Organochlorine knockdown-resistance (kdr) association in housefly (Musca domestica): A systematic review and meta-analysis.

Introduction: Although house flies (Musca domestica) do not directly cause disease in humans, they transmit pathogens to them, which provide the basis for many diseases. The main way to deal with this insect is to use insecticides. Due to the resistance from insecticides, the fight against house flies has been hampered. This study aimed to determine the prevalence of knockdown resistance against organochlorine insecticides in house flies worldwide. Methods: This study was conducted via a systematic review and meta-analysis to investigate the prevalence of knockdown resistance against organochlorine insecticides in house flies. Accordingly, by searching the databases of Web of Science, PubMed, Scopus, Proquest, Bioone, and Embase, all published articles were extracted, and reviewed until the end of May 2022. Statistical data analysis was performed using the random-effects model in the meta-analysis, meta-regression, and I2 index. Results: Nine studies entered the meta-analysis process. Based on this, the prevalence of knockdown resistance against organochlorine insecticide in house flies was estimated to be 49.1%. Meta-regression showed that the prevalence of knockdown resistance increased with increasing years of study but decreased with increasing sample size. Conclusion: According to the findings, about 50% of house flies have knockdown resistance against organochlorine insecticide. As a result, it is necessary to adopt effective and combined methods to combat this insect to control it and prevent the transmission of diseases caused by it.

Identification, molecular characterization, and in silico structural analysis of larval salivary glands Netrin-A as a potent biomarker from Lucilia sericata (Diptera: Calliphoridae).

The greenbottle blowfly Lucilia sericata (L. sericata) is increasingly used in larval therapy of chronic wounds. Netrins as bifunctional proteins are in the superfamily of Laminins secreted from larval salivary glands. The Netrin protein has a significant instructive role in axon guidance, causing neuronal outgrowth, angiogenesis, and cell migration. It seems to be crucial in wound healing and acts as a potential biomarker in diagnosing some clinical diseases. This survey aimed to identify molecular features and analyze in silico structural configuration of Netrin-A in L. sericata larvae. The larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericata Netrin-A (LSN-A) was then identified using rapid amplification of circular DNA ends (RACE) and rapid amplification of genomic ends (RAGE). Parts of the Netrin-A gene, including the middle, 3'-, and 5'-ends, were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its estimated molecular weight was 78.6 kDa. Sequencing of this fragment and its BLAST analysis revealed laminin-based high (95%) similarity with the mRNA sequence of Lucilia cuprina Netrin-A. The 3-D structure of Netrin-A drawn by SWISS-MODEL exhibited its partial resemblance to the reference molecule Netrin-1 of Homo sapiens. This study supports the molecular and structural analyses of LSN-A protein, which could lead to wound treatment. Ultimately, it can be an effective candidate to ameliorate injury. Our next attempt is to produce LSN-A recombinant protein for use in biomedical sciences.

Nanoliposomes containing limonene and limonene-rich essential oils as novel larvicides against malaria and filariasis mosquito vectors.

BACKGROUND: Mosquito-borne diseases such as malaria and encephalitis are still the cause of several hundred thousand deaths annually. The excessive use of chemical insecticides for transmission control has led to environmental pollution and widespread resistance in mosquitoes. Botanical insecticides' efficacies improvement has thus received considerable attention recently. METHODS: The larvicidal effects of three essential oils from the Citrus family and limonene (their major ingredient) were first investigated against malaria and filariasis mosquito vectors. An attempt was then made to improve their efficacies by preparing nanoliposomes containing each of them. RESULTS: The larvicidal effect of nanoformulated forms was more effective than non-formulated states. Nanoliposomes containing Citrus aurantium essential oil with a particle size of 52 ± 4 nm showed the best larvicidal activity (LC50 and LC90 values) against Anopheles stephensi (6.63 and 12.29 µg/mL) and Culex quinquefasciatus (4.9 and 16.4 µg/mL). CONCLUSION: Due to the green constituents and high efficacy of nanoliposomes containing C. aurantium essential oil, it could be considered for further investigation against other mosquitoes' populations and field trials.

