Vol 10 No 3 (2016)

Original Article

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    Background: Rhipicephalus (Boophilus) annulatus tick is one of the most important ectoparasite of cattle. Re­cently, several laboratories in the world have been concentrated on immunizing cattle against tick using various types of tissue extracts of ticks. The aim of this study was to evaluate the effect of immunization of cattle with tick salivary gland extract on biological parameters of ticks and humoral immune responses of cattle.
    Methods:
    Fourteen more dominant protein bands identified as immunogenic by Western-blot analysis were eluted from polyacrylamide gel. Test and control groups were injected three times with eluted proteins and sterile PBS (pH= 7.2) respectively with equivalent amount of adjuvant. After four weeks a tick challenge was performed. Fi­nally, biological parameters of collected engorged female ticks were recorded and humoral immune responses to immunization measured by ELISA.
    Results:
    The results indicated immunization of cattle resulted in reduction in mean tick counts, attachment, en­gorgement weights, feeding index, egg mass weight, hatchability and fertility index (respectively 63.1%, 62.6%, 30.2%, 36.4%, 40%, 78.7% and 13.3%) and increased duration of feeding, pre-oviposition and incubation period of eggs (respectively 8.6%, 45 and 31.34%). All changes were statistically significant (P< 0.05). Results showed an increase in antibody production of test group from the first week after immunization. The antibody level was boosted following tick infestation.
    Conclusion:
    This investigation indicates that immunization of cattle with these antigens could induce a protective immune response against Rh. (B.) annulatus tick that would be expected to provide a safe non-chemical means of tick control.

  • XML | PDF | downloads: 865 | views: 975 | pages: 292-304

    Background: The aim of this study was to design pictorial key and taxonomic literature of Culex pipiens complex in Iran.
    Methods:
    Larvae were collected using standard dipping methods in 13 randomly selected areas of Bushehr, Hame­dan, Kerman, Khorasan-e-Razavi, Khuzistan, Mazandaran, Tehran, Sistan and Baluchistan and Yazd Provinces from April 2009 to October 2010. The data were analyzed using SPSS Ver. 11.5.
    Results:
    Culex pipiens larvae were identified based on the Seta 1 of the abdominal segments III–IV in north and central parts of Iran. This diagnostic character had some variation among the Cx. quinquefasciatus collected from south of the country. The identification value of intersection of costa, subcosta and bifurcation of R2+3 of female veins, was calculated as 90–100 % for Cx. pipiens. This diagnostic character was varied among the Cx. quinquefas­ciatus specimens. The male genitalia found as the main characters to distinguish of Cx. quinquefasciatus from Cx. pipiens.
    Conclusion:
    It is necessary more studies on the behavior and genetic variations of Cx. pipiens complex in Iran.

  • XML | PDF | downloads: 661 | views: 1148 | pages: 305-316

    Background: Mosquitoes transmit serious human diseases, causing millions of death every year. Vector control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. Nine different locally available medicinally important plants suspected to posse larvicidal property were screened against fourth instar larvae of Aedes aegypti and Anoph­eles stephensi to a series of concentrations of the methanolic extracts.
    Methods:
    Susceptibility tests on Ae. aegypti and An. stephensi were conducted using standard WHO methods. The larvae of two mosquito species were exposed to methanolic extracts and mortality counts were made after 24 hours of exposure as per WHO method. Larvae of Ae. aegypti were more susceptible than that of An. stephensi.
    Results:
    Among the nine plant species tested, Annona reticulata leaf extract was more effective against Ae. aegypti larvae with LC50 and LC90 values of 95.24 and 262.64 ppm respectively and against An. stephensi larvae 262.71 and 636.94 ppm respectively. The least efficacy was in Cosmos bipinnatus with LC50 and LC90 values of 442.6 and 1225.93 ppm against Ae. aegypti and LC50 and LC90 values of 840.69 and 1334.01 ppm of Thespesia populnea against An. stephensi.
    Conclusion:
    The crude methanolic extract of the An. reticulata with good larvicidal efficacy could be considered for further characterization to control mosquito vectors instead of chemical insecticides. High efficacy found in An. re­ticulata extract will be considered for further studies to isolate the bioactive compound.

