Bioefficacy of bendiocarb WP80 in vector-borne and zoonotic diseases areas in borderline of Iran and Pakistan.

Malaria and leishmaniasis are the public health problems in southern Iran. The main activity of vector control is indoor residual spraying using pyrethroids, using different insecticides as larviciding and impregnated bednets. The aim of study was to evaluate the biological assays of bendiocarb wettable powder (WP) at different surfaces of wall. The residual effect of bendiocarb WP80 at 400 milligram/meter square (mg/m2) was evaluated on various local surfaces of rooms such as mud and plaster as well as thatch roofs and wooden. World Health Organization standard cones using contact bioassays were carried out using laboratory reared sugar-fed, 48-72 h old females of Anopheles stephensi. Contact bioassays were carried out on sprayed surfaces for 150 days. Contact bioassay on surfaces treated with bendiocarb WP80 at different surfaces was estimated about 2 months. Fumigant tests of bendiocarb WP80 at 400 mg/m2 revealed 50-93.83% mortality with 1 month persistency. The results showed that carbamate insecticide could be used as rotation with pyrethroids for malaria vector control. Monitoring and evaluation of environmental toxicology of pesticides is important for decision making for choosing appropriate pesticides for disease vector control.

Wash resistance and bio-efficacy of Olyset® Plus, a long-lasting insecticide-treated mosquito net with synergist against malaria vector, Anopheles stephensi.

OBJECTIVE: To determine the wash resistance of Olyset® Plus using World Health Organization Pesticide Evaluation Scheme standard washing procedure and to assess the value of knock down and mortality rates of Anopheles stephensi at different regimens of long lasting insecticide treated nets washings. METHODS: The study was conducted at the Bioassay Laboratory of Culicidae Insectary, School of Public Health, Tehran University of Medical Sciences, Iran. The net was made of polyester impregnated with permethrin and piperonyl butoxide at a ratio of 2:1. The washing resistance was assessed using Le Chat® soap and a shaker incubator set at a speed of 155 r/min, 30 °C for 10 min. The cone bioassay test was carried out according to World Health Organization recommended guideline with tolerant field strain of female Anopheles stephensi to pyrethroids. RESULTS: The knockdown and mortality rates of female mosquitoes exposed to Olyset® Plus from un-washed nets to 2 washings were 79.7% and 88.8% respectively. Mortality was dropped to zero while active ingredient estimated 0.532 µg/100 cm2 to 0.481 µg/100 cm2 after 15 washings. A positive correlation was seen between residues of permethrin on nets, knockdown rate and mortality rate of female Anopheles stephensi exposed to different regimes of washed Olyset® Plus (r = 0.954, P = 0.001). CONCLUSIONS: It is recommended that a preliminary survey conducted on resistance level of Anopheles vectors before the distribution of Olyset® Plus in malaria endemic communities.

Evaluation of Deltamethrin in Combination of Piperonyl Butoxide (PBO) against Pyrethroid Resistant, Malaria Vector, Anopheles stephensi in IRS Implementation: an Experimental Semi-Filed Trial in Iran.

BACKGROUND: The aim of this study was to evaluate different concentrations of deltamethrin combined with formulated piperonyl butoxide (PBO) synergist on various surfaces against the wild strain of Anopheles stephensi, the main malaria vector in Southern Iran under semi-field condition. METHODS: Four concentrations of deltamethrin WG 25% (Tagros) and PBO 800EC-UV (Endura) were prepared and sprayed on the pre-designed surfaces in accordance with WHO alliance line of the IRS Micronair®. The WHO's recommended bioassay kit and method was used during this study. RESULTS: Comparing the mortality rate of mosquitoes, the results showed a significant difference between months after treatment of IRS (Indoor Residual Spraying) (P< 0.05) but didn't show any significant differences between days during the first and second months (P> 0.05).Statistical test revealed a significance difference between mortality rate of mosquitoes in exposing to concentrations of 1 and 4 (P< 0.05) which demonstrated effect of synergizing PBO on mortality rate. CONCLUSION: This research as the first semi-field trial on deltamethrin added to different concentrations of formulated PBO for IRS, indicates that deltamethrin+10X PBO is more effective than other concentrations. Therefore, using synergists can be suggested as a new tool for prevention of pyrethriod resistance, although more studies are recommended.

Discrimination of Paederus fuscipes and Paederus littoralis by mtDNA-COI PCR-RFLP.

