Exploratory investigation of Q fever in apparently healthy meat sheep flocks in Belgium.

Q fever is a cosmopolitan disease affecting both humans and many animal species. Although sheep are often implicated in human Q fever outbreaks, the disease remains largely underestimated in meat sheep flocks. In order to fulfil this gap, a preliminary study was performed aiming to investigate the serological and molecular aspects of infection with Coxiella burnetii among meat sheep flocks in Belgium. Five Belgian sheep flocks were recruited for this work. Indirect ELISA was used, and in addition, real-time PCR was performed on samples of milk, rectal and vaginal swabs, to understand the dynamics of bacterial shedding. Despite the low overall apparent seroprevalence of 1.39% (95% CI: 0.04-7.5), a high rate of bacterial shedding was found, with 27.7% of tested sheep (N = 72) with a positive result to PCR, especially through the rectal and vaginal routes and in seronegative animals. Furthermore, Coxiella burnetii DNA was detected in 26.76% of seronegative animals. It can be concluded that an overall good clinical condition of the sheep cannot be used to exclude the presence of C. burnetii in a flock. Furthermore in the diagnosis of Q fever in sheep, serology alone was not a sensitive diagnostic tool. On the contrary, molecular biology allowed to detect bacterial shedding, which is an essential element in order to assess the risk due to the contact with shedding animals. At the light of these results, the role of meat sheep flocks in the epidemiology of Q fever in Belgium needs to be better understood.

Serogroups and genotypes of Leptospira spp. strains from bovine aborted foetuses.

Leptospirosis is a global disease of animals, with potential major economic impact on livestock industry and important zoonotic capacities. The disease represents a major challenge in the developing countries as humans and animals frequently live in close association. The serovar Hardjo of Leptospira whose primary host is cattle has been studied extensively, but few data exist on other current circulating or emerging serotypes. To better understand the disease in cattle and how to prevent and/or control it, it is necessary to identify the genotype and the serotype of circulating Leptospira. This study presents results of several investigations performed on a historical Belgian collection of congenital jaundice in bovine aborted foetuses coming from the leptospirosis emerging episode of 2014 (Delooz et al., Transboundary and Emerging Diseases, 62, 2015, 124). The results revealed that L. Grippotyphosa and L. Australis were the most prevalent serogroups with, respectively, 17/42 and 13/42 positive microscopic agglutination test (MAT) during this emerging event associated with the same clinical pattern. The study also confirms that congenital jaundice is associated with L. kirscheneri and L. interrogans and provides the genotyping of DNA obtained from these two species.

Bayesian assessment of two competitive enzyme-linked immunosorbent assays for the detection of bovine viral diarrhoea virus antibodies in bovine sera.

Infections due to bovine viral diarrhoea virus (BVDV) are endemic in most cattleproducing countries throughout the world and bovine viral diarrhoea is considered a transboundary disease. The key elements of a BVDV control programme are vaccination, biosecurity, elimination of persistently infected (PI) animals and surveillance. The aim of this study was to assess the sensitivity (Se) and the specificity (Sp) of two commercial competitive enzyme-linked immunosorbent assays (ELISAs) based on selected immune-dominant BVDV proteins: the non-structural protein NS3 (p80) and the recombinant envelope glycoprotein E0(Erns). Both tests were used on individual serum samples from randomly sampled young bovines in southern Belgium in order to detect specific BVDV antibodies. The Se and Sp were assessed using a Bayesian approach and were estimated, respectively, at 97.2% (with 95% credibility interval [Cr I]: 95.1-99.8) and 98.7%(95% Cr I: 96.6-99.9) for the first test and 95.8% (95% Cr I: 91.1-99.7) and 96.1%(95% Cr I: 95.1-97.7) for the second test. The results obtained with the two tests were not significantly different. In addition, using both ELISAs, the current BVDV exposure among young bovines in southern Belgium was estimated at 23.3% (95%Cr I: 20.6-26.2). Combining virological testing of all newborns to detect PI animals with regular serological testing of young stock using ELISAs is recommended in the surveillance of BVDV.

