Vol 10 No 2 (2016)

Original Article

  • XML | PDF | downloads: 611 | views: 920 | pages: 127-140

    Background: Crimean Congo hemorrhagic fever virus (CCHFV) is a member of the Bunyaviridae family and Nairovirus genus. It has a negative-sense, single stranded RNA genome approximately 19.2 kb, containing the Small, Medium, and Large segments. CCHFVs are relatively divergent in their genome sequence and grouped in seven distinct clades based on S-segment sequence analysis and six clades based on M-segment sequences. Our aim was to obtain new insights into the molecular epidemiology of CCHFV in Iran.
    Methods:
    We analyzed partial and complete nucleotide sequences of the S and M segments derived from 50 Iranian patients. The extracted RNA was amplified using one-step RT-PCR and then sequenced. The sequences were ana­lyzed using Mega5 software.
    Results:
    Phylogenetic analysis of partial S segment sequences demonstrated that clade IV-(Asia 1), clade IV-(Asia 2) and clade V-(Europe) accounted for 80 %, 4 % and 14 % of the circulating genomic variants of CCHFV in Iran respectively. However, one of the Iranian strains (Iran-Kerman/22) was associated with none of other sequences and formed a new clade (VII). The phylogenetic analysis of complete S-segment nucleotide sequences from selected Ira­nian CCHFV strains complemented with representative strains from GenBank revealed similar topology as partial sequences with eight major clusters. A partial M segment phylogeny positioned the Iranian strains in either associa­tion with clade III (Asia-Africa) or clade V (Europe).
    Conclusion:
    The phylogenetic analysis revealed subtle links between distant geographic locations, which we pro­pose might originate either from international livestock trade or from long-distance carriage of CCHFV by infected ticks via bird migration.

  • XML | PDF | downloads: 559 | views: 1271 | pages: 141-147

    Background: Leishmaniasis is an important public health disease in many developing countries as well in Iran. The main objective of this study was to investigate on leishmania infection of wild caught sand flies in an endemic focus of disease in Esfarayen district, north east of Iran.
    Methods
    : Sand flies were collected by sticky papers and mounted in a drop of Puri’s medium for species identification. Polymerase chain reaction techniques of kDNA, ITS1-rDNA, followed by restriction fragment length polymorphism were used for identification of DNA of Leishmania parasites within infected sand flies.
    Results
    : Among the collected female sand flies, two species of Phlebotomus papatasi and Phlebotomus salehi were found naturally infected with Leishmania major. Furthermore, mixed infection of Leishmania turanica and L. major was observed in one specimen of P. papatasi. Sequence analysis revealed two parasite ITS1 haplotypes including three L. major with accession numbers: KJ425408, KJ425407, KM056403 and one L. turanica. (KJ425406). The haplotype of L. major was identical (100%) to several L. major sequences deposited in GenBank, including isolates from Iran, (Gen Bank accession nos.AY573187, KC505421, KJ194178) and Uzbekistan (Accession no.FN677357).
    Conclusion
    : To our knowledge, this is the first detection of L. major within wild caught P. salehi in north- east of Iran.

  • XML | PDF | downloads: 537 | views: 1249 | pages: 148-158

    Background: Several compounds obtained from plants have potential insecticidal, growth deterrent or repellent characteristics. The control of hard ticks by non-chemical substances was targeted in this study.
    Methods:
    The effect of 36 materials on in-vitro ticks was studied, including 2 absolute controls (water only or ab­solute ethyl alcohol only), 6 conventionally used spinosad preparations (aqueous solutions), 12 Nigella sativa (N. sativa) preparations (aqueous and alcoholic solutions), and 12 Thyme preparations (aqueous and alcoholic solutions). The engorged ticks were tested in-vitro for mortality and oviposition ability using the studied materials.
    Results:
    The final mortality after 48 hours of application in N. sativa aqueous preparations began from 10.0% con­centration, 1.0% to 100% by concentration preparations ≥10%. In addition, N. sativa alcoholic preparations began from 50.0% concentration, 2 % to 100% by concentration ≥5%. Meanwhile, Thyme aqueous and alcoholic prepa­rations began from 70.0% concentration, 5% to 90% by concentration 10–20%. Additionally, spinosad aqueous preparations and both of control preparations (Water and Alcohol) resulted in no mortality. All differences were sta­tistically significant. The oviposition was stopped in N. sativa (aqueous ≥10% and alcoholic ≥5%) and in spinosad (aqueous≥25%). The aqoues dilution of the used matters killed B. annulatus larvae beginning from the concentration 5%.
    Conclusion:
    Nigella sativa alcohol 20% was the best of studied preparations being the lowest concentration (20%) that could achieve the highest lethal (100%) effect in shortest time (12 hours). Moreover, Thyme oil and spinosad could not kill 100% of adult but did on larvae.

