Q Fever Dairy Herd Status Determination Based on Serological and Molecular Analysis of Bulk Tank Milk

Ruminants are recognized as the main reservoirs of Coxiella burnetii. EFSA highlighted the lack of knowledge about Q fever prevalence in many European countries. A cross‐sectional study was carried out in randomly selected dairy herds (n = 109) from central Portugal to screen for C. burnetii infection and to correlate it with herd factors. Bulk tank milk (BTM) samples from cattle (n = 45) and small ruminant (n = 64) herds were tested by ELISA and PCR. The apparent seroprevalence of Q fever was estimated in 45.9% (95% CI: 36.3–55.7) being higher in small ruminants (51.6; 95% CI: 39.6–63.4) than in cattle (37.8; 95% CI: 25.1–52.4). The shedding of C. burnetii in BTM was detected in 11.9% (95% CI: 7.1–19.4) of BTM, and it was higher in cattle (20%; 95% CI: 10.9–33.8) than in sheep and mixed herds (6.3%; 95% CI: 2.5–15). A high bacterial load (≥ 3 × 10³ bacteria/ml) was observed in 85% of PCR‐positive BTM. A significant correlation was found between the bacterial load and positive samples on ELISA (P < 0.001). Antibody positivity was significantly associated with the increased herd size (P < 0.01) and the occurrence of abortion (P < 0.05), whereas the shedding of C. burnetii was significantly associated with the report of infertility (P < 0.05). The results highlight that serological and molecular methods in combination are a useful tool to screen for Q fever and to clarify the herd infection status. The shedding of C. burnetii through milk is important, especially in dairy cattle, and thus, the role of milk as a potential source of infection among dairy workers should not be neglected. To our knowledge, this is the first study reporting C. burnetii infection in dairy livestock in Portugal showing that Q fever is significant in dairy herds, leading to economic losses and being a risk for public health, which highlights the need of implementation of control measures.     

Clinical Indicators of Exposure to Coxiella burnetii in Dairy Herds

Q fever is a zoonosis occurring worldwide in livestock. Often neglected in differential diagnoses, Q fever can persist in herds causing financial losses in the long run. In ruminants, well‐known manifestations of Q fever are abortion, stillbirth, delivery of weak offspring and premature delivery. In cattle, Q fever is frequently asymptomatic and/or under‐reported. The use of new methodologies in veterinary clinical epidemiology is of prime importance to find accurate clinical indicators of exposure to C. burnetii at herd level. A retrospective randomly cross‐sectional survey was conducted to estimate the seroprevalence of Q fever in southern Belgium by means of an ELISA test performed on the bulk tank milk (n = 206 cattle herds). At the same time, a questionnaire was accomplished allowing the investigation of presumptive clinical signs observed at herd level during the previous twelve months for dairy cows. A multivariate logistic regression analysis was used to identify abortion and irregular repeat breeding as two indicators associated with Q fever exposure in dairy herds. In addition, a bootstrapped quantile regression revealed that the average score of putative clinical signs related to Q fever was significantly more important in exposed versus non‐exposed herds. A classification and regression tree (CART) analysis confirmed the importance of the average clinical score and the irregular repeat breeding as main splitters, considering or not each clinical sign separately. Considering herd clinical patterns, instead of taking each clinical sign separately, seems to be more useful to differentiate herds at risk of Q fever exposure.     

The Effect of Wind on Coxiella burnetii Transmission Between Cattle Herds: a Mechanistic Approach

