Human seroprevalence to Coxiella burnetii (Q fever) in Northern Ireland

Despite the widespread prevalence of infection with Coxiella burnetii, there have been few large population-based studies examining the epidemiology of this infection. The aim of this study was to examine the distribution and determinants of C. burnetii past infection in Northern Ireland (NI). Coxiella burnetii phase II specific IgG antibodies were measured by enzyme-linked immunosorbent assay in stored serum from 2,394 randomly selected subjects, aged 12-64, who had participated in population-based surveys of cardiovascular risk factors performed in 1986 and 1987. The overall prevalence of C. burnetii antibody positivity was 12.8%. The prevalence of sero-positivity was slightly higher in males than that in females (14.3% versus 11.2%, P = 0.02). Sero-positivity was low in children (<10%), increasing to 19.5% and 16.4% in males and females, respectively, in the 25-34 age group and subsequently remaining fairly steady with increasing age. Sero-positivity among farmers, at 48.8%, was significantly higher than the general population. More sero-positive than sero-negative women had a history of a miscarriage or still-birth (19.5% versus 9.8%, P < 0.001). In conclusion, this study demonstrated a high prevalence of evidence of past C. burnetii infection in NI. Associations between past C. burnetii infection and age, sex, social class, occupation and reproductive history were seen. We estimate that 20% of Q fever infections in NI occur in farmers.     

Serological evidence of Coxiella burnetii infection in beef cattle in Queensland

Background Queensland has the highest incidence of Q fever in Australia. The aim of this study was to undertake a cross‐sectional seroprevalence survey of Coxiella burnetii, the causative agent of Q fever, in beef cattle in Queensland. Methods Serum samples were tested by ELISA for both phase II and phase I antigens of the organism using an Australian isolate. Blood samples were collected at an abattoir that processes beef cattle originating from northern and north‐western Queensland, in addition to blood samples taken from beef cattle across Queensland as part of a second survey. Results Seropositivity was 16.8% (95% confidence interval 16.7–16.8%). Conclusion Evidence of C. burnetii infection in beef cattle has public health implications for occupational exposure of primary producers and veterinarians and for the proximity of beef cattle properties to residential areas in regional Queensland. This study is the first known investigation of C. burnetii seroprevalence in beef cattle in Queensland and the first known use of an Australian C. burnetii isolate for screening using both phase II and phase I antigens.     

Q热贝纳柯克斯体TaqMan荧光定量PCR检测方法的建立

根据Q热贝纳柯克斯体(Coxiella burnetii)插入IS1111序列设计引物和探针,建立快速检测Q热的TaqMan实时荧光定量PCR方法。以梯度稀释含有目的扩增片段的重组质粒作为标准品,进行定量PCR反应。结果显示,该方法能够检测出10个拷贝数的阳性质粒;标准曲线相关系数为0.995,扩增效率为103%;结核分枝杆菌(M.tuberculosis)、衣原体(C.psittaci)、布鲁氏菌(Brucella.spp)及牛血液的核酸样本特异性检测结果均为阴性。本研究建立的TaqMan荧光定量PCR法灵敏度高、特异性好,对Q热的检测与鉴定中具有良好的应用前景。     

Epidemiology and clinical features of human infection with Coxiella burnetii in Denmark during 2006-07