  • XML | PDF | downloads: 575 | views: 702 | pages: 317-329

    Background: Geographic distribution of West Nile virus (WNV) is heterogeneous in Iran by a high circulation in the southern-western areas. The objective of our study was to determine environmental and climatic factors associ­ated with the risk of WNV equine seropositivity in Iran.
    Methods:
    Serological data were obtained from a serosurvey conducted in equine population in 260 districts in Iran. The climate and environmental parameters included in the models were distance to the nearest wetland area, type of stable, Normalized Difference Vegetation Index (NDVI), annual mean temperature, humidity and precipitation.
    Results:
    The important risk factors included annual mean temperature, distance to wetlands, local and seasonal NDVI differences. The effect of local NDVI differences in spring was particularly notable. This was a normalized difference of average NDVI between two areas: a 5 km radius area centered on the stable and the 5–10 km sur­rounding area.
    Conclusion:
    The model indicated that local NDVI’s contrast during spring is a major risk factor of the transmission of West-Nile virus in Iran. This so-called oasis effect consistent with the seasonal production of vegetation in spring, and is associated to the attractiveness of the local NDVI environment for WNV vectors and hosts.  

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    Background: There are nearly 1000 species of Phlebotomine sand flies in 6 genera, of which only two, Phlebotomus in the old world and Lutzomyia in the new world are medically important. Globally, leishmaniasis prevalent in 98 countries and affects estimated 12 million people with almost two million new cases per year. Some rural areas of Azarshahr District in East Azarbaijan Province have been reported to be endemic for visceral leishmaniasis. This study is the first attempt to determine the species diversity and density in a new focus of visceral leishmaniasis in Azarshahr District, East Azarbaijan Province, Iran.
    Methods:
    Sand flies were collected form indoor and outdoor biweekly using sticky traps. Diversity index of the collected sand flies within different villages were estimated by the Shannon-Weaver.
    Results:
    The activity of the sand flies extended from April to October with one peak in August. Diversity of sand flies within study area were estimated as 0.917, 1.867, 1.339, 1.673, and 1.562 in Almalodash, Jaragil, Segaiesh, Amirdizaj, and Germezgol Vvillages, respectively.
    Conclusion:
    Identifying the diversity and seasonal abundance of the collected species is of importance for prediction of the period of maximum risk for leishmaniasis transmission and for the successful implementation of a control program. Species diversity is one of the most important factors in ecological studies.

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    Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formula­tions of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longi­palpa F. under laboratory and field conditions.
    Methods:
    Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml-1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs us­ing different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer.
    Results:
    Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough path­ogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in- water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sun­flower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses.
    Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising al­ternative for cockroach control.  

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    Background: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mos­quitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cyto­plasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases.
    Methods:
    Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these en­dosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene.
    Results
    : Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phyloge­netic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world.
    Conclusion:
    It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evo­lutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population repro­ductive incompatibility pattern and its usefulness as a bio-agent control measure.

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    Background: The aim of the present study was to determine the prevalence of ectoparasite infestations in stray dogs in Mazandaran, Gilan and Qazvin Provinces in fall and winter in 2013(December to March).
    Methods:
    Seventy dogs in 2013, from these Provinces were examined for ectoparasite infestation and diagnosis of them based on parasitological methods and identification keys was done.
    Results:
    The rate of infestation in these areas was 100%, 68.5% and 93.3% respectively. Fleas were the most com­mon ectoparasites on dogs in this study followed by lice, ticks, flies and mites respectively. The isolated arthropods were fleas in 77.5%, lice in 50%, ticks in 8.6%, flies in 6.8% and mites in 5.1% of infested dogs. The ectoparasite of the dogs included 4 flea species: Ctenocephalides canis (29.8%), C. felis (19.9%), Pulex iritans (2.9%) and Xenopsi­ella cheopis (0.7%), 1 louse species: Trichodectes canis (41.3%), 1 tick species: Rhipicephalus sanguinus (0.7%), 1 fly species: Hippobosca sp. (1.1%) and 1 mite species: Sarcoptes scabiei (3.6%).
    Conclusion:
    Fleas and lice were the most common ectoparasites in stray dogs of the studied area. Some ectopara­sites transmit important human disease, therefore regular monitoring of them is a major concern to control the ar­thropods and arthropods-borne diseases.