Monitoring of synthetic insecticides resistance and mechanisms among malaria vector mosquitoes in Iran: A systematic review.

BACKGROUND: In Iran, the prospect of malaria control relies mainly on insecticides used against the genus Anopheles (Diptera: Culicidae) as important vectors of malaria, arboviruses, and so on. Only eight out of 30 malaria mosquito vectors (Anopheles species) have been examined for insecticide resistance in Iran. This study aimed to review articles related to the incremental trend in insecticide resistance and their mechanisms among anopheline malaria vectors in Iran. METHODS: A literature review was conducted based on such search engines as Iran doc, Web of Science, SID, PubMed, Scopus, and Google Scholar websites using the following keywords: "Anopheles," "Malaria," "Resistance," "Vectors," "Insecticide Resistance," and "Iran" for data collection. Published papers in English or Persian covering 1980 to 2020 were reviewed. RESULTS: A total of 1125 articles were screened, only 16 of which were filtered to be pertinent in this review. While most of the mosquito vectors of malaria, such as Anopheles stephensi, were resistant to DDT, dieldrin, malathion, and becoming less susceptible to deltamethrin and other synthetic pyrethroid insecticides, few like Anopheles fluviatilis s. l. were susceptible to all insecticides. A disseminating trend in insecticide resistance among different anopheline mosquito vector species was evident. Metabolic and insecticide target-site resistance mechanisms were involved with organochlorines and pyrethroids, respectively. CONCLUSIONS: Insecticide resistance is becoming a severe scourge to the effectiveness of vector-borne disease management measures. This event is especially critical in developing and marginalized communities that applied chemical-based vector elimination programs for malaria; therefore, it is crucial to monitor insecticide resistance in malaria vectors in Iran using biochemical and molecular tools.

Solid-lipid nanoparticles (SLN)s containing Zataria multiflora essential oil with no-cytotoxicity and potent repellent activity against Anopheles stephensi.

Malaria is still a global health concern with more than 400,000 death annually. Personal protection using mosquitoes' repellent is an effective prevention strategy, especially in endemic areas. The toxic effects of synthetics repellents and their adverse effects on fabricated goods have made the development of green repellent critical. In this study, ingredients of Zataria multiflora essential oil (ZMEO) were identified using GC-MS analysis. Solid-lipid nanoparticles containing ZMEO (1%) were prepared (SLN-ZMEO) using the high-pressure homogenizer method. The repellent activity of ZMEO and SLN-ZMEO was investigated using Klun and Debboun method and compared together. Besides, their cytotoxicity on a human skin normal cell line (HFFF2) was evaluated. Five major components of ZMEO were carvacrol (27.05%), thymol (26.452%), γ-terpinene (15.144%), o-cymene (13.584%), and α-pinene (9.483%). The SLN-ZMEO showed a spherical shape with a particle size of 134 ± 7 nm. Moreover, their polydispersity index (PDI), zeta potential and entrapment efficiency were determined as 0.24 ± 0.1, - 9.82 ± 0.95 mV and 64.6 ± 3.8%, respectively. Interestingly, the protection time of nanoformulation (93 ± 5 min) was three times longer than that of the non-formulated essential oil (29 ± 2 min). Interestingly, both samples did not show cytotoxicity on HFFF2. Therefore, the prepared nanoformulation can be used as a green and potent repellent.

Prevalence of allergenic arthropods in domestic dwellings of referrals to an asthma and allergy clinic in the Islamic Republic of Iran.

