Leishmania major Infection in Synanthropic Rodents: Evidence for the Urbanization of Zoonotic Cutaneous Leishmaniasis (ZCL) in Southern Iran.

Cutaneous leishmaniasis is of particular importance in southern Iran. This study aimed to investigate the infection of rodents with Leishmania major in an urban area of Fars Province, located in southern Iran. Rodents were trapped and samples from the liver, spleen, and skin were collected. Impression smears were prepared from these tissues and any skin lesions and were examined microscopically. In addition, a portion of the samples were preserved for subsequent DNA extraction. A total of 41 rodents belonging to three species were caught from 10 trapping stations in gardens or houses within the area. The caught rodent species were Rattus rattus (n = 25, 60.97%), Mus musculus (n = 15, 36.58%), and Meriones persicus (n = 1, 2.5%). Leishmania amastigotes were seen in the spleen tissue smear of 6 (2.43%) of the rodents, including 4 of R. rattus and 2 of M. musculus. Skin lesions were observed on the muzzles of two R. rattus and one M. musculus. Samples taken from these lesions tested positive for Leishmania infection. Leishmania DNA was detected in 18 (43.9%) rodents, including 11 R. rattus, 6 M. musculus, and one M. persicus, based on DNA sequencing of the ITS2 gene and PCR of the kDNA. Phylogenetic reconstruction revealed that the parasite infecting the rodents was L. major. The detection of Leishmania infection in these rodents in urban areas raises concerns about the urbanization of cutaneous leishmaniasis caused by L. major. This urbanization poses unique challenges for control and prevention efforts.

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.

Mosquito surveillance and the first record of morphological and molecular-based identification of invasive species Aedes (Stegomyia) aegypti (Diptera: Culicidae), southern Iran.

Ae. aegypti is an important vector for transmission of some dangerous arboviral diseases, including Dengue Fever. The present study was conducted (from August 2017 to January 2020) to survey the fauna of Culicine mosquitoes, emphasizing the existence of this invasive species in oriental parts of the country located near the Persian Gulf. Different sampling methods were used to collect all life stages of the mosquito. After morphological identification, a molecular study based on Cytochrome Oxidase (COI) gene-specific primers was performed. Then, the COI gene was sequenced via the Sanger method. A total of 4843 adults and 11,873 larvae were collected (8 species of Culex, one species of Culiseta, and 5 species of Aedes). Fifty-five Ae. aegypti specimens (8 adults and 47 larvae) were identified. Based on the biology and ecological requirements of Ae. aegypti, the possibility of the permanent establishment of this species in the tropical climate of the region is very likely. Considering the detection of this invasive vector mosquito species in Iran and the high incidence of some arboviral diseases in the neighboring countries, and continuous movements of the settlers of these areas, potential outbreaks of arboviral diseases can be predicted. Planning and implementing an immediate surveillance and control program of the vector mosquito is vital to prevent the permanent establishment of this invasive vector mosquito species in southern Iran.

In vitro evaluation of CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania major as a primary step to control leishmaniasis.

Cutaneous leishmaniasis (CL) is caused by intracellular obligate parasites (Leishmania spp.) carried by the blood-sucking of female sandflies and transmitted between mammalian hosts. Despite the high incidence and prevalence of Leishmania cases in many countries, it has been a neglected tropical disease. The current treatment approaches are limited by the complications such as loss of fertility and drug resistance. It is, therefore, essential to find new medicines to treat leishmaniasis. CRISPR/Cas9 as a powerful genome-editing tool provides the opportunity to create precise genetic manipulation to investigate the molecular basis of different leishmaniasis cases. Therefore, our main goal was to evaluate the CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania majorin vitroto challenge for using CRISPR/Cas9 as a therapeutic CL through the reduction of L. major pathogenicity by manipulating the GP63 gene. In this study, L. major parasites were transfected with CRISPR/Cas9 vectors constructed by electroporation and then added to macrophage cells on RPMI. The effect of CRISPR/Cas9 constructs on GP63 mutation, viability, and status of L. major was investigated by counting phagocytic parasites into macrophages and DNA sequence analysis. Our data validate that the use of CRISPR/Cas9 in L. major creates a new stop codon and disrupts the frame sheet of the gene by creating a new insertion (thymine), which prevents its expression. In addition, the parasite count was significantly different in the case and control of infected macrophages (P < 0.05). This study shows the successfully targeted manipulation of the L. major GP63 gene via the adaptation of the CRISPR/Cas9 editing tool. The manipulation of GP63 revealed a reduction in the infection load compared to wild-type parasite infection. Therefore, more studies are necessary for this field to help achieve a new method for the prevention and treatment of CL disease.

