Molecular Detection of Spotted Fever Group Rickettsia (Rickettsiales: Rickettsiaceae) in Ticks of Iran.

Ticks are reservoir hosts of pathogenic Rickettsia in humans and domestic animals. Most pathogenic Rickettsia species belong to the spotted fever group (SFG). The present study aimed to determine the tick species infected with Rickettsia based on the genus-specific 23S ribosomal ribonucleic acid (rRNA), 16S rRNA, and citrate synthase (gltA) gene fragments. A total of 61 tick specimens were selected for molecular assay and 12 samples for sequencing. Phylogenetic analysis was conducted using neighbor-joining and Bayesian inference methods. Argas persicus, Haemaphysalis sulcata, Ha. inermis, and Hyalomma asiaticum were infected by spotted fever Rickettsia. The SFG is the main group of Rickettsia that can be detected in the three genera of ticks from Iran.

Molecular Detection of Anaplasma marginale and Anaplasma ovis (Rickettsiales: Anaplasmataceae) in Ixodid Tick Species in Iran.

The present study was conducted as the first molecular detection of Anaplasma species in tick samples based on the sequencing of major surface proteins 4 (msp4) gene fragments in different parts of Iran. A total of 130 tick specimens were collected from Hormozgan, Lorestan, and Guilan, Iran, within 2015 to 2017. Hyalomma asiaticum, Hyalomma dromedarii, Rhipicephalus sanguineus, and Rhipicephalus (Boophilus) species were identified in different geographical regions. An amplicon of 464-bp msp4 of Anaplasma was amplified using polymerase chain reaction in various tick species. Three sequences, including one Anaplasma marginale from R. (Boophilus) species and two Anaplasma ovis from Rhipicephalus sanguineus, were obtained after sequencing. It is concluded that bovine and ovine anaplasmosis agents are present in tick samples in Iran. The use of the gene families of six major surface proteins for the detection of various Anaplasma species is recommended.

Evidence for the presence of Ctenocephalides orientis in livestock dwellings in northwest Iran.

Fleas are important vectors of diseases such as murine typhus, tularaemia, hymenolepiasis and plague. The presence of active foci and history of human- and flea-transmitted plague in northwest Iran prompted the present group to collect and identify fleas from human and livestock dwellings across West Azerbaijan Province. Adult fleas were collected and identified using routine taxonomic keys. Species designation was confirmed by sequencing the cytochrome oxidase subunit I (COI). Of the collected specimens (n = 989), 104 were collected off-host (30 from human dwellings and 74 in light traps) and the rest were found on hosts (107 on animals and 778 by human bait). Of these fleas, 394 (40%) were male and 595 (60%) were female. The collected specimens belonged to the species Ctenocephalides canis, Ctenocephalides felis, Ctenocephalides orientis and Pulex irritans (all: Siphonaptera: Pulicidae). The amplified COI fragment, in addition to confirming the morphological identification of species, showed good efficacy in separating the different species in the phylogenetic analysis. In addition to the identification of fleas from human and livestock dwellings using morphological and molecular characteristics, the current paper represents the first report of the presence of C. orientis in northwest Iran. This finding suggests that changing climate conditions may have expanded the distribution of this species.

Molecular detection of Crimean-Congo haemorrhagic fever virus in ticks collected from infested livestock populations in a New Endemic Area, South of Iran.

OBJECTIVE: Crimean-Congo haemorrhagic fever (CCHF) is a viral zoonotic disease with potentially fatal systemic effects on man. We aimed to determine the presence of CCHF virus among collected ticks from domestic livestock from October 2012 to September 2013. METHODS: A total of 1245 hard and soft ticks were collected from naturally infested ruminants in Marvdasht County, Fars Province, south of Iran. Nine tick species and one unidentified species in four disparate genera were detected. A total of 200 ticks were randomly selected and analysed by reverse transcription-polymerase chain reaction (RT-PCR) for the presence of CCHF virus genome. RESULTS: The viral genome was detected in 4.5% (9 samples) of the studied tick population. The infected ticks belonged to the species of Hyalomma marginatum' Hyalomma anatolicum and Rhipicephalus sanguineus. The viruses detected in these three tick species were clustered in the same lineage as Matin and SR3 strains in Pakistan and some other Iranian strains. These results indicate that the ticks were wildly infected with a genetically closely related CCHF virus in the region. CONCLUSION: Regular controls and monitoring of livestock to reduce the dispersion of ticks and providing information to those involved in high-risk occupations are urgently required.

