Biodiversity of culicid mosquitoes in rural Neka township of Mazandaran province, northern Iran.

BACKGROUND & OBJECTIVES: This paper presents the results of a study on ecological parameters including diversity indices applied on populations of Culicidae mosquitoes (Diptera: Culicidae) from three rural areas (Darvishan, Chalmardi and Khairabad) of Neka township in Mazandaran province, northern Iran. METHODS: Adult and larval stages of mosquitoes were systematically sampled to obtain data required for measuring diversity of mosquitoes in these areas. Alfa and beta indices were analyzed to compare mosquito diversity. A total of 5270 specimens belonging to four genera and 14 different species were collected and identified. RESULTS: Evaluation of dominance structure of species showed that culicid mosquitoes of Neka rangelands include one eudominant species, two dominant species, two subdominant species, two rare species and eight subrare species when Heydemann classification was used. INTERPRETATION & CONCLUSION: Data analysis revealed a higher diversity of mosquitoes in Khairabad with a low degree of interspecific dominance, while Darvishan and Chalmardi recorded the lowest observed diversity and a high degree of interspecific dominance. The cluster analysis based on Jaccard's index indicated the relationship between the categories and indicates that Darvishan and Khairabad are the closest categories in their specific composition.

Insecticide resistance and synergism of three field-collected strains of the German cockroach Blattella germanica (L.) (Dictyoptera: Blattellidae) from hospitals in Kermanshah, Iran.

The development of insecticide resistance in the German cockroach, Blattella germanica (L.) is a serious problem in controlling this medically important household pest. The insecticide resistance status in three hospital-collected strains of the German cockroach using four commonly used insecticides from different classes (permethrin, cypermethrin, bendiocarb and chlorpyrifos) was detected by topical bioassay method and preliminary information on possible involvement of monooxygenases in permethrin resistant strains employing synergist piperonyl butoxide (PBO) was obtained. For each insecticide, four to six concentrations resulting in >0% and <100% mortality were used. Three to six replicates of 10 cockroaches per concentration were conducted. For synergism studies, 100 µg PBO per gram body weight of cockroach as the maximum sublethal dose was administered to the first abdominal segment 1 h before insecticide treatment. The differences between LD50 (µg/g) values were considered statistically significant only when the 95% confidence intervals did not overlap. The resistance ratio and synergism ratio were calculated for each insecticide. All three hospital-collected strains of the German cockroach showed different levels of resistance to permethrin and cypermethrin based on resistance ratios compared with SUS strain. Permethrin and cypermethrin resistance ratios ranged from 11.61 to 17.64 and 11.45 to 26.45 at LD50 levels, respectively. Low to moderate levels of bendiocarb resistance and low level of chlorpyrifos resistance were also observed in the hospital-collected strains under study. The synergist piperonyl butoxide (PBO) significantly enhanced the toxicity of permethrin to all strains with different degrees of synergist ratio, 2.45-, 1.87-, 2.51- and 2.38-fold, suggesting monooxygenase involvement in permethrin resistance.

Laboratory and semi-field evaluation of long-lasting insecticidal nets against leishmaniasis vector, Phlebotomus (Phlebotomus) duboscqi in Kenya.

BACKGROUND & OBJECTIVES: Phlebotomine sandflies are vectors of leishmaniases and other diseases. Long-lasting insecticidal nets (LLINs) as possible tools for control have not been widely tested against them. The objective of this study was to determine the efficacy of Olyset Net and PermaNet LLINs alongside a local brand, K-O Tab treated net (Supanet) against Phlebotomus duboscqi female sandflies. METHODS: Four replicates of unwashed and 20x washed Olyset Nets and PermaNets, K-O Tab-treated and untreated Supanet and 'no net' treatments were evaluated against sandflies within the laboratory by tunnel tests and in semi-field conditions in the greenhouse model for their efficacy. RESULTS: All bednets allowed entry of P. duboscqi sandflies and subsequent blood-feeding. Olyset net's blood feeding inhibition was significantly higher than that of Supanet in the laboratory but not in semi-field condition. Of the LLINs, only Olyset net had sandflies that could not feed significantly more than those of Supanet. Additionally, no significant efficacy difference was observed between LLINs washed 20x and unwashed ones. The only significant difference noted in number of sandflies that were found dead or paralyzed within bednets in the semi-field condition was between Olyset and K-O Tab treated Supanet. In the laboratory, unwashed Olyset had a significantly higher number of sandflies killed than all other bednet treatments. CONCLUSION: Olyset net use in areas where sandflies are nuisance biters and/or disease vectors could be more beneficial in preventing sandfly bites than other tested bednets. It is recommended that mesh sizes of LLINs should be smaller for control of sandflies than those used for control of mosquitoes.

Application of predictive degree day model for field development of sandfly vectors of visceral leishmaniasis in northwest of Iran.

