TY - JOUR T1 - The AMR-ARRAY: A modular bead array detecting β-lactam, (fluoro) quinolone, colistin, aminoglycoside and macrolide resistance determinants in Gram-negative bacteria. JF - J Microbiol Methods Y1 - 2022 A1 - Michaël Timmermans A1 - Samuel Latour A1 - Pieter-Jan Ceyssens A1 - Cristina Garcia-Graells A1 - Carole Kowalewicz A1 - David Fretin A1 - Denis, Olivier A1 - P Wattiau A1 - Cécile Boland KW - Aminoglycosides KW - Anti-Bacterial Agents KW - beta-Lactams KW - Colistin KW - Drug Resistance, Bacterial KW - Escherichia coli KW - Gram-Negative Bacteria KW - Macrolides KW - Quinolones AB -

The aim of this study was to develop a highly multiplexed bead array to detect genes and/or mutations frequently associated with resistance to antimicrobials of the β-lactam, (fluoro)quinolone, colistin, macrolide and aminoglycoside families in Enterobacteriaceae such as Escherichia coli, Shigella spp. and Salmonella spp. Ligase Chain Reaction and the Luminex® technology were combined in a 53-plex assay designed to target selected genetic markers with 3 internal controls. The AMR-ARRAY consistently detected resistance determinants as compared to phenotypically expressed resistance for 94.7% (856/904) of the assessed resistances. When compared to resistance profiles inferred from whole genome sequencing results, the AMR-ARRAY showed a selectivity and specificity of 99.3% and 100%, respectively. The strong features of the AMR-ARRAY are (i) its competitive cost, currently 18€/sample (ii) its wide analytical scope, currently 50 markers covering 5 antimicrobial families, (iii) its robust and user-friendly design consisting in a single-tube assay conducted in 4 successive steps (iv) its relatively short turnaround time, less than 8 h (v) its ability to detect allelic variability at critical SNPs (vi) its open access and easily upgradable design, with probes sequences, procedure and software source code freely available. The use of the AMR-ARRAY as a screening method in official antimicrobial resistance monitoring could improve the granularity of the collected data and pinpoint remarkable isolates harbouring unusual resistance determinants thereby enabling fit-for-purpose selection of isolates for Whole Genome analysis.

VL - 196 M3 - 10.1016/j.mimet.2022.106472 ER - TY - JOUR T1 - An in‐house 45‐plex array for the detection of antimicrobial resistance genes in Gram‐positive bacteria JF - MicrobiologyOpen Y1 - 2022 A1 - Carole Kowalewicz A1 - Michael Timmermans A1 - David Fretin A1 - P Wattiau A1 - Cécile Boland KW - Enterococcus KW - optrA KW - poxtA KW - staphylococcus KW - vanA KW - vanL VL - e1341 M3 - 10.1002/mbo3.1341 ER - TY - JOUR T1 - Belgian bulk tank milk surveillance program reveals the impact of a continuous vaccination protocol for small ruminants against Coxiella burnetii. JF - Transbound Emerg Dis Y1 - 2021 A1 - Wiebke Jansen A1 - Mickael Cargnel A1 - Samira Boarbi A1 - Ingeborg Mertens A1 - Van Esbroeck, Marjan A1 - David Fretin A1 - Marcella Mori AB -

Endemic Q fever in small ruminants remains an ongoing challenge for veterinary and human public health agencies. Though surveillance programs are implemented in Belgium, infection patterns and vaccination profiles, driving variables, as well as geographical clustering were not presented until now. Based on data from a decade of bulk tank milk analysis between 2009 and 2019, shedding in dairy goat herds declined from 16% (8/50) to 6% (10/162), whereas seroprevalence remained between 32% and 40%. Merely up to two shedding dairy sheep flocks were detected until 2019; seroprevalence peaked in 2017 (43%, 12/28) and declined thereafter. The number of animals in the holding influenced significantly (p = .048) the likelihood of shedding, whereas other established risk factors such as uncovered manure, high abortion rates and diversified farm structure could not be confirmed to significantly affect infection on Belgian herd level. Intermittent, incomplete and unsynchronized vaccinated herds shed Coxiella burnetii significantly more often and longer (p < .001) than continuously, complete and synchronized vaccinated herds. Spatial analyses revealed restricted but matching, homogenous clusters with ≤35 km diameter, concentrated in the coastal region close to the border to the Netherlands from 2009 to 2012, and broadened, heterogeneous clusters with ≥45 km diameter between 2014 and 2016 spreading south-west. Though the majority of human cases was notified in this region, the animal clusters could not be allied with Q fever cases. The impact of environmental factors as well as the role of wildlife, rodents and ticks on the transmission between flocks and to humans remains to be elucidated to harness additional epidemiological drivers of Q fever in Belgium. In conclusion, attempts to reduce the burden of Q fever in Belgium should particularly focus on the timely, complete and synchronized vaccination of flocks, including the breeding sire, and particularity in high-risk areas.

M3 - 10.1111/tbed.14273 ER - TY - RPRT T1 - La résistance antimicrobienne chez les E. coli BLSE et Commensales, Campylobacter spp., Salmonella spp., Staphylococcus aureus résistants à la méthicilline (MRSA) et Enterococcus faecalis et faecium isolés des populations d’animaux producteurs d’aliments Y1 - 2021 A1 - Cristina Garcia-Graells A1 - François Bricteux A1 - Cécile Boland A1 - Carole Kowalewicz A1 - Koenraad Van Hoorde A1 - David Fretin A1 - Katelijne Dierick KW - AMR KW - Campylobacter KW - e.coli KW - Enterococcus KW - Salmonella KW - staphylococcus AB -

En Belgique, l’AFSCA surveille la situation de la résistance antimicrobienne (AMR) à la fois dans les denrées alimentaires que chez les animaux producteurs d’aliments (production primaire). La résistance chez les bactéries zoonotiques Salmonella et Campylobacter et chez les Staphylococcus aureus résistants à la méthicilline (MRSA), ainsi que la résistance chez les bactéries indicatrices Escherichia coli, Enterococcus faecalis et Enterococcus faecium ont été surveillées en 2020. Les surveillances des Salmonella, Campylobacter et MRSA ont été effectuées uniquement chez la volaille en 2020. De même, une surveillance spécifique comprenait l'évaluation des niveaux d’E. coli producteurs présumés de bêta-lactamases à spectre étendu (BLSE)-/AmpC-/carbapénémases provenant d'animaux destinés à l'alimentation et des viandes provenant de ces catégories d’animaux a été aussi réalisée. La résistance microbiologique a été évaluée à l'aide des valeurs seuils épidémiologiques (ECOFF).

