Vol 14 No 3 (2020)

Original Article

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    Background: Malaria is a major public health problem and life threatening parasitic vector-borne disease. For the first time, we established and report the molecular mechanism responsible for Anopheles gambiae s.l. resistance to pyrethroids and DDT from Yamaltu Deba, Southern Guinea Savanna, Northern-Nigeria.
    Methods: The susceptibility profile of An. gambiae s.l. to four insecticides (DDT 4%, bendiocarb 0.1%, malathion 5% and deltamethrin 0.05%) using 2–3 days old females from larvae collected from study area between August and Novem­ber, 2018 was first established. Genomic DNA was then extracted from 318 mosquitoes using Livak DNA extraction protocol for specie identification and kdr genotyping. The mosquitoes were identified to species level and then 96 geno­typed for L1014F and L1014S kdr target site mutations.
    Results: The mosquitoes were all resistant to DDT, bendiocarb and deltamethrin but fully susceptible to malathion. An. coluzzii was found to be the dominant sibling species (97.8%) followed by An. arabiensis (1.9%) and An. gambiae s.s (0.3%). The frequency of the L1014F kdr mutation was relatively higher (83.3%) than the L1014S (39%) in the three species studied. The L1014F showed a genotypic frequency of 75% resistance (RR), 17% heterozygous (RS) and 8% susceptible (SS) with an allelic frequency of 87% RR and 13% SS while the L1014S showed a genotypic frequency of RR (16%), RS (38%) and SS (46%) with an allelic frequency of 40% RR and 60% SS, respectively.
    Conclusion: This study reveals that both kdr mutations present simultaneously in Northern-Nigeria, however contribution of L1014F which is common in West Africa was more than twice of L1014S mutation found in East Africa.

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    Background: Bedbugs are blood feeding ectoparasites of humans and several domesticated animals. There are scar­city of information about the bed bugs population throughout Iran and only very limited and local studies are availa­ble. The aim of this study is to assess the phylogenetic relationships and nucleotide diversity using partial sequences of cytochrome oxidase I gene (COI) among the populations of tropical bed bugs inhabiting Iran.
    Methods: The bedbugs were collected from cities located in different geographical regions of Iran. After DNA ex­traction PCR was performed for COI gene using specific primers. Then DNA sequencing was performed on PCR products for the all 15 examined samples.
    Results: DNA sequencing analysis showed that the all C. hemipterus samples were similar, despite the minor nu­cleotide variations (within the range of 576 to 697bp) on average between 5 and 10 Single nucleotide polymorphisms (SNPs). Subsequently, the results were compared with the database in gene bank which revealed close similarity and sequence homology with other C. hemipterus from other parts of the world.
    Conclusion: In conclusion, this study has demonstrated the ability of the COI gene to differentiate between the C. hemipterus populations from a few different locations in Iran. The current research is the first report of phylogenetic and genetic species diversity analysis conducted on C. hemipterus in Iran. These results provided basic information for further studies of molecular epidemiology, public health and pest control operators in Iran.

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    Background: Organophosphate pesticides are of concern to the drinking water supply and high risks on human health.
    Methods: An occurrence survey was performed in the spring and summer of 2016 for 6 months to assess the preva­lence of organophosphate pesticides in the Hablehrood River that is located in Semnan Province. Besides, 10 kinds of organophosphate pesticides were sampled in 5 stations. Moreover, were measured by Gas Chromatography (GC) was supported by an electron capture detector (ECD) and Turbochrom software. In all stations, Diazinon was detect­ed in the spring and summer, but Malathion only in the spring.
    Results: The highest concentration of Diazinon was observed at the Mahmoud Abad station in spring (0.94ppb) and the Bonekooh station in the summer (0.93ppb). The highest and lowest concentrations of Malathion were detected in Mahmoud Abad (0.35ppb), and Gache station, respectively.
    Conclusion: The concentration of pesticides in BoneKooh and Mahmood Abad was higher than the standard.

