymph of experimental groups than the control and in the tissues the values were significantly higher when compared with control. ALT enzyme activity in haemolymph of experimental groups was higher than controls while its activity in tissue was lower. AST enzyme activity was higher in haemolymph and tissue of experimental groups than in controls.