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BACKGROUND:Phelebotumus papatsi is considered to be the main vector of zoonotic cutaneous leishmaniasis as well as sand fly fever in Iran. There are several measures for vector control, with emphasis on insecticides. The objective of this study was to determine the baseline susceptibility of this vector to the commonly used insecticides in an endemic focus of the disease in central Iran. P. papatasi collected from the field were used for susceptibility status. Its baseline susceptibility to DDT and pyrethroids was assessed on about 6866 specimens collected from Badrood rural district, Esfahan Province, Iran, during the summer of 2010. The LT50 and LT90 values were measured according to the World Health Organisation (WHO) test using probit analysis and regression lines. RESULTS: Results of tests against female P. papatasi revealed LT50 values to DDT 4%, permethrin 0.75%, deltamethrin 0.1%, cyfluthrin 0.15% and lambdacyhalothrin 0.05% of 1104.97, 182.35, 26.79, 15.42 and 1.48 s respectively. The figures for male P. papatasi were 973.51, 59.5, 4.4, 2.65 and 1.5. CONCLUSION: The results of this study provide a guideline for implementation of different vector control measures. Furthermore, guidelines are needed for monitoring and evaluation of insecticide susceptibility tests against sand flies. Copyright © 2012 Society of Chemical Industry

Zoonotic cutaneous leishmaniasis (ZCL) due to Leishmania major is a great public health problem in the Old World. Leishmania major is widely distributed in populations of rodents in arid and savannah regions. In this study, seasonal variation of natural infection with Leishmania parasites in Rhombomys opimus (Rodentia: Gerbillinae) population of an endemic focus of ZCL in Iran was monitored. The study was conducted from October 2007 to October 2008 in the central part of the country. Nested polymerase chain reaction (PCR) assay was used for the detection and identification of Leishmania parasites, and the results were confirmed by PCR-restriction fragment length polymorphism (RFLP). The results showed that Leishmania infection rate was 55.8% (29 out of 52 gerbils) using nested PCR. The highest and lowest Leishmania infection rates were observed in fall and summer, respectively. Gerbils that were found to be infected only with L. major were 5.8%, and that with Leishmania turanica were 23.1%. A mixed natural infection was seen in the rodents with L. major and L. turanica (21.2%), with L. major and L. gerbilli (1.9%), and with all the three species (3.9%). Leishmania major infection alone was seen in fall and winter whereas mixed infection of L. major and L. turanica was observed in all seasons except in summer. Leishmania turanica infection was observed throughout the year. It is concluded that L. major, L. gerbilli, and L. turanica circulate in the population of R. opimus in central part of Iran. Leishmania major infection is usually accompanied by L. turanica in naturally infected gerbils with the highest rate in fall. It is recommended that the role of L. turanica in the epidemiology and transmission of ZCL should be reconsidered.

The annual incidence of cutaneous leishmaniasis (CL) in Iran rose by 43% over a five year period, from 2002 to 2006; most of these cases were caused by Leishmania major. Two complementary standard and nested polymerase chain reactions (PCRs) were used to detect parasites within their natural vector, Phlebotomus papatasi. Twelve different sand fly species were morphologically identified. The most abundant species (31.3%) was P. papatasi. Leptomonads were found in nine (2.4%) phlebotomines. Twenty (5.3%) sand fly species were found positive for Leishmania-genus DNA using standard PCR. The infection rate of this species was 5% and 7% by microscopic and molecular methods, respectively.

Discrimination of Leishmania infantum and L. donovani, the members of the L. (L.) donovani complex, is important for diagnosis and epidemiological studies of visceral leishmaniasis (VL). We have developed two molecular tools including a restriction fragment length polymorphisms of amplified DNA (PCR-RFLP) and a PCR that are capable to discriminate L. donovani from L. infantum. Typing of the complex was performed by a simple PCR of cysteine protease B (cpb) gene followed by digestion with DraIII. The enzyme cuts the 741-bp amplicon of L. donovani into 400 and 341 bp fragments whereas the 702 bp of L. infantum remains intact. The designed PCR species-specific primer pair is specific for L. donovani and is capable of amplifying a 317 bp of 3' end of cpb gene of L. donovani whereas it does not generate an amplicon for L. infantum. The species-specific primers and the restriction enzyme were designed based on a 39 bp insertion/deletion (indel) in the middle of the cpb gene. Both assays could differentiate correctly the two species and are reliable and high-throughput alternatives for molecular diagnosis and epidemiological studies of VL in various foci.

Field studies were carried out in Iran to evaluate the effect of various factors (washing, sun, smoke, dust and dirt) on the residual insecticidal activity of PermaNet (a brand of long-lasting insecticidal net), and on nets conventionally treated with deltamethrin (K-O Tab), using bioassay tests. Thirty-two nets were washed five or 15 times, and eight nets were not washed at all. Nets were washed vigorously in cold tap water (17 degrees C, pH 8.9) with a detergent. Hand rubbing continued for 3min. After washing, some nets were exposed to dense smoke from a dung-hay fire for 3min and were also left exposed to the dusty wind between washes. One group of nets was exposed to the sunlight for the full 3-d interval between washes; another was exposed to sunlight for just 3h after each wash; two other groups were kept in the shade. There was a significantly greater loss of activity in nets exposed to the sun throughout the 3-d interval between washes: that is, for a total of 15 to 45 d. However, short sunlight exposure (maximum 3h between washes) during drying did not have any effect. We did not find any significant effect of exposure to dirt, dust and smoke after washing. It is concluded that the effect of sun is much smaller than that of washing, and that drying nets for a few hours in the sun is not harmful.