BACKGROUND: Allergenic arthropods are crucial agents in inducing medically important respiratory diseases like asthma and the inflammation of the respiratory tract worldwide. AIMS: This study was conducted to determine the prevalence of all arthropods in the dwellings of people referred to the asthma and allergy clinic in Shiraz. METHODS: This was was a cross-sectional descriptive study. Participants were 100 allergic patients who had tested positive (roach- and mite-sensitive). Mites were collected from their houses using a vacuum cleaner; other arthropods were caught with sticky traps. Direct observation and flotation methods were used and the samples were stored in 70% ethanol. Morphological characteristics were identified using valid taxonomic keys. RESULTS: Overall, 624 specimens were identified belonging to 14 orders (4 orders of mites: Astigmata, Cryptostigmata, Prostigmata and Mesostigmata; and 10 other arthropod orders: Diptera, Coleoptera, Hymenoptera, Thysanura, Thysanoptera, Entomobryomorpha, Blattodea, Siphonaptera, Pscoptera and Isopoda). The 2 most numerous species collected were Musca domestica and Dermanyssus gallinae. CONCLUSION: A small number of dwellings were infested with cockroaches; none were infested with the common house dust mites. The allergies induced in these patients could likely be attributed to other arthropods that are not considered main allergens in asthma and allergy clinics in the Islamic Republic of Iran. Health surveillance and prevention of infestation for these arthropods could have an immense impact on the control of the allergenic arthropod community, prevention of respiratory diseases, and personal health in Shiraz.

Comparative efficacy of three pediculicides to treat head lice infestation in primary school girls: a randomised controlled assessor blind trial in rural Iran.

BACKGROUND: Head lice infestation (Pediculosis) is one of the most important health challenges particularly in primary school-aged children. It is often present among 6-11-year-old students in various tropical and temperate regions of the world. The aim of this study was to examine epidemiologic indices and comparative analysis of two pyrethroid-based and one non-chemical pediculicide products on head lice treatment of primary school girls in a rural setting of Fars province, south Iran, as part of a randomized controlled assessor blind trial. METHODS: Before treatment, infested students were screened using plastic detection combs to find live head lice. Three independent parallel groups, each with about 25 participants (#77) were eventually twice with a week apart treated with either 1% permethrin, 0.2% parasidose (d-phenothrin) or 4% dimeticone lotion preparations. In each case, a questionnaire form was completed on epidemiologic factors. Data were registered after a fortnight from primary scalp treatment and re-inspection on days 2, 6, 9 and 14. Data analyses were performed using Chi-square test with a P-value < 0.05 being taken as statistically significant. RESULTS: From 3728 inspected students, 87 (2.33%) girls were infested with head lice, Pediculus humanus capitis De Geer, 1778. Ten students dropped out pertaining to exclusion criteria. No significant correlation was found between head lice infestation level and hair length, hair style, itching, nationality, age, settlement site and baths; but there was a significant relationship between age and hair style (P = 0.027). The efficacy values on each of the above re-inspection days from each of the three treatments were 81, 74, 70 and 63% for permethrin; 83, 92, 100 and 100% for dimeticone; and 96, 88, 96 and 92% for d-phenothrin; respectively. A quartile difference in efficacy of permethrin relative to dimeticone on day 14 represented the scale of head lice resistance to permethrin treatment. There were significant statistical differences in case re-inspection days 9 (P = 0.008) and 14 (P = 0.003) post treatment. Only two dropout cases, one non-compliant and the other lost before the second-week treatment, from permethrin trial were observed following two applications a week apart. CONCLUSIONS: Dimeticone lotion had the fullest efficacy (100%) among all treatments. This high cure rate was attributed to the low level of infestation and the extent of patients' involvement. Parasidose swiftly ameliorated the infested cases by the second day since initial treatment. Female third grade students were the most infested cohort. TRIAL REGISTRATION: Current Controlled Trials- IRCT2016041627408N1 , Dated: 21-08-2017.

Malaria preventive behaviors among housewives in suburbs of Bandar-Abbas City, south of Iran: interventional design based on PRECEDE model.

Suburban areas suffer from unfavorable conditions as a result of poor health standards, low income, illness, and slight levels of education. One of the most important components in malaria elimination programs is health education. Bandar-Abbas is a malaria endemic city. Therefore, this study aimed to promote malaria preventive behaviors among housewives in suburbs of Bandar-Abbas City, Iran, through a PRECEDE model-based intervention program. This quasi-experimental study was carried out on 172 housewives under the coverage of four healthcare centers in Bandar-Abbas in 2016. The participants were randomly divided into two experimental and control groups, based on the inclusion criteria. The data collection tools included ademographic questionnaire, aresearcher-made questionnaire based on the educational phase of the PRECEDE model and achecklist for assessing malaria preventive behaviors. After the pre-test, the educational intervention was implemented on the intervention group at the first stage and the same questionnaire was administered as the post-test for both groups two months after the intervention. In this study, after implementing the intervention using the PRECEDE educational program, asignificant difference was found between the intervention and control groups in the mean scores for predisposing factors (knowledge and attitude) (P< 0.001), enabling factors (P< 0.001) and reinforcing factors (P< 0.001), which contributed to the improvement of preventive behaviors (P< 0.001). The PRECEDE model-based health education was effective in increasing knowledge and attitude, enabling and reinforcing factors, and promoting malaria preventive behaviors in the studied endemic areas.

