Preliminary monitoring of knockdown resistance (kdr) mutation in Anopheles stephensi: insights from a malarious area in Southeastern Iran.

BACKGROUND: Anopheles stephensi is recognized as the main malaria vector in Iran. In recent years, resistance to several insecticide classes, including organochlorine, pyrethroids, and carbamate compounds, has been reported for this medically important malaria vector. The main objective of the present study was to evaluate the insecticide susceptibility status of An. stephensi collected from the southern part of Iran, and to clarify the mechanism of resistance, using bioassay tests and molecular methods comparing the sequence of susceptible and resistant mosquitoes. METHODS: Mosquito larvae were collected from various larval habitats across six different districts (Gabrik, Sardasht, Tidar, Dehbarez, Kishi and Bandar Abbas) in Hormozgan Provine, located in the southern part of Iran. From each district standing water areas with the highest densities of Anopheles larvae were selected for sampling, and adult mosquitoes were reared from them. Finally, the collected mosquito species were identified using valid keys. Insecticide susceptibility of An. stephensi was tested using permethrin 0.75%, lambdacyhalothrin 0.05%, deltamethrin 0.05%, and DDT 4%, following the World Health Organization (WHO) test procedures for insecticide resistance monitoring. Additionally, knockdown resistance (kdr) mutation in the voltage-gated sodium channel (vgsc) gene was sequenced and analysed among resistant populations to detect possible molecular mechanisms of observed resistance phenotypes. RESULTS: The susceptibility status of An. stephensi revealed that resistance to DDT and permethrin was found in all districts. Furthermore, resistance to all tested insecticides in An. stephensi was detected in Gabrik, Sardasht, Tidar, and Dehbarez. Analysis of knockdown resistance (kdr) mutations at the vgsc did not show evidence for the presence of this mutation in An. stephensi. CONCLUSION: Based on the results of the current study, it appears that in An. stephensi from Hormozgan Province (Iran), other resistance mechanisms such as biochemical resistance due to detoxification enzymes may be involved due to the absence of the kdr mutation or non-target site resistance. Further investigation is warranted in the future to identify the exact resistance mechanisms in this main malaria vector across the country.

Knowledge and Practices of Women Regarding Malaria and Its Prevention: A Community-Based Study in an Area under Malaria Elimination Programme in Iran.

Background: Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) are two core interventions for control and prevention of malaria infection. This descriptive cross-sectional study aimed to determine the knowledge and practices of women regarding malaria and its preventive measures in a malarious area in the Jask County, southeast of Iran. Methods: Data was collected from 400 households through a two-stage random cluster sampling method in six villages. Pre-tested structured questionnaire was used to collect data. During the survey, visual observations were also made by the interviewers to check use of LLINs and IRS. The data were analyzed by SPSS software version 21. Results: The majority of households had a high level of knowledge about symptoms (98%, n=392) and transmission (74.5%, n=298) of malaria. The main preventive measures against malaria were: LLINS (39.5%, n=158), IRS (32%, n=128), and screens on doors/windows (16.7%, n=67). In addition, participants with primary or higher educational level reported that they practice more malaria preventive measures, compared to those who were illiterate (OR=3.3, 95% CI= 1.6-6.6, p=0.0001, Table 6). In spite of positive perceptions about IRS and LLINs, only 35.5% (n=142) and 17% (n=68) of households used IRS and LLINs, respectively. Conclusion: According to the results of this study, despite a high level of knowledge about malaria and its preventive measures, a small number of participants use LLINs and IRS for malaria prevention. Therefore, implementation of effective educational interventions is recommended to improve preventive practices against malaria in the study area.

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.

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.

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.

Significant decline of malaria incidence in a low socioeconomic area in the southeast of Iran: 10 years field assessment during malaria elimination programme.

Although malaria burden and its active foci have sharply declined after the implementation of elimination programme since 2010, it is still considered as a major public health problem in southeast Iran. This descriptive-analytical study aimed to determine 10-years of malaria epidemiological trends in Bashagard County. Data were collected from 7 selected malarious region of the county based on active and passive surveillance of clinical cases. For diagnosis of malaria, the examination of microscopic slides and rapid diagnostic test, were used. In total, 237 malaria cases were found from 2009 to 2018. Plasmodium vivax was the dominant parasite species and identified in 232 (97.9%) individuals. Males were infected more than females and the majority of malaria cases (67.4%) were recorded from rural areas. Although about 98% of malaria cases were indigenous, they have decreased form 200 cases in 2009 to zero indigenous transmission in 2018. During the study period, malaria cases had decreased significantly by about 99% and the incidence rate had declined from 5.47/1000 cases in 2009 to 0.002/1000 in 2018. The incidence of malaria, especially indigenous cases, in Bashagard County has decreased dramatically in the past 10 years. However, there is still probability of malaria re-introduction and outbreak in the county due to climatical and geographical conditions. Therefore, it is necessary to implement an active surveillance system to detect and treat malaria cases quickly, during the elimination phase in this county.