  • XML | PDF | downloads: 552 | views: 849 | pages: 159-167

    Background: The aim of this study was to investigate the effect of Hemiscorpius lepturus venom on leukocytes and the leukocyte subgroups in peripheral blood of rat.
    Methods:
    In this experimental study, sixty N-Mari rats were divided into three groups of 20 rats. Then the rats in each group were divided into four subgroups based on the blood sampling time that was 2, 6, 24 and 48 hours after the venom injection, respectively. The control group did not receive anything, however, the first and the second ex­perimental groups received 0.1 and 0.01mg/kg of venom, subcutaneously. In accordance with a designated four sam­pling times, the blood sampling was carried out in three groups. After RBC lysis, the leukocytes and leukocyte sub­populations were determined and counted using appropriate hematological standard methods.
    Results:
    The leukocyte and the neutrophil count at two (P<0.05), six (P<0.01) and 24 (P<0.05) hours after the venom injection showed a significant decline compared with the control group, this decrease was significant at the dose of 0.1 mg/kg until 48 hours after the venom injection (P<0.05). The lymphocyte count showed a significant decline throughout the all hours of the experiment, compared with the control group (P<0.05).
    Conclusion:
    Leukocytes are probably affected by the cytotoxicity effect of the H. lepturus venom in a dose-dependent manner. This could be a wakeup call for the medical staff to perform quick and accurate treatment in the least time possible.

  • XML | PDF | downloads: 545 | views: 742 | pages: 168-177

    Background: This comprehensive study was conducted on multi-purpose one-humped camel (Camelus dromedarius) sera and ticks to assess the epidemiological aspects of the Crimean-Congo hemorrhagic fever virus (CCHFV) in northeast Iran.
    Methods
    : From May 2012 to January 2013, eleven cities were randomly selected in the Khorasan Provinces of Iran as “clusters,” and at least 14 one-humped camels were sampled from each area. Reverse transcriptase polymerase chain reaction was used for the detection of the CCHFV genome in ticks. Sera were analyzed using specific enzyme-linked immunosorbent assay tests.
    Results:
    Four hundred and eighty ixodid ticks were collected, and the genome of the CCHFV was detected in 49 (10.2%) out of 480 ticks. The CCHFV genome was detected in two out of four tick species, and in tick samples from three cities in Khorassan-e-Jonoobi. All three provinces, and six out of eleven cities, were CCHFV-specific IgG-positive. In total, nine (5.3%) out of 170 one-humped camels were IgG-positive. The highest rate of IgG-positive samples was found in Nehbandan (16.67%).
    Conclusion:
    Continued surveillance and strictly enforced importation and quarantine practices should be imple­mented to prevent human exposure and the on-going dispersal of infected ticks and livestock in these regions. It is recommended that acaricides be used to prevent CCHF transmission to humans, and to reduce the tick population. In addition, care should be taken by abattoirs workers and people who work with one-humped camels.