Sandflies species composition, activity, and natural infection with Leishmania, parasite identity in lesion isolates of cutaneous leishmaniasis, central Iran.

Zoonotic cutaneous leishmaniosis (ZCL) is a crucial public health challenge in Iran. Sandflies feed on reservoir rodents' blood infected with Leishmania parasite and transmit it to other hosts. This study was conducted to find out the composition and monthly activity of sandflies as well as to identify the protozoan pathogens (Leishmania/Crithidia) by polymerase chain reaction (PCR) in an emerging ZCL focus of Abarkooh, Yazd province, Iran, in 2016. A cross-sectional study was done in rural areas of Abarkooh. From April to November 2016, sticky traps were used indoor and outdoor to capture sandflies once every fortnight. Their composition and monthly activity were recorded. Following identification of sandflies and DNA extraction from them, PCR was used to identify their parasite and match it against samples taken from ZCL confirmed and suspected patients' lesions. After collection, a total of 2045 sandflies (779 indoor, 1266 outdoor) were identified to species level. Sandfly activity started early April in this area with two active peaks (one late May and the other late August) terminated about mid-November. Seven Phlebotomus species and three Sergentomyia species were identified. The most and the least abundant species were P. papatasi (40.1%) and P. alexandri (0.09%), respectively. Using PCR, only 6% (12:200) of P. papatasi sandflies were infected with Leishmania parasite. No Crithidia was detected in either sandflies or human lesions (176 specimen). Based on the highest abundance both indoor and outdoor of P. papatasi, this sandfly was considered the main vector of ZCL in this area. The capture of P. caucasicus, P. mongolensis, and P. ansarii from rodent burrows showed these species were likely involved in pathogen transmission in reservoir rodents' burrows.

Predictive determinants of scorpion stings in a tropical zone of south Iran: use of mixed seasonal autoregressive moving average model.

BACKGROUND: More than 1.2 million scorpion stings occur annually worldwide, particularly in tropical regions. In the absence of proper medical care, mortality due to venomous scorpion stings is an important public health issue. The aim of the present study is to explore the temporal trend of scorpionism with time series models and determine the effective factors on this event using regression models. METHODS: A retrospective cross sectional study was conducted on 853 scorpion stung patients. They were referred to Haji-Abad Hospital of Hormozgan University of Medical Sciences (HUMS), south Iran, from May 2012 to July 2016. A linear model to describe and predict the monthly trend of scorpion sting cases is fit with autoregressive moving average (ARMA) model. RESULTS: Of 853 victims, 384 (45%) patients were female and 30.2% of them lived in urban areas. The mean (± SD) age of patients was 30.1 (± 19.6) years and the most affected age group was 20-29 years (21.8%). Most victims were unemployed people and farmers (54.7%) followed by housewives (30.2%). The majority of the stings occurred indoors (53.7%), between midnight and 6 a.m. (29.2%), in the summer (44.2%), and the most affected limbs were hands and legs (81.2%). Patient genders and occasions of being stung by scorpions were significantly different between outdoors and indoors (p < 0.001). Scorpion stings due to Odontobuthus doriae were significantly higher than due to other species in urban and rural patients (p = 0.04). Mixed seasonal ARMA at lag 12, ARMA (1, 1) × (0, 1), was selected as the best process for monthly trend of data. Regression results indicated that significant climate factors associated with scorpion stings are temperature (p < 0.001) and relative humidity (p = 0.002). CONCLUSIONS: Scorpion stings have a noticeable effect on tropical rural populations, mainly farmers. Two effective climate factors associated positively and negatively with scorpion sting cases are temperature and relative humidity, respectively. The results of time series and regression models to predict the trends and determinants of scorpion stings are almost the same.

