Elodie Rousset

Coxiella burnetii Shedding Routes and Antibody Response after Outbreaks of Q Fever-Induced Abortion in Dairy Goat Herds

ABSTRACT Q fever is a zoonosis caused by Coxiella burnetii, a bacterium largely carried by ruminants and shed into milk, vaginal mucus, and feces. The main potential hazard to humans and animals is due to shedding of bacteria that can then persist in the environment and be aerosolized. The purpose of this study was to evaluate shedding after an outbreak of Q fever abortion in goat herds and to assess the relationship with the occurrence of abortions and antibody responses. Aborting and nonaborting goats were monitored by PCR for C. burnetii shedding 15 and 30 days after the abortion episodes. PCR analysis of all samples showed that 70% (n = 50) of the aborting and 53% (n = 70) of the nonaborting goats were positive. C. burnetii was shed into vaginal mucus, feces, and milk of 44%, 21%, and 38%, respectively, of goats that aborted and 27%, 20%, and 31%, respectively, of goats that delivered normally. Statistical comparison of these shedding results did not reveal any difference between these two groups. PCR results obtained for the vaginal and fecal routes were concordant in 81% of cases, whereas those for milk correlated with only 49% of cases with either vaginal or fecal shedding status. Serological analysis, using enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and complement fixation tests, showed that at least 24% of the seronegative goats shed bacteria. Positive vaginal and fecal shedding, unlike positive milk shedding, was observed more often in animals that were weakly positive or negative by ELISA or IFA. Two opposite shedding trends were thus apparent for the milk and vaginal-fecal routes. Moreover, this study showed that a nonnegligible proportion of seronegative animals that delivered normally could excrete C. burnetii.

The Importance of Ticks in Q Fever Transmission: What Has (and Has Not) Been Demonstrated?

Q fever is a widespread zoonotic disease caused by Coxiella burnetii, a ubiquitous intracellular bacterium infecting humans and a variety of animals. Transmission is primarily but not exclusively airborne, and ticks are usually thought to act as vectors. We argue that, although ticks may readily transmit C. burnetii in experimental systems, they only occasionally transmit the pathogen in the field. Furthermore, we underscore that many Coxiella-like bacteria are widespread in ticks and may have been misidentified as C. burnetii. Our recommendation is to improve the methods currently used to detect and characterize C. burnetii, and we propose that further knowledge of Coxiella-like bacteria will yield new insights into Q fever evolutionary ecology and C. burnetii virulence factors.

Efficiency of a phase 1 vaccine for the reduction of vaginal Coxiella burnetii shedding in a clinically affected goat herd.

The main route of human Q fever infections is inhalation of infectious aerosols. The bacterial agent, Coxiella burnetii, is frequently found throughout domestic ruminants. Females constitute potential shedders of C. burnetii through vaginal mucus, faeces and milk. Q fever is also a common cause of abortion, especially in goats. Massive load of bacteria is associated with placentas and aborted foetuses. In experimental conditions, the Coxevac inactivated phase 1 vaccine (CEVA Sante Animale, Libourne, France) was efficient, and dramatically reduced abortion and excretion of bacteria [1]. The aim of the present study was to evaluate the Coxevac vaccination impact on bacterial shedding in goats affected by Q fever in natural conditions.

Coxiella burnetii vaginal shedding and antibody responses in dairy goat herds in a context of clinical Q fever outbreaks