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    Background: Tropical Theileriosis caused by Theileria annulata is a tick-borne disease which transmitted by the ix­odid tick members of the genus Hyalomma. Studies on different aspects of disease require to access infective sporozoite of parasite which produced by tick vector. This study was carried out to establish of T. annulata life cycle to achieve T. annulata infected ticks.
    Methods: Laboratory rabbit and calf were used for rearing of Hyalomma anatolicum different instars. Unfed nymphs were fed on T. annulata infected calf. Clinical signs, Giemsa stained smears and Polymerase Chain Reaction (PCR) methods were used for detection of infection in blood and tick specimens. Susceptible calf was used for confirmation of sporozoites maturation and infectivity in bioassay test.
    Results: Hyalomma anatolicum two and three-host strategies of life cycle was lasted 90 and 116 days respectively. The PCR confirmed T. annulata infection in blood and tick samples. Maturation of T. annulata sporozoites was confirmed in bioassy test. First clinical symptom of disease was seen earlier in the case of transmission of disease through feeding of live ticks in comparison with blood injection method.
    Conclusion: Complete life cycle of T. annulata was done and confirmed by clinical signs, microscopic examination, molecular methods and bioassay test. According to published reports to date, this is the first report of establishment of H. anatolicum tick infection with T. annulata using susceptible calf under controlled conditions in Iran.

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    Background: The incidence and area of arbovirus infections is increasing around the world. It is largely linked to the spread of the main arbovirus vectors, invasive mosquito of the genus Aedes. Previously, it has been reported that Aedes aegypti reemerged in Russia after a 50-year absence. Moreover, in 2011, Ae. albopictus was registered in the city of Sochi (South Russia, Black Sea coast) for the first time. In 2013, Asian Ae. koreicus was found in Sochi for the first time.
    Methods: Mosquitoes were collected using the following methods: larvae with a dip net, imago on volunteers and using bait traps. The mosquitoes were identified using both morphology and sequencing of the second internal transcribed spacer of the nuclear ribosomal RNA gene cluster.
    Results: In August 2016, Ae. koreicus larvae and imago and a single male of Ae. aegypti were found on the southern coast of the Crimean Peninsula, where they were not registered before. Newly obtained DNA sequences were registered in GenBank with the accession numbers MF072936 and MF072937.
    Conclusion: Detection of invasive mosquito species (Ae. aegypti and Ae. koreicus) implies the possibility of their area expansion. Intensive surveillance is required at the Crimean Peninsula to evaluate the potential for the introduction of vector-borne diseases.

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    Background: Phlebotomus kandelakii and Phlebotomus perfiliewi transcaucasicus sand flies are the vectors of vis­ceral leishmaniasis in Iran. The aim of this study was to evaluate the susceptibility of Ph. kandelakii and Ph. perfil­iewi trans­caucasicus, collected from an endemic focus of visceral leishmaniasis (VL) in the northwest of Iran, to different select­ed insecticides.
    Methods: Sand flies were collected from the villages of Meshkinshahr and Germi Counties using light traps and aspira­tors from May to October 2019. The sand flies were identified as Ph. kandelakii and Ph. perfiliewi transcauca­sicus us­ing standard identification keys. Susceptibility test was carried out against DDT (4%), Malathion (5%), Propoxur (0.1%) and Lambda-cyhalothrin (0.05%) according to the WHO standard method. MedCalc statistical software was employed to calculate LT50 and LT90 and to compare the chances of sand flies surviving the exposure to the studied in­secticides.
    Results: A total of 1,278 female specimens were used for the susceptibility tests. Out of which 1,063 samples were used for the test and 215 for the control group. The estimated LT50 values for DDT (4%), Malathion (5%), Propoxur (0.1%), and Lambda-cyhalothrin (0.05%) for Ph. kandelakii were 15.1, 13.4, 15.4 and 5.8 minutes respectively, and for Ph. per­filiewi transcaucasicus were 11.9, 15.6, 15.9 and 5.8 minutes  respectively.
    Conclusion: This susceptibility studies revealed different LT50 values for different insecticides efficient against Ph. kandelakii and Ph. perfiliewi transcaucasicus. The regular monitoring for the resistance of Ph. kandelakii and Ph. per­filiewi transcaucasicus sand flies seems necessary in diseases control programs in this area.

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    Background: Ticks are vectors of a wide variety of pathogens that can be transmitted to humans, and tick-borne diseases are a significant public health issue worldwide. The present study was carried out on the hard tick infestation of livestock transported to Rafsanjan slaughter house in the southeast of Iran.
    Methods: A cross-sectional survey was carried out biweekly from April to September 2016 to determine tick infestation of the meat-producing animals. All the livestock included in our study were thoroughly inspected for the presence of hard ticks on different parts of their bodies.
    Results: A total of 258 hard ticks were collected from the body of livestock hosts. The ticks that were sampled were classified into two genera and five species: Hyalomma marginatum, Hy. anatolicum, Hy. asiaticum, Hy. dromedarii, and Rhipicephalus sanguineus. Hyalomma dromedarii was the most abundant species in the study area. More than 50 percent of the sampled ticks were collected from the body of camels brought to the slaughter house however molecular analysis showed no Crimean Congo Hemorrhagic Fever (CCHF) virus infection in tick specimens. The Sex ratio of the sampled hard ticks shows that female tick infestation was more common among the study livestock.
    Conclusion: Due to the crucial role of hard ticks in the transmission of different pathogens to humans, additional investigations are necessary to determine the risk of consumption of infested meat-producing animals in the study area.