There is a consensus that wind plays a key role in the transmission of Coxiella burnetii, the causative agent of Q fever, between ruminants and from ruminants to humans. However, no observational study so far has focused on the mechanisms associated with this airborne transmission. This study applied a mechanistic epidemiological approach to investigate the processes underlying the wind effect and to assess its influence on the risk for a dairy herd to become C. burnetii infected. Ninety‐five dairy cattle herds located in the Finistère department (western France) were subjected to samplings of bulk tank milk and indoor dust every 4 months over a 1‐year period to determine their C. burnetii status using PCR tests. A total of 27 incident herd‐periods (negative‐tested on both PCR tests and becoming positive‐tested at least once at the subsequent sampling time) and 71 negative herd‐periods were retained for analysis. Using logistic regression, we assessed the effect of (i) the cumulated number of bacteria in herds located under the main wind direction and (ii) the mean wind speed in this area, on a given herd's risk of becoming incident. Compared to herds in areas with low wind speed (≤5.5 m/s), the risk was significantly higher (OR = 3.7) in herds in areas with high wind speed (>5.5 m/s) and high bacterial load (>10), whereas it was not significantly different from unity in other situations. In agreement with our assumptions, C. burnetii transmission to a previously infection‐free herd occurs only when (i) the wind transporting from infected sources and (ii) the load in the contaminated particles/aerosols generated are high enough to act jointly.     

Swab cloths as a tool for revealing environmental contamination by Q fever in ruminant farms

Q fever is a zoonotic abortive disease of ruminants mostly transmitted by inhalation of aerosols contaminated by Coxiella burnetii. Clusters of cases or even epidemics regularly occur in humans but, to date, there is no consensus about the best way to carry out outbreak investigations in order to identify potential farms at risk. Although environmental samples might be useful during such investigations, there are few baseline data on the presence of C. burnetii in the environment of ruminant farms. We thus investigated dust samples from cattle, sheep and goat farm buildings in order to (a) estimate C. burnetii detection frequency and bacterial loads in the environment, and (b) determine whether this environmental contamination is associated with series of abortions attributed to Q fever. We considered 113 herds with a recent abortive episode potentially related (n = 60) or not (n = 53) to C. burnetii. Dust was sampled using a swab cloth and tested by a quantitative PCR method targeting the IS1111 gene. Coxiella burnetii DNA was detected on 9 of 50 cattle farms, 13 of 19 goat farms and 30 of 40 sheep farms. On 16 cloths, bacterial loads were higher than 10⁸ genome equivalents, levels as high as in infectious materials such as placentas and aborted foetuses. Overall, the probability of detecting C. burnetii DNA was higher on small ruminant farms than cattle farms, in herds suspected of Q fever and in large herds. We conclude that swab cloths are a putative indicator of contamination of ruminant farms by C. burnetii.     

Coxiella burnetii Infections in Small Ruminants and Humans in Switzerland

The recent Q fever epidemic in the Netherlands raised concerns about the potential risk of outbreaks in other European countries. In Switzerland, the prevalence of Q fever in animals and humans has not been studied in recent years. In this study, we describe the current situation with respect to Coxiella (C.) burnetii infections in small ruminants and humans in Switzerland, as a basis for future epidemiological investigations and public health risk assessments. Specific objectives of this cross‐sectional study were to (i) estimate the seroprevalence of C. burnetii in sheep and goats, (ii) quantify the amount of bacteria shed during abortion and (iii) analyse temporal trends in human C. burnetii infections. The seroprevalence of C. burnetii in small ruminants was determined by commercial ELISA from a representative sample of 100 sheep flocks and 72 goat herds. Herd‐level seroprevalence was 5.0% (95% CI: 1.6–11.3) for sheep and 11.1% (95% CI: 4.9–20.7) for goats. Animal‐level seroprevalence was 1.8% (95% CI: 0.8–3.4) for sheep and 3.4% (95% CI: 1.7–6) for goats. The quantification of C. burnetii in 97 ovine and caprine abortion samples by real‐time PCR indicated shedding of >10⁴ bacteria/g in 13.4% of all samples tested. To our knowledge, this is the first study reporting C. burnetii quantities in a large number of small ruminant abortion samples. Annual human Q fever serology data were provided by five major Swiss laboratories. Overall, seroprevalence in humans ranged between 1.7% and 3.5% from 2007 to 2011, and no temporal trends were observed. Interestingly, the two laboratories with significantly higher seroprevalences are located in the regions with the largest goat populations as well as, for one laboratory, with the highest livestock density in Switzerland. However, a direct link between animal and human infection data could not be established in this study.     