 

Chez Campylobacter jejuni isolés à partir de la viande de volaille une diminution de la résistance est remarquée pour les fluoro(quinolones) (60%) et la tétracycline (40%). De même une augmentation du nombre d’isolats sensibles à tous les antibiotiques testés a été remarquée en 2020. Les taux de résistance ont également été déterminés pour Campylobacter coli isolés à partir de la viande de volaille. Le profil de résistance observé est similaire à celui de C. jejuni bien que le niveau de prévalence de la résistance soit supérieur chez C. coli que C. jejuni. La surveillance de Campylobacter jejuni dans la matière fécale de poulets de chair montre un profil de résistance similaire à ceux retrouvés dans les viandes de volaille. Ce profil de résistance comprend les fluoro(quinolones) et la tétracycline et a été retrouvé dans 60% des isolats.

 

Chez Salmonella spp. provenant de carcasses de volailles à l’abattoir, le sérovar prédominant est Infantis, comme dans les années précédentes, et le taux de multirésistance est très élevé (88.52%). Un profil commun de résistance à l’ampicilline, sulfaméthoxazole, tétracycline et fluoro(quinolones) se retrouve fréquemment chez ce sérotype. La résistance aux céphalosporines de troisième génération a été détectée exclusivement dans un isolat de S. Minnesota.

 

Une surveillance spécifique d’E. coli productrices de BLSE, AmpC ou de carbapénémases a été menée dans les populations d’animaux, volailles, porcs d’engraissement et bovins de moins d’un an et dans les viandes fraiches de ces 3 populations d’animaux. Comme attendu, le taux de prévalence le plus élevé est retrouvé chez la volaille (79%), suivi des bovins de moins d’un an (76%) et des porcs d’engraissement (44%). En ce qui concerne les viandes fraiches des 3 catégories d’animaux précédentes, le taux le plus élevé a été retrouvé chez la viande de volaille (50.4%), suivi de la viande de bœuf (2.68%) et de la viande de porc (2.02%). Malgré une prévalence très basse chez la viande de bœuf et de porc, les isolats montrent une forte multirésistance. Aucun isolat n’a été isolé de la surveillance spécifique aux carbapénémases présumées et aucun n’a été détecté résistant au méropénème.

 

Chez les E. coli indicateurs isolés à partir de matière fécale dans les trois populations d’animaux producteurs de viande à l’abattoir, le taux de résistance aux antibiotiques de dernier choix (colistine, tigécycline et méropénème) était très faible ou inexistant. A noter, la diminution progressive de la résistance aux fluoro(quinolones) chez la volaille (50%), les bovins (20%) et les porcs d’engraissement (<10%). En ce qui concerne les céphalosporines de troisième génération, une résistance <10% a été retrouvée pour les trois catégories d’animaux.

 

La surveillance des Staphylococcus aureus résistants à la méthicilline (MRSA) chez les animaux producteurs de denrées alimentaires a été réalisée chez la volaille en 2020. En effet, la résistance présente chez nos animaux est importante à évaluer puisque des échanges de MRSA et potentiellement des résistances associées de l’animal vers l’homme, et inversement, ont été décrits. Cette année, la prévalence globale observée chez la volaille reste faible (3.8%, 4/106 échantillons, IC95% [0.1-7.4%]) et stable depuis 2011. Les 4 échantillons positifs pour MRSA ont été typés génétiquement comme appartenant au complexe clonal CC398, type génétique caractéristique des clones MRSA d’origine animale (LA-MRSA) ainsi qu’au spa-type t011, déjà rapporté chez la volaille en Belgique lors des surveillances précédentes (2011, 2014 et 2017) et connu dans la littérature pour être associé au complexe clonal CC398 ou aux LA-MRSA (Hetem et al.,2013, Köck et al., 2013, Vandendriessche et al, 2013, Sharma et al., 2016). Toutes les souches MRSA isolées en 2020 étaient résistantes à la céfoxitine et à la pénicilline, comme attendu, ainsi qu’à la tétracycline, ce qui est également caractéristique des LA-MRSA (Crombé et al., 2013). Un profil de résistance différent a été observé pour chacune des MRSA isolées, et 3 MRSA sur 4 étaient multirésistantes en 2020. Aucune souche n’était résistante à la vancomycine, au linézolide ou à la mupirocine.

 

La surveillance des Enterococcus faecalis et Enterococcus faecium, organisée en Belgique chez les animaux producteurs de denrées alimentaires entre 2011 et 2013, et reprise en 2019, a continué. Elle permet de faire un état des lieux de la prévalence de ces bactéries commensales indicatrices et de compléter l’image de la situation de la résistance aux antimicrobiens au sein de nos élevages, complémentairement à la surveillance des E. coli indicatrices. Les entérocoques sont en effet aussi considérés comme des réservoirs de gènes de résistances aux antibiotiques, présents à la fois chez l’homme et chez les animaux. En 2020, les prévalences des espèces d’entérocoques par catégorie animale étaient similaires à celles observées en 2019. En effet, Enterococcus faecium a été plus fréquemment isolée qu’E. faecalis au sein des échantillons de poules reproductrices (88.2%), poules pondeuses (79.2%), veaux (71.7%) et porcs (67.8%). A l’inverse, Enterococcus faecalis a été isolée plus fréquemment qu’E. faecium au sein des échantillons de dindes (88.1%) et de poulets de chair (66.1%). Les tests de susceptibilité antimicrobienne réalisés cette année ont montré que, dans l’ensemble, les pourcentages de résistance observés chez Enterococcus faecalis et Enterococcus faecium au sein des différentes matrices animales étudiées semblent stables depuis 2019. Les résistances à la tétracycline, à l’érythromycine et à la quinupristine/dalfopristine étaient les résistances les plus observées, à la fois chez E. faecalis et E. faecium, en taux variable selon la matrice animale. Malgré une diminution significative de la résistance à la tétracycline (-11.2%) observée chez E. faecium isolées de porcs depuis 2019, celle-ci restait extrêmement élevée à élevée en 2020, à la fois chez E. faecalis et E. faecium isolées de poulets de chair, dindes, poules reproductrices, veaux et porcs. La résistance au chloramphénicol était particulièrement élevée chez E. faecalis isolées de veaux (43.2%) et de porcs (23.7%), bien que significativement diminuée chez les veaux depuis 2019. Particulièrement observée chez les souches isolées de poulets de chair et de dindes (30.6% et 24.1% respectivement), la résistance à l’ampicilline n’a été observée qu’au sein des E. faecium en 2020 alors qu’également observée chez E. faecalis isolées de poulets de chair, veaux et poules reproductrices en 2019. La résistance à la daptomycine a significativement diminué depuis 2019 chez E. faecium isolées de poulets de chair, poules reproductrices, poules pondeuses et veaux mais des taux modérés de résistance ont été observés en 2020 chez E. faecium isolées de poulets de chair et de dindes.  Des souches résistantes au linézolide ont également été observées en 2020, à savoir 7 E. faecalis isolées de veaux (n=5) et porcs (n=2) et 5 E. faecium isolées de poulets de chair (n=2), porcs (n =1) et veaux (n=2). Une souche E. faecalis isolée de veaux en 2020 était résistante à la vancomycine. En conclusion, au sein d’une même matrice animale, les résistances antimicrobiennes observées peuvent varier selon l’espèce bactérienne étudiée. Ainsi, les E. faecalis isolées de porcs et veaux avaient tendance à accumuler un plus grand nombre de résistances antimicrobiennes, tandis que les E. faecium isolées de poulets de chair et de dindes sont celles qui en accumulaient le plus.  Les souches E. faecalis et E. faecium isolées de poules pondeuses étaient les souches présentant le moins de résistances. Enfin, la multirésistance (résistance à au moins 3 familles d’antimicrobiens), était principalement observée chez les poulets de chair (78.2% d’E. faecalis et 68.8% d’E. faecium), les dindes (66.9% d’E. faecalis et 51.0% d’E. faecium) et les veaux (78.2% d’E. faecalis).