DNA-based detection of Leishmania and Crithidia species isolated from humans in cutaneous and post-kala-azar dermal leishmaniasis from Shiraz and Kharameh, southern Iran.

BACKGROUND & OBJECTIVES: Leishmania major and L. tropica are the main pathogens of cutaneous leishmaniasis (CL) in several rural and some urban regions of Iran, respectively. The aim of this study was to detect Leishmania species, and update the distribution data of these species in humans suspected to CL in two endemic foci in southern Iran. METHODS: From March 2016 to March 2017, 276 positive samples from of 350 suspected cases were diagnosed and compared by different diagnostic methods, viz. microscopy, culture, and PCR. In PCR assay, four different gene identifications were performed including minicircle kDNA, and cysteine protease B genes for Leishmania detection, and glyceraldehyde-3-phosphate dehydrogenase, and internal transcribed spacer 1 genes for Crithidia detection. RESULTS: In total, 68% (235/350) and 65.3% (177/271) of patients suspected of leishmaniasis were positive by microscopy and cultivation methods. In PCR assay, L. major, and L. tropica were detected in 86.2% (238/276), and 13.1% (36/276) of CL cases, respectively. Also, dermal L. infantum strain was isolated from 0.7% (2/276) of post-kala-azar dermal leishmaniasis patients. In addition, Crithidia fasciculata was detected in two CL patients chronically infected with L. major. INTERPRETATION & CONCLUSION: It appears that the epidemiology of CL has changed during the last decades and can complicate the control strategy aspects of CL in southern Iran. Therefore, more epidemiological, ecological, and gene polymorphism studies are needed to understand the pathogenic role of these species in human, as a main host of leishmaniasis in Iran.

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.

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.

First report on the molecular phylogenetics and population genetics of Aedes aegypti in Iran.

BACKGROUND: Aedes aegypti, the primary vector of various human arboviral diseases, is a significant public health threat. Aedes aegypti was detected in Iran in 2018, in Hormozgan province, but comprehensive information regarding its genetic diversity and origin within the country remains scarce. This study aimed to determine the origin and genetic diversity of Ae. aegypti in southern Iran. METHODS: Aedes aegypti mosquitoes were collected from Bandar Abbas City, Hormozgan Province, southern Iran, between May and July 2022. Specimens were morphologically identified. Origin and assess genetic diversity were assessed based on the mitochondrial DNA-encoded cytochrome c oxidase subunit I (mtDNA-COI) gene. RESULTS: BLAST (basic local alignment search tool) analysis confirmed the accuracy of the morphological identification of all specimens as Ae. aegypti, with 100% similarity to GenBank sequences. Calculated variance and haplotype diversity were 0.502 and 0.00157, respectively. Among the 604 examined nucleotide sequences, only a single site was non-synonymous. Total nucleotide diversity and average pairwise nucleotides were determined as 0.00083 and 0.502, respectively. Fu and Li's D test values were not statistically significant. Strobeck's S statistic value was 0.487, and Tajima's D value was 1.53395; both were not statistically significant (P > 0.10). CONCLUSIONS: Phylogenetic analysis revealed two distinct clades with minimal nucleotide differences and low haplotype diversity, suggesting the recent establishment of Ae. Aegypti in the southern region of Iran. The phylogenetic analysis also indicated an association between Ae. aegypti populations and mosquitoes from Saudi Arabia and Pakistan.

Cloning, expression and molecular analysis of recombinant Netrin-A protein of Lucilia sericata Meigen (Diptera: Calliphoridae) larvae.