Predictive determinants of scorpion stings in a tropical zone of south Iran: use of mixed seasonal autoregressive moving average model

Abstract Background: More than 1.2 million scorpion stings occur annually worldwide, particularly in tropical regions. In the absence of proper medical care, mortality due to venomous scorpion stings is an important public health issue. The aim of the present study is to explore the temporal trend of scorpionism with time series models and determine the effective factors on this event using regression models. Methods: A retrospective cross sectional study was conducted on 853 scorpion stung patients. They were referred to Haji-Abad Hospital of Hormozgan University of Medical Sciences (HUMS), south Iran, from May 2012 to July 2016. A linear model to describe and predict the monthly trend of scorpion sting cases is fit with autoregressive moving average (ARMA) model. Results: Of 853 victims, 384 (45%) patients were female and 30.2% of them lived in urban areas. The mean (± SD) age of patients was 30.1 (± 19.6) years and the most affected age group was 20-29 years (21.8%). Most victims were unemployed people and farmers (54.7%) followed by housewives (30.2%). The majority of the stings occurred indoors (53.7%), between midnight and 6 a.m. (29.2%), in the summer (44.2%), and the most affected limbs were hands and legs (81.2%). Patient genders and occasions of being stung by scorpions were significantly different between outdoors and indoors (p 0.001). Scorpion stings due to Odontobuthus doriae were significantly higher than due to other species in urban and rural patients (p = 0.04). Mixed seasonal ARMA at lag 12, ARMA (1, 1) x (0, 1), was selected as the best process for monthly trend of data. Regression results indicated that significant climate factors associated with scorpion stings are temperature (p 0.001) and relative humidity (p = 0.002). Conclusions: Scorpion stings have a noticeable effect on tropical rural populations, mainly farmers. Two effective climate factors associated positively and negatively with scorpion sting cases are temperature and relative humidity, respectively. The results of time series and regression models to predict the trends and determinants of scorpion stings are almost the same.

Predictive determinants of scorpion stings in a tropical zone of south Iran: use of mixed seasonal autoregressive moving average model

Background: More than 1.2 million scorpion stings occur annually worldwide, particularly in tropical regions. In the absence of proper medical care, mortality due to venomous scorpion stings is an important public health issue. The aim of the present study is to explore the temporal trend of scorpionism with time series models and determine the effective factors on this event using regression models. Methods: A retrospective cross sectional study was conducted on 853 scorpion stung patients. They were referred to Haji-Abad Hospital of Hormozgan University of Medical Sciences (HUMS), south Iran, from May 2012 to July 2016. A linear model to describe and predict the monthly trend of scorpion sting cases is fit with autoregressive moving average (ARMA) model. Results: Of 853 victims, 384 (45%) patients were female and 30.2% of them lived in urban areas. The mean (± SD) age of patients was 30.1 (± 19.6) years and the most affected age group was 20-29 years (21.8%). Most victims were unemployed people and farmers (54.7%) followed by housewives (30.2%). The majority of the stings occurred indoors (53.7%), between midnight and 6 a.m. (29.2%), in the summer (44.2%), and the most affected limbs were hands and legs (81.2%). Patient genders and occasions of being stung by scorpions were significantly different between outdoors and indoors (p < 0.001). Scorpion stings due to Odontobuthus doriae were significantly higher than due to other species in urban and rural patients (p = 0.04). Mixed seasonal ARMA at lag 12, ARMA (1, 1) x (0, 1), was selected as the best process for monthly trend of data. Regression results indicated that significant climate factors associated with scorpion stings are temperature (p < 0.001) and relative humidity (p = 0.002). Conclusions: Scorpion stings have a noticeable effect on tropical rural populations, mainly farmers. Two effective climate factors associated positively and negatively with scorpion sting cases are temperature and relative humidity, respectively. The results of time series and regression models to predict the trends and determinants of scorpion stings are almost the same.(AU)

Faunal Distribution and Seasonal Bio-Ecology of Naturally Infected Sand Flies in a New Endemic Zoonotic Cutaneous Leishmaniasis Focus of Southern Iran.

