Alain Joly

Prevalence of Coxiella burnetii infection in domestic ruminants: A critical review

Reliable detection of Coxiella burnetii is a critical point for the control of the spread of this zoonotic disease (Q fever), ruminants being considered as the main source for human infection as confirmed by the recent human outbreak in the Netherlands since 2007. Considering both public and animal health, providing consolidated prevalence data could be relevant within the decision process of public policy makers or producers organizations. The objective of this study was to conduct a critical review of the literature focused on the prevalence of C. burnetii infection at animal, herd and within-herd levels in cattle, goat and sheep. A qualitative assessment of the 69 selected publications, based on the analysis of the sampling frame and testing procedures, was also performed. While the number of publications increased recently, major methodological issues were still evidenced. These critical issues were related to (i) the absence of description of the sampling strategy and (ii) the lack of sensitivity of the testing procedure. The lack of well designed studies makes not possible to estimate accurately the current prevalence of the infection. Nevertheless, the literature review reported the detection of C. burnetii infection in the all 5 continents with a wide range whatever the species. The apparent prevalence was slightly higher in cattle (20.0% and 37.7% of mean apparent prevalence at animal and herd level respectively) than in small ruminants (around 15.0% and 25% respectively for animal and herd level in sheep and goat). The present conclusions and the current situation support the persistent need of conducting well designed studies, aiming at estimating the true prevalence of C. burnetii infection in the three main domestic ruminant species.

Prevention of Coxiella burnetii shedding in infected dairy herds using a phase I C. burnetii inactivated vaccine

The main objective of this study was to assess the efficacy of a monovalent inactivated vaccine containing phase I Coxiella burnetii to prevent Coxiella shedding in susceptible dairy cows within infected herds in comparison to a placebo. A total of 336 dairy cows and heifers, from six spontaneously infected herds, were followed over a 1-year period. Before treatment (i.e. vaccination or placebo), the C. burnetii infection status of the cows was determined on the basis on PCR results on milk, vaginal mucus and faeces and serological analysis performed 2 weeks apart. A cow was considered susceptible (i.e. non-infected) when all results were negative, and was considered infected otherwise. The allocation of treatments was performed randomly within pregnant and non-pregnant cows. After treatment (D0), the animals were subject to systematic sampling (milk, vaginal mucus and faeces) on D90, D180, D270 and D360 to detect putative shedding. In addition, the same samples were taken within 15 days after calving. An animal was considered as a shedder at a given time t, if at t, it was found PCR-positive on at least one test taken among the samples (milk, vaginal mucus and faeces). The effect of the treatment on the probability for an initially susceptible animal of becoming shedder was assessed using survival analysis techniques (Cox regression model). Almost all heifers were detected as susceptible before treatment. When vaccinated while not pregnant, an animal had a five times lower probability of becoming a shedder than an animal receiving placebo. An animal which was vaccinated while pregnant had a similar probability of becoming shedder as an animal receiving the placebo. There was no significant farm effect in this multi-centric trial. These results highlight the value of implementing vaccination, if possible, in non-infected herds. In infected herds, the vaccination should be implemented in quite all presumably susceptible animals, i.e. at least the heifers. The vaccination of the dairy cows should be performed when the within-herd seroprevalence is low, i.e. in herds where the infection has not spread widely yet.

Shedding and serological patterns of dairy cows following abortions associated with Coxiella burnetii DNA detection.

To describe both shedding and serological patterns following abortions detected as being associated with Coxiella burnetii (Cb), 24 cows experiencing an abortion due to Cb were followed over a one month period. Samples taken on the day of abortion (D0) were followed 3-fold by weekly samplings from day 14 (D14) to D28 after the abortion. Milk and vaginal mucus were collected at each weekly sampling and tested using real-time PCR while blood samples were collected 2-fold on D21 and D28 and tested using ELISA. We found a very short duration of C. burnetii shedding in vaginal mucus after abortion, highlighting the need to collect samples as rapidly as possible following an abortion to avoid false negative results. In contrast with previous results, concomitancy of vaginal and mucus shedding was frequent, especially for cows shedding a high bacterial load on DO leading to the hypothesis that the clinical onset of the infection influences the modalities of Cb shedding. Lastly, serological results indicating a lack of sensitivity to detect Cb shedder cows (especially for cows for which Ct values were high) suggest that ELISA is not a useful tool to diagnose abortions at the individual level.

