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.

Determination of the feeding behavior of Phlebotomus sergenti using multiplex PCR and tent-baited traps in a new focus of Anthroponotic Cutaneous Leishmaniasis in the southeast of Iran.

OBJECTIVES: This study aimed to determine feeding behaviors of Phlebotomus sergenti Parrot, in a new focus of Anthroponotic Cutaneous Leishmaniasis in Bam County, southeast Iran. METHODS: Two methods were used to determine the feeding behavior of Phlebotomus sergenti. In the first method, blood-fed sand flies were captured using a mouth aspirator in human and animal dwellings and consequently, blood meal identification was made using Multiplex PCR. The results were used for calculating Host Feeding Index (HFI) and Forage Ratio (FR) parameters. In the second method, human (Homo sapiens), goat (Capra aegagrus), cattle (Bos taurus), chicken (Gallus gallus) and dog (Canis lupus) were used as baits in tent-baited traps to determine the feeding behavior of Phlebotomus sergenti. RESULTS: Multiplex PCR analysis revealed that the most frequent blood in the stomack of sand flies' were from chicken, but the calculation of the FR revealed that this species prefers canine and poultary blood as meal. Human and animal tent-baited traps revealed that most Phlebotomus sergenti were attracted to chicken rather than the other hosts. CONCLUSIONS: Sand flies are attracted to animals for various reasons such as eating blood, mating on their bodies and laying eggs on their feces. Molecular methods are effective and accurate methods to determine the type of host that sandfly fed on, but they do not show host preferences. The results of the molecular analysis, along with the calculation of HFI and FR, can determine the preferred host of sand flies. The current study revealed that dogs, the secondary reservoir of ACL in Iran, is the first preferred host of Phlebotomus sergenti.

Susceptibility of Culiseta longiareolata to insecticides in West Azerbaijan province, northwestern Iran.

Mosquitoes are involved in the transmission of a wide range of diseases and among them, Culiseta longiareolata, acts as a vector of avian malaria, tularemia and several arboviruses like West Nile fever. The current study was conducted to determine the susceptibility of C. longiareolata against deltamethrin 0.05% (pyrethroids), fenitrothion 1.00% (organophosphate) and bendiocarb 0.10% (carbamate). Mosquitoes were collected from different parts of Urmia county, the capital of West Azerbaijan. In the current study, 443 empty 3 - 5 days old adult female C. longiareolata were used for susceptibility tests. The susceptibility status of C. longiareolata was determined using WHO's recommended procedure. The results indicated higher mortality rates of Culiseta longiareolata against studied insecticides. Culiseta longiareolata, was resistant to all studied insecticides (deltamethrin, mortality rate: 62.50%, fenitrothion, mortality rate: 35.96% and bendiocarb, mortality rate: 20.16%). The current study represents the first report of susceptibility status of C. longiareolata to fenitrothion and bendicarb in Iran. Considering the climatic diversity and geographical conditions of northwestern Iran, reports of resistance of other mosquitoes' of this region, the susceptibility status of medically and veterinary important mosquitoes should be given serious attention.

New Record of Spiders (Arachnida: Araneae) from Bashagard District, Southern Iran, with the Report of a Medically Important Species.

Spiders are the largest order of arachnids with some medically important species. Considering that no comprehensive research has been conducted on the fauna and distribution of Araneae in Bashagard County (Hormozgan Province) so far, the present investigation has been carried out on these essential issues in this deprived tropical region. Fifteen sampling locations were selected based on the extent of each area and considering climatic characteristics. Samplings were carried out at least four times in each region from February 2017 to September 2018, covering up to a radius of 500 meters from the defined locations. Specimens were collected using the hand collection method, pitfall trapping, and aspirator sampling. All collected samples were preserved in 70% ethanol and were identified using valid taxonomic keys. Of all the 390 collected samples, 134 identifiable specimens were considered for morphological identification. 11 species belonging to 10 genera and seven families were identified. Immature specimens were identified only at the family level. Wadicosa fidelis was the most distributed and abundant species in the area, with 13 localities and 84.33% of all identifiable samples. Plexippus minor is a new record for the spider fauna of Iran. Moreover, Loxosceles rufescens, a medically important species, Artema doriae, and Eusparassus mesopotamicus were reported for the first time from Hormozgan province. Despite collecting only one specimen, of L. rufescens, due to the lack of proper medical facilities and transportation systems in the area, health staff must be alert about this medically important species and warn residents about its potential dangers.

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.

First molecular-based detection of SARS-CoV-2 virus in the field-collected houseflies.