BACKGROUND: Linear dermatitis is endemic in Iran where most cases occur in the Caspian Sea coast and Fars province. The disease is caused by beetles of the genus Paederus which are active from early spring to beginning of autumn although its incidence rises from May to August. The classic taxonomy of Paederus spp. is based on the male genitalia that is very complex and needs expertise. In this study, we report a DNA-based method to discriminate Paederus fuscipes and Paederus littoralis (=syn: P. lenkoranus, P. ilsae). METHODS: Type specimens were collected from north and south of Iran. Molecular typing of the species was performed using restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified fragments of mtDNA-COI. RESULTS: Sequence analyses of the data obtained in this study showed significant DNA polymorphisms. There were 89 substitutions between COI sequences of the two species. The mtDNA-COI fragment comprises several useful species-specific restriction sites comprising HaeIII that could result in distinctively different species-specific PCR-RFLP profiles. The HaeIII enzyme cuts the 872 bp PCR amplicon of P. littoralis into 737 and 100 bp and two small nonvisible bands whereas it does not cut P. fuscipes amplicon into fragments. CONCLUSION: This study demonstrates that molecular typing is useful method and allows one to differentiate between two species and is recommended for discrimination of other Paederus species, which morphologically are indistinguishable or very difficult to be distinguished.

Diversity of Sand Flies (Diptera: Psychodidae) in Endemic Focus of Visceral Leishmaniasis in Azar Shahr District, East Azarbaijan Province, North West of Iran.

BACKGROUND: There are nearly 1000 species of Phlebotomine sand flies in 6 genera, of which only two, Phlebotomus in the old world and Lutzomyia in the new world are medically important. Globally, leishmaniasis prevalent in 98 countries and affects estimated 12 million people with almost two million new cases per year. Some rural areas of Azarshahr District in East Azarbaijan Province have been reported to be endemic for visceral leishmaniasis. This study is the first attempt to determine the species diversity and density in a new focus of visceral leishmaniasis in Azarshahr District, East Azarbaijan Province, Iran. METHODS: Sand flies were collected form indoor and outdoor biweekly using sticky traps. Diversity index of the collected sand flies within different villages were estimated by the Shannon-Weaver. RESULTS: The activity of the sand flies extended from April to October with one peak in August. Diversity of sand flies within study area were estimated as 0.917, 1.867, 1.339, 1.673, and 1.562 in Almalodash, Jaragil, Segaiesh, Amirdizaj, and Germezgol Vvillages, respectively. CONCLUSION: Identifying the diversity and seasonal abundance of the collected species is of importance for prediction of the period of maximum risk for leishmaniasis transmission and for the successful implementation of a control program. Species diversity is one of the most important factors in ecological studies.

The insecticidal effect of diatomaceous earth against adults and nymphs of Blattella germanica.

OBJECTIVE: To evaluate the insecticidal effect of diatomaceous earth (DE) against adults and nymphs of Blattella germanica. METHODS: This cross sectional study has been done on the laboratory strain of German cockroaches. Two stages, nymph and adult, were exposed to six dose rates of the DE, 2.5, 5, 10, 15, 20 and 25 g/m(2), at 24, 48 and 72 h exposure period. Mortality (number of dead cockroaches) was assessed after 24 h. Other exposed specimens were transferred to the beakers contained food and water for counting the retard mortality rate after 1 week. RESULTS: Increasing in dose rates of DE increased mortality rate, so that the lowest and highest mortality rates were observed in 2.5 and 25 g/m(2), respectively. The results of the statistical analysis showed no significant difference in the lethality of 50% of DE plus water on the German cockroach nymphs. CONCLUSIONS: Due to the resistance of German cockroach against organochloride, organophosphorus, carbamate and pyrethriodes insecticides, it is suggested to use DE for insect's control.

Biodiversity of Aquatic Insects of Zayandeh Roud River and Its Branches, Isfahan Province, Iran.

BACKGROUND: Aquatic insects are the major groups of arthropods that spend some parts of their life cycle in the water. These insects play an important role for transmission of some human and animal diseases. There is few information about the aquatic insects fauna of Iran. METHODS: To study the aquatic insects fauna, adult, nymphal and larval collections were carried out from different habitats using the standard technique in Zayandeh Roud River, Isfahan Province,central Iran, during summer 2011. RESULTS: In total, 741 speimens of aquatic insects were collected and morphologically identified. They include 7 families and 12 genera representing 2 Orders. The order of Diptera (92.31%) and Coleoptera (7.69%). The families Culicidae, Syrphidae and Chironomidae from Diptera order, Gyrinidae, Dytiscidae, Haliplidae, Hydrophilidae from Coleoptera order were identified. CONCLUSION: Some aquatic insects play an important role for transmission of human and animal diseases. These insects also are important for biological control. Therefore ecological study on aquatic insects can provide information about ecology of insects in an area for any decision making.