Query (Q) fever was virtually unknown in Denmark in 2005, when, after the introduction of new sensitive diagnostic methods for Coxiella burnetii, an increasing number of positive cattle created concern among people with frequent exposure. This led to a dramatic rise in examinations for Q fever among humans in the following 2 years. The aim of our study was to assess indication for testing and symptoms in individuals with serological signs of infection with C. burnetii. We performed a case-review study of seropositives among all humans tested for Q fever in 2006-07 in Denmark. Seropositive cases were categorized with acute infection: 4-fold increase in immunoglobulin G (IgG) phase II or concomitant IgM phase II ≥ 1 : 256 and IgG phase II ≥ 1 : 1024; and previous infection: IgG phase II ≥ 1 : 1024. A borderline result was defined as: IgG phase II = 1 : 512. Physicians completed a questionnaire retrospectively. Of the 1613 people tested, 177 (11%) were seropositive [37 (2%) acute infection, 140 (9%) previous infection], 180 had a borderline result. Among 127 seropositives responders, 31% were tested due to symptoms compatible with Q fever after a possible exposure to C. burnetii, 64% were asymptomatic and were tested following relevant exposure only; 64% were males, 43% farmers, 39% veterinarians, 84% had been exposed to cattle. The most frequently reported symptoms were asthenia (25%), myalgia (21%), fever (17%) and headache (13%). About two-thirds of seropositives reported asymptomatic infections, and were tested for Q fever because of concern for occupational exposure to cattle. One-third of the seropositives reported symptoms consistent with Q fever, the majority being mild. Our study provided important evidence that increased requests for Q fever testing in 2006-07 arose from heightened public awareness of the disease, and not from an outbreak of clinical disease. Nonetheless, Q fever should be considered endemic in Denmark.     

Serological evidence of Coxiella burnetii infection in dogs in a regional centre

OBJECTIVE Investigate the seroprevalence of the causative agent of Q fever, Coxiella burnetii in domestic dogs in the Townsville region, North Queensland, Australia. METHOD Blood samples were collected from dogs attending veterinary clinics for routine procedures. RESULTS An overall seropositivity of 21.8% (95% confidence interval (CI) 21.6-22.1%) was observed. A retrospective study of samples collected in the same region during 1984-85 was also performed, with an overall seropositivity of 16.0% (95% CI 15.9-16.2). CONCLUSION Evidence of C. burnetii infection in domestic dogs may have public health implications for dog owners, as well as veterinarians because of occupational exposure. This study is the first known investigation of C. burnetii seroprevalence in dogs in Queensland.     

Seroprevalence and Factors Associated with Coxiella burnetii Infection in Small Ruminants in Baringo County,Kenya

To improve estimates of C. burnetii epidemiology in Kenya, a survey was undertaken in small ruminants in Baringo County, where acute cases of Q fever in humans had been reported in 2014. From 140 household herds selected, 508 (60.5%) goats and 332 (39.5%) sheep were included and an indirect ELISA assay for C. burnetii IgG antibodies performed. In addition, epidemiological information at both herd and animal level was collected. Generalized mixed‐effects multivariable logistic model using herd as the random effect was used to determine variables correlated to the outcome. Overall seroprevalence was 20.5% (95% CI: 17.8%, 23.3%). Goats had 26.0% (95% CI: 22.2%, 30.0%) compared to sheep 12.2% (95% CI: 8.7%, 16.0%). Nomadic pastoralism, goats and older animals (>1 year) were associated with greater risk of C. burnetii seropositivity (P = ≤0.05). Heterogeneity in C. burnetii seropositivity was observed across the sublocations (P = 0.028). Evidence of C. burnetii exposure in small ruminants revealed poses a potential risk of exposure to the people living in close proximity to the animals. We recommended integrated animal–human surveillance and socio‐economic studies for C. burnetii, to aid our understanding of the risk of transmission between the animals and humans, and in the design of prevention and control strategies for the disease in the region.     

Q Fever (Coxiella burnetii) Knowledge and Attitudes of Australian Cat Breeders and Their Husbandry Practices

A Q fever outbreak in a small animal veterinary hospital, associated with a cat caesarean section, initiated a cat seroprevalence study (n = 712) that found circulating antibodies to Coxiella burnetii was highest in cattery‐confined breeding cats (9.3%). These findings stimulated interest about potential sources of C. burnetii infection for cats and humans associated with cats. Cat breeders are potentially a group at increased risk of C. burnetii infection, and this study sought to identify potential risk factors. A cross‐sectional online survey was conducted targeting all domestic cat breeders registered with an affiliate member body in Australia in 2015. Responses from 177 cat breeders across Australia were analysed. Forty per cent of responding cat breeders had not heard of Q fever. Raw meat was fed as an integral constituent of the diet by 89% of respondents. Eighty per cent of respondents allowed queens access to the home for parturition, and assistance of queens and resuscitation of kittens at the time of birth were reported by 97% of respondents. Respondents who perceived some level of exposure to Q fever through their breeding activities were three times less likely to perform mouth‐to‐snout resuscitation (OR 0.3 95% CI 0.1–0.9; P = 0.034) than those who did not perceive a risk of exposure. Similarly, respondents who perceived Q fever as a risk through breeding activities were close to eight times more likely to use personal protective equipment during parturition (OR 7.7 95% CI 1.5–39.9; P = 0.015) than those who did not. Husbandry practices of cat breeders that may increase the risk of C. burnetii transmission require further targeted investigations to assess the contribution of these risk factors to the acquisition of disease. Concurrent education forums are recommended to inform Australian cat breeders of the aetiopathogenesis of Q fever.     