  • XML | PDF | downloads: 446 | views: 1022 | pages: 178-191

    Background: The irritant effect of some insecticides can cause a proportion of mosquitoes to leave the sprayed rooms before acquiring a lethal dose, so the repeated contact al sub-lethal dose may lead to extent the resistance.
    Methods: Larvae and pupae of Culex pipiens complex were collected in mass from open canals of waste water in capital city Tehran and reared to obtain the first generation at laboratory. Sugar-fed 2–3 days female mosquitoes were used for the experiments and compared with laboratory strain. The irritability tests of insecticides impregnated pa­pers were measured in plastic conical exposure chambers placed which implemented at controlled conditions ac­cording  to  the  method  described  by WHO .Number of take-offs were counted during 15  minutes of exposure  time.
    Results:
    DDT had the most irritancy effect against field population of Cx. pipiens. DDT, permethrin and deltamethrin was moderately irritable against laboratory strain, whereas, addition to three previous insecticides, malathion, cyfluthrin and propoxur should be also considered as moderately irritable insecticides for field population of. Irritability level of etofenprox, fenithrothion, bendiocarb, and lambdacyhalothrin did not differ from control group.
    Conclusion:
    The irritability response of mosquitoes may have a negative impact on control measures. Periodical execution of irritability tests with insecticides that routinely used in vector control program is highly recommended.

  • XML | PDF | downloads: 510 | views: 743 | pages: 192-200

    Background: The species of ectoparasites that live on a specific host in a geographical region form an ectoparasite community. Species abundance distributions describe the number of individuals observed for each different species that is encountered within a community. Based on properties of the species abundance distribution, the expected total number of species present in the community can be estimated.
    Methods:
    Preston’s lognormal distribution model was used to fit the expected species abundance distribution curve. Using the expected species abundance distribution curve, we estimated the total number of expected parasite species present and the amount of species that were likely missed by our sampling in the field.
    Results:
    In total, 8040 ectoparasites (fleas, sucking lice, gamasid mites and chigger mites) were collected from 431 Norway rats (Rattus norvegicus) from a localized area in southwest China. These ectoparasites were identified to be 47 species from 26 genera in 10 families. The majority of ectoparasite species were chigger mites (family Trombicu­lidae) while the majority of individuals were sucking lice in the family Polyplacidae. The expected species abun­dance distribution curve demonstrated the classic pattern that the majority of ectoparasite species were rare and that there were a few common species. The total expected number of ectoparasite species on R. norvegicus was estimated to be 85 species, and 38 species were likely missed by our sampling in the field.
    Conclusions:
    Norway rats harbor a large suite of ectoparasites. Future field investigations should sample large num­bers of host individuals to assess ectoparasite populations.

  • XML | PDF | downloads: 548 | views: 1091 | pages: 201-210

    Background: One of the best ways to control the malaria disease and to be protected human against Anopheles mos­quito biting is the use of repellents. Throughout repellents, herbal ones may be an appropriate and safe source for protection.
    Methods
    : Chemical constituents of Achillea vermiculata and Satoreja hortensis were determined by using gas chromatography-mass spectrometry. Efficacy and the protection time of these plants were assessed on Anopheles stephensi under the laboratory condition.
    Results
    : The mean assessed protection time and efficacy for A. vermiculata was 2.16 and 3.16 hours respectively and the obtained ED50 and ED90 for this plant was 5.67 and 63 µl/cm2 respectively. The figured for S. hortensis was 4.16 and 5 hours respectively.  ED50 and ED90 for this plant were 5.63 and 45.75µl/cm2 respectively.
    Conclusion
    : Results of investigation showed that S. hortensis plant has an acceptable protection time, therefore, this plant could be considered as a good herbal repellent against anopheles mosquitoes.

  • XML | PDF | downloads: 449 | views: 719 | pages: 211-221

    Background: The aim of the current study was to develop a marking technique as an internal marker to mark post blood meal mosquitoes by using stable phosphate isotope 32P and determine the optimal concentration of it.
    Methods
    : An isotonic physiological saline solution, containing different concentration of radioactive isotope 32P-labeled disodium phosphate (Na2H32PO4) was injected into rabbits via the jugular vein in the laboratory. Emerged Cx. pipiens were marked after feeding on rabbit. At the same time, the labeled conditions of emerged Cx. pipiens were also measured by placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside.
    Results
    : According to the label condition of Cx. pipiens after taking blood and the effect of different dosage Na2H32PO4 on rabbit health, the optimal concentration of radioactive isotope was determined, that is, 0.1211 mCi/kg. By placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside, the emerged mosquitoes were also labeled. Therefore, feeding mosquitoes on the animal injected with radioactive Na2H32PO4 was more prac­tical for detecting and tracing mosquitoes.
    Conclusion:
    The method was less time-consuming, more sensitive and safer. This marking method will facilitate post-bloodmeal studies of mosquitoes and other blood-sucking insects.