Serological and molecular epidemiology of Crimean-Congo hemorrhagic fever in Ghaemshahr county in Mazandaran province; Iran.

Crimean-Congo Hemorrhagic Fever (CCHF) is a tick-borne viral hemorrhagic fever disease which is known as an endemic disease within some provinces of Iran. Ticks play an important role in transmission of the disease. As vector and reservoir, ticks transmit CCHF virus from livestock to human. The current study reports the presence of CCHFV in Ghaemshahr county of Mazandaran province, in north of Iran based on the evidences obtained from ELISA and RT- PCR. Based on our results, IgG antibodies against CCHFV were detected in 4(4.8%) out of 84 sheep sera samples. Forty sera were obtained from people who were in close contact with the examined sheep, none of which had IgG antibodies against CCHFV. Using RT-PCR, we confirmed the existence of CCHFV genome in 1.7% of hard tick samples. Sequence analysis demonstrated that CCHFV genomes isolated from ticks were 100% identical to those isolated from the corresponding livestock. This study confirms the presence of the virus in this region; so people in close contact with livestock and health care workers should be alerted.

Wild rodents and their ectoparasites in Baluchistan area, southeast of Iran.

During 2008-2009 a total of 67 individuals of rodents, Tetera indica, Meriones hurrianae, Meriones libycus and Gerbillus nanus were trapped in three areas, Bampor, Daman and Qasre Qand from Iranshahr and Nikshahr districts. There is a significant difference between comparative abundance of four species (P<0.05). A total of 1422 ectoparasites collected including 299 mites (21%), 127 fleas (8.9%), 972 lice (68.4%) and 24 ticks (1.7%). Significant findings amongst the ectoparasites is the lice group with three species identified, Laelaps accuninata, Andralaelaps hermophrodita and Paracheylaellaps pyriformis being the first record in the study areas. All four captured genera of rodents are known as main/ potential reservoir hosts of zoonotic cutaneous leishmaniasis. The migration habit of rodents may affect the spatial distribution of parasitic ticks and their transmitted diseases like CCHF, which has been reported in recent years from Sistan and Baluchestan province. Monitoring of rodent populations and their ectoparasites will help to predict the potential of zoonotic arthropod-borne diseases.

A survey of Crimean-Congo haemorrhagic fever in livestock and ticks in Ardabil Province, Iran during 2004-2005.

Crimean-Congo haemorrhagic fever (CCHF) is a viral haemorrhagic fever caused by the CCHF virus. It is mainly transmitted to humans and animals by ticks. In recent y, large numbers of livestock have been transported across the border areas of Ardabil Province resulting in an outbreak of CCHF in the adjacent districts. A comprehensive study was carried out to assess the epidemiological aspects of the disease in this province. In the study area, 130 ticks were collected from randomly selected villages and classified into 9 species of hard tick and 2 species of soft tick. All ticks were analyzed for the presence of CCHF virus genome using gel-based and real-time reverse transcriptase polymerase chain reactions (RT-PCR). The results showed CCHF infection in almost 28% of ticks collectively. Also, of 56 livestock sera, around 39% were IgG-positive. The presence of anti-CCHF virus IgG antibodies and the CCHF virus genome in ticks points to a great hidden threat of an outbreak in these districts. Those in high-risk professions in this province should be informed and trained on the risk of CCHF with urgency.

Hard Ticks on Domestic Ruminants and their Seasonal Population Dynamics in Yazd Province, Iran.