Serosurvey and Observational Study of US Army Veterinary Corps Officers for Q Fever Antibodies from 1989 to 2008

Since World War II, the military has experienced outbreaks of Q fever among deploying units including recent case reports of Q fever in US military personnel returning from serving in the Middle East during Operation Iraqi Freedom and Operation Enduring Freedom. Occupational exposure and prevalence of Q fever among US Army Veterinary Corps officers have not been examined. A retrospective serosurvey and observational study of 500 military veterinarians were conducted using archived serum specimens from military veterinarians who entered and served between 1989 and 2008 and were tested for exposure to Coxiella burnetii. Corresponding longitudinal health‐related, demographic, medical and deployment data were examined. A total of 69 (13.8%) individuals at military entry and 85 (17%) had late career positive titres. A total of 18 (3.6%) individuals showed seroconversion. Women were more likely to be seropositive after military service [prevalence ratio (PR) 1.96; 95% confidence interval (CI) 1.15–3.35] and were also more likely to seroconvert (incidence rate ratio 3.55; 95% CI 1.19–12.7). Women who deployed to Operation Iraqi Freedom were more likely to be seropositive (PR 3.17; 95% CI 1.03–9.71). Veterinarians with field service and pathology specialties had the highest incidence rates (7.0/1000 PY; 95% CI 4–12 and 3–19, respectively). This is the first report documenting US military veterinarians' exposure to C. burnetii. Military veterinarians are at risk prior to service, with moderate number of new cases developing during service and most maintaining titres for long periods of time. Women consistently demonstrated higher seroprevalence and incidence levels. As increasing numbers of women enter the veterinary profession and subsequently the US Army, this may warrant close monitoring. This study likely underestimates exposure and risk and does not address chronic health effects, which may be valuable to explore in future health studies.     

Risk factors associated with self-reported Q fever in Australian wildlife rehabilitators: Findings from an online survey

Australian wildlife rehabilitators (AWR) are at increased risk of developing Q fever, a serious zoonotic disease caused by the intracellular bacterium Coxiella burnetii. Previous studies have suggested that Australian wildlife may be a potential C. burnetii infection source for humans. However, a recent serological survey of AWR found no association between C. burnetii exposure and direct contact with any wildlife species. To further explore the potential risk that wildlife may pose, this study aimed to identify associations between self-reported Q fever in AWR and risk factors for exposure to C. burnetii. An online cross-sectional survey was implemented in 2018 targeting AWR nationwide. Risk factors for self-reported Q fever were determined using multivariable logistic regression. Medically diagnosed Q fever was self-reported in 4.5% (13/287) of unvaccinated respondents. Rehabilitators who self-reported medically diagnosed Q fever were significantly more likely to: primarily rehabilitate wildlife at a veterinary clinic (OR 17.87, 95% CI: 3.09–110.92), have domestic ruminants residing on the property where they rehabilitate wildlife (OR 11.75, 95% CI: 2.91–57.42), have been educated at a High School/Technical and Further Education level (OR 10.29, 95% CI: 2.13–84.03) and be aged >50 years (OR 6.61, 95% CI: 1.60–38.35). No association was found between self-reported Q fever and direct contact with wildlife. These findings support previous work suggesting that AWR are at increased risk of C. burnetii infection and may develop Q fever potentially via exposure to traditional infection sources including livestock, other domestic animals, or contaminated environments, in association with their rehabilitation practices and lifestyle. Although Q fever vaccination is recommended for AWR, vaccine uptake is low in this population. Future studies should aim to determine the level of Q fever awareness and identify barriers to Q fever vaccination in this at-risk group. The difficulty in accessing the AWR population also highlights the need for a national centralized AWR database.     