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    Background: Aedes aegypti mosquito is responsible for transmitting human diseases like dengue and chikungunya. Personal or space protection with insect repellents is a practical approach to reducing human mosquito contact, thereby minimizing disease transmission. Essential oils are natural volatile substances from plants used as protective measure against blood-sucking mosquitoes.
    Methods:
    Twenty-three essential oils were evaluated for their repellent effect against Ae. aegypti female mosquito in laboratory conditions using Y-tube olfactometer. 
    Results:
    The essential oils exhibited varying degree of repellency. Litsea oil showed 50.31%, 60.2 %, and 77.26% effective mean repellency at 1 ppm, 10 ppm and 100 ppm respectively, while DEET exhibited 59.63%, 68.63%, 85.48% and DEPA showed 57.97%, 65.43%, and 80.62% repellency at respective above concentrations. Statistical analysis revealed that among the tested essential oils, litsea oil had effective repellency in comparison with DEET and DEPA against Ae. aegypti mosquito at all concentration. Essential oils, DEET and DEPA showed significant repellence against Ae. aegypti (P< 0.05) at all 3 concentration tested.
    Conclusion:
    Litsea oil exhibited effective percentage repellency similar to DEET and DEPA. The essential oils are natural plant products that may be useful for developing safer and newer herbal based effective mosquito repellents.

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    Background: Crimean-Congo Hemorrhagic Fever (CCHF) is a feverous and hemorrhagic disease endemic in some parts of Iran and caused by an arbovirus related to Bunyaviridae family and Nairovirusgenus. The main virus reser­voir in the nature is ticks, however small vertebrates and a wide range of domestic and wild animals are regarded as reservoir hosts. This study was conducted to determine the infection rate of CCHF virus in hard ticks of Sarpole-Zahab County, Kermanshah province, west of Iran.
    Methods: From total number of 851 collected ticks from 8 villages, 131 ticks were selected randomlyand investi­gated for detection of CCHF virus using RT-PCR.
    Results: The virus was found in 3.8% of the tested ticks. Hyalommaanatolicum, H.asiaticum and Rhipicephalus sanguineus species were found to have viral infection, with the highest infection rate (11.11%) in Rh. sanguineus.
    Conclusion: These findings provide epidemiological evidence for planning control strategies of the disease in the study area.

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    Background: The aim of this study was to assess community knowledge and behavioral impact of the social mobili­zation and communication strategy applied in a dengue high-risk area in Sri Lanka.
    Methods:
    A group of adults visiting selected primary care facilities in Colombo district were interviewed to collect socio-demographicdata, attributes of knowledge regarding dengue and the responsive behaviors adopted by them following the dengue control program though the media and social marketing campaigns. These attributes were clas­sified as `good ', `fair,' or `poor' by developing a composite scale for analysis and interpretation of data.
    Results:
    The primary source of information was television in the majority. The overall knowledge of the disease, vector and control methods was poor. The overall level of contribution to dengue control activities was good.
    Conclusion:
    Awareness of the disease and its complications had not contributed to favorable behavior changes.  While the social mobilization and behavior change campaign in Sri Lanka had low impact on knowledge and be­haviors, a better understating of community perceptions of DF and how these perceptions are formulated within the social and cultural context; would be useful to improve uptake. This knowledge would be valuable for program plan­ners to strengthen dengue control activities in SL and other similar settings across the region.

