Detection of Various Rickettsial Species in Ticks Collected from Small Ruminants in Western Iran.

Background: Most of the rickettsioses are transmitted by ticks, and often overlooked by the medical profession, but are clinically important as they cause major human diseases. Recent studies have shown the existence of some rickettsial species in Iran, but very little information is available about the status of rickettsial epidemiology and ecology. This study investigated the presence of Rickettsia spp. in ticks and ruminants in western of Iran by molecular methods. Materials and Methods: 250 blood samples were collected from sheep and goats, as well as 244 ticks were collected opportunistically from ruminants in the Kurdistan province. The collected samples were tested using a real-time quantitative PCR (qPCR) assay targeting the Rickettsia 16SrRNA gene. Rickettsia spp. positive by the qPCR were further amplified by conventional PCR of the gltA and OmpA genes. These ampliqons were further analyzed by sequencing. Results: The ticks species collected in this study included Rhipicephalus sanguineus, Rh. turanicus, Haemaphysalis concinna, and Dermacentor marginatus. In total, DNA of Rickettsia spp. was detected in 131 collected ticks (53.7%). Of the positives, Rickettsia slovaca (59.2%) and Ri. hoogstraalii (16.3%) were the most common species identified followed by Ri. raoultii, Ri. massiliae, Ri. sibirica, and Ri. conorii subsp. israelensis. In contrast, there were no positives observed in the blood samples collected from ruminants. Conclusion: The results indicate the presence of rickettsial species in ticks. The detection of these pathogens is significant because they cause clinical disease in humans. The results support the notion that the Iranian public health system needs to be more aware of these diseases.

Clinical, hematological and molecular evaluation of piroplasma and Anaplasma infections in small ruminants and tick vectors from Kurdistan province, western Iran.

Tick-borne haemoparasite infections are a major challenge in small ruminant (SR) production across tropical areas. The present study evaluated the prevalence of Theileria, Babesia and Anaplasma in SRs and their tick vectors and estimated the association between pathogen prevalence with clinical hematological findings among SR populations in Kurdistan province, western Iran. In total, 250 blood samples and 250 tick species (one per animal) were collected from SR populations, along with clinical and hematological examinations. Microscopy of blood smears and molecular analysis were performed to detect potential infection with Theileria, Babesia and Anaplasma. Moreover, haemoparasites were explored in the isolated ticks using semi-nested PCR. Based on microscopy, the prevalence of Theileria, Anaplasma and Babesia infections was 91.2%, 23.2% and 2.4%, respectively. Semi-nested PCR analysis of blood samples demonstrated 86.8%, 78.8% and 14% prevalence for T. ovis, A. ovis and B. ovis, respectively. Dermacentor marginatus and Rhipicephalus turanicus were predominant isolated tick vectors from SR, while D. marginatus was the most contaminated tick in all investigated counties. There were, also, a statistically significant association between the estimated molecular prevalence rates with semi-yellow conjunctiva (A. ovis), body temperature (T. ovis and A. ovis), heart rate (T. ovis and B. ovis), mean white blood cell count (T. ovis and A. ovis), mean red blood cell count (T. ovis and B. ovis), as well as mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in all haemoparasite infections. Future studies are recommended to reveal the epidemiology of such infections in SRs in Iran.

Determination of seroprevalence of brucellosis in livestock and high-risk population in Kurdistan, Western Iran.

BACKGROUND: Brucellosis is one of the most prevalent zoonotic diseases. Using serological tests are valid and rapid methods that could be used in the detection of the history of getting brucellosis. Considering that Iran is an endemic country for brucellosis, we aimed to investigate the rate of seroprevalence of brucellosis among livestock and human in Kurdistan province. MATERIAL AND METHOD: Serum sampling was performed from 51 slaughterhouse workers, veterinarians, and husbandry workers, along with 260 livestock (80 cattle, 120 sheep, and 60 goats). Rose Bengal plate test (RBPT), and Enzyme-linked immunosorbent assay (ELISA) were used for livestock and the anti-Brucella IgG antibody was evaluated in human participants. RESULTS: The seroprevalence (based on ELISA assay) in sheep, goats, and cows was 5.8%, 5%, and 1.2%, respectively. Also, the rate of anti-Brucella IgG was 3.9% among human participants. DISCUSSION: the current study, provided some valuable information on the seroprevalence of brucellosis in animal and human participants from the west of Iran. Considering the effects of brucellosis on causing reproductive disorders, including abortion, placental retention, andendometritis controlling the infection could have a significant impact on terms of economy.

The epidemiological survey of Coxiella burnetii in small ruminants and their ticks in western Iran.

BACKGROUND: Q fever is one of the most important zoonotic diseases caused by Coxiella burnetii. Although Q fever is an endemic disease in Iran, epidemiological data on C. burnetii infection are not yet complete in reservoirs and vectors in some parts of Iran. This survey investigated C. burnetii infection in small ruminants (sheep and goat blood samples) and their ticks in western Iran (Kurdistan province) in 2020. The presence of C. burnetii DNA was identified in these samples by targeting the IS1111 gene using the quantitative PCR (qPCR) method. RESULTS: Out of 250 blood samples (232 sheep and 18 goats), C. burnetii was detected in two samples (0.8%) belonging to the sheep (0.9%). In addition, 34 of 244 collected ticks (13.9%) from infested animals (244) were positive for C. burnetii infection. The highest prevalence of infection was found in Dermacentor marginatus (18.3%) and Haemaphysalis concinna (12.5%). CONCLUSIONS: The present study showed that ticks could have a possible role in the epidemiology of Q fever in Iran.