Large scale assessment of the effect associated with bovine viral diarrhoea virus infection on fertility of dairy cows in 6149 dairy herds in Brittany (Western France)

This study aimed at quantifying on a large scale the risk of both three-week-return-to-service and late-return-to-service for dairy cows according to the BVDV-infection status of the herds. Three-week- and late-returns-to-service were defined as a repeat service occurring respectively from 19 to 25 days and after 25 days following a first or a second service. The level of BVDV-specific-antibodies in bulk tank milk were measured four times at four months intervals by a blocking ELISA test to define five BVDV-infection-herd-statuses: (i) presumed non-infected herds for a long time, (ii) presumed not-recently-infected herds, (iii) presumed past-infected-recently-recovered herds, (iv) presumed past-steadily-infected herds, (v) presumed recently-infected herds. A total of 150,854 AIs from 122,697 cows in 6149 herds was included in the analysis. The risk of return-to-service was assessed using logistic regression and survival analysis models. The BVDV-infection-herd-status was not significantly associated with the risk of 3-week-return-to-service, but significantly with that of late-return-to-service. Cows in herds presumed past-infected-recently-recovered, past-steadily-infected, or recently-infected had a significantly higher risk of late-return-to-service (relative risk of 1.03, 1.11, 1.12, respectively) compared with cows in herds presumed not-recently-infected. Risk of late-return-to-service was not significantly different in cows from herds presumed non-infected for a long time or not-recently-infected. In conclusion, BVDV-infection was found to mainly increase the risk of embryonic and foetal death.

Effectiveness of vaccination and antibiotics to control Coxiella burnetii shedding around calving in dairy cows

Effectiveness of phase 1 vaccine, combined or not with tetracycline, to control Coxiella burnetii vaginal shedding at calving in cows was assessed through a 13 months study in 22 Q fever clinically affected commercial dairy herds. Four medical strategies implemented at herd level but randomly assigned to cows (vaccination, vaccination and tetracycline, tetracycline, nothing) were compared. There was no significant interaction effect between vaccination and antibiotherapy. Tetracycline used once at drying off was associated with a lower risk of being detected shedder at calving (OR=0.40, CI 95% [0.21-0.75]), but had no significant effect on the bacterial load shed. Vaccination did not significantly prevent shedding but was significantly (OR=0.15, CI 95% [0.03-0.85]) associated with lower bacterial load shed. Thus, vaccination using a phase 1 vaccine and antibiotherapy using tetracycline is associated with a decrease in shedding in dairy cows and could contribute to reduce the bacterial load generated in the environment. To the best of our knowledge, this is the first study providing useful information for evidenced-based and rational use of medical strategy combining antibiotic and vaccination in infected dairy cattle herds.

Increase in the occurrence of abortions associated with exposure to the Bluetongue virus serotype 8 in naïve dairy herds

The transplacental transmission capacity demonstrated for Bluetongue virus serotype 8 (BTV-8) in cattle probably is associated with an increased occurrence of abortions. The objectives of this study were to quantify the effect of BTV-8 exposure on the occurrence of abortions in previously naive dairy cow herds under natural infection conditions, and to determine a possible risk period during pregnancy associated with this increase. Two criteria were considered in order to estimate the occurrence of abortion: late return-to-service after a first artificial insemination (AI), and short gestations. A late return-to-service was defined as a return taking place 90 to 200 days after a first AI. These criteria were compared between cows in herds exposed during the 2007 epizootic in France and cows in herds that were not exposed. To determine the risk period during a pregnancy, variations in the occurrence of abortions were quantified according to the stage of the pregnancy during which the exposure took place. Survival analyses were used to estimate the risk of increased occurrence of abortion associated with BTV-8 exposure, adjusted by the principal factors known to influence the risk of abortion. Exposure to the BTV-8 virus under natural conditions in previously naive dairy herds notified after clinical suspicion during the 2007 epizootic was associated with an increase in the occurrence of abortions, regardless of the stage of pregnancy. The at-risk gestation period depended on the criteria used to detect abortions. The mean effect of BTV-8 exposure in the ensemble of detected outbreaks corresponded to an increase of 6.7% in late return-to-service. BTV-8 exposure during the first 3 mo of gestation was associated with a 15% increase in late return-to-service for cows with no return-to-service at 90 days, while this increase was 6% for exposure starting from the third month of gestation (in outbreaks detected in September). BTV-8 exposure from the third month of gestation was associated with a 1.9% increase of short gestations. The effect of exposure was more pronounced for outbreaks detected early in the epizootic compared with those detected later.