BACKGROUND: Cutaneous leishmaniasis is a major health problem in Iran in spite of implementation of control program. This infectious disease caused morbidity in less than 27000 people in 2010. This study was set to determine some ecological aspects of sand flies in Fasa district, Fars Province, southern Iran during 2011-2012. METHODS: A total of 4792 sand flies were captured by means of sticky paper and CDC miniature light traps in 10 selected villages from the beginning to the end of the active season, from which 1115 specimens were captured for abundance study and 3677 specimens captured for monitoring monthly activities in Fasa. After species identification, extracted DNA was processed for detection of Leishmania parasite infection in sand flies. RESULTS: Twelve species (6 Phlebotomus, 6 Sergentomyia) were identified. The most common sand fly was P. papatasi (82.4%) which represented 86.6% of sand flies from indoors and 82.7% from outdoors. The monthly activity of the species extended from April to the end of November. There were two peaks in the density curve of this species, one in June and the second in September. Natural infection to L. major was detected in P. papatasi (25 out of 130 sand flies, 19.2%). CONCLUSION: Phlebotomus papatasi is considered as a main vector of zoonotic cutaneous leishmaniasis in Fasa, Fars Province, south of Iran.

Molecular detection of Leishmania parasites and host blood meal identification in wild sand flies from a new endemic rural region, south of Iran.

Zoonotic Cutaneous Leishmaniosis (ZCL) remains the most crucial vector-borne public health disease particularly in endemic rural parts of Iran. The main aim of this study is to identify wild sand flies (Diptera: Psychodidae), determine their infection rate, and differentiate their host blood meal sources using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Sand fly populations were caught with sticky paper traps from 10 different villages in the county of Darab, Fars province, southern Iran. Following their species identification, they were used in one step PCR to determine their infection with Leishmania spp. parasites. They were then subjected to PCR-RFLP protocol to identify and differentiate their blood meal sources. Two genera of Phlebotomus and Sergentomyia comprising 13 species of sand flies were identified in this region. From a total of 150 parous female sand flies, encompassing 4 different medically important species, 7 specimens (4.7%) including 6 Phlebotomus papatasi and 1 Phlebotomus bergeroti were infected with Leishmania major. Molecular data indicated that about 32% of female sand flies fed on man, while nearly 43% fed on rodent and canine hosts. Molecular detection is an efficient way of differentiating the source of blood meals in female sand flies feeding on different vertebrate hosts. It is suggested that P. papatasi is not highly anthropophagic and appears to be an opportunistic feeder on man. This species is, however, the primary vector of ZCL in this region.

In vitro efficacy of ethanolic extract of Artemisia absinthium (Asteraceae) against Leishmania major L. using cell sensitivity and flow cytometry assays.

Leishmaniasis is one of the most neglected human diseases with an estimated global burden ranking second in mortality and fourth in morbidity among the tropical infections. Chemotherapy involving the use of drugs like glucantime is the mainstay treatment in endemic areas of Iran. Drug resistance is increasingly prevalent, so search for alternative therapy is gathering pace. Medicinal herbs, like wormwood Artemisia, have chemical compounds effective against a number of pathogens. In this study, the efficacy of ethanol extract from Artemisia absinthium (Asteraceae) against Leishmania major L. was investigated in vitro. The outcome of different effective doses (1-40 mg/ml) of ethanol extracts from this medicinal herb, A. absinthium, on a standard Iranian parasite strain of L. major was examined. The L. major promastigote cell sensitivity and mortality or viability effects due to the addition of herbal extract were measured using the MTT assay and the flow cytometry technique, respectively. There was complete agreement between the two assays. The lethal concentration (LC50) was measured as 101 mg/ml. Some contrasting relationships between the medicinal herb concentrations and the viability of parasites were observed; so that there was an increased multiplication of the parasite at low concentrations of the drug, but an anti-parasitic apoptotic effect was seen at high concentrations of A. absinthium. It was concluded that there might be one or more chemical constituents within the herbal extract of wormwood which at high concentration controlled cell division and affected the relevant activity within the only one giant mitochondrion in this flagellate parasite. At low doses, however, it showed the opposite effect of leading to mitotic cell divisions.

