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1.  Brucella melitensis in France: Persistence in Wildlife and Probable Spillover from Alpine Ibex to Domestic Animals 
PLoS ONE  2014;9(4):e94168.
Bovine brucellosis is a major zoonosis, mainly caused by Brucella abortus, more rarely by Brucella melitensis. France has been bovine brucellosis officially-free since 2005 with no cases reported in domestic/wild ruminants since 2003. In 2012, bovine and autochthonous human cases due to B. melitensis biovar 3 (Bmel3) occurred in the French Alps. Epidemiological investigations implemented in wild and domestic ruminants evidenced a high seroprevalence (>45%) in Alpine ibex (Capra ibex); no cases were disclosed in other domestic or wild ruminants, except for one isolated case in a chamois (Rupicapra rupicapra). These results raised the question of a possible persistence/emergence of Brucella in wildlife. The purpose of this study was to assess genetic relationships among the Bmel3 strains historically isolated in humans, domestic and wild ruminants in Southeastern France, over two decades, by the MLVA-panel2B assay, and to propose a possible explanation for the origin of the recent bovine and human infections. Indeed, this genotyping strategy proved to be efficient for this microepidemiological investigation using an interpretation cut-off established for a fine-scale setting. The isolates, from the 2012 domestic/human outbreak harbored an identical genotype, confirming a recent and direct contamination from cattle to human. Interestingly, they clustered not only with isolates from wildlife in 2012, but also with local historical domestic isolates, in particular with the 1999 last bovine case in the same massif. Altogether, our results suggest that the recent bovine outbreak could have originated from the Alpine ibex population. This is the first report of a B. melitensis spillover from wildlife to domestic ruminants and the sustainability of the infection in Alpine ibex. However, this wild population, reintroduced in the 1970s in an almost closed massif, might be considered as a semi-domestic free-ranging herd. Anthropogenic factors could therefore account with the high observed intra-species prevalence.
doi:10.1371/journal.pone.0094168
PMCID: PMC3986073  PMID: 24732322
2.  Exposure of Wild Boar to Mycobacterium tuberculosis Complex in France since 2000 Is Consistent with the Distribution of Bovine Tuberculosis Outbreaks in Cattle 
PLoS ONE  2013;8(10):e77842.
The Eurasian wild boar (Sus scrofa) is increasingly considered as a relevant actor in the epidemiology of animal tuberculosis (TB). Therefore, monitoring TB in wild boar becomes a key tool for establishing comprehensive control schemes for this disease. To estimate the exposure of free living wild boar to Mycobacterium tuberculosis complex (MTC) in France, a bovine-purified protein derivative based ELISA was used to test 2,080 archived serum samples of hunter-harvested animals in 58 French “départements”. Two cut-off values were used for diagnostic interpretation: 0.2, recommended by the manufacturer (specificity: 96.43%; sensitivity: 72.6%), and 0.5 (specificity: 100%; sensitivity: 64%). During the same period, at the 0.2 cut-off, global true seroprevalence was 5.9% (IC95%: 4.3%-7.7%) and 76% of the sampled “départements” had seropositive wild boar, including seven cattle TB-free “départements. At the 0.5 cut-off, global true seroprevalence was 2.2% (IC95%: 1.5-3.2) and positive wild boar belonged to 21% of the “départements”. All but one of these positive “départements” had reported at least one cattle TB outbreak since 2000. A good consistence between seropositive wild boar and TB outbreaks in cattle was found, especially at the 0.5 cut-off value (the mean distance to the nearest cattle TB outbreak was 13km and 27km for seropositive and seronegative wild boar, respectively; P<0.05). The use of an ELISA to detect MTC antibodies in wild boar has permitted the description of the geographic distribution of MTC contact in wild boar in France. Our results suggest that the ELISA could be used as a first screening tool to conduct TB surveillance in wild boar at a population level. High-risk wild boar populations (e.g. overabundant) could be tested and if identified positive by ELISA they should be surveyed in detail by combining pathology and culture.
doi:10.1371/journal.pone.0077842
PMCID: PMC3805591  PMID: 24167584
3.  Assessment of the diagnostic sensitivity and specificity of an indirect ELISA kit for the diagnosis of Brucella ovis infection in rams 
Background
Brucella ovis causes an infectious disease responsible for infertility and subsequent economic losses in sheep production. The standard serological test to detect B. ovis infection in rams is the complement fixation test (CFT), which has imperfect sensitivity and specificity in addition to technical drawbacks. Other available tests include the indirect enzyme-linked immunosorbent assays (I-ELISA) but no I-ELISA kit has been fully evaluated.
The study aimed to compare an I-ELISA kit and the standard CFT. Our study was carried out on serum samples from 4599 rams from the South of France where the disease is enzootic. A Bayesian approach was used to estimate tests characteristics (diagnostic sensitivity, Se and diagnostic specificity, Sp). The tests were then studied together in order to optimise testing strategies to detect B. ovis.
Results
After optimising the cut-off values in order to avoid doubtful results without deteriorating the concordance between the results of the two tests, the I-ELISA appeared to be slightly more sensitive than CFT (Se I-ELISA = 0.917 [0.822; 0.992], 95% Credibility Interval (CrI) compared to Se CFT = 0.860 [0.740; 0.967], 95% CrI). However, CFT was slightly more specific than I-ELISA (Sp CFT = 0.988 [0.947; 1.0], 95% CrI) compared to Sp I-ELISA =0.952 [0.901; 1.0], 95% CrI).
The tests were then associated with two different interpretation schemes. The series association increased the specificity of screening and could be used for pre-movement testing in rams from uninfected flocks. The parallel association increased sequence sensitivity, thus appearing more suitable for eradicating the disease in infected flocks.
Conclusions
The high sensitivity and acceptable specificity of this I-ELISA kit support its potential interest to avoid the limitations of CFT. The two tests could also be used together or combined with other diagnostic methods such as semen culture to improve the testing strategy. The choice of test sequence and interpretation criteria depends on the epidemiological context, screening objectives and the financial and practical constraints.
doi:10.1186/1746-6148-8-68
PMCID: PMC3391974  PMID: 22640401
Brucella ovis; Diagnostic tests; CFT; I-ELISA; Sensitivity; Specificity; Bayesian approach

Results 1-3 (3)