Objectives: Lucilia sericata (Diptera: Calliphoridae) is used in larval therapy for wound healing. Netrin-A is an enzyme secreted from the salivary glands of these larvae, and has a central role in neural regeneration and angiogenesis. This study aimed to produce the recombinant Netrin-A protein from Lucilia sericata larvae by the baculovirus expression vector system in the Sf9 insect cell line. Methods: The coding sequence of Netrin-A was cloned, amplified in the pTG19 vector, and then cloned in the pFastBac HTA vector. It was then transformed into DH10Bac, and the recombinant Bacmid was subsequently transfected into Sf9 cells. The recombinant Netrin-A was purified by Ni-NTA agarose. The evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay. Results: The molecular weight of this protein was 52 kDa with 404 amino acids. The signal peptide was located between amino acids 24 and 25. The concentration of Netrin-A was calculated to be 48.8 µg/ml. It reaffirmed the characterized gene codes of Lucilia sericata Netrin-A in a previous study. Conclusions: The generation of recombinant Netrin-A could be used in larval therapy, and as a biomarker in certain diseases. The netrin-A of Lucilia sericata was unprecedentedly cloned and expressed in a eukaryotic cell line. Given that this larva is FDA-approved, and non-pathogenic, it conduces to research on the development of maggot therapy in future.

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.

Calomyscid Rodents (Rodentia: Calomyscidae) as a Potential Reservoir of Zoonotic Cutaneous Leishmaniasis in a Mountainous Residential Area in the Plateau of Iran: Inferring from Molecular Data of kDNA and ITS2 Genes of Leishmania Major.

Cutaneous leishmaniasis (CL), a neglected tropical disease, is an important health problem in Fars Province, southern Iran. Fars, the fourth most populous Province in Iran, is the center of both anthroponotic and zoonotic cutaneous leishmaniasis (ZCL). Rodents, the reservoir of Leishmania major, play an important role in transmitting ZCL. In the present study, we report Leishmania infection in calomyscid rodents for the first time in mountainous residential areas of Shiraz, the capital of Fars Province, in southern Iran. Rodents were trapped in urban mountainous areas. The skin, liver, and spleen of rodents were examined microscopically for Leishmania infection. In addition, DNA was extracted from the tissues and they were evaluated for Leishmania infection by targeting the kDNA and subsequent sequencing of the nuclear rDNA internal transcribed spacer two (ITS2) region. DNA of L. major was detected in the spleen and liver of calomyscid rodents. Molecular evolution based on DNA-sequencing of the ITS2 gene confirmed the taxonomic situation of the parasite as L. major. Our findings suggest the eco-epidemiological importance of calomyscid rodents in the foci of leishmaniasis in the mountainous residential area on the plateau of Iran. These rodents may play a role in the transmission of leishmaniasis in a residential area and could be considered a potential reservoir for CL.

Absence of Asymptomatic Malaria Reservoirs in an Area with a Previous History of Local Malaria Transmission: A Successful Experience in Line with the Malaria Elimination Program in Iran.

Background: Asymptomatic malaria is a major challenge to be addressed in the implementation of the malaria elimination program. The main goal of the malaria surveillance system in the elimination phase is to identify reliably all the positive cases of malaria reliably (symptomatic and asymptomatic) in the shortest possible time. This study focused on the monitoring of asymptomatic malaria reservoirs in areas where local transmission had been previously established. Methods: It was a case-study approach that was conducted in the Anarestan area. A total of 246 residents and immigrants living in the area at the age range of 4-60 years old were randomly selected to be tested for malaria by microscope, RDT, and nested-PCR techniques. The inclusion criterion for participants to be entered into the study was the absence of specific symptoms of malaria. Moreover, participants who have been taking antimalarials for the last month were excluded from the study. Results: The results indicated no positive cases of asymptomatic malaria among the participants tested by all methods. Conclusion: The results of this study have shown that, without concerns for asymptomatic parasitic patients, a malaria elimination program has been successfully implemented within the studies area. In addition, the findings emphasized the existence of a strong malaria surveillance system in this area.

First molecular detection of SARS-CoV-2 virus in cockroaches.

