Molecular detection of Coxiella burnetii in Kope cheese and cattle milk in West Azerbaijan, Iran.

There are few studies on Coxiella burnetii (Cb) as a causative agent of Q fever in dairy products in Iran. The prevalence of Cb was studied by polymerase chain reaction (PCR) method in Kope (pot) cheese and cattle milk collected from West Azerbaijan province, Iran. A total number of 240 Kope cheese and 560 milk samples were collected during the year 2020. All samples were subjected to PCR based on transposable gene IS1111. The results showed that 12.50% (95.00% confidence interval (CI): 9.00 - 16.10%) of Kope cheese and 13.00% (95.00% CI: 10.00 - 17.30%) of milk samples were positive for Cb. There was a significant difference in cheese and milk contaminations with Cb among the defined age groups as well as regional and seasonal variations. It was concluded that Kope cheese and cattle milk are important sources of Cb and should be considered as important risk factors in the epidemiology of Q fever disease in public health.

Prevalence and Molecular Characterization of Coxiella burnetii in Cattle, Goats, and Horses in the Republic of Korea.

Coxiella burnetii is an obligate intracellular zoonotic bacterium with a global distribution. This study was conducted to investigate the prevalence of C. burnetii in different animals and to assess the potential role of these species as reservoirs of infection and transmission to humans. A total of 592 blood samples (105 beef cattle, 61 dairy cattle, 110 Korean native goats, 83 Boer goats, and 233 horses) were collected in the Republic of Korea (ROK). The C. burnetii DNA was detected from blood samples using the transposon-like repetitive region (IS1111) by PCR method. The results showed that 22.7% of the Korean-native goats, 16.4% of the dairy cattle, 15.2% of the beef cattle, 6.0% of the Boer goats, and 5.2% of the horses were positive for C. burnetii. Significant differences were found between the animal species. The univariable binary logistic regression analysis revealed that the risk of contracting C. burnetii was significantly high by 5.4-fold in Korean-native goats (95% confidence interval [CI]: 2.60%-11.27%, p = 0.000), 3.6-fold in dairy cattle (95% CI: 1.48%-8.82%, p = 0.005), and 3.3-fold in beef cattle (95% CI: 1.51%-7.28%, p = 0.003) compared with horses. A phylogenetic tree based on the IS1111 gene revealed that our sequences had 92.2%-99.9% similarity and were clustered with those detected in humans, cattle, goats, dogs, rodents, and ticks. C. burnetii circulating in the ROK exhibits genetic variation. To the best of our knowledge, this is the first study to identify C. burnetii DNA in a horse in the ROK. These results suggest that cattle, goats, and horses can be potential reservoirs for C. burnetii and play an important role in the transmission of infection. Further studies should assess the pathogenicity of C. burnetii circulating in the ROK.

Coxiella burnetii in camels (Camelus dromedarius) from Algeria: Seroprevalence, molecular characterization, and ticks (Acari: Ixodidae) vectors.

Q fever is a widespread zoonotic disease caused by Coxiella burnetii that most commonly infects not only a variety of mammals but also arthropods and in particularly ticks. The aim of this study was to detect C. burnetii infection in camels including ixodid ticks using serological and molecular assays. Between July 2018 to June 2019, blood samples from 184 male and female camels (Camelus dromedarius) were collected from 3 regions of South-East Algeria and serum samples were tested for antibodies against Coxiella burnetii using indirect enzyme-linked immunosorbent assay (ELISA) kit. The positive sera and a total of 60 ticks were tested by quantitative PCR (qPCR) for detection of C. burnetii with primers and probes specific to the transposon-like repetitive region (IS1111 gene). Positive samples were genotyped by amplification and sequencing of partial sequences based on the IS1111 gene. The seroprevalence of antibodies against C. burnetii was 75.5%. Statistical analysis pointed out three potential risk factors associated with Q fever infection: geographic location, age class and season. No positive DNA of camel blood sample was observed. However, five Hyalomma dromedarii, one H. impeltatum and one H. excavatum tick species were detected positive for Coxiella burnetii DNA by qPCR, with an overall prevalence rate of 11.66% (7/60). The revealed Algerian strains by phylogenetic and comparative analysis of the IS1111 nucleotide sequences were clustered with several pathogenic C. burnetii strains isolated from ticks, human, and cattle located in Tunisia, Greece and in some Mediterranean countries, respectively. The study results clearly indicate that camels and their ticks in Algeria may play an important role as a reservoir for C. burnetii and can be considered as a significant source of Q fever transmission to other animal species and humans.

Molecular Investigation of the Status of Ticks on Infected Cattle for Coxiella burnetii in India.