Coxiella burnetii Shedding by Farmed Red Deer (Cervus elaphus)

Wildlife and notably deer species – due to the increasing relevance of deer farming worldwide – may contribute to the maintenance of Coxiella burnetii, the causal agent of Q fever. Currently, there are no precedents linking exposure to deer species with human Q fever cases. However, a human case of Q fever was recently diagnosed in a red deer (Cervus elaphus) farm, which led us to investigate whether deer could be a source for environmental contamination with C. burnetii and ascertain the implication of C. burnetii in reproductive failure in the farm. Blood serum and vaginal swabs were collected from hinds either experiencing or not reproductive failure and tested to detect the presence of antibodies and DNA, respectively, of C. burnetii, Chlamydia abortus, Neospora caninum and Toxoplasma gondii. Serology and PCR results suggest C. burnetii was the primary cause of the reproductive failure. We identified vaginal shedding of C. burnetii in hinds, confirming red deer as a source of Q fever zoonotic infection.     

Liver fluke (Fasciola hepatica) co‐infection with bovine tuberculosis in cattle: A prospective herd‐level assessment of herd bTB risk in dairy enterprises

Co‐infection of tuberculosis (TB) and helminths is recognized as a significant problem in regions where such pathogens are endemic and chronic cases exist. Co‐infection can modulate the immune system leading to interference with diagnostic tests, increased pathological impacts and pathogen persistence. However, research has found that such interactions between pathogens can be context and species specific. Recent studies have suggested that liver fluke, Fasciola hepatica, infection may impact on immunological responses and diagnostics for bovine tuberculosis (bTB; caused by Mycobacterium bovis) in cattle. Where evidence of such interaction exists, there would be an onus on policy makers to adjust eradication programs to minimize impacts. We assessed the association between herd‐level bTB breakdown risk and seasonal variation in liver fluke exposure based on 5,753 bulk tank milk (BTM) samples from 1,494 dairy herds across Northern Ireland. BTM was tested by an IDEXX antibody specific enzyme‐linked immunosorbent assay (ELISA) using the ‘f2’ antigen as a detection agent. The ELISA determined the result based on a sample to (known) positive ratio (S/P%) from which binary status and categories of exposure were derived. Associations were tested using multivariable random effects models. Models predicting bTB risk were not improved with the inclusion of liver fluke exposure levels. Variations in modelling liver fluke exposure (S/P%, binary, categories of exposure) and bTB risk (skin test breakdowns, post‐mortem confirmed breakdowns, breakdown size and lag effects) also failed to support associations (neither positive nor negative) between the pathogens at herd‐level. These results, along with previously published animal‐level data from Northern Ireland, suggest that the nexus between bTB and F. hepatica may have small size effects at the population‐level. However, our results also highlight the high prevalence of F. hepatica in cattle in our study population, and therefore we cannot fully discount the potential hypothesis of population‐level depression of immune response to M. bovis due to co‐infection.     

Prevalence and risk factors analysis of bovine tuberculosis in cattle raised in mixed crop‐livestock farming system in Tigray region,Ethiopia

Bovine tuberculosis (BTB) is a disease of animal and public health importance in developing countries. In rural Ethiopia, there is potential for a shift in the epidemiologic of this disease driven by transformation of dairy industry. This includes gradual change from the traditional mixed crop‐livestock husbandry practice to a semi‐intensification system. It is therefore, essential to document the prevalence and risk factors of BTB to continuously update the designing and implementation of control and prevention strategies. Here, we present findings of a cross‐sectional study on the prevalence and associated risk factors of BTB among cattle reared under mixed crop‐livestock farming system in Tigray region, Ethiopia. A multistage purposive sampling approach was used to select districts, villages, herds and individual cattle. A total of 1,357 cattle from 310 herds were examined for BTB infection using a comparative intradermal tuberculin skin test (CIDT). Questionnaires were used to gather data on herd structure and herd management practices. A multilevel logistic mixed effect model was used to determine risk factors after accounting for clustering effect at three levels (village, herd and individual animal). Overall prevalence of BTB was 4.3% (95% CI = 3.4–5.6), with the highest prevalence recorded in Alamata district (5.6%) and lowest in Korem (1.6%). Multilevel logistic mixed effect model analysis identified exotic breed (OR = 3, p = 0.014), closed barn (OR = 2.6, p = 0.018), large herd size (OR = 2.6, p = 0.05) and purchase of cattle (OR = 2.1, p = 0.027) as important risk factors for BTB. Taken together, these findings suggest that the current dairy development program centred on the introduction of exotic and or crossed animals could have contributed to changing epidemiological situations of BTB in the study area.     