Coronavirus is one of the main pathogens that primarily targets the human respiratory system. There are several ways to transmit this virus, such as direct contact or droplets spread by coughing or sneezing, and direct contact with fomites and surfaces is another way. This cross-sectional study was conducted in Shiraz, southern Iran, in 2021. 5 locations, including 3 hospitals and 2 dormitories, were selected for the survey. The cockroaches were collected from selected locations and transferred to the Laboratory of Medical Entomology at Shiraz University of Medical Sciences. All specimens were identified morphologically. The external and gastrointestinal washouts of collected samples with sterile phosphate-buffered saline separately were used for molecular analysis. An RT-qPCR assay, which suggests the possible insect­borne transmission, was used. External and gastrointestinal washout of B. germanica from Dastgheyb Dormitory and P. americana from Ali-Asghar Hospital were positive for contamination with the SARS-CoV-2. Cockroaches spread the virus in the environment and contaminate human food and various surfaces of buildings. Their role will be more important in crowded places such as hotels, lodging houses, restaurants, and hospitals; vector control programs should be carried out with more accuracy in such places.

Identification and frequency distribution of Leishmania (L.) major infections in sand flies from a new endemic ZCL focus in southeast Iran.

Zoonotic cutaneous leishmaniasis (ZCL) is an important public health challenge in Iran. It is often caused by the protozoan parasite, Leishmania major. This pathogen is principally transmitted by the infectious bites of adult female phlebotomine sand flies which belong to the subgenus Phlebotomus (Phlebotomus) Rondani and Berte (Diptera: Psychodidae). A recent outbreak of clinical ZCL cases in the rural district of Jask, Hormozgan province, southern Iran prompted the identification of sand flies naturally infected with L. major using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). A total of 8,123 wild sand flies were caught using sticky paper traps indoors and outdoors in seven villages of Jask from March 2007 to February 2008. About 120 trap-nights per month were carried out during peak seasonal density. Eight sand fly species of two distinct genera were morphologically identified. The most abundant species, Phlebotomus papatasi (≈60%) and Phlebotomus salehi (17%), had simultaneous population peaks in May and October. The anthropophilic index of P. papatasi was approximately twice that of P. salehi. PCR reactivity of L. major infections in P. salehi was weaker than those in P. papatasi. This is discussed with regard to their role in the natural transmission cycle of ZCL. This study provided the first PCR and ELISA evidence on P. salehi as a vector of L. major parasites in a new endemic region of ZCL in southeast Iran.

Investigating composition and production rate of healthcare waste and associated management practices in Bandar Abbass, Iran.

The objective of this study was to identify the composition and production rate of healthcare waste and associated management practices in healthcare centres in Bandar Abbas, southern Iran. A total of 90 centres, including 30 physician offices, 30 dental offices and 30 clinics were selected in random way. Two samples in summer and two samples in winter were taken and weighed from each selected centre at the end of successive working day on Mondays and Tuesdays. Results showed that the mean of daily production rate for each clinic, dental and physician office were 2125.3, 498.3 and 374.9 g, respectively. Domestic-type and potentially infectious waste had the highest and chemical and pharmaceutical waste and sharps had the lowest percentages in all centres. Questionnaire results indicated that there were no effective activity for waste minimization, separation, reuse and recycling in healthcare centres and management of sharps, potentially infectious and other hazardous waste was poor.

Therapeutic Potential of Bee and Scorpion Venom Phospholipase A2 (PLA2): A Narrative Review.

Venomous arthropods such as scorpions and bees form one of the important groups with an essential role in medical entomology. Their venom possesses a mixture of diverse compounds, such as peptides, some of which have toxic effects, and enzymatic peptide Phospholipase A2 (PLA2) with a pharmacological potential in the treatment of a wide range of diseases. Bee and scorpion venom PLA2 group III has been used in immunotherapy, the treatment of neurodegenerative and inflammatory diseases. They were assessed for antinociceptive, wound healing, anti-cancer, anti-viral, anti-bacterial, anti-parasitic, and anti-angiogenesis effects. PLA2 has been identified in different species of scorpions and bees. The anti-leishmania, anti-bacterial, anti-viral, and anti-malarial activities of scorpion PLA2 still need further investigation. Many pieces of research have been stopped in the laboratory stage, and several studies need vast investigation in the clinical phase to show the pharmacological potential of PLA2. In this review, the medical significance of PLA2 from the venom of two arthropods, namely bees and scorpions, is discussed.