Les infections par le virus de la diarrhée virale bovine (VDVB) sont endémiques dans la plupart des pays d'élevage du monde et la diarrhée virale bovine estune maladie transfrontalière. Les éléments d'un programme de contrôle de la VDVB sont la vaccination, la biosécurité, l'élimination d'animaux porteurs d'une infection persistante et la surveillance. L'objectif de cette étude était d'évaluer la sensibilité (Se) et la spécificité (Sp) de deux kits commerciaux ELISA (épreuve immuno-enzymatique) par compétition basés sur une sélection de protéines immunodominantes du VDVB, la protéine non structurale NS3(p80) et la glycoprotéine d'enveloppe recombinante E0 (Erns). Les deux kits étaient testés sur des échantillons individuels de sérum collectés demanière aléatoire chez de jeunes bovins dans le sud de la Belgique afin de détecter les anticorps VDVB spécifiques. L'analyse bayésienne montrait une Se de 97,2 % (intervalle de crédibilité de 95 % [ICr] de 95,1 à 99,8) et une Sp de 98,7 (ICr 95 % de 96,6 à 99,9) pour le premier kit et une Se de 95,8 (ICr 95 % de91,1 à 99,7) et une Sp de 96,1 (ICr 95 % de 95,1 à 97,7) pour le deuxième. Les différences n'étaient pas significatives. De même, l'application des deux ELISA montrait que l'exposition actuelle des jeunes bovins du sud de la Belgique au VDVB s'élevait à 23,3 % (ICr 95 % de 20,6 à 26,2). L'association de tests virologiques effectués chez tous les nouveau-nés afin de détecter des animaux à infection persistante et de tests sérologiques de routine par ELISA chez les jeunes animaux est recommandée pour la surveillance de VDVB.

Las infecciones por el virus de la diarrea viral bovina (BVDV), considerada enfermedad transfronteriza, son endémicas en la mayoría de los países del mundo que albergan producción bovina. Los principales elementos de todo programa de lucha contra este virus son la vacunación, la seguridad biológica, la eliminación de los animales con infección persistente y la vigilancia. Los autores describen un estudio encaminado a evaluar la sensibilidad y especificidad de dos ensayos inmunoenzimáticos (ELISA) comerciales basados en sendas proteínas inmunodominantes del virus: la proteína no estructural NS3 (p80) y la glucoproteína recombinante de envoltura E0 (Erns). Ambas pruebas fueron aplicadas a sueros procedentes de una muestra aleatoria de bovinos jóvenes del sur de Bélgica con el fin de detectar anticuerpos específicos contra el virus. Empleando un método de estadística bayesiana se calcularon la sensibilidad y la especificidad, que resultaron, respectivamente, de un 97,2% (intervalo de credibilidad [I Cr] al 95%: 95,1­99,8) y un 98,7% (I Cr 95%: 96,6­99,9) en el caso dela primera prueba y de un 95,8% (I Cr 95%: 91,1­99,7) y un 96,1% (I Cr 95%: 95,1­97,7) en el caso de la segunda. Los resultados obtenidos con una y otra prueba no diferían significativamente entre sí. Además, utilizando ambas técnicas ELISA se calculó que la exposición actual al virus de los bovinos jóvenes del sur de Bélgica se cifraba en un 23,3% (I Cr 95%: 20,6­26,2). Para las tareas de vigilancia del BVDV se recomienda combinar el análisis virológico de todos los recién nacidos ­ para detectar animales con infección persistente ­ con la realización periódica de pruebas serológicas en el ganado joven con la técnica ELISA.

Q Fever Serological Survey and Associated Risk Factors in Veterinarians, Southern Belgium, 2013.

A sero-epidemiological survey was organized among veterinarians working in Southern Belgium to estimate the seroprevalence of Q fever and the risk factors associated with exposure. A total of 108 veterinarians took part to this cross-sectional study, with a majority practicing with livestock animals. The overall seroprevalence was 45.4%, but it increased to 58.3% among veterinarians having contact with livestock. Three main serological profiles were detected (relatively recent, past and potentially chronic infections). The contact with manure during the prior month was the risk factor associated with seropositivity after multivariate logistic regression analysis. Classification and regression tree analysis identified the age as the most predictive variable to exclude potentially chronic infection in apparently healthy seropositive veterinarians. In conclusion, livestock veterinarians practicing in Southern Belgium are highly exposed to Q fever, a neglected zoonosis for which serological and medical examinations should be envisaged in at risk groups.

Laboratory Findings Suggesting an Association Between BoHV-4 and Bovine Abortions in Southern Belgium.