 

PB - LNR AMR Sciensano CY - Brussels, Belgium ER - TY - JOUR T1 - Large diversity of linezolid-resistant isolates discovered in food-producing animals through linezolid selective monitoring in Belgium in 2019 JF - Journal of Antimicrobial Chemotherapy Y1 - 2021 A1 - Michaël Timmermans A1 - Bert Bogaerts A1 - Kevin Vanneste A1 - Sigrid C.J. De Keersmaecker A1 - Nancy Roosens A1 - Carole Kowalewicz A1 - Guillaume Simon A1 - Maria A Argudín A1 - Deplano, Ariane A1 - Hallin, Marie A1 - P Wattiau A1 - David Fretin A1 - Denis, Olivier A1 - Cécile Boland VL - 77 CP - 1 M3 - 10.1093/jac/dkab376 ER - TY - JOUR T1 - Large diversity of linezolid-resistant isolates discovered in food-producing animals through linezolid selective monitoring in Belgium in 2019. JF - J Antimicrob Chemother Y1 - 2021 A1 - Michaël Timmermans A1 - Bert Bogaerts A1 - Kevin Vanneste A1 - Sigrid C.J. De Keersmaecker A1 - Nancy Roosens A1 - Carole Kowalewicz A1 - Guillaume Simon A1 - Argudín, M Angeles A1 - Deplano, Ariane A1 - Hallin, Marie A1 - P Wattiau A1 - David Fretin A1 - Denis, Olivier A1 - Cécile Boland AB -

BACKGROUND: Linezolid is a critically important antibiotic used to treat human infections caused by MRSA and VRE. While linezolid is not licensed for food-producing animals, linezolid-resistant (LR) isolates have been reported in European countries, including Belgium.

OBJECTIVES: To: (i) assess LR occurrence in staphylococci and enterococci isolated from different Belgian food-producing animals in 2019 through selective monitoring; and (ii) investigate the genomes and relatedness of these isolates.

METHODS: Faecal samples (n = 1325) and nasal swab samples (n = 148) were analysed with a protocol designed to select LR bacteria, including a 44-48 h incubation period. The presence of LR chromosomal mutations, transferable LR genes and their genetic organizations and other resistance genes, as well as LR isolate relatedness (from this study and the NCBI database) were assessed through WGS.

RESULTS: The LR rate differed widely between animal host species, with the highest rates occurring in nasal samples from pigs and sows (25.7% and 20.5%, respectively) and faecal samples from veal calves (16.4%). WGS results showed that LR determinants are present in a large diversity of isolates circulating in the agricultural sector, with some isolates closely related to human isolates, posing a human health risk.

CONCLUSIONS: LR dedicated monitoring with WGS analysis could help to better understand the spread of LR. Cross-selection of LR transferable genes through other antibiotic use should be considered in future action plans aimed at combatting antimicrobial resistance and in future objectives for the rational use of antibiotics in a One Health perspective.

M3 - 10.1093/jac/dkab376 ER - TY - JOUR T1 - Phylogeography of Human and Animal Strains: Genetic Fingerprinting of Q Fever in Belgium. JF - Front Cell Infect Microbiol Y1 - 2021 A1 - Sara Tomaiuolo A1 - Samira Boarbi A1 - Fancello, Tiziano A1 - Michel, Patrick A1 - Damien Desqueper A1 - Gregoire, Fabien A1 - Jozefien Callens A1 - David Fretin A1 - Devriendt, Bert A1 - Cox, Eric A1 - Marcella Mori KW - Animals KW - Belgium KW - Cattle KW - Cattle Diseases KW - Coxiella burnetii KW - DNA Fingerprinting KW - Europe KW - Goat Diseases KW - Goats KW - Humans KW - Phylogeography KW - Q Fever KW - Sheep KW - Sheep Diseases AB -