This is the first report of SARS-CoV-2 detection on field-collected Musca domestica housefly surface and tissue samples using the high-sensitive PCR assay which suggests the possible insect-borne transmission. The study was conducted in Shiraz city, southern Iran, in May and Jun 2020. Adult flies were sampled at the outdoor areas of two hospitals treating COVID-19 patients. Fly samples were first washed twice to remove the insect surface attached to SARS-CoV-2 virions. After that, the disinfected fly samples were homogenized. Fly surface washout and homogenate samples were tested using Taq Man real-time PCR assay for the SARS-CoV-2 virus. In a total of 156 houseflies, 75% of samples from the body washout samples were positive for SARS-CoV-2. Strikingly, 37% of the homogenized specimens were positive for the SARS-CoV-2, suggesting the possible infection of the insects or uptake of the virion to the insect metabolism. The other possibility is the houseflies up took the blood or blood fluids of the patients and the RNA of the SARS-CoV-2 survived in the insect body without replicating. Our preliminary findings suggest that the houseflies could transmit SARS-CoV-2 as a mechanical or biological vector especially during the warm seasons while increasing the population and activity of houseflies.

A comprehensive entomological survey and evaluation of the efficacy of different therapies on a suspected delusional parasitosis case.

OBJECTIVE: Delusional parasitosis (DP) is one type of psychotic disorders. It is a multifactorial disorder with different etiologies. Given that very little attention is paid to entomological survey in these cases, a comprehensive study was designed and performed for the first time in Iran on a suspected DP case and its home from Shiraz during 2018-2019. METHODS: In this study, entomological survey, dermatological studies, and psychological and psychiatric interventions were done respectively on a 40-year-old man who was referred to as a suspected case of DP. RESULTS: No medical importance arthropods were collected from the patient's home. He was not infected with lice and other ectoparasites. Skin samples were negative for scabies, but he was infected with Demodex folliculorum at low level. Psychiatric studies showed that he was a secondary DP case with depression. No effect and partial remission were reported after treatment with risperidone (2-6 mg/d) and olanzapine (5 mg/d), respectively. Therapeutic effects of hypnotism were also not satisfactory. CONCLUSIONS: In Iran, the most important challenges these patients faced with are as follows: the absence of a specific referral center, patients resort to self-treatment or traditional methods, and lack of a certain duration and dosage of antipsychotic for these cases. Comprehensive clinical trials should be done on this rare syndrome at the national level for better understanding the epidemiological profile of DP and finding the best method of treatment for Persian community.

Molecular Characterisation of Cytochrome Oxidase I and Internal Transcribed Spacer 2 Fragments of Culiseta longiareolata.

Objective: Among the mosquitoes, Culiseta longiareolata plays a notable role in the transmission of avian malaria, tularemia and arboviral diseases, including West Nile fever. We conducted this study to characterise the cytochrome oxidase I (COI) and internal transcribed spacer 2 (ITS2) fragments of Cs. longiareolata in northwestern Iran to determine the classification status of this species. Methods: The COI and ITS2 fragments from six populations of Cs. longiareolata were amplified, sequenced and analysed. For phylogenetic analysis, the evolutionary history was estimated using the Tamura-Nei-based Maximum Likelihood approach. Results: Thirteen sequences (six for ITS2 and seven for COI) from six populations of Cs. longiareolata were acquired and deposited into the GenBank. Phylogenetic analysis showed that the COI sequences from the current study cluster together with the same species from other parts of the world. Moreover, the ITS2 sequences of the current study and sequences retrieved from the GenBank, despite intraspecies variation, fall into a distinct clade with acceptable bootstrap values. Conclusion: Notable genetic variations were observed between various Cs. longiareolata populations based on the evaluations of ITS2 and COI fragments. By conducting such studies, the exact classification status of this species can be determined.

Identification of symbiotic bacteria in the midgut of the medically important mosquito, Culiseta longiareolata (Diptera: Culicidae).

OBJECTIVE: The potential use of symbiotic bacteria for the control of mosquito-borne diseases has attracted the attention of scientists over the past few years. Culiseta longiareolata is among the medically important mosquitoes that transmit a wide range of vector-borne diseases worldwide. However, no extensive studies have been done on the identification of its symbiotic bacteria. Given the role of this species in the transmission of some important diseases and its widespread presence in different parts of the world, including northwestern parts and the West Azerbaijan Province in Iran, a knowledge about the symbiotic bacteria of this species may provide a valuable tool for the biological control of this mosquito. Accordingly, the present study was conducted to isolate and identify the cultivable isolates bacterial symbionts of Culiseta longiareolata using 16S rRNA fragment analysis. RESULTS: The midguts of 42 specimens of Cs. longiareolata were dissected, and the bacteria were cultured on agar plates. After the purification of the bacterial colonies, 16srRNA region amplification and gene sequence analysis were performed, and the sequences were confirmed by biochemical methods. In the present study, 21 isolates belonging to the genera Acinetobacter, Aerococcus, Aeromonas, Bacillus, Carnobacterium, Klebsiella, Morganella, Pseudomonas, Shewanella and Staphylococcus were identified.