Epidemiologic profile of oriental sore caused by Leishmania parasites in a new endemic focus of cutaneous leishmaniasis, southern Iran.

Cutaneous leishmaniasis (CL) is the first and most important vector-borne zoonotic disease transmitted by sand flies in Iran. As a parasitic disease in the Old World, it is a complex zoonosis with multiple vertebrate hosts and arthropod vectors of pathogenic flagellate protozoan in the genus of Leishmania in different parts of its range. Phlebotomine sand flies are proven as vectors of this parasite which can be transmitted through the bite of an infected female sand fly distributed in almost all parts of Iran. This research performed on all CL patients as that were registered into special forms by physicians and experts during the study period 2006-2013 in the county town of Fasa, Iran. Data were analyzed by Chi square test using SPSS 17 statistics software. Overall, 1,908 patients (59.18 %) lived in rural and 1,316 (40.82 %) lived in urban areas. All ages were between 1 and ≥30 year. The most frequent age group was ≥20 years (54.6 %). Sex ratio of patients was almost 1:1 (1,561; 48.42 % male vs. 1,663; 51.58 % female). Most of them (66.84 %) had wet lesions and those with dry lesions were less frequent (33.16 %). There was a significant difference between the frequencies of these two groups (P < 0.05). Hand ulcers were the most prevalent part of body (43.24 %). The highest prevalence rate (35.14 %) of lesions occurred in autumn. The unstable trend of this disease in different years and its relatively high disease burden affecting all age groups in Fasa with respect to other counties in Iran showed that it was most likely an endemic disease in this region.

Natural transovarial and transstadial transmission of Leishmania infantum by naïve Rhipicephalus sanguineus ticks blood feeding on an endemically infected dog in Shiraz, south of Iran.

BACKGROUND: The visceral leishmaniasis parasite, Leishmania infantum, is naturally transmitted through the bites of phlebotomine sand flies. Alternative routes of transmission are questioned. The main aim is to verify the passage of L. infantum kDNA in ticks, Rhipicephalus sanguineus, blood feeding on a parasitemic dog in Shiraz, south of Iran. METHODS: A total of 180 Leishmania-free ticks collected from fields and bred on lab rodents, were divided into eight groups and allowed to feed on a dog (Canis familiaris) for fixed periods of time. These and all third generation stages of ticks were checked for L. infantum kDNA using conventional PCR protocol. RESULTS: The infection rate was significantly higher in female than male ticks (p=0.043). The rates were higher among nymphs (25/60; 42%) than adult ticks (37/120; 30.8%). The kDNA of L. infantum was not detected in ticks 24 h post-feeding. It was, however, positive among the second to fourth groups of nymphs (4/10; 40%, 10/20; 50% and 11/20; 55%) and adult (12/30; 40%, 14/30; 46.6% and 11/30; 36.6%) ticks. Eggs and unfed larvae recovered from the third and fourth adult groups (2 weeks, 4 weeks) were 100% PCR-positive. The data revealed the passage of L. infantum kDNA in nymphs and adults of brown dog tick following fixed time intervals post blood feeding on an infected dog. CONCLUSIONS: The natural transovarial and transstadial passage of kDNA through ticks was shown by PCR.

Faunal distribution of fleas and their blood-feeding preferences using enzyme-linked immunosorbent assays from farm animals and human shelters in a new rural region of southern Iran.

Blood sucking insects, such as fleas, are responsible for the transmission of many infectious disease-causing agents which impose an intolerable burden on the health of people living particularly in endemic parts of the world. Fleas (Insecta: Siphonaptera) are found in many parts of the world including Iran. Both adult male and female fleas are obligatory ectoparasites. They are one of the main public health concerns as a result of their nuisance or the potential to act as vectors of a number of medically-important pathogens. The current study was conducted to examine the geographical distribution and fauna of fleas and their anthropophagic index in part of Fars province, southern Iran. This study was the first to be done in Iran. A total of 20 villages were randomly selected. From October 2011 to May 2012, adult fleas were collected by direct hand catch from human to animal shelters. Overall 848 fleas, most of which were blood-fed, were captured from the floor or the body of farm animal hosts (cattle, sheep, goat and hens). Only two different genera of fleas were identified, the main species (99.76 %) was human flea, Pulex irritans. The village of Shamsabad was the most heavily infested area. P. irritans had an anthropophagic index of 15 % using indirect enzyme-linked immunosorbent assays (ELISA). It could be concluded that P. irritans is widely distributed in this area. Based on their blood feeding activity, fleas thus posed a serious health threat to residents and their economically important livestock in this part of Iran.