Assessing the within-herd prevalence of Coxiella burnetii milk-shedder cows using a real-time PCR applied to bulk tank milk

Thirty-seven bulk tank milk (BTM) and individual milk samples of all contributing cows were tested for Coxiella burnetii detection by a real-time PCR assay and used to assess the relationship between the BTM PCR-response and (i) the within-herd prevalence of milk-shedder cows and (ii) the proportion of heavy milk-shedder cows. The within-herd prevalence of milk-shedder cows (i) was found to be significantly higher in herds with a positive BTM and (ii) increased significantly with the estimated titre in Coxiella burnetii obtained in positive BTM. The proportion of heavy milk-shedder cows among the milk-shedder cows increased significantly with an increased estimated titre in Coxiella burnetii in positive BTM. Therefore, a real-time PCR assay applied to BTM samples collected repeatedly over time appears to be a valuable tool to assess on a larger scale the status of herds towards Coxiella shedding, and to evaluate the efficiency of control actions aimed at controlling and/or preventing Coxiella shedding in dairy herds.     

Coxiella burnetii seroprevalence in unvaccinated veterinary workers in Australia: Evidence to support Q fever vaccination

Q fever (caused by Coxiella burnetii) is a serious zoonotic disease that occurs almost worldwide. Occupational contact with animals increases the risk of exposure, and Q fever vaccination is recommended for veterinary workers in Australia. This study aimed to investigate C. burnetii seroprevalence among unvaccinated veterinary workers in Australia and determine factors associated with a positive serological result. During 2014 and 2015, convenience sampling at veterinary conferences and workplace vaccination clinics was undertaken. Participants completed a questionnaire and provided a blood sample for C. burnetii serology. Participants were predominantly veterinarians (77%), but veterinary support staff, animal scientists, and administration workers also participated. Blood samples (n = 192) were analysed by an immunofluorescence assay and considered positive where the phase I or phase II IgG titre was ≥1/50. Seroprevalence was 19% (36/192; 95% CI 14%–25%). A positive serological result was significantly associated with (a) working in outer regional/remote areas (odds ratio [OR] 6.2; 95% CI 1.9–20.8; reference = major cities; p = .009) and (b) having spent more than 50% of total career working with ruminants (OR 4.8; 95% CI 1.7–13.5; reference = <15% of career; p = .025). These findings confirm an increased risk of exposure to C. burnetii compared to the general population, providing new evidence to support Q fever vaccination of veterinary workers in Australia.     

Transmission of Coxiella burnetii to cage mates using murine animal model

Aerosols from the products of abortions of infected animals with Coxiella burnetii were known to be the main source of infection in humans with this bacterium. However, little is known about the excretion of C. burnetii in the feces and urine of infected mice, or the dynamic of transmission between infected and healthy mice. To investigate whether C. burnetii can be excreted in the feces and urine of infected mice and whether transmission to uninfected cage mates occurs, male mice were inoculated with C. burnetii using a “whole body aerosol system” and feces and urine were collected different time points post-infection from these mice. One hour post exposure to aerosols, uninfected mice was placed with infected mice and the transmission was monitored using blood, and organs biopsies collected after sacrifice of contact mice different time points post-contact. Bacterial DNA was not detected in the feces and urine of infected mice at 3, 7, 14 and 28 days post-inoculation suggesting that C. burnetii was not excreted in the feces and urine and consequently they cannot be source of contamination. However, based on the positive PCR results for lungs, blood, spleen, tracheal lymph nodes and cervical lymph nodes, some of the contact animals were considered contaminated at 8 days post-contact. These results indicated that transmission of C. burnetii to contact animals occurs, and it is unlikely that feces and urine act as source of this transmission. Further experiments are needed to clarify the exact mode of contamination.     