This study, carried out in three goat herds, was aimed at describing individual responses to Q fever infection in an abortive context, focusing on both antibody and shedding levels. Seroprevalence of Coxiella burnetii (Cb) infection and vaginal shedding of 1083 goats were investigated using ELISA and realtime qPCR assays, respectively. At the end of the outbreaks, a seroprevalence of 45.0% was found, and vaginal shedding appeared massive with levels above 10(4) Cb per swab in 42.3% of the whole population and above 10(6) Cb per swab for 90.9% of aborted goats. Susceptible animals (i.e. seronegative nonshedders) were unfrequent (31.2%), most of them being kids (94.7%). Seronegative females were predominant among nonshedders and conversely seropositive ones, predominant among high shedders (above 10(6) Cb per swab). Nevertheless, at least 43.3% of seronegative goats shed bacteria confirming the need of interpreting serology on a herd scale. The subsequent farrowing period was characterized by a significant reduction in the number of clinical cases. Females that had already aborted were more often involved than others. Shedding quantities remained high, particularly for primiparous does, mainly when facing infection for the first time. Thus, Q fever control must be based on both preventive measures directed to the preherd and environmental precautions.

Assessment of vaccination by a phase I Coxiella burnetii-inactivated vaccine in goat herds in clinical Q fever situation

A study was carried out to assess the efficacy of vaccination, using a phase I Coxiella burnetii-inactivated vaccine (Coxevac®; CEVA), within three goat herds experiencing Q fever abortions waves. The stratification of the population (n = 905) was based on parity and on infection status related to both serological and qPCR vaginal shedding results. Control (n = 443) and vaccinated (n = 462) groups were established in each farm. Vaccination was administered to does before mating and to kids after active immunity acquisition (at least 3–4 months old). The vaccine effectiveness was analyzed at subsequent farrowing on both clinical incidence and vaginal shedding at the delivery day. Among the 231 animals considered as susceptible, that is, seronegative nonshedders, about 90% were infected whatever the group, showing that vaccination did not prevent infection under high infection exposure. Fortunately, vaccination induced an overall decrease in shedding levels. A significant average difference between groups was estimated to 1.16 log(10) bacteria per swab for primiparous and even higher (1.81 log(10)) for initially susceptible ones. Thus, in a clinical context, vaccination should be implemented first in renewal animals. Indeed, young animals are those which best respond to vaccination by significantly reducing C. burnetii burden and, conversely, which excrete bacteria most massively if not vaccinated.

Circulation of Coxiella burnetii in a Naturally Infected Flock of Dairy Sheep: Shedding Dynamics, Environmental Contamination, and Genotype Diversity

ABSTRACT Q fever is a worldwide zoonosis caused by Coxiella burnetii. Domestic ruminants are considered to be the main reservoir. Sheep, in particular, may frequently cause outbreaks in humans. Because within-flock circulation data are essential to implementing optimal management strategies, we performed a follow-up study of a naturally infected flock of dairy sheep. We aimed to (i) describe C. burnetii shedding dynamics by sampling vaginal mucus, feces, and milk, (ii) assess circulating strain diversity, and (iii) quantify barn environmental contamination. For 8 months, we sampled vaginal mucus and feces every 3 weeks from aborting and nonaborting ewes (n = 11 and n = 26, respectively); for lactating females, milk was obtained as well. We also sampled vaginal mucus from nine ewe lambs. Dust and air samples were collected every 3 and 6 weeks, respectively. All samples were screened using real-time PCR, and strongly positive samples were further analyzed using quantitative PCR. Vaginal and fecal samples with sufficient bacterial burdens were then genotyped by multiple-locus variable-number tandem-repeat analysis (MLVA) using 17 markers. C. burnetii burdens were higher in vaginal mucus and feces than in milk, and they peaked in the first 3 weeks postabortion or postpartum. Primiparous females and aborting females tended to shed C. burnetii longer and have higher bacterial burdens than nonaborting and multiparous females. Six genotype clusters were identified; they were independent of abortion status, and within-individual genotype diversity was observed. C. burnetii was also detected in air and dust samples. Further studies should determine whether the within-flock circulation dynamics observed here are generalizable.