BACKGROUND & OBJECTIVES: Temperature plays a significant role in insect's development where a rise in temperature, accelerates the insect's metabolic rates, increases egg production and makes blood feeding more frequent. It also shortens the time period required for the development of pathogens within insects. Visceral leishmaniasis (VL) is one of the most important vector-borne diseases transmitted by different sandfly species. In this study, a phenological model was used to estimate the number of generations, peak activity and temporal variability of sandflies in the main VL foci in northwest Iran. METHODS: Development requirements of different life stages of a Phlebotomus papatasi laboratory colony were measured and were subjected to the formula for calculation of accumulated degree day (ADD) for field sandflies using the online soft (UC IPM), using horizontal cut-off method and single triangle model. Sandflies population dynamics was monitored in the field during the seasonal activity in the region and its association with the ADD was tested using SAS software. RESULTS: Populations of sandflies accommodated well with the amount of accumulated degree days (ADD) in the region. During the seasonal activity, a total of 639 ADD were produced which was enough to support one complete life cycle and growth of the next generation up to late larval instar. Larvae of the second generation hibernate through winter and the first adult population appears in the mid to late June of the next year when they receive at least 182 ADD from the beginning of the spring. The highest population density of sandflies was observed in early August, followed by a rapid decrease in early September, with the adult population disappearing completely in late September. This is the first degree day model related to sandflies in the most important VL foci of Iran. INTERPRETATION & CONCLUSION: Further studies in various regions with variable climate are recommended in order to better estimate and understand the development time, population dynamics and activities of the vectors which in turn could be used in proper implementation of effective vector control programmes.

Propetamphos resistance in Rhipicephalus bursa (Acari, Ixodidae).

Hard tick infestation is an important problem in both traditional and industrial animal husbandry as well as in veterinary and public health. Several acaricides have been used to control tick infestation in Iran. Poor control has been attributed to acaricide resistance in tick populations although this has not been demonstrated experimentally. In this study, susceptibility status to propetamphos (Blotic) of 2-3-week-old larvae of Rhipicephalus bursa, one of the most common tick species in Iran, was evaluated using the FAO recommended larval packet test (LPT) method with some modifications. Log dosage probit analysis of propetamphos dose-response regression lines produced a resistance ratio based on LC(50) in the most resistant strain of approximately 103-fold. Biochemical assays suggested that the tick strains contained multiple insecticide resistance mechanisms involving elevation of esterases, GSTs, MFOs and alteration of acetylcholinesterase. In light of this data, practical implications for pest and pesticide resistance management strategies are discussed.

Electronic mosquito repellents for preventing mosquito bites and malaria infection.

BACKGROUND: Electronic mosquito repellents (EMRs) are marketed to prevent mosquitoes biting and to prevent malaria. OBJECTIVES: To assess whether EMRs prevent mosquito bites, and to assess any evidence of an effect on malaria infection. SEARCH STRATEGY: In August 2006, we searched the Cochrane Infectious Diseases Group Specialized Register, CENTRAL, MEDLINE, EMBASE, LILACS, Cambridge Scientific Abstracts, and the Science Citation Index. We also checked conference proceedings, contacted international specialist centres and EMR manufacturers, and checked reference lists. SELECTION CRITERIA: Field entomological studies, which controlled for geographic site, time, and attractiveness of human participants, of EMRs for preventing mosquito bites; and randomized and quasi-randomized controlled trials of EMRs to prevent malaria infection. DATA COLLECTION AND ANALYSIS: Two authors assessed trial quality, and extracted and analysed the data. MAIN RESULTS: Ten field entomological studies met the inclusion criteria. All 10 studies found that there was no difference in the number of mosquitoes caught from the bare body parts of the human participants with or without an EMR. No randomized or quasi-randomized controlled trials on the efficacy of EMR on malaria infection were found. AUTHORS' CONCLUSIONS: Field entomological studies confirm that EMRs have no effect on preventing mosquito bites. Therefore there is no justification for marketing them to prevent malaria infection.

Insect glutathione transferases and insecticide resistance.

Glutathione transferases (GSTs) are a diverse family of enzymes found ubiquitously in aerobic organisms. They play a central role in the detoxification of both endogenous and xenobiotic compounds and are also involved in intracellular transport, biosynthesis of hormones and protection against oxidative stress. Interest in insect GSTs has primarily focused on their role in insecticide resistance. GSTs can metabolize insecticides by facilitating their reductive dehydrochlorination or by conjugation reactions with reduced glutathione, to produce water-soluble metabolites that are more readily excreted. In addition, they contribute to the removal of toxic oxygen free radical species produced through the action of pesticides. Annotation of the Anopheles gambiae and Drosophila melanogaster genomes has revealed the full extent of this enzyme family in insects. This mini review describes the insect GST enzyme family, focusing specifically on their role in conferring insecticide resistance.

Field issues related to effectiveness of insecticide-treated nets in Tanzania.