Bovine Viral Diarrhoea Virus (BVDV) in Dairy Cattle: A Matched Case–Control Study

Bovine viral diarrhoea virus (BVDV) causes one of the most important diseases of cattle in terms of economic costs and welfare. The aims were to estimate herd prevalence and to investigate the factors associated with antibodies in bulk tank milk (BTM) in dairy herds through a matched case–control study. To estimate herd prevalence, BTM samples were randomly selected (n = 314) from a population (N = 1604). The true prevalence of BVDV was 24.3% (CI 95% = 20.1–29.3%). For the case–control study, BVDV antibody‐positive herds (high antibody titres) were classified as cases (n = 21) and matched (n = 63) by milk production with herds presenting low antibody titres (ratio of 1 : 3). Three multivariable models were built: 1) full model, holding all 21 variables, and two models divided according to empirical knowledge and similarity among variables; 2) animal factor model; and 3) biosecurity model. The full model (model 1) identified: age as a culling criteria (OR = 0.10; CI 95% = 0.02–0.39; P < 0.01); farms that provided milk to other industries previously (OR = 4.13; CI 95% = 1.17–14.49; P = 0.02); and isolation paddocks for ill animals (OR = 0.14; CI 95% = 0.01–0.26; P = 0.02). The biosecurity model revealed a significant association with the use of natural mating (OR = 9.03; CI 95% = 2.14–38.03; P < 0.01); isolation paddocks for ill animals (OR = 0.06; CI 95% = 0.05–0.83; P = 0.03); years providing milk for the same industry (OR = 0.94; CI 95% = 0.91–0.97; P = 0.02); and direct contact over fences among cattle of neighbouring farms (OR = 5.78; CI 95% = 1.41–23.67; P = 0.04). We recommend the application of grouping predictors as a good choice for model building because it could lead to a better understanding of disease–exposure associations.     

Control of Bovine Brucellosis from Persistently Infected Holdings Using RB51 Vaccination with Test‐and‐Slaughter: A Comparative Case Report from a High Incidence Area in Portugal

Bovine brucellosis due to Brucella abortus infection causes significant reproductive and production losses in cattle and is a major zoonosis. Eradication of this disease has proved difficult to achieve in Portugal where it still occurs in some regions despite an ongoing national eradication programme. In 2004, the Alentejo region, a major cattle producing area, reported one of the highest levels of bovine brucellosis in the country, especially in one divisional area. In that area, bovine brucellosis was particularly problematic in a holding of ten herds, the largest extensive cattle unit in the country, which remained infected despite an extensive test‐and‐slaughter programme and depopulation of five herds. A 5‐year programme of RB51 vaccination with biannual test‐and‐slaughter was thus implemented in 2004. The apparent animal seroprevalence decreased from 19% (646/3,400) to 3% (88/2930) on the third herd‐level test and remained below 0.8% (27/3324) after the fourth test. After the tenth test, the holding had a prevalence of 0.1% (2/2332) and only one herd remained positive with a within‐herd prevalence of 1.1% (2/177). The results were compared to all other herds (n = 10) in the divisional area that were also persistently infected but were subject only to test‐and‐slaughter before being depopulated. In these herds, the strategy of test‐and‐slaughter did not reduce the prevalence, which remained significantly higher than the vaccinated group (median = 0.48% and 8.5% in vaccinated versus non‐vaccinated herds; Wilcoxon rank sum test; P < 0.01). The success of this pilot programme in continental Portugal provided a valuable case study to the official veterinary services by illustrating the value of RB51 vaccination with parallel testing and improved biosecurity as a comprehensive and sustainable strategy for bovine brucellosis control in persistently infected herds.     