Short Communication

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    Background: Mosquitoes are blood sucking arthropods and serve as vectors of many diseases causing serious health problems to human beings. Culex quinquefasciatus and Aedes aegypti were responsible for Filariasis and Dengue. Syn­thetic pesticides were effective against mosquitoes as well as main sources of environmental pollution and most of them are immunosuppressant. Botanicals were widely used as insecticides, growth disruptors, repellents, etc. The aim of this research was to determine larvicidal properties of powdered leaf, Elytraria acaulis against late third or early fourth in­star larvae of Cx. quinquefasciatus and Ae. aegypti.
    Methods: Larvae of Cx. quinquefasciatus and Ae. aegypti were tested at various concentrations of 100, 120, 140, 160, 180 and 200mg/100ml and mortality was recorded after 24h. The LC50 values of the E. acaulis leaf powder were calcu­lated by Probit analysis.
    Results: The plant powder exhibited strong larvicidal activity against Cx. quinquefasciatus with LC50 value of 116.07mg/100ml against Ae. aegypti 124.25mg/100ml respectively. The result indicated that the plant powder of E. acaulis showed potential larvicidal activity against Cx. quinquefasciatus and Ae. aegypti.
    Conclusion: The overall findings of the present investigation suggested that the E. acaulis highly effective against Cx. quinquefasciatus and Ae. aegypti larvae. Elytraria acaulis may be used as an alternative to synthetic chemical pesticides for control of vectors to reduce vector borne diseases and did not harm to total environment.

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    Background: Different forms of leishmaniasis are significant infectious diseases in Iran. While, Rudbar County of Gui­lan Province has been introduced as a new cutaneous leishmaniasis focus, there are few published data about the phlebotomine sand flies (Diptera: Psychodidae) of the province.
    Methods: To study the phlebotomine fauna of Rudbar County, the sampling was performed in 12 collection sites by light traps, sticky traps and manual aspirators throughout August–December 2015. Sand flies were removed from the sticky traps, rinsed in acetone and stored in 80% ethanol along with the collections of light traps and hand catch­es.
    Results: In total, 2186 sand flies were collected and ten species representing two genera were morphologically identi­fied: Phlebotomus (Adlerius) halepensis (0.27%), Ph. (Larroussius) kandeladii (0.10%), Ph. (Lar.) neglectus (0.91%), Ph. (Lar.) perfiliewi (53.88%), Ph. (Lar.) tobbi (43.45%), Ph. (Paraphlebotomus) sergenti (0.82%), Ph. (Phlebotomus) papatasi (0.10%), Sergentomyia (Parrotomyia) baghdadis (0.27%), Se. (Sintonius) clydei (0.05%), Se. (Sin.) tiberiadis (0.10%). The species Ph. halepensis, Ph. neglectus, Ph. perfiliewi, Se. baghdadis, Se. clydei and Se. tiberiadis were reported for the first time in Guilan Province. This study also verified the presence of Ph. neglec­tus (Ph. major krimen­sis as a synonym and morphotype) in Iran. Moreover, the taxonomy of the subgenus Larrous­sius of the province was discussed.
    Conclusion: The prevalence of suspected or proven cutaneous and visceral leishmaniasis vectors is noteworthy. The study of ecology of sand flies and detecting the exact vectors of leishmaniasis and phlebotomine fever by molecular specific tests in Guilan Province are recommended.

Case Report

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    Myiasis is infection with a fly larva, usually occurring in tropical and subtropical areas. A 32-years-old immunocom­pro­mised woman with fever, gastrointestinal pain, cramps, vomiting; and fatigue was referred to Ghaem Hospital in Mash­had in July 2018. Entomological characterisation of cephaloskeletons, posterior spiracles of the excreted larvae and gen­ital dissection of male fly were diagnosed as Sarcophaga (Liopygia) argyrostoma. Accidental intestinal myiasis caused by unplanned intake of dipterous larvae by contaminated food staff or water. Patients with immune deficiency may be more at risk for the threat of maggot’s infestation, so in such cases, hospital standard should be more in atten­tion. To best of our knowledge, S. argyrostoma has not been reported before in the indexed literature from Iran.