Q fever is a zoonotic disease caused by the bacteria Domestic ruminants are the primary source for human infection, and the identification of likely contamination routes from the reservoir animals the critical point to implement control programs. This study shows that Q fever is detected in Belgium in abortion of cattle, goat and sheep at a different degree of apparent prevalence (1.93%, 9.19%, and 5.50%, respectively). In addition, and for the first time, it is detected in abortion of alpaca (), raising questions on the role of these animals as reservoirs. To determine the relationship between animal and human strains, Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) (n=146), Single-Nucleotide Polymorphism (SNP) (n=92) and Whole Genome Sequencing (WGS) (n=4) methods were used to characterize samples/strains during 2009-2019. Three MLVA clusters (A, B, C) subdivided in 23 subclusters (A1-A12, B1-B8, C1-C3) and 3 SNP types (SNP1, SNP2, SNP6) were identified. The SNP2 type/MLVA cluster A was the most abundant and dispersed genotype over the entire territory, but it seemed not responsible for human cases, as it was only present in animal samples. The SNP1/MLVA B and SNP6/MLVA C clusters were mostly found in small ruminant and human samples, with the rare possibility of spillovers in cattle. SNP1/MLVA B cluster was present in all Belgian areas, while the SNP6/MLVA C cluster appeared more concentrated in the Western provinces. A broad analysis of European MLVA profiles confirmed the host-species distribution described for Belgian samples. genotyping (WGS) further identified the spacer types and the genomic groups of Belgian strains: cattle and goat SNP2/MLVA A isolates belonged to ST61 and genomic group III, while the goat SNP1/MLVA B strain was classified as ST33 and genomic group II. In conclusion, Q fever is widespread in all Belgian domestic ruminants and in alpaca. We determined that the public health risk in Belgium is likely linked to specific genomic groups (SNP1/MLVA B and SNP6/MLVA C) mostly found in small ruminant strains. Considering the concordance between Belgian and European results, these considerations could be extended to other European countries.

VL - 10 M3 - 10.3389/fcimb.2020.625576 ER - TY - JOUR T1 - Laboratory Diagnosis of Bovine Abortions Caused by Non-Maintenance Pathogenic Leptospira spp.: Necropsy, Serology and Molecular Study Out of a Belgian Experience. JF - Pathogens Y1 - 2020 A1 - Gregoire, Fabien A1 - Bakinahe, Raïssa A1 - Petitjean, Thierry A1 - Samira Boarbi A1 - Delooz, Laurent A1 - David Fretin A1 - Saulmont, Marc A1 - Marcella Mori AB -

Bovine leptospirosis is a bacterial zoonotic disease caused by pathogenic spp. The pathology and epidemiology of this infection are influenced by the numerous existing serovars and their adaptation to specific hosts. Infections by host-maintained serovars such as Hardjo are well documented, unlike those from the incidental ones. In July 2014, an emerging phenomenon of an increased incidence of icteric abortions associated with leptospiral infection occurred in southern Belgium. First-line serological analyses targeting cattle-adapted serovars failed at initial diagnosis. This study provides a comprehensive description of laboratory findings-at the level of necropsy, serology and molecular diagnosis-regarding icteric and non-icteric abortions (n = 116) recorded during this time (years 2014-2015) and associated with incidental infection by serovars such as Grippotyphosa, Australis and Icterohaemorrhagiae. Based on these tests, a diagnostic pathway is proposed for these types of infection in cattle to establish an affordable but accurate diagnosis in the future. These investigations add insights into the understanding of the pathogenesis of bovine leptospirosis associated with serovars classically described as non-maintenance.

VL - 9 CP - 6 M3 - https://www.mdpi.com/2076-0817/9/6/413 ER - TY - JOUR T1 - Overview of spatio-temporal distribution inferred by multi-locus sequence typing of Taylorella equigenitalis isolated worldwide from 1977 to 2018 in equidae JF - Vet Microbiol. Y1 - 2020 A1 - F Duquesne A1 - A Merlin A1 - I Pérez-Cobo A1 - K Sedlák A1 - Melzer, F A1 - G Overesch A1 - David Fretin A1 - W Iwaniak A1 - MF Breuil A1 - U Wernery A1 - J Hicks A1 - M Agüero-García A1 - N Frías-Serrano A1 - E San Miguel-Ibanez A1 - E Patrasová A1 - AS Waldvogel A1 - K Szulowski A1 - M Joseph A1 - J Jeeba A1 - J Shanty A1 - P Varghese A1 - A Hans A1 - S Petry KW - Contagious equine metritis KW - Infectious equine disease KW - MLST KW - Taylorellaequigenitalis AB -

The accurate identification of Taylorella equigenitalis strains is essential to improve worldwide prevention and control strategies for contagious equine metritis (CEM). This study compared 367 worldwide equine strains using multilocus sequence typing according to the geographical origin, isolation year and equine breed. The strains were divided into 49 sequence types (STs), including 10 described for the first time. Three major and three minor clonal complexes (CCs), and 11 singletons, were identified. The genetic heterogeneity was low (0.13 STs/strain) despite the wide diversity of geographical origins (n = 16), isolation years (1977-2018) and equine breeds (n = 18). It was highest outside Europe and in the 1977-1997 period; current major STs and CCs already existed before 1998. Previous data associated the major CC1 with the first CEM outbreaks in 1977-1978 in the United Kingdom, Australia and the United States, and revealed its circulation in France. Our study confirms its circulation in France over a longer period of time (1992-2018) and its distribution in Spain and Germany but not throughout Europe. In addition to CC1, relationships between non-European and European countries were observed only through ST4, ST17 and ST30. Within Europe, several STs emerged with cross-border circulation, in particular ST16 and ST46 from the major complexes CC2 and CC8. These results constitute a baseline for monitoring the spread of CEM outbreaks. A retrospective analysis of a higher number of strains isolated worldwide between 1977 and the early 2000s would be helpful to obtain an exhaustive picture of the original CEM situation.

VL - 242 M3 - 10.1016/j.vetmic.2020.108597 ER - TY - JOUR T1 - Bayesian Evaluation of Three Serological Tests for Detecting Antibodies against spp. among Humans in the Northwestern Part of Ecuador. JF - Am J Trop Med Hyg Y1 - 2019 A1 - Jorge Ron-Román A1 - Ron-Garrido, Lenin A1 - Abatih, Emmanuel A1 - Maritza Celi-Erazo A1 - Laura Vizcaíno-Ordóñez A1 - Jaime Calva-Pacheco A1 - Pablo González-Andrade A1 - Berkvens, Dirk A1 - Washington Benítez-Ortíz A1 - Jef Brandt A1 - David Fretin A1 - Saegerman, Claude KW - ADOLESCENT KW - Adult KW - Aged KW - Agglutination Tests KW - Animals KW - Antibodies, Bacterial KW - Bayes Theorem KW - Brucella abortus KW - Brucellosis KW - Cattle KW - cross-sectional studies KW - Ecuador KW - Edetic Acid KW - Enzyme-Linked Immunosorbent Assay KW - Epidemiological Monitoring KW - Female KW - Humans KW - Male KW - middle aged KW - prevalence KW - Rose Bengal KW - Sensitivity and Specificity AB -