Coxiella burnetii in wild mammals: A systematic review

Coxiella burnetii is a multi‐host bacterium that causes Q fever in humans, a zoonosis that is emerging worldwide. The ecology of C. burnetii in wildlife is still poorly understood and the influence of host, environmental and pathogen factors is almost unknown. This study gathers current published information on different aspects of C. burnetii infection in wildlife, even in species with high reservoir potential and a high rate of interaction with livestock and humans, in order to partially fill the existing gap and highlight future needs. Exposure and/or infection by C. burnetii has, to date, been reported in 109 wild mammal species. The limited sample size of most of the existing studies could suggest an undervalued prevalence of C. burnetii infection. Knowledge on the clinical outcome of C. burnetii infection in wildlife is also very limited, but currently includes reproductive failure in waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus niger), dama gazelle (Nanger dama) and water buffalo (Bubalus bubalis) and placentitis in the Pacific harbor seal (Phoca vitulina richardsi), Steller sea lion (Eumetopias jubatus) and red deer (Cervus elaphus). The currently available serological tests need to be optimised and validated for each wildlife species. Finally, there is a huge gap in the research on C. burnetii control in wildlife, despite of the increasing evidence that wildlife is a source of C. burnetii for both livestock and humans.     

Coxiella burnetii (Q‐Fever) Seroprevalence in Prey and Predators in the United Kingdom: Evaluation of Infection in Wild Rodents,Foxes and Domestic Cats Using a Modified ELISA

Coxiella burnetii, the agent of Q‐fever, is recognized as a worldwide zoonosis with a wide host range and potentially complex reservoir systems. Infected ruminants are the main source of infection for humans, but cats and other mammals, including wild rodents, also represent potential sources of infection. There has been a recent upsurge of reported cases in humans, domestic ruminants and wildlife in many parts of the world, and studies have indicated that wild brown rats may act as true reservoirs for C. burnetii and be implicated in outbreaks in livestock and humans. However, investigation of reservoir systems is limited by lack of validated serological tests for wildlife or other non‐target species. In this study, serum samples from 796 wild rodents (180 bank voles, 309 field voles, 307 wood mice) 102 wild foxes and 26 domestic cats from three study areas in the UK were tested for the presence of antibodies to C. burnetii using a commercial indirect ELISA kit modified for use in multiple wildlife species. Test thresholds were determined for each species in the absence of species‐specific reference sera using a bi‐modal latent class mixture model to discriminate between positive from negative results. Based on the thresholds determined, seroprevalence in the wild rodents ranged from 15.6% to 19.1% depending on species (overall 17.3%) and was significantly higher in both foxes (41.2%) and cats (61.5%) than in rodents. This is the first report to quantify seroprevalence to C. burnetii in bank voles, field voles, wood mice, foxes and cats in the UK and provides evidence that predator species could act as indicators for the presence of C. burnetii in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both livestock and humans, and the high seroprevalence in domestic cats highlights the potential zoonotic risk from this species.     

Modelling the variation in skin‐test tuberculin reactions,post‐mortem lesion counts and case pathology in tuberculosis‐exposed cattle: Effects of animal characteristics,histories and co‐infection