Risk factors associated with scabies infestation among primary schoolchildren in a low socio-economic area in southeast of Iran.

BACKGROUND: Scabies is a neglected tropical disease caused by the mite Sarcoptes scabiei that burrows under the skin. It is a major health problem in tropical areas, largely affecting children. Scabies is common and highly contagious and in schoolchildren spreads quite rapidly, due to overcrowding and close contact within the schools. This study aimed to determine the risk factors associated with scabies infestation among primary schoolchildren in Bashagard County, one of the low socio-economic areas in southeast of Iran. METHODS: To conduct this community-based, descriptive, and cross-sectional study, four primary schools were randomly selected in the Bashagard County. All students in these schools were selected and examined for scabies. Clinical examination and sociodemographic profile of students were assessed using a pre-tested structured questionnaire. Chi-square test, and binary logistic regression were used to analyse the factors associated with scabies infestation by SPSS version 21 software. RESULTS: Out of 480 studied schoolchildren, 15 cases of scabies with a prevalence of 3.1 % were observed. The frequency of infestation in males was 1.6 % and it was 4.7 % in females. Independent factors associated with a high risk of scabies infestation in unadjusted analysis were being student of grade 5-6 (cOR = 13.12, 95 % CI 2.92-58.89, p = 0.0001), low educational level of father (cOR = 4.37, 95 % CI 0.97-19.59, p = 0.036), low educational level of mother (cOR = 4.14, 95 % CI 1.92-18.57, p = 0.045), joblessness of father (cOR = 14.77, 95 % CI 4.97-43.89, p = 0.0001), employment of mother (cOR = 5.28, 95 % CI 1.38-20.16, p = 0.007), large family size (cOR = 3.34, 95 % CI 1.05-10.64, p = 0.031), use of shared articles (cOR = 33.37, 95 % CI 10.82-102.90, p = 0.0001), and absence of bathroom in the house (cOR = 11.77, 95 % CI 2.16-63.94, p = 0.0001). CONCLUSIONS: Results of this study confirmed that scabies is still one of the most important health problems in the primary schools of the Bashagard County. Low socioeconomic status and personal hygiene of the schoolchildren were the most important factors influencing the prevalence of scabies. Improvement of socioeconomic conditions and implementation of appropriate educational programs and active surveillance system to quickly detect and treat scabies cases are necessary in order to reduce the prevalence of scabies in schoolchildren in this area.

Species composition, seasonal abundance and distribution of potential anopheline vectors in a malaria endemic area of Iran: field assessment for malaria elimination.

BACKGROUND: Despite decreases in incidence, malaria remains a major public health challenge in south and southeast Iran, where vector control is considered one of the main strategies for elimination of the disease. The efficacy of this strategy depends on understanding malaria vector ecology, which varies by species. This study was conducted to determine the species composition, seasonal abundance and distribution of potential anopheline vectors in Bashagard County, one of the important malaria-endemic areas in south Iran. METHODS: In this cross-sectional study, four typical foothill and mountainous villages were selected by simple random sampling. Anopheline mosquitoes were collected by the standard dipping method for larvae and total catch for adults. Anopheline specimens were morphologically identified using taxonomic keys. Statistical analyses were performed using SPSS ver.20 software. RESULTS: In total, 1211 anopheline specimens, including 1055 (87.12%) larvae and 156 (12.88%) adults, were collected and identified. They consisted of 9 species: Anopheles moghulensis (27.89%), Anopheles dthali (18.91%), Anopheles culicifacies (16.60%), Anopheles stephensi (15.38%), Anopheles turkhudi (9.83%), Anopheles superpictus (9.66%), Anopheles apoci (1.40%), Anopheles fluviatilis (0.17%), and Anopheles sergentii (0.08%). The most prevalent species in adult catches were An. culicifacies (44.23%), An. dthali (21.15%) and An. stephensi (16.03%), and the most prevalent species caught as larvae were An. moghulensis (31.94%), An. dthali (18.85%) and An. stephensi (15.26%). Most of the anopheline species were distributed in different topographical areas and two proven malaria vectors, An. culicifacies and An. superpictus, were significantly associated with altitude and collected more frequently in the foothill regions. Most of the anopheline species were present almost throughout the year with a major peak in April and a smaller peak in October. CONCLUSION: The results of this study revealed that there are five malaria vectors in Bashagard County and some of them are more abundant in areas with special topographic features and are reproductively active throughout the year. These findings can be applied to successful planning and providing effective control measures in problematic areas during the malaria elimination phase in Iran.

Species Composition of Sand Flies (Diptera: Psychodidae) and Modeling the Spatial Distribution of Main Vectors of Cutaneous Leishmaniasis in Hormozgan Province, Southern Iran.