Brucellosis is an important but neglected zoonosis that causes serious economic losses both in livestock and human populations. The aim of the present study was to estimate the true prevalence of brucellosis together with diagnostic sensitivity and specificity of three serological tests in humans of the northwestern part of Ecuador using a Bayesian approach adjusted for the dependencies among the multiple tests to avoid any misinterpretation. In addition, the causal agent responsible for human brucellosis was also identified. Using a total of 3,733 samples collected from humans in this area between 2006 and 2008, the prevalence of human brucellosis and the diagnostic test characteristics of the Rose Bengal fast agglutination test (RBT), Wright's slow agglutination test with ethylenediaminetetraacetic acid disodium salt dehydrate (EDTA) (SAT-EDTA), and indirect ELISA (iELISA) were estimated using a Bayesian approach. The estimated true prevalence of human brucellosis was 1% (credibility interval: 0.4-1.6). The sensitivities of iELISA and RBT were higher than and similar (95.1% and 95.0%, respectively) to those of SAT-EDTA (60.8%). Even though all tests indicated a high specificity (> 99.0%), the specificity of SAT-EDTA was highest (99.9%). The circulating strain in this study area was identified to be biotype 4 based on culture and microbiological characterization. The RBT and the iELISA are recommended for estimating the true prevalence of human brucellosis and/or for surveillance programs following their high sensitivities and specificities. The proposed strategy supports evidence-based medicine for clinicians and policy-makers to ensure appropriate preventive and control program of brucellosis worldwide.

VL - 100 CP - 6 M3 - 10.4269/ajtmh.18-0622 ER - TY - JOUR T1 - Evaluation of mycobacteria-specific gamma interferon and antibody responses before and after a single intradermal skin test in cattle naturally exposed to M. avium subsp. paratuberculosis and experimentally infected with M. bovis. JF - Vet Immunol Immunopathol Y1 - 2018 A1 - Virginie Roupie A1 - Elena Alonso-Velasco A1 - Sarah Van Der Heyden A1 - Holbert, Sébastien A1 - Duytschaever, Lucille A1 - Patricia Berthon A1 - Iris Van Dosselaer A1 - Willem Van Campe A1 - Laurent Mostin A1 - Franck Biet A1 - S. Roels A1 - Huygen, Kris A1 - David Fretin KW - Animals KW - Antibody Formation KW - Cattle KW - Interferon-gamma KW - Interferon-gamma Release Tests KW - Male KW - Mycobacterium avium subsp. paratuberculosis KW - Mycobacterium bovis KW - Paratuberculosis KW - Real-Time Polymerase Chain Reaction KW - Tuberculin Test KW - Tuberculosis, Bovine AB -

This study reports on the diagnostic potential of IFN-γ release assays and serology for Mycobacterium bovis in six naturally M. avium subsp. paratuberculosis (Map) exposed bulls of which four were intratracheally infected with a Belgian field strain of M. bovis. Heparinized blood, serum and fecal samples were collected at regular time intervals for mycobacteria-specific IFN-γ release assays, antibody analysis and for Map culture respectively. Single intradermal skin test (SIT) with bovine tuberculin (PPD-B) was performed on day 115 and animals were sacrificed on day 133 after M. bovis infection. Organs were collected and stored for histopathological examination, modified Ziehl-Neelsen staining and bacteriological analysis of M. bovis and Map by culture and RT-PCR. Prior to infection five animals showed positive IFN-γ responses to avian PPD (PPD-A) and four were positive in Map PCR (IS900) on faeces. Three M. bovis infected animals reacted as early as day 14 with sustained higher PPD-B than PPD-A specific IFN-γ responses, whereas the fourth animal (with the strongest PPD-A response prior to infection) showed sustained higher PPD-B specific IFN-γ levels only a day 56 after infection. Two of the infected animals had a sustained positive IFN-γ response to the ESAT-6/CFP-10/TB7.7 (QuantiFERON-TB Gold) peptide cocktail as early as day 14, among which the animal with the initial high PPD-A response. Later during infection, positive responses were found to ESAT-6 peptides in three infected bulls and to CFP-10 peptides in all four infected bulls. One of the control animals reacted intermittently to the ESAT-6/CFP10/TB7.7 cocktail. Prior to SIT, weak but positive MPB83/MBP70 specific antibody responses were detected in two of the infected bulls. All four M. bovis infected bulls reacted with a positive skin test and showed, as reported by others, increased mycobacteria specific IFN-γ production and increased positive responses in MPB83/MBP70 specific serology after SIT. At autopsy, M. bovis lesions were detected in all four experimentally infected bulls. Our results indicate that in Map exposed cattle, M. bovis diagnosis using IFN-γ assays needs a combination of PPD-B/A and ESAT-6/CFP10 for early and optimal sensitivity and that sensitivity of MPB83/MBP70 serodiagnosis is dramatically increased by prior skin testing. Map exposure did not interfere with the development of SIT in M. bovis infected animals, but resulted in a false positive M. bovis specific IFN-γ and antibody response after SIT in one of the two control animals (which remained negative in skin-test).

VL - 196 M3 - 10.1016/j.vetimm.2017.12.007 ER - TY - JOUR T1 - Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh. JF - Zoonoses Public Health Y1 - 2017 A1 - Rahman, A K M A A1 - Saegerman, C A1 - Berkvens, D A1 - Melzer, F A1 - Neubauer, H A1 - David Fretin A1 - Abatih, E A1 - Dhand, N A1 - Ward, M P KW - Animals KW - Bangladesh KW - Brucella abortus KW - Brucellosis KW - DNA, Bacterial KW - Humans AB -

To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh.

VL - 64 CP - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28068003?dopt=Abstract M3 - 10.1111/zph.12344 ER - TY - JOUR T1 - Detection and characterization of Brucella spp. in bovine milk in small-scale urban and peri-urban farming in Tajikistan. JF - PLoS Negl Trop Dis Y1 - 2017 A1 - Lindahl-Rajala, Elisabeth A1 - Hoffman, Tove A1 - David Fretin A1 - Godfroid, Jacques A1 - Sattorov, Nosirjon A1 - Boqvist, Sofia A1 - Lundkvist, Åke A1 - Magnusson, Ulf KW - Agriculture KW - Animals KW - Brucella KW - Cattle KW - DNA, Bacterial KW - DNA-Directed RNA Polymerases KW - Humans KW - milk KW - prevalence KW - Real-Time Polymerase Chain Reaction KW - Sequence Analysis, DNA KW - Suburban Population KW - Tajikistan KW - Urban Population AB -