Insecticide-treated nets (ITNs) impregnated with pyrethroid insecticides have become one of the most promising interventions to prevent malaria in highly endemic areas. Despite the large body of experience documenting their health impact and the best way to distribute them, some key practical issues remain unresolved. For example, the duration of effective life of a net under field conditions is unknown. The most important factor affecting net effectiveness is the issue of regular re-treatment with insecticide. Washing is also an important determinant of insecticide longevity in the field. Trials were undertaken to provide some essential field information on ITNs within the site of an extended ITN programme in the Morogoro region of Tanzania. It was found that 45% of all nets were in bad condition (defined as more than seven large holes). It is concluded that an effective 'life' for polyester nets is 2-3 years. Further, two-thirds of the 20% of nets that were reported as having been re-treated within the last 12 months had less than 5 mg/m(2) of insecticide. According to the World Health Organization this is insufficient to be effective. People reported that they washed their nets four to seven times per year, usually with soap. Observations showed that such washing does not harm the nets and that the wash-water was unlikely to have an impact on the environment. Finally, bioassays were carried out with Anopheles gambiae on polyester netting with 0.5, 2, 5, 10 and 30 mg/m(2) of deltamethrin, alphacypermethrin and lambdacyhalothrin to assess the effectiveness of pyrethroids. The results confirmed that even with low insecticide concentrations, nets can still provide partial protection.

Molecular evidence for a kdr-like pyrethroid resistance mechanism in the malaria vector mosquito Anopheles stephensi.

The mosquito Anopheles stephensi Liston (Diptera: Culicidae) is the urban vector of malaria in several countries of the Middle East and Indian subcontinent. Extensive use of residual insecticide spraying for malaria vector control has selected An. stephensi resistance to DDT, dieldrin, malathion and other organophosphates throughout much of its range and to pyrethroids in the Middle East. Metabolic resistance mechanisms and insensitivity to pyrethroids, so-called knockdown resistance (kdr), have previously been reported in An. stephensi. Here we provide molecular data supporting the hypothesis that a kdr-like pyrethroid-resistance mechanism is present in An. stephensi. We found that larvae of a pyrethroid-selected strain from Dubai (DUB-R) were 182-fold resistant to permethin, compared with a standard susceptible strain of An. stephensi. Activities of some enzymes likely to confer pyrethroid-resistance (i.e. esterases, monooxygenases and glutathione S-transferases) were significantly higher in the permethrin-resistant than in the susceptible strain, but the use of synergists--piperonyl butoxide (PBO) to inhibit monooxygenases and/or tribufos (DEF) to inhibit esterases--did not fully prevent resistance in larvae (permethrin LC50 reduced by only 51-68%), indicating the involvement of another mechanism. From both strains of An. stephensi, we obtained a 237-bp fragment of genomic DNA encoding segment 6 of domain II of the para type voltage-gated sodium channel, i.e. the putative kdr locus. By sequencing this 237 bp fragment, we identified one point mutation difference involving a single A-T base change encoding a leucine to phenylalanine amino acid substitution in the pyrethroid-resistant strain. This mutation appears to be homologous with those detected in An. gambiae and other insects with kdr-like resistance. A diagnostic polymerase chain reaction assay using nested primers was therefore designed to detect this mechanism in An. stephensi.

Quantification of pyrethroid insecticides from treated bednets using a mosquito recombinant glutathione S-transferase.

Recombinant glutathione S-transferase (agGST1-6) from the malaria vector mosquito Anopheles gambiae Giles (Diptera: Culicidae) was expressed in Escherichia coli using a pET3a vector system. The expressed enzyme was biochemically active with reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). Activity of agGST1-6 with GSH and CDNB was inhibited to different degrees by both alpha-cyano and non-alpha-cyano pyrethroid insecticides. This inhibition was used to develop an assay for quantification of pyrethroids. Standard curves of insecticide concentration against percentage of enzyme inhibition or volume of iodine solution were established by spectrophotometry and iodine volumetric titration, respectively, for permethrin and deltamethrin. These assays allowed estimation of pyrethroid concentrations both spectrophotometrically and visually. For the residue assay of each insecticide, a cut-off point of 50% of the initial pyrethroid impregnation concentration was used, which should differentiate between biologically active and inactive treated bednets. The cross-reactivity of the primary permethrin photodegradants (3-phenoxyalcohol and 3-phenoxybenzoic acid) with the recombinant agGST1-6 was assayed in the same system. No agGST1-6 inhibition by the insecticide metabolites was observed, suggesting that the system is unaffected by primary permethrin metabolites and will accurately measure insecticide parent compound concentrations. The estimated pyrethroid insecticide concentrations, given spectrophotometrically and by iodine titration assay, were comparable to those obtained by direct HPLC quantification of residues extracted from bednets. Hence, it should be relatively easy to adapt this method to produce a test kit for residue quantification in the field.