The insecticidal effect of diatomaceous earth against adults and nymphs of Blattella germanica

Objective: To evaluate the insecticidal effect of diatomaceous earth (DE) against adults and nymphs of Blattella germanica.Methods:cockroaches. Two stages, nymph and adult, were exposed to six dose rates of the DE, 2.5, 5, 10, 15, 20 and 25 g/m2, at 24, 48 and 72 h exposure period. Mortality (number of dead cockroaches) was assessed after 24 h. Other exposed specimens were transferred to the beakers contained food and water for counting the retard mortality rate after 1 week. This cross sectional study has been done on the laboratory strain of German Results: Increasing in dose rates of DE increased mortality rate, so that the lowest and highest mortality rates were observed in 2.5 and 25 g/m2, respectively. The results of the statistical analysis showed no significant difference in the lethality of 50% of DE plus water on the German cockroach nymphs.Conclusions:Due to the resistance of German cockroach against organochloride organophosphorus, carbamate and pyrethriodes insecticides, it is suggested to use DE for insect’s control.

Wash Resistance and Bioefficacy of Alpha-cypermethrin Long Lasting Impregnated Nets (LLIN-Interceptor(®)) against Anopheles stephensi using Tunnel Test.

BACKGROUND: The Long-lasing insecticide impregnated nets (LLINs) is considered as an effective tools for malaria vector control. The aim of this study was to evaluate the residual efficacy of alpha-cypermethrin long lasting impregnated nets (LLIN-Interceptor(®)) against Anopheles stephensi using tunnel test. METHODS: The wash-resistance of Interceptor(®) nets were assessed under laboratory conditions using tunnel test. Females of An. stephensi were released into the tunnel and then they were provided blood meals from guinea pigs. Bed nets were washed according to the standard procedure up to 20 times. The bioefficacy indicators such as inhibition of bloodmeal from experimental animal, knockdown, irritancy rate, survival rate, entry index and mortality were calculated. RESULTS: It induced 90-100% mortalities in the population of An. stephensi up to 15 washes. The KT50 values reduced from 73.47 to 26.30 minutes in unwashed in comparison to one washed, respectively. The mean of mortality rate of blood-feeding inhibition and entry indexes was reached to 91.6%±2.8, 87.0±3.4 and 24.9±2.8 respectively after 20 washing. CONCLUSION: This net could provide a good personal protection against malaria vectors and could induce relatively high mortality, inhibit the blood-feeding as well as reduce the entry rates of female mosquitoes even after several washes.

Notes on the Iran Caddisflies and Role of Annulipalpian Hydropsychid Caddisflies as a Bio-monitoring Agent.

BACKGROUND: Eco-faunistic studies are inevitable step in environmental researches. Aquatic organisms like caddisflies are known as biological indicators for water quality assessment and water resource management. They have special role for energy flow in the freshwater habitats as food web and food chain among aquatic creatures. METHODS: In addition to an extensive literature review on Iran Caddisflies, a field study was carried out in Lavasan river flows in north east of Tehran to collect aquatic insects using D-frame nets and or direct search on stone beneath. The water quality was measured using analytical method. RESULTS: Literature revealed record of 62 trichopterid species in the country comprising 14 families. The most abundant species belonged to the Hydropsychidae. Herein we report presence of the Annulipalpian Hydropsyche sciligra H Malicky, 1977 in the study area. Habitat water quality of H. sciligra resembled human drinkable water. However presence of snail, Physa acuta and fish Capoeta buhsei in the water sampling area indicated inferior quality. CONCLUSION: From ecological point of view caddisfly larvae are predators of most important medical vectors like mosquitoes, blackflies and midges. Also they are useful and important indicator for monitoring physicochemical effects in the nature, so that they can be used for bio-monitoring program. From medical point of view, wing hairs or other body parts of caddisflies can be inhalant and contact allergens in Trichopterists and in sensitive individuals who come in contact.

Organophosphorous pesticides in surface water of Iran.

This research aims to evaluate the presence and distribution of pesticides in Babolrood River of Mazandaran Province in Iran. Mean diazinon levels in surface water ranged from 77.6 to 101.6 µg L(-1) with maximum level of 768.9 µg L(-1) and mean malathion levels ranged from 55.7 to 75.9 µg L(-1) with maximum level of 506.6 µg L(-1). The residues of malathion and diazinon pesticides in all of the stations, 2 weeks after spraying, were more than allowed limits.

Organophosphorous residue in Liza aurata and Cyprinus carpio.