Brucellosis is one of the most common zoonoses globally, and Central Asia remains a Brucella hotspot. The World Health Organization classifies brucellosis as a neglected zoonotic disease that is rarely in the spotlight for research and mainly affects poor, marginalized people. Urban and peri-urban farming is a common practice in many low-income countries, and it increases the incomes of families that are often restrained by limited economic resources. However, there is a concern that the growing number of people and livestock living close together in these areas will increase the transmission of zoonotic pathogens such as Brucella. This study investigates the presence of Brucella DNA in bovine milk in the urban and peri-urban area of Dushanbe, Tajikistan. Brucella DNA was detected in 10.3% of 564 cow milk samples by IS711-based real-time PCR. This finding is concerning because consumption of unpasteurized dairy products is common in the region. Furthermore, Brucella DNA was detected in the milk of all seropositive cows, but 8.3% of the seronegative cows also showed the presence of Brucella DNA. In addition, sequence analysis of the rpoB gene suggests that one cow was infected with B. abortus and another cow was most likely infected with B. melitensis. The discrepancies between the serology and real-time PCR results highlight the need to further investigate whether there is a need for implementing complementary diagnostic strategies to detect false serological negative individuals in Brucella surveillance, control, and eradication programmes. Furthermore, vaccination of cattle with S19 in addition to vaccination of small ruminants with Rev 1 might be needed in order to control Brucella infections in the livestock population but further research focusing on the isolation of Brucella is required to obtain a comprehensive understanding of the Brucella spp. circulating among the livestock in this region.

VL - 11 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28296882?dopt=Abstract M3 - 10.1371/journal.pntd.0005367 ER - TY - JOUR T1 - Exploring the Diversity of Field Strains of Brucella abortus Biovar 3 Isolated in West Africa. JF - Front Microbiol Y1 - 2017 A1 - Sanogo, Moussa A1 - David Fretin A1 - Thys, Eric A1 - Saegerman, Claude AB -

Brucellosis is one of the most widespread bacterial zoonotic diseases in the world, affecting both humans and domestic and wild animals. Identification and biotyping of field strains of Brucella are of key importance for a better knowledge of the epidemiology of brucellosis, for identifying appropriate antigens, for managing disease outbreaks and for setting up efficient preventive and control programmes. Such data are required both at national and regional level to assess potential threats for public health. Highly discriminative genotyping methods such as the multiple locus variable number of tandem repeats analysis (MLVA) allow the comparison and assessment of genetic relatedness between field strains of Brucella within the same geographical area. In this study, MLVA biotyping data retrieved from the literature using a systematic review were compared using a clustering analysis and the Hunter-Gaston diversity index (HGDI). Thus, the analysis of the 42 MLVA genotyping results found in the literature on West Africa [i.e., from Ivory Coast (1), Niger (1), Nigeria (34), The Gambia (3), and Togo (3)] did not allow a complete assessment of the actual diversity among field strains of Brucella. However, it provided some preliminary indications on the co-existence of 25 distinct genotypes of Brucella abortus biovar 3 in this region with 19 genotypes from Nigeria, three from Togo and one from Ivory Coast, The Gambia, and Niger. The strong and urgent need for more sustainable molecular data on prevailing strains of Brucella in this sub-region of Africa and also on all susceptible species including humans is therefore highlighted. This remains a necessary stage to allow a comprehensive understanding of the relatedness between field strains of Brucella and the epidemiology of brucellosis within West Africa countries.

VL - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28713359?dopt=Abstract M3 - 10.3389/fmicb.2017.01232 ER - TY - JOUR T1 - Imported human brucellosis in Belgium: Bio and molecular typing of bacterial isolates, 1996-2015. JF - PLoS One Y1 - 2017 A1 - Hanot Mambres, Delphine A1 - Samira Boarbi A1 - Michel, Patrick A1 - Bouker, Nora A1 - Escobar-Calle, Luisa A1 - Damien Desqueper A1 - Fancello, Tiziano A1 - Van Esbroeck, Marjan A1 - Godfroid, Jacques A1 - David Fretin A1 - Marcella Mori KW - Bacterial Typing Techniques KW - Belgium KW - Brucella KW - Brucellosis KW - DNA, Bacterial KW - History, 20th Century KW - History, 21st Century KW - Humans KW - Minisatellite Repeats KW - Risk Factors AB -

OBJECTIVES: The aim of this study was to characterize by classical biotyping and Multi-Locus variable number tandem repeats (VNTR) Analysis (MLVA) all Brucella spp. derived from human cases in Belgium from 1996 to 2015. Final goals were to determine the species and biovar, to trace-back on genetic grounds the origin of each strain when patient history and risk factors were missing, and to survey for particular trends at the national level.

METHODS: A total of 37 Brucella strains, isolated from 37 patients in Belgium, were analyzed by both classical biotyping and MLVA, and the genetic patterns compared to those of human strains isolated worldwide.

RESULTS: Classical biotyping revealed that isolates were mainly Brucella melitensis. Most of them belonged to biovar 3, the most abundant biovar in the Mediterranean region. MLVA confirmed that Brucella melitensis is too diverse in VNTRs to be able to make clusters associated to each biovar, but it allowed retrieving precious epidemiological information. The analysis highlighted the imported nature of the strains from all over the world with a dominant part from the Mediterranean countries. Findings of the MLVA11 testing were in line with the travel history of patients coming from Italy, Turkey, Lebanon and Peru. The analysis was particularly useful because it suggested the geographical origin of the infection for 12/16 patients for whom no case history was available.

CONCLUSION: Classical biotyping and MLVA analysis are not exclusive but remain complementary tools for Brucella melitensis strain surveillance. MLVA11 is sufficient for Brucella-free countries such as Belgium to trace the geographical origin of infection, but complete MLVA16 is needed to search for links with endemic areas.