Seroprevalence of Q fever in naturally infected dairy cattle herds

Coxiella burnetii is the causal agent of Q fever, a worldwide spread zoonosis. Prevention of C. burnetii shedding in cattle is critical to control the spread of the pathogen between animals, and from animals to humans. Vaccination with a phase 1 vaccine has been shown to be effective in preventing shedding when implemented in still susceptible animals, even in infected cattle herds. The identification of these animals (dairy cows and nulliparous females) as targets for vaccination consequently is crucial. Hygiene measures conventionally also are implemented, but their relative impact on C. burnetii diffusion remains unknown. The objectives of this study therefore were to (i) describe the distribution of the within-herd apparent seroprevalence among cows and nulliparous females and (ii) to explore the association between management practices and herd characteristics on the one hand, and these seroprevalences on the other. In a sample of 100 naturally and clinically infected dairy herds, blood samples were taken systematically from all nulliparous females (older than 12 months) and cows, and serologically tested. Information on herd characteristics and management practices were collected through a questionnaire filled in by each farmer. The variation in within-herd seroprevalence among cows and the risk for a herd of having at least one seropositive nulliparous female were investigated using multivariate (linear and logistic respectively) regression models. Median within-herd seroprevalence was 0.32 (Q1=0.22; Q3=0.43). We observed a low to null (median=0.01; Q1=0; Q3=0.10) within-herd seroprevalence in nulliparous females contrary to a high value (median=0.42) and variability (Q1=0.28; Q3=0.56) in cows. Only a few herd characteristics and management practices were found to be related to seroprevalence. Within-herd seroprevalence in cows was found to be significantly (P<0.10) higher in herds (i) with a number of cows<46, (ii) with seasonal calving, and (iii) with grazing or contact through the fence with other ruminant herds. The risk of having at least one seropositive nulliparous female was increased in herds (i) with seasonal calving and (ii) where the foetus and/or the placenta of aborted cows were not systematically removed. Our findings support, in addition to the implementation of high level of hygiene measures, the relevance of vaccination (at least in nulliparous females) as a method to control the spread of C. burnetii within an infected herd, as vaccination is effective in susceptible animals and given that nulliparous females are mostly not infected even in infected herds.

Quantification and at-risk period of decreased fertility associated with exposure to bluetongue virus serotype 8 in naïve dairy herds

The detrimental effect of bluetongue virus serotype 8 (BTV-8) on fertility was quantified in seroconverting cows. Although the effect on individual cows provides information regarding the potential biological burden of infection, losses at a herd level are also dependent on the proportion of infected cows within the herd. The objectives of this study were to quantify the average effect of BTV-8 exposure in field conditions on the fertility of dairy cows in previously naïve herds, and to determine the at-risk period of decreased fertility related to the date of detection of the disease in the herd. The effect of BTV-8 exposure on fertility was assessed using the 90-d-return-to-service rates after the first artificial insemination (AI) calculated for cows in exposed herds (during the 2007 epizootic in France) and compared with that for cows in unexposed herds. Only herds with a confirmed detection that were reported after clinical suspicion were included. To determine the at-risk period of decreased fertility, variations of fertility in exposed herds were quantified according to the time interval between the date of AI for individual cows and the date that disease was detected in the herd. Survival analyses were used to assess the risk of decreased fertility associated with BTV-8 exposure, adjusting for the main factors known to influence fertility. The episode at risk for decreased fertility depended on the month of disease detection in the herd. For herds detected early in the epizootic, fertility was decreased for cows inseminated from 1 mo before to 1 mo after the date of disease detection in the herd. Depending on time interval between the date of AI of cows and the date of detection in the herd, the increase of return-to-service rate associated with BTV-8 exposure varied from 8 to 21 percentage points of 90-d return to service. The episode of decreased fertility is likely due to a combination of the effect of the infection at different stages of conception and early pregnancy and the delayed exposure of cows due to the spreading of the virus within herds. For herds detected during the second half of the epizootic, fertility was decreased for cows inseminated more than 2 mo before detection, which suggests a delay in the detection of clinical signs following virus introduction in the herd. No correlation was observed between the effect of BTV-8 exposure on fertility and the incidence of BTV-8 in the local geographical area. Given the duration of the period that cows were at risk for decreased fertility and the magnitude of the effect, the average BTV-8 exposure in naïve herds led to major losses.

A side effect of decreased fertility associated with vaccination against bluetongue virus serotype 8 in Holstein dairy cows

Inactivated virus vaccines have been widely used to control bluetongue after introduction of serotype 8 of the bluetongue virus (BTV) in northern Europe in 2006. To evaluate vaccination, quantitative knowledge of its possible side effects is needed. One current adverse reaction with inactivated vaccines is a rise in body temperature, which could reduce cow reproductive performance. The objective of this study was to quantify a possible side effect of vaccination on fertility before the implantation of the embryo of dairy cows under field conditions. The study was performed on herds that were not exposed to BTV. Fertility was assessed by return-to-service following artificial insemination (AI). Biological assumptions for a possible side effect of vaccination were conception failure and embryonic death. Associations between return-to-service rates and vaccine injections were assessed using mixed-logistic regression models and survival analysis. Two models were considered: a 3-week-return-to-service model comparing cows vaccinated between 3 days before and 16 days after AI and unvaccinated cows (assuming an effect on conception failure or early embryonic death), and a 90-day-return-to-service model comparing cows vaccinated between 3 days before and 42 days after AI and unvaccinated cows (assuming an effect on conception failure, early or late embryonic death). Only cows receiving a second vaccine injection between 2 and 7 days after AI had a significantly higher risk of 3-week-return-to-service (RR=1.19 [1.07-1.33]). This corresponds to an increase of return-to-service by 4 percentage points. A side effect of vaccination could be due to early embryonic death. The slight side effect on fertility associated with vaccination was low compared to effects of BTV-8 exposure on fertility.