Disease Risk Surface for Coxiella burnetii Seroprevalence in White‐Tailed Deer

Coxiella burnetii is considered a re‐emerging zoonosis in many countries. The bacterium is enzootic in livestock and wildlife in the United States, and environmental contamination is widespread. Despite the potential for exposure, the estimated prevalence of Q fever in humans and animals is not well elucidated, and reported human infections in the United States are relatively rare. Zoonotic transmission of the bacterium is usually associated with abortions in domestic ruminants, but other modes of transmission, such as contact with infected blood and/or milk during field dressing of infected wildlife, have not been thoroughly investigated. Studies of zoonotic pathogen transmission between animal reservoir hosts and humans are usually established in response to documented emergence or re‐emergence of a zoonosis in a particular locale, and, as such, the prevalence of infection in wildlife is largely unknown for many zoonotic pathogens, including C. burnetii. The objective of this study was to create a disease risk surface for C. burnetii seroprevalence in wild white‐tailed deer (Odocoileus virginianus) in New York State. Blood samples were collected from hunter‐harvested deer from across New York State in 2009 and 2010. The samples were processed and tested for the presence of anti‐C. burnetii antibodies via indirect microimmunofluorescence assays using phase II C. burnetii strain RSA439. Overall, 14.50% of the tested white‐tailed deer were C. burnetii phase II seropositive. The dual Kernel density estimation method was used to create a smoothed disease risk surface, which revealed variation in seroprevalence ranging from 0% to 32.0%. Areas of higher seroprevalence were detected in four discrete areas of Central New York and in one additional area in the southwest corner of the northern part of the state. This suggests certain locales where humans may be at increased risk for exposure to the bacterium secondary to contact with potentially infected deer.     

Seroprevalence of Coxiella burnetii in Australian dogs

The role of dogs in the transmission of Coxiella burnetii to humans is uncertain, and extensive seroprevalence studies of dogs have not been previously conducted in Australia. This study determined C. burnetii exposure in four diverse canine subpopulations by adapting, verifying and comparing an indirect immunofluoresence assay (IFA) and an enzyme‐linked immunosorbent assay (ELISA) used to detect anti‐C. burnetii antibodies in humans. Canine serum samples (n = 1223) were tested with IFA from four subpopulations [breeding establishments; household pets; free‐roaming dogs in Aboriginal communities; shelter dogs]. The proportions of seropositive dogs were as follows: breeding (7/309, 2.3%), household pets (10/328, 3%), Aboriginal communities (21/321, 6.5%) and shelters (5/265, 1.9%). Dogs from Aboriginal communities were 2.8 times (CI 1.5–5.1; P < 0.001) more likely to be seropositive than dogs from other populations. The ELISA was used on 86 of 1223 sera tested with IFA, and a Cohen's Kappa coefficient of 0.60 (CI 0.43–0.78) indicated good agreement between the two assays. This study has established that Australian dogs within all four subpopulations have been exposed to C. burnetii and that a higher seroprevalence was observed amongst free‐roaming dogs associated with Aboriginal communities. As C. burnetii recrudesces during pregnancy and birth products contain the highest concentration of organism, individuals assisting at the time of parturition, those handling pups shortly after birth as well as those residing in the vicinity of whelping dogs are potentially at risk of developing Q fever. However, the identification of active antigen shed in excreta from seropositive dogs is required in order to accurately define and quantify the public health risk.     

DNA microarray-based detection of Coxiella burnetii,the causative agent of Q fever

Background

An easy-to-handle microarray assay based on the cost-effective ArrayTube™ platform has been designed for the rapid and unequivocal identification of Coxiella burnetii, the causative agent of Q fever. The gene targets include the chromosomally coded markers icd, omp/com1, and IS1111 as well as the plasmid coded markers cbbE and cbhE.

Results

A representative panel comprising 50 German C. burnetii isolates and 10 clinical samples was examined to validate the test. All tested isolates harboured plasmid QpH1 and were correctly identified, corresponding to 100% sensitivity. The assay’s limit of detection was 100 genome equivalents (GE) for icd, omp/com1, cbbE and cbhE and 10 GE for IS1111. Assay specificity was 100% as determined by analysing a panel of 37 non-Coxiella strains.