  • XML | PDF | downloads: 518 | views: 852 | pages: 222-237

    Background: The Malaria Early Warning System is defined as the use of prognostic variables for predicting the occurrence of malaria epidemics several months in advance. The principal objective of this study was to provide a malaria prediction model by using meteorological variables and historical malaria morbidity data for malaria-en­demic areas in south eastern Iran.
    Methods:
    A total of 2002 locally transmitted microscopically confirmed malaria cases, which occurred in the Minab district of Hormozgan Province in Iran over a period of 6 years from March 2003 to March 2009, were analysed. Meteorological variables (the rainfall, temperature, and relative humidity in this district) were also assessed. Monthly and weekly autocorrelation functions, partial autocorrelation functions, and cross-correlation graphs were examined to explore the relationship between the historical morbidity data and meteorological variables and the number of cases of malaria. Having used univariate auto-regressive integrated moving average or transfer function models, significant predictors among the meteorological variables were selected to predict the number of monthly and weekly malaria cases. Ljung-Box statistics and stationary R-squared were used for model diagnosis and model fit, respec­tively. Results: The weekly model had a better fit (R2= 0.863) than the monthly model (R2= 0.424). However, the Ljung-Box statistic was significant for the weekly model. In addition to autocorrelations, meteorological variables were not significant, except for different orders of maximum and minimum temperatures in the monthly model.
    Conclusions:
    Time-series models can be used to predict malaria incidence with acceptable accuracy in a malaria early-warning system. The applicability of using routine meteorological data in statistical models is seriously limited.

  • XML | PDF | downloads: 465 | views: 963 | pages: 238-245

    Background: Latrodectism, a syndrome caused by Latrodectus genus, is one of the clinical problems that occur predominantly in north east of Iran. Nowadays antivenom therapy has become the most useful treatment for animal bites; however there is still a controversy about route and time of antivenom administration in spider bite. The aim of the present study was to determine the efficacy of specific antivenom in neutralizing hepatic and renal symptoms 24 h after Latrodectus dahli envenomation.
    Methods:
    We selected a group of male New Zealand white rabbits, weighing 2±0.3 kg. The L. dahli venom (0.5 mg/kg) was injected subcutaneously. Specific antivenom (2.5 ml, I.V) was injected 24 h following venom injection. Blood sampling was performed before and 24 h after venom injection, as well within 24, 48 and 72 h after antivenom administration. Serum levels of (aspartate amino transferase (AST) alanine amino transferase (ALT), alkaline phosphatase (ALP), urea, bilirubin, creatinine and albumin were determined in all the sam.
    Results:
    Latrodectus dahli venom caused significant increase (P< 0.05) in all foresaid serum parameters. Antivenom reversed the AST, ALP, creatinine, urea and bilirubin to normal levels, but failed about ALT level, also non-significant decrease was observed in albumin levels.
    Conclusion:
    Antivenom administration 24 h after venom injection can greatly reverse symptoms caused by venom. Future studies in human beings should be conducted to assess the protection against the specific-Latrodectus antivenom.

  • XML | PDF | downloads: 478 | views: 989 | pages: 246-252

    Background: Ehrlichiosis is an emerging tick-borne zoonotic disease caused by the family of Anaplasmatacea. Re­cently, outbreak of human monocytic ehrlichiosis was reported in northern part of Iran. Besides, serological evidence of canine monocytic ehrlichiosis caused by Ehrlichia canis was reported from southeastern of Iran but the epidemi­ology of this disease is almost undetermined in Iran. The present study was designed to use PCR for detection of Ehrlichia spp. in tick infested household dogs and determination of risks of disease transmission to dog’s owners.
    Method:
    Blood samples were prepared from 100 tick infested household dogs after complete clinical examination. Complete cell blood count was done for each sample. DNA extraction was done and PCR was carried out by a com­mercial kit afterwards. Regarding to PCR results, blood samples were collected from owners and family members who were exposed to infected and non-infected dogs. A similar method was utilized for DNA extraction and PCR in human samples.
    Result:
    Ehrlichial DNA was detected by PCR in six percent of Rhipicephalus sanguineus tick pools and 9% of the examined dogs. No positive sample was detected among the 67 examined human bloods.
    Conclusion:
    Ehrlichiosis could be considered as an emerging canine disease but owning a dog should not be consid­ered a major risk factor for ehrlichiosis in humans. Further serological and molecular studies in different parts of Iran are required to clarify the epidemiology of ehrlichiosis in canine, ticks, and human population.