With a growing international focus on emerging diseases that might impact on human health there is a recognition that we need to gain more understanding of the ecology of diseases such as avian influenza, paramyxoviruses and viral vector borne diseases (e.g. Ross River Virus, West Nile Virus). Many zoonotic diseases have a wildlife reservoir with several significant diseases occurring in a range of avian species. New Zealand has a unique avifauna and, to date, the information we have on the diseases of, and the disease risks to or from, these species is limited This presentation will outline some of the projects currently underway in New Zealand with specific reference to test development and surveillance initiatives for wildlife diseases.

Background Identification of the microflora of the sand fly gut and the environmental distribution of these bacteria are important components for paratransgenic control of Leishmania transmission by sand flies. Methods Biotic and abiotic bacterial communities of four compartments of a hyper-endemic focus of Zoonotic Cutaneous Leishmaniasis (ZCL) were investigated using 16S ribosomal DNA sequencing and phylogenetic tree construction. These compartments include Phlebotomus papatasi’s gut, skin and intestinal tract of great gerbil Rhombomys opimus, the gerbil nest supplies, and plant food sources of the vectors and reservoirs. Results Sequence homology analysis using nine available 16S rDNA data bases revealed 40, 24, 15 and 14 aerobic bacterial species from the vector guts, the gerbil bodies, the gerbil nests, and the plants, respectively. The isolated bacteria belong to wide ranges including aerobic to facultative anaerobic, pathogen to commensals, sand fly oviposition inducers, land to air and ocean habitats, animal and human probiotics, and plant growth-promoting rhizobacteria. Matching data analysis suggested that the adult P. papatasi gut bacteria could be acquired from three routes, adult sugar feeding on the plant saps, adult blood feeding on the animal host, and larval feeding from nest supplies. However, our laboratory experiment showed that none of the bacteria of the reservoir skin was transmitted to female sand fly guts via blood feeding. The microflora of sand fly guts were associated with the sand fly environment in which the predominant bacteria were Microbacterium, Pseudomonas, and Staphylococcus in human dwellings, cattle farms, and rodent colonies, respectively. Staphylococcus aureus was the most common bacterium in sand fly guts. Presence of some sand fly ovipoisition inducers such Bacillus spp. and Staphylococcus saprophyticus support association between gut flora and oviposition induction. Conclusions Results of this study showed that Bacillus subtilis and Enterobacter cloacae particularly subsp. dissolvens are circulated among the sand fly guts, the plants, and the sand fly larval breeding places and hence are possible candidates for a paratransgenic approach to reduce Leishmania transmission.

A study was made in rural region of Shahrood city, Semnan province in the central of Iran during 2005 to investigate of vectors of zoonotic cutaneous leishmaniasis. Sticky traps and an aspirator were used for collecting of sandflies. Three species of Phlebotomus papatasi, P. caucasicus and Sergentomyia sintoni were collected and identified and the first species was dominant (53%). Nested PCR method were employed for identifying of isolated parasites of dissected female of sandflies. Among the dissected sand flies 3 out of 24 (12.5%) Phlebotomus papatasi and 2 out of 48 (4.2%) Phlebotomus caucasicus were found naturally infected with promastigotes. Species-specific amplification ofGiemsa staind promastigote slides revealed specific PCR production of Leishmania major DNA in the infected P. papatasi and P. caucasicus sand flies. Having high prevalence and infection rate provide enough evidence to incriminate of P. papatasi as the main and proven vector of cutaneous leishmaniasis to human in the region and the species of P. caucasicus play the second role for maintenance of disease between rodents.

Background: In domestic and per domestic area,insecticides such as DDT, malathion, fenitrothion, propoxur and, more recently, synthetic pyrethroids such as deltamethrin and lambda-cyhalothrin, have been successfully used to control sand flies in many countries. The present study reports the results of time-mortality bioassay to DDT 4%, lambda-cyhalothrin 0.05%, permethrin 0.75%, cyfluthrin 0.15% and deltamethrin 0.05% in recently colonized Phlebotomus papatasi populations in Iran. Methods: The insecticide susceptibility status of P. papatasi laboratory population was assessed during 2016–2017, following the standard WHO technique for mosquito (WHO, 2013) based on diagnostic dose. Sand flies collected from rural area of Badrood (Matin Abad), Natanz County, Esfahan Province, using aspirator. Results: Susceptibility test to DDT and pyrethroids was assessed on 3534 laboratory-reared P. papatasi (1746 females and 1788 males). The LT50 and LT90 values were measured using probit analysis and regression lines. The test results against males of P. papatasi revealed that LT50 values to DDT 4%, Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15% and Lambdacyhalothrin 0.05% were 439.28, 108.90, 97.75, 5.00 and 57.84 seconds. The figures for females were 641.62, 136.15, 146.44, 8.71 and 72.69 seconds , respectively. Conclusion: According to presented results, the reared population of sand flies collected from a hyper-endemic region of Esfahan Province is still susceptible to prethroids and Resistance candidate to DDT 4%.