VL - 12 CP - 4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28384245?dopt=Abstract M3 - 10.1371/journal.pone.0174756 ER - TY - JOUR T1 - Reproductive Disorders and Leptospirosis: A Case Study in a Mixed-Species Farm (Cattle and Swine). JF - Vet Sci Y1 - 2017 A1 - Marcella Mori A1 - Bakinahe, Raïssa A1 - Philippe Vannoorenberghe A1 - Jo Maris A1 - de Jong, Ellen A1 - Marylène Tignon A1 - Martine Marin A1 - Damien Desqueper A1 - David Fretin A1 - Isabelle Behaeghel KW - Leptospirosis AB -

Animal leptospirosis, exempt in rodents, manifests as peculiar biology where the animal can function, simultaneously or not, as a susceptible host or reservoir. In the first case, clinical symptoms are likely. In the second case, infection is subclinical and manifestations are mild or absent. Mild clinical symptoms encompass reproductive failure in production animals for host-adapted sp. serovars. This work presents a study on sp. infection in a mixed-species (bovine and swine) farm with documented reproductive disorders in the cattle unit. A long calving interval (above 450 days) was the hallmark observed in cows. Some cows (2/26 tested) presented a high titre of antibodies against sp. serogroup Sejroe, but the overall within-herd prevalence was low (11.5% and 7.7% for cut-off titres of 1:30 and 1:100, respectively). The in-herd prevalence of leptospirosis in the sow unit (determined for 113/140 animals) was high when using a lowered cut-off threshold (32.7% vs. 1.8% for cut-off titre of 1:30 and 1:100, respectively). In this unit, the most prevalent serogroup was Autumnalis. The final diagnostic confirmation of sp. maintenance within the farm was obtained through detection by PCR of sp. DNA in an aborted swine litter. Despite the fact that a common causative infective agent was diagnosed in both species, the direct link between the two animal units was not found. Factors such as drinking from the same water source and the use of manure prepared with the swine slurry might raise suspicion of a possible cross-contamination between the two units. In conclusion, this work suggests that leptospirosis be included in the differential diagnosis of reproductive disorders and spontaneous abortions in production animals and provides data that justify the use of a lowered threshold cut-off for herd diagnosis.

VL - 4 CP - 4 M3 - 10.3390/vetsci4040064 ER - TY - JOUR T1 - Seroprevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Francisella tularensis Infections in Belgium: Results of Three Population-Based Samples. JF - Vector Borne Zoonotic Dis Y1 - 2017 A1 - De Keukeleire, Mathilde A1 - Vanwambeke, Sophie O A1 - Cochez, Christel A1 - Heyman, Paul A1 - David Fretin A1 - Deneys, Véronique A1 - Luyasu, Victor A1 - Kabamba, Benoît A1 - Robert, Annie KW - Adult KW - Aged KW - Anaplasma phagocytophilum KW - Anaplasmosis KW - Belgium KW - Borrelia burgdorferi KW - Female KW - Francisella tularensis KW - Humans KW - Lyme Disease KW - Male KW - middle aged KW - Risk Factors KW - Seroepidemiologic Studies KW - Tularemia AB -

To estimate the seroprevalence of Borrelia burgdorferi (Bb), Anaplasma phagocytophilum (Ap), and Francisella tularensis (Ft) in Belgium, we tested sera from three population-based samples in which exposure to pathogen is assumed to vary: 148 samples from workers professionally exposed, 209 samples from rural blood donors, and 193 samples from urban blood donors. Sera were tested using ELISA or the immunofluorescence assay test. The seroprevalence of Bb was 5.4% in workers professionally exposed, 2.9% in rural blood donors, and 2.6% in urban blood donors, which is similar to other studies. The fraction of negative results decreases significantly from urban blood donors and rural blood donors to workers. Regarding the seroprevalence of Ap, the cutoff titer of 1:64 recommended by the manufacturer may be set too low and produces artificially high seroprevalence rates. Using a cutoff titer of 1:128, the seroprevalence of Ap was estimated at 8.1% for workers professionally exposed, 6.2% for rural blood donors, and 5.7% for urban blood donors. Tularemia sera confirmed the presence of the pathogen in Belgium at 2.0% for workers and 0.5% for rural and urban blood donors. Our study is one of the few providing an estimation of the seroprevalences of Bb, Ap, and Ft in three different populations in Belgium, filling the gap in seroprevalence data among those groups. Our findings provide evidence that the entire Belgian population is exposed to Bb, Ap, and Ft infections, but a higher exposure is noticed for professionals at risk. Education on the risk factors for tick bites and preventive measures for both professionals exposed and the general population is needed.

VL - 17 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/27828762?dopt=Abstract M3 - 10.1089/vbz.2016.1954 ER - TY - JOUR T1 - Trend analysis suggested a change in subspecies among Mycobacterium avium isolated from pigs in Belgium, 1967–2013 JF - Veterinary Record Y1 - 2017 A1 - Karine Soetaert A1 - C. Vluggen A1 - L Duytschaever A1 - J. Denoël A1 - Virginie Roupie A1 - F. Smeets A1 - Nicolas Bruffaerts A1 - Huygen, K. A1 - David Fretin A1 - M. Diels A1 - L. Rigouts A1 - C. Saegerman A1 - Vanessa Mathys VL - 180 CP - 18 M3 - 10.1136/vr.103951 ER - TY - JOUR T1 - Trend analysis suggested a change in subspecies among isolated from pigs in Belgium, 1967-2013. JF - Vet Rec Y1 - 2017 A1 - Karine Soetaert A1 - Duytschaever, L A1 - Denoël, J A1 - Virginie Roupie A1 - Smeets, F A1 - Nicolas Bruffaerts A1 - Huygen, K A1 - David Fretin A1 - Diels, M A1 - Rigouts, L A1 - Saegerman, C A1 - Vanessa Mathys KW - Animals KW - Belgium KW - Genotyping Techniques KW - Mycobacterium avium KW - Swine KW - Swine Diseases KW - Tuberculosis VL - 180 CP - 18 U1 - https://www.ncbi.nlm.nih.gov/pubmed/28283669?dopt=Abstract M3 - 10.1136/vr.103951 ER - TY - JOUR T1 - Virulence and immunogenicity of genetically defined human and porcine isolates of M. avium subsp. hominissuis in an experimental mouse infection JF - PLOS ONE Y1 - 2017 A1 - Nicolas Bruffaerts A1 - Vluggen, Christelle A1 - Virginie Roupie A1 - Duytschaever, Lucille A1 - Christophe Van den Poel A1 - Joseph Noël A1 - Wattiez, Ruddy A1 - Letesson, Jean-Jacques A1 - David Fretin A1 - Rigouts, Leen A1 - Elie Chapeira A1 - Vanessa Mathys A1 - Saegerman, Claude A1 - Huygen, Kris ED - Cloeckaert, Axel KW - Paratuberculosis AB -