Correctly identifying bovine tuberculosis (bTB ) in cattle remains a significant problem in endemic countries. We hypothesized that animal characteristics (sex, age, breed), histories (herd effects, testing, movement) and potential exposure to other pathogens (co‐infection; BVDV , liver fluke and Mycobacterium avium reactors) could significantly impact the immune responsiveness detected at skin testing and the variation in post‐mortem pathology (confirmation) in bTB ‐exposed cattle. Three model suites were developed using a retrospective observational data set of 5,698 cattle culled during herd breakdowns in Northern Ireland. A linear regression model suggested that antemortem tuberculin reaction size (difference in purified protein derivative avium [PPD a ] and bovine [PPD b ] reactions) was significantly positively associated with post‐mortem maximum lesion size and the number of lesions found. This indicated that reaction size could be considered a predictor of both the extent (number of lesions/tissues) and the pathological progression of infection (maximum lesion size). Tuberculin reaction size was related to age class, and younger animals (<2.85 years) displayed larger reaction sizes than older animals. Tuberculin reaction size was also associated with breed and animal movement and increased with the time between the penultimate and disclosing tests. A negative binomial random‐effects model indicated a significant increase in lesion counts for animals with M. avium reactions (PPD b− PPD a <  0) relative to non‐reactors (PPD b− PPD a =  0). Lesion counts were significantly increased in animals with previous positive severe interpretation skin‐test results. Animals with increased movement histories, young animals and non‐dairy breed animals also had significantly increased lesion counts. Animals from herds that had BVDV ‐positive cattle had significantly lower lesion counts than animals from herds without evidence of BVDV infection. Restricting the data set to only animals with a bTB visible lesion at slaughter (n  = 2471), an ordinal regression model indicated that liver fluke‐ infected animals disclosed smaller lesions, relative to liver fluke‐negative animals, and larger lesions were disclosed in animals with increased movement histories.     

Seroprevalence of canine neosporosis and bovine viral diarrhoea in dairy cattle in selected regions of Kenya

Neospora caninum is the causative agent for canine neosporosis (CN), a disease of potential zoonotic importance causing reproductive losses in cattle while causing neuromuscular disease in dogs. Bovine viral diarrhoea on the other hand is caused by the bovine viral diarrhoea virus (BVDV) and is one of the most important reproductive diseases of cattle worldwide. In Kenya, these infections are of economic importance due to the losses they cause in farms in which they are diagnosed or are subclinical. Such losses include reduced milk production, reduced conception, early embryonic deaths and abortions which lead to reproductive wastage. This study was conducted between April 2017 and July 2018 and determined the seroprevalence of neoporosis and BVD in select dairy herds in Kenya. Kakamega, Nandi and Makueni Counties from where dairy farms were purposively sampled were used. Serum samples were collected from randomly selected dairy animals aged at least 2 years in the selected farms and screened for BVDV and CN antibodies. Seroprevalence of N. caninum was 24.1% (n = 552) and BVD, 52.3% (n = 545) across all the counties. Co‐infection where antibodies against the two infective agents were present was in 14.6% (n = 541) animals. Chi‐square tests of association between prevalence and county were significant for BVD (p = .000) but not for neosporosis (p = .626). Further chi‐square tests of association between the two infections were not significant (p = .105) neither were the associations of BVD (p = .575) and neosporosis (p = .626) on pregnancy status. These two diseases are rarely investigated as causes of bovine infertility. Detection of antibodies in the studied dairy herds underpins the need for enhanced surveillance by laboratories and for further studies to understand associated risk factors to formulate effective control strategies in dairy cattle to forestall abortions and production and reproduction losses.     

Large‐Scale Cross‐Sectional Serological Survey of Schmallenberg Virus in Belgian Cattle at the End of the First Vector Season

A cross‐sectional survey was conducted in the Belgian cattle population after the first period of infection of the emerging Schmallenberg virus. A total number of 11 635 cattle from 422 herds sampled between 2 January and 7 March 2012 were tested for the presence of Schmallenberg‐specific antibodies using an ELISA kit. Between‐herd seroprevalence in cattle was estimated at 99.76% (95% CI: 98.34–99.97) and within‐herd seroprevalence at 86.3% (95% CI: 84.75–87.71). An Intraclass Correlation Coefficient of 0.3 (P < 0.001) was found, indicating that the correlation between two animals within a herd with respect to their serological status was high. Those results corroborate the conclusion that the Schmallenberg virus was widespread in Belgium during winter 2011. Seroprevalence was shown to be statistically associated to the animal's age (P < 0.0001): with 64.9% (95% CI: 61.34–68.3) estimated for the 6–12 months of age, 86.79% (95% CI: 84.43–88.85) for the 12–24 months of age and 94.4% (95% CI: 93.14–95.44) for the animals older than 24 months. Based on the results of the described serological survey, we can conclude that after the first Schmallenberg virus episode, almost every Belgian cattle has already been in contact with the virus. In consequence, the vast majority of the host animals should have developed post infection protective immunity against the virus.     