Cutaneous Leishmaniasis (CL) is one of the main neglected vector-borne diseases in the Middle East, including Iran. This study aimed to map the spatial distribution and species composition of sand flies in Hormozgan Province and to predict the best ecological niches for main CL vectors in this area. A database that included all earlier studies on sand flies in Hormozgan Province was established. Sand flies were also collected from some localities across the province. Prediction maps for main vectors were developed using MaxEnt model. A total of 27 sand fly species were reported from the study area. Phlebotomus papatasi Scopoli, Phlebotomus sergenti s.l. Parrot, Phlebotomus alexandri Sinton, Sergentomyia sintoni Pringle, Sergentomyia clydei Sinton, Sergentomyia tiberiadis Adler, and Sergentomyia baghdadis Adler (Diptera: Psychodidae) had the widest distribution range. The probability of their presence as the main vectors of CL was calculated to be 0.0003-0.9410 and 0.0031-0.8880 for P. papatasi and P. sergenti s.l., respectively. The best ecological niches for P. papatasi were found in the central south, southeast, and a narrow area in southwest, whereas central south to northern area had better niches for P. sergenti s.l. The endemic areas are in Bandar-e Jask, where transmission occurs, whereas in Bastak, the cases were imported from endemic foci of Fars province. In conclusion, proven and suspected vectors of CL and VL were recorded in this study. Due to the existence of endemic foci of CL, and favorite ecological niches for its vectors, there is potential risk of emerging CL in new areas.

Household knowledge and practices concerning malaria and indoor residual spraying in an endemic area earmarked for malaria elimination in Iran.

BACKGROUND: Indoor residual spraying of insecticide (IRS) is a key intervention for reducing the burden of malaria infection. Effectiveness and success of this strategy are to a considerable extent dependent on knowledge and practice of the target community regarding the IRS. Iran has entered the malaria elimination phase, and IRS has been considered as the main strategy for malaria vector control. Therefore, this study was conducted to determine the household knowledge and practices about malaria and IRS in Bashagard County, one of the malaria-endemic areas in the southeast of Iran. METHODS: A community-based cross-sectional survey was conducted among 420 households in Bashagard County. The participants who were selected using a two-stage randomized cluster sampling procedure were subjected to a tested structured questionnaire. During the survey, direct observations were made concerning the use of IRS as well as housing conditions. The data were coded and analysed using SPSS version 19. RESULTS: Knowledge levels about malaria as a disease and the mosquito as its vector were high and of equal magnitude (85.5% and 85.4%, respectively), while knowledge levels of IRS were even higher (91.6%). The main source of households' information about malaria and IRS was primarily community health workers (73.3%). Despite positive perceptions towards IRS only 26.7% of respondents had sprayed their houses which is lower than the WHO targeted coverage of 80%. Respiratory disorders and headache (33.3%), food contamination (24.9%), discolouring of inner house walls (17.7%), difficulty in furniture's movement (13.8%), and unpleasant odour (10.4%) were the main reasons for IRS refusal. CONCLUSION: There is a discrepancy between knowledge about symptoms and the transmission route of malaria and control practices related to IRS use. Therefore, IRS campaigns accompanied with education for behaviour change should be considered to ensure householders' participation and cooperation in the IRS programme. Moreover, continuous evaluation and monitoring of IRS as well as conducting more surveys on knowledge, attitude, and practices are recommended to improve malaria control measures and to identify indicators for effective, successful, and sustainable malaria elimination programme.

Prevalence of head lice infestation and pediculicidal effect of permethrine shampoo in primary school girls in a low-income area in southeast of Iran.

BACKGROUND: Head lice infestation is a common public health problem that is most prevalent in primary school children throughout the world, especially in developing countries including different parts of Iran. This study aimed to determine the prevalence and risk factors associated with head lice infestation and pediculicidal effect of 1% permethrin shampoo in primary schools girls of Bashagard County, one of the low socioeconomic areas in southeast of Iran. METHODS: In this interventional study six villages with similar demographical situations were selected and randomly assigned into intervention and control areas. In each area 150 girl students aged 7-12 years were selected randomly and screened for head lice infestation by visual scalp examination. In intervention area, treatment efficacy of 1% permethrin shampoo was evaluated via re-examination for infestation after one, two, and three weeks. Pre-tested structured questionnaire was used to collect data on socio-demographic and associated factors of head lice infestation. RESULTS: The prevalence of head lice infestation was 67.3%. There was significant association between head lice infestation and school grade, family size, parents' literacy, bathing facilities, frequency of hair washing, and use of shared articles (p < 0.05). The effectiveness of 1% permethrin shampoo for head lice treatment was 29.2, 68.9, and 90.3% after the first, second, and third weeks, respectively. CONCLUSION: The head lice infestation is a health problem in primary school girls of Bashagard County. Improvement of socioeconomic status and providing appropriate educational programs about head lice risk factors and prevention can be effective for reduction of infestation in this area. TRIAL REGISTRATION: This trial has been registered and approved by Hormozgan University of Medical Sciences ethical committee (Trial No.764). Trial registration date: March 17 2014.