PURPOSE: In Indian subcontinent, the epidemiological studies on the status of ticks in the transmission of Coxiella burnetii have not been explored comprehensively. The objective of the present study was to investigate the status of ticks for C. burnetii among coxiellosis positive cattle. METHODS: The present study was carried out in three locations of the northern states of India. A total of 1648 tick samples were collected from the tick infested cattle (n = 146) that were tested positive for coxiellosis by indirect serum-ELISA assay and/or the trans-PCR assay. The tick samples were screened using the trans-PCR assay targeting species-specific IS1111 transposase gene of C. burnetii. The sequencing of PCR products was planned to differentiate C. burnetii and Coxiella-like bacteria (CLB). RESULTS: The collected ticks were identified as Rhipicephalus microplus (n = 1049), Hyalomma anatolicum (n = 416), and Hyalomma spp. (n = 183). On molecular investigation, none of the collected tick samples were found to be positive for the IS1111 gene. CONCLUSION: The findings of the present study ruled out the involvement of ticks in circulation of the pathogen within the cattle population that were screened. However, extensive epidemiological studies are needed to conclusively rule out or establish the role of ticks as a competent vector for C. burnetii transmission in cattle and other hosts.

Evidence of Coxiella burnetii in Punjab province, Pakistan.

Coxiella burnetii causes query (Q) fever, an important zoonotic disease with worldwide significance. The role of environment in the ecology of C. burnetti, and its influence on seroconversion in animals has not been elucidated in Pakistan. We carried out a cross-sectional study in Punjab province to (1) determine the prevalence and distribution of C. burnetii in soil using an ISIIII gene-based real time-polymerase chain reaction (RT-PCR) assay, (2) analyze association between the occurrence of C. burnetii in soil and its predictors i.e. soil characteristics (macro- and micro-nutrients) and several likely risk factors including the seroconversion in small ruminants at places where its genome had or had not been detected, and (3) predict homology and genetic diversity of the identified strains using sequences originated from different hosts worldwide. A total of 2425 soil samples from nine districts of Punjab province were processed. C. burnetii DNA was detected in 47 samples (1.94%, 95% CI: ±0.55) originating from 35 villages of studied districts (7.22%, 95% CI: ±2.30). The highest prevalence was found in Attock (7.11%, 95% CI: ±3.36), followed by Lahore (4.83%, 95% CI: ±3.49), Sahiwal (4.70%, 95% CI: ±2.6), Dera Ghazi Khan (2.33%, 95% CI: ±2.02), Faisalabad (1.35%, 95% CI: ±1.18) and Sheikhupura (0.68%, 95% CI: ±0.94). The odds of detecting bacterial DNA in soil was increased with a unit increase in organic matter [2.511 (95% CI: 1.453-4.340), p=0.001] and sodium [1.013 (95% CI: 1.005-1.022), p=0.001], whereas, calcium [0.984 (95% CI: 0.975-0.994), p=0.002] and potassium [0.994 (95% CI: 0.990-0.999), p=0.011] had protective effect where a unit increase in each analyte decreased odds for its occurrence by 1.0% approximately. Likewise, for categorical variables (risk factors), the odds of detecting C. burnetii were higher at locations >500m away from a main road [1.95 (95% CI: 1.06-3.78), p=0.04]. The enzyme-linked immunosorbent assay (ELISA) revealed an increased prevalence of antibodies in sheep (17.9%, 95% CI: ±5.54) compared with goats (16.4%, 95% CI: ±4.34). When determining the association between soil DNA and C. burnetii antibodies in small ruminants, the odds of detecting these antibodies were significant in sheep at the livestock barns [2.81 (95% CI: 1.20-7.37), p=0.02]. The IS1111 gene-based sequence analysis revealed a clustering of the DNA into two distinct groups with much genetic divergence (0.76-68.70%): the first group that contained sequences from Lahore district clustered with human and buffalo origin isolates, whereas the second group that contained the sequences from the remaining study districts clustered with goat-, rodent- and human-origin isolates. This study provides the first evidence of the presence of C. burnetii in the environment in Punjab province, Pakistan. Future studies are needed to ascertain the bacteria's molecular epidemiology over a wide geographical area, type the isolates, and evaluates the potential risks to human populations, particularly farmers and veterinarians.

Isolation of Coxiella burnetii from bovines with history of reproductive disorders in India and phylogenetic inference based on the partial sequencing of IS1111 element.

In the present study, a total of 158 blood samples from 148 bovines and 10 dogs having a history of reproductive disorders were screened for Coxiella burnetii by trans-PCR method. In case of bovines, 6.08% (9/148) blood samples comprised of 4.54% (4/88) cattle and 8.33% (5/60) buffaloes turned out to be positive for C. burnetii DNA while all the samples from dogs (10) were found negative. Of the 9 PCR-positive bovine blood samples, the organism could be isolated only from 3 cases of buffaloes by chick embryo inoculation method. Further, to predict the homology and genetic diversity, the recovered C. burnetii isolates designated as Y1, Y3 and Y7 were partially sequenced for IS1111 gene. On phylogenetic analysis, Y3 and Y7 isolates clustered to a common node away from Y1 isolate. This study may enlighten the nature of circulating C. burnetii isolates in different parts of the world. To the best of our knowledge, this appears to be the first report describing phylogenic analysis of C. burnetii isolates based on IS1111 gene sequence.