Prevalence and molecular typing of Coxiella burnetii in bulk tank milk in Belgian dairy goats, 2009–2013

Q fever, a worldwide zoonosis, is an arousing public health concern in many countries since the recent Dutch outbreak. An emerging C. burnetii clone, genotype CbNL01, was identified as responsible for the Dutch human Q fever cluster cases. Since 2009, Q fever surveillance in the goat industry was implemented by the Belgian authorities. The herd prevalence (December 2009-March 2013) ranged between 6.3 and 12.1%. Genotypic analysis highlighted the molecular diversity of the Belgian strains from goats and identified an emerging CbNL01-like genotype. This follow-up allowed the description of shedding profiles in positive farms which was either continuous (type I) and associated to the CbNL01-like genotype; or intermittent (type II) and linked to other genotypes. Despite the circulation of a CbNL01-like strain, the number of notified Belgian human cases was very low. The mandatory vaccination (in June 2011) on positive dairy goat farms in Belgium, contributed to a decrease in shedding.

Draft Genome Sequences of Six Ruminant Coxiella burnetii Isolates of European Origin

ABSTRACT Coxiella burnetii is responsible for Q fever, a worldwide zoonosis attributed to the inhalation of aerosols contaminated by livestock birth products. Six draft genome sequences of European C. burnetii isolates from ruminants are presented here. The availability of these genomes will help in understanding the potential host specificity and pathogenicity and in identifying pertinent markers for surveillance and tracing.

Outbreak of Q fever, Florac, Southern France, Spring 2007

INTRODUCTION In May 2007, five patients with Q fever-like symptoms were reported in an agricultural educational center in the rural southern French town of Florac. An investigation was undertaken to identify the outbreak source and risk factors for infection, and to implement control measures. MATERIALS AND METHODS We undertook active case finding. Patients were defined as individuals with an unexplained fever of ≥38.5°C who lived in, worked in, or visited Florac between April 1 and June 30, 2007. Patients were confirmed by a positive Q fever serology test. A cross-sectional survey with a seroprevalence component was carried out in the educational center and surrounding area. A standardized questionnaire on known risk factors for the infection was used and serological testing was carried out on finger prick blood specimens from participants. The veterinary services investigated local herds within a 5-mile radius using polymerase chain reaction and serological tests. RESULTS One hundred twenty-two people were included in the cross-sectional survey. Eighteen serologically confirmed acute cases were identified, of whom 12 were from the educational center. The statistical analysis showed an independent association between acute infection and living or working near an area where manure had been spread (p = 0.0.042) and male gender (p = 0.022). Frequenting the educational centers canteen was also associated with infection (p = 0.008) among staff and students. The veterinary investigations identified 11 of the 26 tested flocks of goats and sheep as seropositive for Coxiella burnetii, including 2 ovine flocks located northwest of Florac that had high shedding levels of the bacterium. DISCUSSION The observed excess of cases of Q fever in Florac, an area endemic for this infection, in spring 2007 could be explained by an aerial transmission from infectious ovine flocks situated close to the town. All local herd owners were re-educated about the risks and prevention practices for Q fever.

Evaluation of the recombinant Heat shock protein B (HspB) of Coxiella burnetii as a potential antigen for immunodiagnostic of Q fever in goats.

Coxiella burnetii is an intracellular bacterium that causes a worldwide zoonosis, the Q fever. Currently, to diagnose the infection in ruminants, whole cell antigens-based ELISAs are used. In this study a heat shock protein, the HspB, was evaluated for its ability to be recognized by the goat immune system and its capacity to sign a stage of infection. The htpB gene of C. burnetii was cloned and sequenced. A high identity (>90%) was observed among the htpB genes of four ruminant strains tested. A recombinant protein was expressed in a prokaryotic expression system. The rHspB protein was used to determine the IgG reactivity by ELISA. Sera from experimentally and naturally infected goats were tested. The rHspB is recognized early during the infection course, at 18 days post-infection. Moreover, 80-90% of the animals tested were positive at 39-60dpi. In addition, animals presenting a reactivation of the infection displayed a higher reactivity, statistically significant (p<0.05), than that of the animals in latent infection. These findings suggest that the rHspB could be a good candidate for the development of an ELISA test making possible the detection of recent C. burnetii infection in goats as well as reactivation in those with latent infection.