OBJECTIVE: To determine the amount of azinphos methyl and diazinon residues in two river fishes, Liza aurata and Cyprinus carpio, in the north of Iran. METHODS: This study was done during 2006-2007. In this survey, 152 water and fish samples from Gorgan and Qarasu rivers, north of Iran, were investigated. Sampling was done in three predetermined stations along each river. Organophosphorus compounds (OPs) were extracted from the fishes and the water of rivers. After extraction, purification and concentration processes, the amount and type of insecticides in water and fish samples were determined by high performance thin layer chromatography (HPTLC). RESULTS: There was a significant difference in the residue of the insecticides in the water and fish samples between summer and other seasons in the two rivers. The highest amount of insecticides residue was seen during summer. In both rivers, the amount of diazinon and azinphos methyl residues in the two fishes was more than 2 000 mg/L in summer. There was no significant difference in insecticides residue between the fishes in two rivers. The diazinon residue was higher than the standard limits in both rivers during the spring and the summer, but the residual amount of azinphos methyl was higher than the standard limits only during the summer and only in Qarasu River. CONCLUSIONS: It can be concluded that the amount of OPs in the water and the two fishes, Liza aurata and Cyprinus carpio, is higher than the permitted levels.

Bioassay evaluation on the efficacy of α-cypermethrin impregnated into long lasting insecticide treated nets using Anopheles stephensi

OBJECTIVE@#To evaluate the bioefficacy of α-cypermethrin impregnated into long lasting insecticide treated nets (LLITNs-INTERCEPTOR®) against main malaria vector, Anopheles stephensi (An. stephensi).@*METHODS@#The effectiveness of bed net impregnated with α-cypermethrin (INTERCEPTOR®) with washing was evaluated. The washing procedure and bioassay tests were carried out according to the WHO-recommended methods. Malaria vector, An. stephensi was exposed to impregnated bed net for three minutes and then mortality measured after 24 h recovery period. Knockdown was also measured according to the logarithmic times.@*RESULTS@#Result of cone bioassay method showed that bioefficacy of α-cypermethrin decreased from 100% in unwashed to 15% in 20 washes. KT(50) was measure as one minute in one wash and increased to 40 min in 20 washes.@*DISCUSSION@#Findings of this study provide guideline for malaria vector control authorities and people using pyrethroid-impregnated bed nets.

Laboratory evaluation of dimethyl phthalate against Anopheles stephensi and Culex pipiens.

Two repellents includes Iranian and Merck dimethyl phthalate (DMP) were evaluated against Anopheles stephensi and Culex pipiens, using American Society for Testing and Material (ASTM) standard ED 951-83 procedure, a free choice method based on the variable dose-fixed time. Also a modified of ASTM method we used for determination of effective dosages of the repellents. In ASTM method there were no significant differences between the two repellents (Iranian and Merck's DMP) as indicated by the ED50 and ED95 values (p > 0.05). But, there were significant differences in repellent sensitivity between An. stephensi and Cx. pipiens at the ED95 level. In modified ASTM method there were no significant differences between the two repellents against An. stephensi, as indicated by the ED50 values (p > 0.05). But, there are significant differences between the two repellents against Cx. pipence based on ED50 value. Results of this study showed that the Iranian synthesized DMP has necessary potential and specificity to compare with Merck manufactured product.

Determination of organophosphorus insecticides (malathion and diazinon) residue in the drinking water.

In this study, the amount and origin of the pollution of Karaj river and Amir-Kabir dam, which is the most important supply of Tehrans (capital of Iran) drinking water with malathion and diazinon, have been assessed by High Performance Thin Layer Chromatography (HPTLC) technique. This is a descriptive and cross-sectional study. After initial assessments, four stations were selected for water sampling. In all stations, sampling was done in one day, one week, two weeks, one month, two months and three months after insecticide spraying. Extraction and separating of organophosphorus insecticides from water samples was done by acetone and methylen chloride solvents, using decanter. Then, the residue of malathion and diazinon insecticides was determined using HPTLC technique. The residue of malathion and diazinon in water decreases with increasing the distance and time of spraying. The residues of malathion and diazinon insecticides at station 1 and 2, which were close to spraying places, 1-2 months after spraying was more than allowed limits. But at the ending stations (3 and 4), malathion and diazinon were detectable just in the 1 day and 1 month after spraying, respectively. It is unlikely that Tehran's drinking water be polluted with these two insecticides and the most adverse effect can be seen in the area of spraying. Therefore not only the environment, but also the people in area of Karaj river are at risk of chronic toxicity with malathion and diazinon through consuming polluted water and agriculture products.