BACKGROUND: Ticks are the main vectors for transmission of different pathogens to human and animals. This survey was performed to find out distribution of ticks, which infested the domestic ruminants in Yazd Province, central Iran during year 2008-2009. METHODS: A total number of 30 villages from both mountainous (20%) and plateau (80%) regions of the province were selected randomly. Ticks were colleted from the body of infested animals and transported to the laboratory of Medical Entomology, School of Public Health, Tehran University of Medical Sciences and then were identified to space level using valid identification key. RESULTS: A total of 583 hard ticks were collected. The ticks were classified into three genera and 7 species including: Hyalomma dromedarii (55.92%), Hy. marginatum (13.20%), Hy. anatolicum (9.78%), Hy. detritum (4.98%), Hy. asiaticum (3.94%), Rhipicephalus sanguineus (11.84%), and Dermacentor marginatus (0.34%). The highest seasonal activities occurred in summer. The prevalence of the Ixodidae ticks was more evident in plateaus area in Yazd Province. Among the hosts including: cow, goat, sheep and camel, the ticks that collected from camel was more prevalent. The ratio of male was more than female ticks. Hyalomma. dromedarii was the predominant tick species and accounted for 55.92% of the ticks. CONCLUSION: Some of the collected ticks may play an important role for transmission of vector borne disease to human; therefore, the results of this study will provide a clue for vectors of tick-borne diseases in the region for local authorities for implementation of disease control.

Study on Presence of Borrelia persica in Soft Ticks in Western Iran.

BACKGROUND: A molecular survey was conducted to investigate the presence of pathogenic Borrelia persica species causing the tick borne relapsing fever (TBRF) in Takistan district Qazvin Province, western Iran. METHODS: A number of 1021 soft ticks were collected from 31 villages including previously reported infected and none-infected TBRF cases and individually examined for the presence of B. persica DNA by conventional PCR targeting the 16S rRNA. RESULTS: A total of 1021 soft ticks of three species of Ornithodouros tholozani (120: 11.75%), O. lahorensis (461: 45.15%) and Argas persicus (440: 43.1%) were collected and tested against Borrelia infection. Soft ticks were more prevalent (67%) in infected areas than none infected areas. The rate O. tholozani in infected areas was much greater (29 times) than none infected areas. Ninety seven percent of soft ticks in none infected areas were of O. tholozani. Sixteen (16.7%) ticks of tested (n=95) O. tholozani were infected with B. persica. Three (1.3%) out of 205 soft ticks of O. lahorensis were positive for Borrelia sp., and no infection was observed in A. persicus. TaqI RFLP analysis and sequence analysis of the positive PCR products showed the presence of B. persica. The RFLP analysis showed that the positive ticks of O. lahorensis were infected with unknown Borrelia species. CONCLUSION: This study showed that although there were no TBRF cases in Takisan, but still infected O. tholozani, the known vector of TBRF, presented in the region. Control measures needs to be fulfilled in Thakisan.

Tick infestation rate of sheep and their distribution in abdanan county, ilam province, iran, 2007-2008.

BACKGROUND: Ticks are hematophagous arthropod belonging to the Class of Arachnids. Ticks are also one of the major vectors of pathogens to animal and human. This study was conducted to determine tick infestation rate of sheep in Abdanan during 2007-2008. METHODS: Sampling was performed seasonally in 19 villages during spring 2007 until winter 2008. A total of 1095 sheep were selected and tested for tick infestation. After collection, all ticks were transported to laboratory of Medical Entomology and were identified with appropriate identification keys. RESULTS: Totally, 864 hard ticks were collected. The ticks were classified into two genera and 5 species including: Hyalomma marginatum (44.67%), Hy. anatolicum (43.17%), Hy.asiaticum (6.37%), Hy. dromedarii (5.55%), Heamaphysalis sulcata (0.24%). The highest seasonal activity was observed in spring (36.46 %) and the lowest seasonal was in winter (11.57%). The rate of tick frequency in mountainous region was 48.15% and it was 51.85% in plateau regions. In this study, tick infestation of sheep was 11.41%. CONCLUSION: Hy.marginatum has the more frequent density in the study area.

Study on Ectoparasites of Rhombomys opimus, the Main Reservoir of Zoonotic Cutaneous Leishmaniasis in Endemic Foci in Iran.