Abortions cause heavy economic losses for the bovine sector. The use of a standardized panel of analyses covering a large spectrum of pathogens responsible of abortion in cattle allowed demonstrating the direct involvement of at least one pathogen in 57% of analysed abortions in the southern part of Belgium. This result suggests a margin of improvement in the diagnostic efficacy. In order to evaluate the interest to broaden the list of pathogens included in the panel of analyses, the implication of bovine herpesvirus 4 (BoHV-4) in abortion was assessed by two different studies. In the first study, coupled serology was performed after abortion on 714 dams to identify specific seroconversion against BoHV-4. The overall seroconversion in cows was 19.5%, with a higher frequency in primiparous compared to multiparous females. In addition, the type of breed (beef cattle) and the time period from the fourth quarter 2008 until the last quarter 2009 were significantly related to the seroconversion of cows. The second study investigated the virus ability to infect the foetus. In this study, 368 cases of bovine abortions were specifically tested for BoHV-4, using PCR on foetus tissues and ELISA on dam and foetus sera. The results showed a maternal seroprevalence of 64.7%, a foetal seroprevalence of 0.8% and a PCR prevalence in foetuses of 1.1%, demonstrating the ability of BoHV-4 to infect the foetus.

Serological diagnosis of bovine neosporosis: a Bayesian evaluation of two antibody ELISA tests for in vivo diagnosis in purchased and abortion cattle.

Currently, there are no perfect reference tests for the in vivo detection of Neospora caninum infection. Two commercial N caninum ELISA tests are currently used in Belgium for bovine sera (TEST A and TEST B). The goal of this study is to evaluate these tests used at their current cut-offs, with a no gold standard approach, for the test purpose of (1) demonstration of freedom of infection at purchase and (2) diagnosis in aborting cattle. Sera of two study populations, Abortion population (n=196) and Purchase population (n=514), were selected and tested with both ELISA's. Test results were entered in a Bayesian model with informative priors on population prevalences only (Scenario 1). As sensitivity analysis, two more models were used: one with informative priors on test diagnostic accuracy (Scenario 2) and one with all priors uninformative (Scenario 3). The accuracy parameters were estimated from the first model: diagnostic sensitivity (Test A: 93.54 per cent-Test B: 86.99 per cent) and specificity (Test A: 90.22 per cent-Test B: 90.15 per cent) were high and comparable (Bayesian P values >0.05). Based on predictive values in the two study populations, both tests were fit for purpose, despite an expected false negative fraction of ±0.5 per cent in the Purchase population and ±5 per cent in the Abortion population. In addition, a false positive fraction of ±3 per cent in the overall Purchase population and ±4 per cent in the overall Abortion population was found.

Congenital jaundice in bovine aborted foetuses: an emerging syndrome in southern Belgium.

Southern Belgium faces an unusual recent increase of icteric bovine aborted foetuses. In the necropsy room, the majority of foetuses presented jaundice and splenomegaly. Despite a wide range of analyses, no definitive cause of abortion has yet been established but some analysis results support the leptospirosis hypothesis. This first description of cases will help veterinary practitioners to recognize more cases and to conduct those to the laboratory for future investigations.

Follow-up of the Schmallenberg Virus Seroprevalence in Belgian Cattle.

Schmallenberg virus (SBV), which emerged in Northwestern Europe in 2011, is an arthropod-borne virus affecting primarily ruminants. Based on the results of two cross-sectional studies conducted in the Belgian ruminant population during winter 2011-2012, we concluded that at the end of 2011, almost the whole population had already been infected by SBV. A second cross-sectional serological study was conducted in the Belgian cattle population during winter 2012-2013 to examine the situation after the 2012 transmission period and to analyse the change in immunity after 1 year. A total of 7130 blood samples collected between 1st January and 28 February 2013 in 188 herds were tested for the presence of SBV-specific antibodies. All sampled herds tested positive and within-herd seroprevalence was estimated at 65.66% (95% CI: 62.28-69.04). A statistically significant decrease was observed between the beginning and the end of 2012. On the other hand, age-cohort-specific seroprevalence stayed stable from 1 year to the other. During winter 2012-2013, calves between 6 and 12 months had a seroprevalence of 20.59% (95% CI: 15.34-25.83), which seems to be an indication that SBV was still circulating at least in some parts of Belgium during summer-early autumn 2012. Results showed that the level of immunity against SBV of the animals infected has not decreased and remained high after 1 year and that the spread of the virus has slowed down considerably during 2012. This study also indicated that in the coming years, there are likely to be age cohorts of unprotected animals.

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.

Seroprevalence of Toxoplasma gondii in domestic sheep in Belgium.

Even though infected sheep are a potential source of Toxoplasma gondii infection in humans, information is lacking concerning the seroprevalence of T. gondii infection in sheep in Belgium. We examined 3170 serum samples for anti-Toxoplasma IgG in sheep by total lysate antigen (TLA) enzyme-linked immunosorbent assay (ELISA). IgG to T. gondii was demonstrated in 87.4% of the tested sheep and in 96.2% of the 209 tested flocks. The seroprevalences in Antwerp (65.2%) and Wallonia (68.6%) are statistically lower than in the other regions in Belgium (96.7-97.8%) (P<0.05). The present study is the first report that analyzed the prevalence of T. gondii infection in sheep in Belgium and confirms the high prevalence of Toxoplasma-specific IgG antibodies in the sheep population.