Bionomics of Phlebotomine Sand Flies in Different Climates of Leishmaniasis in Fars Province, Southern Iran.

Background: Phlebotomus and Sergentomya are distributed in the old-world regions and transmit leishmaniases through mammalian and reptile hosts. Cutaneous leishmaniasis (CL) is one of the most important diseases in Iran. Iranian sand flies belong to three Oriental, Palearctic, and Afrotropical Regions. Fars Province is located in Palearctic, but southern parts are affected Oriental region situations on phglebotomine population variety. Therefore, a comprehensive study was required on the vectors of the disease in this Province. Methods: Regarding the approved role of the environmental factors in creating the fauna and distribution of living creatures, the "de martonne climate method" was used, and the climate was noticed as an important environmental factor for the determination of vector distribution. Accordingly, 14 sampling sites were selected from 10 foci in different climates of Fars Province. 19648 sand flies were collected from the studied areas in this Province during 2016. Results: Phlebotomus papatasi and Se. antennata were the most frequent species, which were caught from in/outdoor areas. Phlebotomus sergenti and Ph. alexandri were caught from both Palearctic and Oriental zones but were more prevalent in the cold semi-arid climate of the Palearctic zone. Moreover, they were not caught from the hot desert and summer Mediterranean climates of the Oriental zone. Conclusions: It seems that Ph. papatasi as the main vector of CL could be well distributed in different climates in Fars. Moreover, some species like Ph. sergenti preferred especial climates in the Palearctic zone. Therefore, these data could be helpful to control leishmaniases more efficiently.

The Fauna, Diversity, and Bionomics of Culicinae (Diptera: Culicidae) in Hormozgan Province, Southern Iran.

Hormozgan Province is one of the important foci of malaria in Iran. In addition to malaria pathogens, mosquitoes also transmit the pathogens that cause dirofilariasis and West Nile fever in the province. Also, the threat of emerging aedine-borne viruses that cause infections, such as Chikungunya, dengue, and Zika, is noticeable. There is little information about the fauna and bionomics of Culicinae in the province. The present investigation aimed to study the fauna, diversity, and bionomics of culicines. The study was conducted from September 2016 to April 2017 in four counties of Bandar Abbas, Bandar Khamir, Bashagard, and Jask. In total, 3,236 larvae and 1,901 adults including 16 culicine species were collected. The larvae of Culiseta longiareolata (Macquart) (25.65%), Culex pipiens Linnaeus (16.62%), and Cx. quinquefasciatus Say (16.16%) were most abundant and Cx. hortensis Ficalbi (0.09%) was least abundant. Among adults, Cx. laticinctus Edwards (33.19%), Cx. quinquefasciatus (31.09%), and Cx. pipiens (11.99%) were the most prevalent species and Cs. longiareolata (0.47%), Aedes caballus (Theobald) (0.90%), and Cx. bitaeniorhynchus Giles (0.90%) were the least prevalent species. The pairwise similarities of fauna of the counties and different collecting methods and diversity indices were investigated. More ecological data, especially on host preference, seasonality, and larval habitat characteristics, are needed as a basic knowledge for any intervention measures using integrated vector management.

Molecular Characterization and In Silico Analyses of Maurolipin Structure as a Secretory Phospholipase A 2 (sPLA2) from Venom Glands of Iranian Scorpio maurus (Arachnida: Scorpionida).