Seroepidemiological and molecular investigation of spotted fever group rickettsiae and Coxiella burnetii in Sao Tome Island: A One Health approach

Spotted fever group rickettsiae (SFGR) and Coxiella burnetii are intracellular bacteria that cause potentially life‐threatening tick‐borne rickettsioses and Q fever respectively. Sao Tome and Principe (STP), small islands located in the Gulf of Guinea, recently experienced a dramatic reduction in the incidence of malaria owing to international collaborative efforts. However, unexplained febrile illnesses persist. A One Health approach was adopted to investigate exposure to SFGR and C. burnetii in humans and examine the diversity of these bacteria in ticks parasitizing domestic ruminants. A cross‐sectional human serological study was conducted in Agua Grande district in Sao Tome Island from January to March 2016, and ticks were collected from farmed domestic ruminants in 2012 and 2016. In total, 240 individuals varying in age were randomly screened for exposure to SFGR and C. burnetii by indirect immunofluorescence assay. Twenty of 240 individuals (8.3%) were seropositive for SFGR (4 for Rickettsia africae and 16 for R. conorii) and 16 (6.7%) were seropositive for C. burnetii. Amblyomma astrion were collected exclusively in 2012, as were A. variegatum in 2016 and Rickettsia spp. were detected in 22/42 (52.4%) and 49/60 (81.7%) respectively. Sequence analysis of multiple gene targets from Rickettsia spp. detected in ticks suggests the presence of a single divergent R. africae strain (Sao Tome). While no ticks were found positive for C. burnetii, Coxiella‐like endosymbionts were detected in nearly all ticks. This is the first study in STP to provide serological evidence in humans of SFGR and C. burnetii and additional molecular evidence in ticks for SFGR, which may be responsible for some of the unexplained febrile illnesses that persist despite the control of malaria. Future epidemiological studies are needed to confirm the occurrence and risk factors associated with SFG rickettsioses and Q fever in both humans and animals.     

Molecular prevalence of Chlamydia and Chlamydia‐like bacteria in Tunisian domestic ruminant farms and their influencing risk factors

Chlamydia and Chlamydia ‐like bacteria are well known to infect several organisms and may cause a wide range of diseases, particularly in ruminants. To gain insight into the prevalence and diversity of these intracellular bacteria, we applied a pan‐Chlamydiales real‐time PCR to 1,134 veterinary samples taken from 130 Tunisian ruminant herds. The true adjusted animal population‐level prevalence was 12.9% in cattle, against 8.7% in sheep. In addition, the true adjusted herd‐level prevalence of Chlamydiae was 80% in cattle and 25.5% in sheep. Chlamydiales from three family‐level lineages were detected indicating a high biodiversity of Chlamydiales in ruminant herds. Our results showed that Parachlamydia acanthamoebae could be responsible for bovine and ovine chlamydiosis in central‐eastern Tunisia. Multivariable logistic regression analysis at the animal population level indicated that strata and digestive disorders variables were the important risk factors of bovine and ovine chlamydiosis. However, origin and age variables were found to be associated with bovine and ovine chlamydiosis, respectively. At the herd level, risk factors for Chlamydia positivity were as follows: abortion and herd size for cattle against breeding system, cleaning frequency, quarantine, use of disinfectant and floor type for sheep. Paying attention to these risk factors will help improvement of control programs against this harmful zoonotic disease.     