     

Short Communication

  • XML | PDF | downloads: 578 | views: 745 | pages: 253-258

    Background: Dengue is amongst the most serious mosquito-borne infectious disease with hot spots in tropical and subtropical parts of the world. Unfortunately, no licensed vaccine for the disease is currently available in medicine markets. The only option available is the management of dengue vector mosquito, Aedes aegypti (Diptera: Culicidae).
    Method:
    Predatory potential of five odonate nymphs namely Anax parthenope, Bradinopyga geminate, Ischnura forcipata, Rhinocypha quadrimaculata, and Orthetrum sabina were evaluated against the 4th instar larvae of the den­gue vector mosquito, Aedes aegypti, under laboratory conditions. The consumption of the mosquito larvae was eval­uated at three water volume levels viz., 1 liter, 2 liter and 3 liter.
    Results:
    The number of Ae. aegypti larvae consumed varied significantly among the five species, and at different levels of water volume (P< 0.01). However, the interaction between odonate nymphs and the water volumes was statistically non-significant (P> 0.05). Ischnura forcipata consumed the highest number of Ae. aegypti larvae (n=56) followed by A. parthenope (n=47) and B. geminate (n=46). The number of larvae consumed was decreased with in­creasing search area or water volume, and the highest predation was observed at 1-liter water volume.
    Conclusion:
    The odonate nymphs could be a good source of biological agents for the management of the mosquitoes at larval stages. 

  • XML | PDF | downloads: 460 | views: 828 | pages: 259-267

    Background: Present investigation is based on the clinicopatho-biochemical alteration related to natural tick borne bovine babesiosis in Punjab state.
    Methods
    : Blood samples from jugular vein of 542 bovines (cattle 466 and buffaloes 76) having history of tick in­festation, fever, hemoglobinurea or anemia were collected and tested for Babesia bigemina by blood smear examina­tion and PCR targeting 18S rRNA gene to distinguish clinically and subclinically infected groups. Further the hemato-biochemical parameters were correlated with the status of infection.
    Results:
    Overall, of the 542 samples tested 16.42 % were positive by PCR while only 1.66 % by blood smear exami­nation. The trend of molecular prevalence was found to decrease from north-eastern towards western Punjab. Analy­sis of the hematobiochemical alterations showed significant decrease in the levels of RBC, Hb, PCV, and MCV with significant increase in TBIL, MCH and MCHC levels.
    Conclusion:
    As the transmission of B. bigemina is transovarian, presence of even few infected Rhipicephalus (Booph­ilus) microplus ticks on a subclinically infection can be the nidus of infection for whole herd, causing severe economic losses, at the same time significantly affecting the physiology of carrier animal.

Case Report

Review Article

  • XML | PDF | downloads: 536 | views: 825 | pages: 113-126

    Visceral leishmaniasis is a deadly parasitic disease that is transmitted via the bite of a female sand fly, Phlebotomus argentipes. The highest burden of this disease is in northern India. In 2005, India embarked on an initiative with Ne­pal, Bangladesh, and the World Health Organization to eliminate visceral leishmaniasis by 2015. With the goal of 1 case in 10,000 people still unmet, it is prudent to evaluate the tools that have been used thus far to reduce vector numbers and cases of the disease. Herein, we present a review of studies conducted on vector-control strategies in India to combat visceral leishmaniasis including indoor residual spraying, insecticide-treated bed nets, environmental modification, and feed-through insecticides. This review suggests that the quality of indoor residual spraying may enhance control measures while a combination of spraying, nets, and feed-through insecticides would best confront the diverse habitats of P. argentipes.