Conclusions

The present array is a rational assembly of established and evaluated targets for the rapid and unequivocal detection of C. burnetii. This array could be applied to the screening of vaginal swabs from small ruminants; screening of environmental samples e.g. on farms or screening of human samples.     

Coxiella burnetii shedding and environmental contamination at lambing in two highly naturally-infected dairy sheep flocks after vaccination

Abortion due to Coxiella burnetii was confirmed in the 2007/08 season in two naturally-infected dairy sheep flocks. Proportion of C. burnetii shedders and bacterial loads in vaginal mucus were high among aborted or lambed ewes, as was within-flock seroprevalence. Before the next reproductive season (2008/09) 75% of ewes and 50% of replacement lambs were vaccinated (Coxevac, CEVA Santé Animale) keeping the remaining as untreated controls. Compared with the previous year results when abortion outbreak started, a great reduction in the percentage of abortions, in the number of shedders and in the bacterial burden excreted by the ewes was found in both flocks. However, seroconversion in non-vaccinated yearlings from both flocks and the presence of C. burnetii DNA in bioaerosols taken at sheep premises at lambing indicated that infection was still active. No differences were observed between vaccinated and control groups in terms of proportion of C. burnetii shedders. These results suggest that optimal results of vaccination in heavily infected flocks may not be obtained in a short-term period.     

Real‐time quantitative PCR‐based detection of Coxiella burnetii in unpasteurized cow's milk sold for human consumption

Coxiella burnetii is a zoonotic pathogen with a worldwide distribution that is responsible for Q fever in humans. It is a highly infectious bacterium that can be transmitted from cattle to humans through the consumption of unpasteurized milk. We report the molecular identification of C. burnetii in raw cow's milk being sold directly for human consumption in Brazil without official inspection or pasteurization. One hundred and twelve samples of raw milk were analysed by real‐time quantitative PCR (qPCR), and C. burnetii was detected in 3.57% (4/112) of the samples at a concentration ranging from 125 to 404 bacteria per millilitre. The identification of this zoonotic pathogen in raw milk sold directly for human consumption is a public health concern since C. burnetii can be transmitted through the oral route. This result indicates that health education and other preventive measures should be officially implemented in Brazil to prevent the spread of infection. To our knowledge, this is the first qPCR‐based detection of C. burnetii in raw milk samples from cows sold in Brazil that do not undergo official inspection or pasteurization.     

Molecular detection of Coxiella burnetii in raw meat intended for pet consumption

The discovery of antibodies against Coxiella burnetii in cattery‐confined breeding cats indicating prior or current exposure (Shapiro et al., 2015) prompted an investigation into possible sources of infection. One hypothesis was that raw meat diets containing reservoir species may provide a source of C. burnetii transmission. The aim of this pilot study was to determine whether C. burnetii DNA was present in raw meat sold exclusively for companion animal consumption. The sample population consisted of raw meat packages (n = 58) of primarily kangaroo origin, with three to four aliquots (50–120 mg) randomly selected from each package. Genomic DNA was extracted from whole tissue in each of these aliquots using a modified protocol. Three quantitative PCR assays were used for the detection of C. burnetii targeting the IS1111 gene, the heat shock operon htpAB and the C. burnetii outer membrane protein‐coding gene, com1. Coxiella burnetii DNA was detected in 25/58 samples (43%) using the IS1111, htpAB and/or com1 PCR assays and confirmed by DNA sequencing. All samples amplifying a product in the com1 assay also amplified a product in the htpAB and IS1111 assays. A total of 17/58 (29%) packets were positive with all three genes, 4/58 (7%) were positive with two genes (IS1111 and htpAB) and 4/58 (7%) were positive with the IS1111 gene only. Coxiella burnetii DNA was five times more likely to be found in offal than skeletal muscle meat samples. All meat samples in which C. burnetii DNA was found were from kangaroo tissues, while samples labelled as non‐kangaroo meat (n = 4) were negative. Multi‐locus variable number of tandem repeat analysis (MLVA) identified three different genotypes of C. burnetii that have all been identified previously from Australian human clinical Q fever cases. Further investigations are required to determine the potential role of certain raw meats in the transmission of C. burnetii to cats and humans.