A Case–control Study of Haemorrhagic Septicaemia in Buffaloes and Cattle in Karachi,Pakistan, in 2012

A retrospective epidemiological case–control study was performed in Karachi, Pakistan, from January to April 2013. The owners of 217 dairy cattle and buffalo farms from six different locations in Karachi were interviewed. The aim of the study was to identify risk factors associated with the presence of haemorrhagic septicaemia (HS). Farms with a history of at least one instance of sudden death in a dairy animal during 2012 and a positive clinical HS diagnosis (made by local veterinarians) were defined as cases. Farms having no history of sudden deaths in 2012 were defined as controls. Univariable analyses were initially conducted, and factors with P ≤ 0.25 were offered to a multivariable logistic regression model to identify putative risk factors. The final multivariable logistic model contained five factors. Vaccination was found to be a protective factor (OR = 0.22) along with the length of time cattle were kept on farm (months). For every extra month cattle were kept, the odds of HS disease were reduced by a factor of 0.9. In contrast, for every extra animal in a herd, the risk of infection increased by a factor of 1.01. Supplying underground water and the presence of foot and mouth disease on the farm increased the risk by 2.90 and 2.37, respectively. To understand the epidemiology of HS in Karachi dairy herds, more in‐depth research is required to study the risk and protective factors identified in this survey and to evaluate risk mitigation strategies, where possible.     

Outbreaks of abortions by Coxiella burnetii in small ruminant flocks and a longitudinal serological approach on archived bulk tank milk suggest Q fever emergence in Central Portugal

Q fever is a worldwide zoonotic infectious disease caused by Coxiella burnetii and sheep and goats are known to be the main reservoir for human infection. This study describes the epidemiological and laboratory findings of C. burnetii outbreaks affecting sheep and goat flocks and also provides the results of a prospective serosurvey in bulk tank milk samples to assess C. burnetii circulation in a population of sheep living in close contact to the human population in Central Portugal. In the epizooties, C. burnetii was identified in tissues of the resulting abortions by qPCR . As for the serological survey, 10.2% (95%CI : 4.5‐19.2) of the 78 bulk tank milk samples collected in 2015 presented IgG antibodies against C. burnetii . The same farms were visited and sampled in 2016 and 25.6% (95%CI : 16.4‐36.8) were positive. This steep increase in the number of anti‐C. burnetii farms between the 2015 and 2016 collections showed to be statistically significant (p  = 0.020) and is strongly suggestive of Q fever emergence in Central Portugal. Measures on animal health and on disease spread control to the human population should be considered.     

Two Years After the Schmallenberg Virus Epidemic in the Netherlands: Does the Virus still Circulate?

Two years after the introduction of the Schmallenberg virus in north‐western Europe, it is unknown whether the virus is still circulating in countries that were the first to be confronted with it. When the population‐level immunity declines in Europe, reintroduction or the re‐emergence of SBV in Europe might eventually result in an outbreak of similar magnitude of that seen in 2011–2012. The Netherlands was part of the primary outbreak region of SBV in 2011. The aim of this study was to determine whether SBV circulated amongst dairy herds in the Netherlands in 2013, and if so, to which extent. For this purpose, the presence of SBV‐specific antibodies in naive cattle was investigated. A total of 394 dairy farms were sampled between October and December 2013 by collecting five serum samples per herd. Antibodies were detected in 1.1% [95% confidence interval (CI): 0.7–1.7)] of the animals. All seropositive animals were single reactors per herd and were at least 8 months old at sampling. As these results were inconclusive in demonstrating freedom of SBV circulation, a more in‐depth investigation was initiated to provide more insight: an additional sample of 20 youngstock within the same age category (including the five initially sampled animals) was collected from 17 of the 21 positive herds and tested for SBV‐specific antibodies. This resulted in 9 antibody‐positive test results of 316 samples. Again, the positive samples were single reactors within the sample obtained from each farm, which is unlikely given the characteristics of SBV. Therefore, assuming the single reactors as false‐positive, this survey showed with 95% confidence that the maximum possible prevalence of herds with SBV circulation in the Netherlands was <1% in 2013.