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    Background: Insecticide resistance is one of the serious problems for German cockroach control program. This study was conducted to determine the bendiocarb and Carbaryl resistance mechanisms in German cockroaches using the piperonyl butoxide (PBO).
    Methods: Bioassay tests were conducted with 4 to 6 different concentrations of both insecticides with four replicates of 10 susceptible strain cockroaches per concentration to determine of discriminative concentration. After determining discriminative concentration, the result was compared to wild strain. The levels of susceptibility and resistance ratio (RR) and synergism ratio (SR) were calculated for each five wild strains. Moreover resistance mechanisms in wild strains were determined using PBO synergist in vivo.
    Results: Hospital strains showed different levels of resistance to bendiocarb and carbaryl compared to susceptible strain. The bendiocarb and carbaryl resistance ratios ranged from 2.11 to 7.97 and 1.67 to 2 at LD50 levels, respec­tively. The synergist PBO significantly enhanced the toxicity of bendiocarb and carbaryl to all strains with different degrees of synergist ratio, 1.31, 1.39, 3.61, 1.78, 1.62 and 2.1 fold for bendiocarb, 1.19, 1.18, 1.12. 1.29, 1.45 and 1.11- fold for carbaryl, suggesting monooxygenase involvement in bendiocarb and carbaryl resistance.
    Conclusion: The synergetic effect of PBO had the highest effect on bendiocarb and resistance level was significantly reduced, which indicates the important role of monoxidase enzyme in creating resistance to Bendiocarb. Piperonyl butoxide did not have a significant synergistic effect on carbaryl and did not significantly break the resistance.

Short Communication

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    Background: Microsporidia are known to infect a wide variety of animals including mosquitoes (Diptera: Cu­licidae). In a recent study on the mosquito fauna of Chahar Mahal and Bakhtiari Province, at the central western part of Iran, a few larvae of Anopheles superpictus were infected with a microsporidium-resembled microorganism. Cur­rent investigation deals with the identification of the responsible microorganism at the genus level.
    Methods: Fresh infected larvae were collected from the field. After determining the species identity they were dis­sected to extract their infective contents. Wet preparations were checked for general appearance and the size of the pathogenic microorganism. Fixed preparations were stained with Geimsa and Ryan-Blue modified Trichrome tech­niques to visualize further morphological characters. The obtained light microscopy data were used in the identifica­tion process.
    Results: The infected larvae were bulged by a whitish material filling the involved segments corresponding to a microsporidium infection. Bottle-shaped semioval spores ranged 4.33±0.19×2.67±0.12 and 4.18±0.43×2.45±0.33 micron in wet and fixed preparations, respectively. They were mostly arranged in globular structures comprised of 8 spores. These data was in favor of a species from the genus Parathelohania in the family Ambliosporidae.
    Conclusion: This is the first report of a microsporidium infection in An. superpictus. The causative agent is diag­nosed as a member of the genus Parathelohania. Further identification down to the species level needs to determine its ultrastructural characteristics and the comparative analysis of ss rRNA sequence data. It is also necessary to un­derstand the detail of the components of the transmission cycle.

Case Report

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    Myiasis is the infection caused by a variety of dipterous (fly) larvae in vertebrate’s tissue (man and domestic or wild animals). Species of Gasterophilus are obligate parasite of horses, donkeys, zebras, elephants and rhinoceroses. There are records worldwide, but mostly, in tropical and subtropical regions. This case report describes a type of gastric myiasis caused by G. intestinalis in an old lion in a zoo in Sistan, southeast Iran. Myiasis in lions is rarely reported and this is the first report of gastric myiasis in lion.

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    Background: Pigeons (Columba livia) have been kept as pet and reared for food in several countries including Iran. Ectoparasites are regarded as the basic causes of retardation in growth, lowered vitality and poor conditions of the birds. Pseudolynchia canariensis a hippoboscidae fly is one of the important ectoparasites of pigeons and is respon­sible for the transmission of pathogens to birds and humans same as pathogenic protozoan Haemoproteus columbae.
    Methods: A herd of domestic pigeons contained 50 pigeons in Shahrekord, southwest Iran was evaluated clinically infested by ectoparasites. Ectoparasites were removed. The samples were collected and then referred to the Laboratory of Parasitology of Shahrekord University, Shahrekord, Iran.
    Results: Usin diagnostic key for diptera fly, these flies were find P. canariensis. This is a rare report of infestation of pigeons herd by P. canariensis in Iran. The infestation rate was 40% that rate of infestation in pipers was more than females and in females was more than males.
    Conclusion: The rate of infested pipers was more than adults that maybe the less potential of pipers in removing of ectoparasites is reason of this higher rate.

Review Article