First phylogenetic analysis of a Crimean-Congo hemorrhagic fever virus genome in naturally infected Rhipicephalus appendiculatus ticks (Acari: Ixodidae).

Crimean-Congo haemorrhagic fever (CCHF) is a potentially fatal systemic viral disease in many parts of the world, including Iran. The nationwide incidence of human CCHF in endemic areas was 870 confirmed cases with 126 deaths (case fatality rate, CFR = 17.6 %) in the decade leading to 2012. The detection of the CCHF virus (CCHFV) genome in tick vectors is of fundamental importance for identifying these ticks as potential reservoirs of CCHFV infection. From May to October 2013, following detection of four new clinical cases resulting in two deaths in the city of Mashhad (northeast Iran), hard ticks were recovered from infested livestock in 40 villages in Khorasan-Razavi province and examined by the microscopic method for species identification. About a quarter of the ticks were then subjected to reverse-transcription polymerase chain reaction (RT-PCR) to detect the CCHFV genome. The PCR products were then sequenced, and their phylogenetic lineages were determined. A total of 407 hard ticks were captured, representing seven different species in two distinct genera. Members of the genus Hyalomma were widely distributed in all but two of the villages studied, and this was also the most frequent (83.3 %) tick genus. Of 105 adult ticks subjected to RT-PCR, four (3.8 %) ticks were found positive for the CCHFV genome. One brown ear tick, Rhipicephalus appendiculatus, was found to be naturally infected for the first time anywhere in the world. Ticks of Hyalomma asiaticum, Hyalomma marginatum, and Rhipicephalus turanicus were also found to be naturally infected with CCHFV. CCHFV found in these four different tick species were clustered in the same lineage with the Matin and SR3 strains from Pakistan and some other strains from Iran, indicating that these tick species were naturally infected with genetically closely related CCHFV in the region. The presence of CCHFV infection in four different hard tick species was confirmed using RT-PCR in northeast Iran. Part of this infection was attributed to Rh. appendiculatus, which is thus a potential new natural vector of CCHFV in Iran. It is also confirmed by phylogenetic analysis that CCHFV in this region is genetically closely related, even in the different tick species.

Nested polymerase chain reaction and sequence- based detection of leishmania infection of sand flies in recently emerged endemic focus of zoonotic cutaneous leishmaniasis, southern iran.

BACKGROUND: Geographical distribution of zoonotic cutaneous leishmaniasis (ZCL) has continuously been extended in recent years in Iran. The Beiza District is one of the newly-emerged endemic foci of ZCL in southern Iran. The main aim of the present study was to detect the vector(s) of ZCL in this area. METHODS: To detect the fauna and vectors of ZCL in this district, sand flies were caught using sticky papers. Seventy randomly selected female sand flies out of 730 were molecularly investigated for Leishmania infection using species-specific nested polymerase chain reaction (PCR) assay between April and October 2010. RESULTS: A total of 2543 sand flies were caught. The fauna was identified as 10 species (five Phlebotomus spp. and five Sergentomyia spp.). Phlebotomus papatasi was the most dominant species both indoors and outdoors (37.55% and 16.35 %, respectively). L. major was detected in 5 out of 48 investigated Phlebotomus papatasi (10.41%). Sequence-based characterization was carried out to confirm the PCR findings. The positive samples were shown to have 75-88% similarity with L. major sequences in GenBank. CONCLUSION: According to the findings of the present study, similar to the other foci of ZCL in Iran, P. papatasi is the proven and primary vector of CL. This study could be drawn upon for future strategy planning in this newly emerged endemic focus.