BACKGROUND: Ectoparasites of mammalian hosts play an important role for transmission of diseases from the host reservoirs to human. The aim of this study was to determine the species composition and infestation parameters for parasitic arthropods associated with Rhombomys opimus. METHODS: Ectoparasites of R. opimus were collected from seven endemic district of ZCL in Iran including Shirvan and Sfaraysen in North Khorasan Province, Kalaleh in Golestan Province, Damghan and Shahrood in Semnan Province, and Badrood and Habibabad in Isfahan Province. The areas of study were mainly desert and plain. Rodents were captured using Sherman life traps during active seasons from May to November 2008. Captured rodents were transported to laboratory and their ectoparasites were picked up using brushing against the fur of the rodents. Ectoparasites were stored in 70% ethanol for their preservation and then identified based on morphological characters. RESULTS: Ectoparasites belonged to one flea species of Xenopsylla nuttalli and one mite species of Ornithonussus bacoti. The flea species with 75.3% was more common than the mite. O. bacoti might play an important role in transmission of rat mite dermatitis among R. opimus colony. CONCLUSION: Results will provide an essential clue for combating zoonotic diseases in the region.

Ectoparasites of rodents captured in bandar abbas, southern iran.

BACKGROUND: Rodents play important role as host of ectoparasites and reservoir of different zoonotic diseases. The aim of this study was to asses the infestation of commensal rodents with ectoparasites in Bandar Abbas, a port city located in the northern part of the Persian Gulf in Iran. METHODS: Rodents were captured using live traps during the study period in year 2007. After transferring the rodents to the laboratory, they were identified and then their ectoparasites were collected and mounted for species identification using appropriate systematic keys. RESULTS: A total of 77 rodents were identified including Rattus norvegicus (74%), R. rattus (16.9%), Mus musculus (7.8%) and one hamster. Among all rodents, 40.3% were found infested with ectoparasites. A total of 69 ectoparasites were collected comprising flea, lice, mite and tick. Two species of fleas; Xenopsylla cheopis and X. astia were identified with higher index of X. astia. Two genera of ticks including Hyalomma sp. and Rhipicephalus sp. were identified. Laelaps nuttalli was the only mite found. The Polyplax spinulosa was considered as lice ectoparasite. CONCLUSION: Among all arthropods collected, flea and lice had the most and the least frequency, respectively. Nearly all rodent species were infested with Xenopsylla. These fleas are important due to their role in plague and murine typhus transmission. Ticks are important due to their role in CCHF (Crimean-Congo Hemorrhagic Fever), theileriosis, babesiosis, anaplasmosis and ehrlichiosis transmission .Monitoring of ectoparaiste infestation is important for preparedness and early warning preparation for possible control of arthropod-borne diseases.

Comparative susceptibility of cypermethrin in Ornithodoros lahorensis Neuman and Argas persicus Oken (Acari: Argasidae) field populations.

The toxicity of cypermethrin was determined in five different soft tick strains of Argas persicus Oken and Ornithodoros lahorensis Neuman by topical application method. The O. lahorensis Bij, O. lahorensis west O1, O. lahorensis Mesh, A. persicus Lor, A. persicus West Ap strains were collected from Bijar, Kurdistan province, Takab, Western Azerbaijan province, Meshkinshar, Ardebil province, Khoramabad, Lorestan province, Takab, Western Azerbaijan province of different areas of Islamic Republic of Iran, respectively during 2004 and 2005. In the topical application bioassay, the average LD50 of O. lahorensis Bij, West O1, Mesh and A. reflexus Lor and West AP strains were 0.03, 0.04, 1.7, 0.7 and 1.7 microg tick(-1), respectively and the steep slopes of dose-response curves indicated that the field population of these soft tick strains were homogenous in response to cypermethrin. Comparison of the resistance ratio of collected strains with susceptible strain showed a resistance ratio of 56.7 and 2.4-folds for cypermethrin in O. lahorensis Mesh and A. reflexus West Ap strains, whereas the O. lahorensis West O1 completely susceptible to cypermethrin.