The venom is a mixture of various compounds with specific biological activities, such as the phospholipase A 2 (PLA2) enzyme present in scorpion venom. PLA2 plays a key role in inhibiting ryanodine receptor channels and has neurotoxic activity. This study is the first investigation of molecular characterization, cloning, and in silico analyses of PLA2 from Iranian Scorpio maurus, named Maurolipin. After RNA extraction from S. maurus venom glands, cDNA was synthesized and amplified through RT-PCR using specific primers. Amplified Maurolipin was cloned in TA cloning vector, pTG19. For in silico analyses, the characterized gene was analyzed utilizing different software. Maurolipin coding gene with 432 base pair nucleotide length encoded a protein of 144 amino acid residues and 16.34 kilodaltons. Comparing the coding sequence of Maurolipin with other characterized PLA2 from different species of scorpions showed that this protein was a member of the PLA2 superfamily. According to SWISS-MODEL prediction, Maurolipin had 38.83% identity with bee venom PLA2 with 100% confidence and 39% identity with insect phospholipase A 2 family, which Phyre2 predicted. According to the three-dimensional structure prediction, Maurolipin with five disulfide bonds has a very high similarity to the structure of PLA2 that belonged to the group III subfamily. The in silico analyses showed that phospholipase A 2 coding gene and protein structure is different based on scorpion species and geographical condition in which they live.

Cutaneous leishmaniasis in Iran: A review of epidemiological aspects, with emphasis on molecular findings.

Leishmania parasites can cause zoonotic cutaneous leishmaniasis (CL) by circulating between humans, rodents, and sandflies in Iran. In this study, published data were collected from scientific sources such as Web of Science, Scopus, PubMed, Springer, ResearchGate, Wiley Online, Ovid, Ebsco, Cochrane Library, Google scholar, and SID. Keywords searched in the articles, theses, and abstracts from 1983 to 2021 were cutaneous leishmaniasis, epidemiology, reservoir, vector, climatic factors, identification, and Iran. This review revealed that CL was prevalent in the west of Iran, while the center and south of Iran were also involved in recent years. The lack of facilities in suburban regions was an aggravating factor in the human community. Some parts of southern Iran were prominent foci of CL due the presence of potential rodent hosts in these regions. Rhombomys opimus, Meriones lybicus, and Tatera indica were well-documented species for hosting the Leishmania species in Iran. Moreover, R. opimus has been found with a coinfection of Leishmania major and L. turanica from the northeast and center of Iran. Mashhad, Kerman, Yazd, and sometimes Shiraz and Tehran foci were distinct areas for L. tropica. Molecular identifications using genomic diagnosis of kDNA and ITS1 fragments of the parasite indicated that there is heterogeneity in leishmaniasis in different parts of the country. Although cutaneous leishmaniasis has been a predicament for the health system, it is relatively under control in Iran.

Title: Leishmaniose cutanée en Iran : une synthèse des aspects épidémiologiques, mettant l'accent sur les découvertes moléculaires. Abstract: Les parasites Leishmania peuvent établir une leishmaniose cutanée zoonotique (LC) en circulant entre les humains, les rongeurs et les phlébotomes en Iran. Dans cette étude, les données publiées ont été collectées à partir de ressources scientifiques telles que Web of Science, Scopus, PubMed, Springer, ResearchGate, Wiley Online, Ovid, Ebsco, Cochrane Library, Google Scholar et SID. Les mots-clés recherchés dans les articles, les thèses et les résumés de 1983 à 2021 étaient leishmaniose cutanée, épidémiologie, réservoir, vecteur, facteurs climatiques, identification et Iran. Cet examen a révélé que la LC est répandue dans l'ouest de l'Iran, tandis que le centre et le sud de l'Iran sont également impliqués ces dernières années. Le manque d'équipements dans les régions suburbaines est un facteur aggravant dans la communauté humaine. Certaines parties du sud de l'Iran sont des foyers importants de LC en raison de la présence d'hôtes potentiels de rongeurs dans ces régions. Rhombomys opimus, Meriones lybicus et Tatera indica sont des espèces bien documentées pour héberger les espèces de Leishmania en Iran. De plus, R. opimus a été trouvé avec une co-infection de Leishmania major et L. turanica au nord-est et au centre de l'Iran. Mashhad, Kerman, Yazd et parfois des foyers de Shiraz et de Téhéran sont des zones distinctes pour L. tropica. Les identifications moléculaires utilisant le diagnostic génomique des fragments d'ADNk et ITS1 du parasite ont indiqué qu'il existe une hétérogénéité dans la leishmaniose dans différentes parties du pays. Bien que la leishmaniose cutanée ait été une situation difficile pour le système de santé, elle est relativement contrôlée en Iran.