[BOUGE-fitness test battery: health-related field-based fitness tests assessment in children and adolescents]. / La condition physique des enfants et adolescents: comment la mesurer en milieu scolaire? L'exemple de la batterie BOUGE.

Physical fitness is an important determinant of global health in children and adolescents. Key components of physical fitness include cardiorespiratory fitness, flexibility, speed, and muscular and endurance strength. The school environment provides a great opportunity to assess the physical fitness level in children and adolescents. The french national program "Bouge... Une priorité pour ta santé! "(Move ... A priority for your health!) aims to assess the physical fitness of children and adolescents aged 6 to 18 years old in French schools. The aim of this paper is to describe the interest to assess different health-related physical fitness components. It presents the chosen tests within said battery for children and adolescents and explains how the battery was conceived. The health-related physical fitness tests included in "BOUGE" were chosen for their validity, reliability, low cost and feasibility for all schools.

Differential diagnosis of neurologically expressed disorders in Western European cattle.

A classification of neurological or neurologically expressed disorders that occur in Western European cattle aged 12 month and over has been established on the basis of aetiology, frequency and conditions of appearance, age and type of animals concerned and the main clinical signs observed. Neurologically expressed disorders have been classified according to different groups of causes: biological, non-biological and non-specific or unknown. Differential diagnosis of neurologically expressed disorders is an essential element in the clinical epidemiological surveillance of bovine spongiform encephalopathy. A growing number of aetiologies are described in the scientific literature. The identification and centralised management of neurological disorders will make it possible, one the one hand, to take account of the inherent variability in the clinical forms encountered and in the diagnostic approaches of the observers and, on the other hand, to identify new risk factors in order to control them.

An association exists between bovine herpesvirus-4 seropositivity and abortion in cows.

The prevalence of cattle seropositive to bovine herpesvirus-4 (BHV-4) is high in Belgium. In Belgian farms, clinical signs associated with BHV-4 infection essentially involve the genital tract and consist mainly of postpartum metritis or metroperitonitis. The role of BHV-4 in abortion has not been definitively demonstrated but epidemiological and experimental facts suggest its involvement. A seroepidemiological investigation was therefore conducted as a case-control study to compare the seroprevalences of BHV-4 infections in the aborted-cow population and in a randomly selected control group in the province of Liège (Belgium). The seroprevalence (17.2%) in aborted cows was significantly higher than that of the control group (10.0%). The odds ratio (OR) was 1.87 (1.06 < 3.30). BHV-4 infection is therefore considered as a risk factor for abortion in cows.

Assessment of the cell-mediated immunity in cattle infection after bovine herpesvirus 4 infection, using an in vitro antigen-specific interferon-gamma assay.

The cell-mediated immunity (CMI) following bovine herpesvirus 4 (BHV4) infection has been poorly investigated in cattle. The in vivo response measured by a delayed type of hypersensitivity (DTH) assay has been reported to be positive in only few animals showing serological evidences of BHV4 infection. We have investigated the CMI following BHV4 infection by an in vitro antigen-specific interferon gamma (IFN-gamma) release assay, as an indicator of an actively acquired immunity to BHV4. Our preliminary results using a partially purified antigen suggest that there was a measurable CMI in 75 out of 168 animals (44.4%) originating from a farm with a clinical history and serological evidences (76.3% seropositivity) of BHV4 infection. If the results of serological tests and BHV4 IFN-gamma test are interpreted in parallel, 81.5% of the animals are classified positive, demonstrating the complementarity of these tests. The specificity of the BHV4 IFN-gamma test was supported by the absence of a measurable CMI in 41 animals originating from a farm with no clinical history or serological evidence of BHV4 infection. In an allied study, we developed a bovine herpesvirus 1 (BHV1) IFN-gamma test. This allowed us to measure the antigen specific IFN-gamma release after stimulation with a mixture of BHV1 and BHV4 antigens. Animals that were classified negative by the BHV4 IFN-gamma test and by the BHV1 IFN-gamma test, were classified negative after stimulation with a mixture of both antigens. Animals that were classified positive by the BHV4 IFN-gamma test or the BHV1 IFN-gamma test, were classified positive after stimulation with a mixture of both antigens. Taken together these results suggest that the in vitro assessment of the CMI after BHV4 infection should be further investigated as a specific and valuable alternative to the DTH assay.