Molecular detection of Francisella tularensis in small ruminants and their ticks in western Iran.

Francisella tularensis is the causative agent of tularemia an infectious zoonotic disease. We attempted the molecular detecting of F. tularensis in small ruminants and ticks attached to these animals in Kurdistan province (the west of Iran). In this study, 250 blood and 244 tick samples were collected from sheep and goats and were tested for F. tularensis ISFtu2 gene detection using the Real Time-TaqMan PCR method. The collected ticks were morphologically classified as Dermacentor marginatus (67.2%), Rhipicephalus turanicus (12.30%), Rhipicephalus sanguineus (10.66%), and Haemaphysalis concinna (9.83%). No positive F. tularensis were identified in animal blood samples. F. tularensis was detected in 2 (0.82%) ticks samples. Positive samples were identified as F. tularensis subsp. holarctica and collected from D. marginatus ticks and Divandareh county. In this study, the presence of F. tularensis in ticks of Kurdistan province was confirmed, the possible role of ticks in the transmission to livestock and human through tick bites in this region should be considered.

Seroepidemiology of leptospirosis in livestock and workers of high-risk occupation in Kurdistan, Iran.

BACKGROUND: Leptospirosis is one of the major zoonotic infectious diseases which could cause disease in both animals and humans. Using ELISA is one of the serological tests that could be used in the detection of leptospirosis. Based on the different reports about the prevalence of leptospirosis in different parts of our country, we aimed to investigate the rate of Leptospira spp, among livestock and human in Kurdistan province. MATERIAL AND METHOD: ELISA assay was performed by ELISA kit (Novatec, Germany) for quantitative detection of anti-Leptospira IgG in human and IgM antibody and total antibodies (IgM and IgG) in serum samples of livestock. RESULTS: In the present study, the seroprevalence in sheep, goats, and cows was (2/30) 6.7% [95% CI 0.8%,22.1%], (1/31) 3.2% [ 95% CI 0.08%,16.7%] and 0%, respectively. Also, the rate of anti-Leptospira was (1/51) 1.9% [95% CI 0.05%,10.4%] among 51 human participants. DISCUSSION: the current study, provided some valuable information on the rate of leptospirosis in animal and human participants from west of Iran, which can be useful in terms of monitoring the disease in the area and helping the health care system to control the roots of bacterial transmission.

Francisella tularensis survey among ranchers and livestock in western Iran.

Tularemia is a zoonotic disease that transmitted to humans and domestic animals by wildlife, especially rodents. There are some evidences of the circulation of F. tularensis in rodents, livestock, human populations, and surface waters in western parts of Iran. In this study, we investigated the exposure of livestock and ranchers to F. tularensis in the endemic regions of western Iran. Blood samples were collected from 289 sheep, 103 cattle, and 51 ranchers in 2018. Animal sera were tested by standard tube agglutination method. The specific IgGs against F. tularensis were evaluated by ELISA in human sera. Moreover, the extracted DNAs from 50 sheep spleen samples were evaluated using TaqMan real-time PCR for the presence of ISFtu2 and FopA genes. All animal sera and spleen samples were negative for tularemia. Of the 51 human samples, two samples were seropositive and one sample showed a borderline status for tularemia. Serologic evidence of F. tularensis in the ranchers but negative results in the livestock indicates different transmission routes in human populations and domestic animals in western Iran. Therefore, drinking contaminated water, contact to wildlife or rodents and arthropod bite should be considered as probable routes in the suspicious areas.

Surveillance of Francisella tularensis in surface water of Kurdistan province, west of Iran.

BACKGROUND: The etiologic agent of tularemia, Francisella tularensis, is transmitted to humans via ingestion of contaminated water or food, arthropods bite, respiratory aerosols, or direct contact with infected animals body fluids or tissues. In the current study, due to the importance of water in transmitting the disease and the report of the disease in different regions of Iran, surface water of Kurdistan province were evaluated for the presence of F.tularensis. MATERIALS AND METHODS: Sampling was carried out in five-counties of Kurdistan province. Sixty-six specimens of surface water were collected. The detection was carried out by targeting ISFtu2 and fopA genes using TaqMan real-time PCR. Moreover, the samples were both cultured and inoculated into NMRI inbreed mice. Spleens of inoculated mice and bacterial isolates were tested by TaqMan real-time PCR. RESULTS: Despite the lack of isolation of F. tularensis, the results of the molecular testing indicate the presence of bacteria in surface water. Molecular positivity of one sample (1.51%) was confirmed using a real-time PCR for both ISFtu2 and fopA genes. Moreover, 4.54% of the samples were positive for ISFtu2. CONCLUSION: Since the in vitro isolation of bacteria from environmental samples is associated with a very low success rate and depends on various environmental parameters, the use of molecular techniques for monitoring of the bacteria in the contaminated areas is fully recommended.