Mycobacterium avium subsp. hominissuis (Mah) represents a health concern for humans and to a lesser extent for pigs, but its zoonotic potential remains elusive. Using multispacer sequence typing (MST) we previously identified 49 different genotypes of Mah among Belgian clinical and porcine isolates, with 5 MSTs shared by both hosts. Using experimental intranasal infection of BALB/c mice, we compared the virulence and immunogenicity of porcine and clinical human isolates with shared genotype or with a genotype only found in humans or pigs. Bacterial replication was monitored for 20 weeks in lungs, spleen and liver and mycobacteria specific spleen cell IFN-γ, IL-10 and IL-17 production as well as serum antibody responses were analyzed. Isolates varied in virulence, with human and porcine isolates sharing MST22 genotype showing a thousand fold higher bacterial replication in lungs and more dissemination to spleen and liver than the human and porcine MST91 isolates. Virulent MST22 type was also associated with progressive suppression of IFN-γ and IL-17 responses, and increased IL-10 production. Whole genome sequencing of the two virulent isolates with MST22 genotype and two avirulent isolates of genotype MST91 and comparison with two well-studied M. avium subsp. hominissuis reference strains i.e. Mah 104 and Mah TH135, identified in the two MST22 isolates nine specific virulence factors of the mammalian cell entry family, that were identical with Mah 104 strain. Despite the obvious limitations of the mouse model, a striking link of virulence and identity at the genome level of porcine and human isolates with the same multisequence type, for which no correlation of place of residence (humans) or farm of origin (pigs) was observed, seems to point to the existence in the environment of certain genotypes of Mah which may be more infectious both for humans and pigs than other genotypes.

VL - 12 CP - 2 M3 - 10.1371/journal.pone.017189510.1371/ ER - TY - JOUR T1 - Coxiella burnetii, agent de la fièvre Q JF - Can J Microbiol Y1 - 2016 A1 - Samira Boarbi A1 - David Fretin A1 - Marcella Mori KW - Coxiella burnetii KW - description KW - epidemiology KW - genomics. KW - Q Fever AB -

Q fever is a zoonosis of worldwide distribution with the exception of New Zealand. It is caused by an
intracellular bacterium, Coxiella burnetii. The disease often goes underdiagnosed because the main manifestation
of its acute form is a general self-limiting flu-like syndrome. The Dutch epidemics renewed attention to this
disease, which was less considered before. This review summarizes the description of C. burnetii (taxonomy,
intracellular cycle, and genome) and Q fever disease (description, diagnosis, epidemiology, and pathogenesis).
Finally, vaccination in humans and animals is also considered

VL - 62 CP - 2 M3 - 10.1139/cjm-2015-0551 ER - TY - Generic T1 - Détermination des sous-espèces et génotypage des souches humaines et porcines de Mycobacterium avium isolées en Belgique Y1 - 2016 A1 - Christelle Vluggen A1 - Karine Soetaert A1 - Dutschaever,L. A1 - J. Denoel A1 - Smeets,F. A1 - Nicolas Bruffaerts A1 - K Huygen A1 - David Fretin A1 - Saegerman,C. A1 - Vanessa Mathys KW - Belgique KW - de KW - EN KW - ET KW - Mycobacterium JF - Belspo RT-project PB - NA CY - NA CP - Belspo U1 - 37202 U2 - 16/03/2016 ER - TY - JOUR T1 - Evaluation of three competitive ELISAs and a fluorescence polarisation assay for the diagnosis of bovine brucellosis. JF - Vet J Y1 - 2016 A1 - Praud, A A1 - Durán-Ferrer, M A1 - David Fretin A1 - Jaÿ, M A1 - O'Connor, M A1 - Stournara, A A1 - Tittarelli, M A1 - I Travassos Dias A1 - Garin-Bastuji, B KW - Agglutination Tests KW - Animals KW - Brucella KW - Brucellosis, Bovine KW - Cattle KW - Complement Fixation Tests KW - Enzyme-Linked Immunosorbent Assay KW - Fluorescence Polarization Immunoassay KW - Rose Bengal KW - Sensitivity and Specificity AB -

Bovine brucellosis is an infectious disease of worldwide public health and economic importance. The usual tests for the diagnosis of this disease include the Rose-Bengal test (RBT), complement fixation test (CFT), serum agglutination test (SAT) and indirect ELISA. New tests such as competitive ELISAs (C-ELISA) and fluorescence polarisation assay (FPA) have been developed. However, C-ELISA may correspond to different protocols and a wide variation may exist in their diagnostic performance. The aim of this study was to evaluate three commercially available C-ELISA kits (C-ELISA1-3) and FPA for the diagnosis of bovine brucellosis and compare test performance with RBT, CFT, indirect ELISA and FPA. Sera submitted to EU laboratories in 2011 from 5111 adult cattle were tested. Individual test sensitivities (Se) and specificities (Sp) were estimated. Threshold assessment using the receiver operating characteristic method was also performed. The most sensitive tests were FPA (99.0%; 95% confidence interval [CI], 97.9-100%), C-ELISA1 (98.4%; 95% CI, 97.0-99.8%) and RBT (97.7%; 95% CI, 95.9-99.3%). The most specific tests were CFT (99.98%; 95% CI, 99.93-100%), SAT (99.98%; 95% CI, 99.93-100%) and RBT (99.89%; 95% CI, 99.79-99.99%). Among the new tests, none of the three C-ELISA kits studied could be recommended as a single screening test because of their low specificity, especially when used in a herd. C-ELISA3 could not be recommended as confirmatory test on individual animals to determine whether false positive serological test results had occurred.

VL - 216 U1 - http://www.ncbi.nlm.nih.gov/pubmed/27687924?dopt=Abstract M3 - 10.1016/j.tvjl.2016.06.014 ER - TY - JOUR T1 - Genotyping and strain distribution of Mycobacterium avium subspecies hominissuis isolated from humans and pigs in Belgium, 2011-2013. JF - Euro Surveill Y1 - 2016 A1 - Vluggen, Christelle A1 - Karine Soetaert A1 - Duytschaever, Lucille A1 - Denoël, Joseph A1 - Fauville-Dufaux, Maryse A1 - Smeets, François A1 - Nicolas Bruffaerts A1 - Huygen, Kris A1 - David Fretin A1 - Rigouts, Leen A1 - Saegerman, Claude A1 - Vanessa Mathys KW - Animals KW - Belgium KW - Genetic Variation KW - Genotype KW - Humans KW - Minisatellite Repeats KW - Mycobacterium avium KW - Phylogeny KW - polymerase chain reaction KW - Polymorphism, Restriction Fragment Length KW - Retrospective Studies KW - Sequence Analysis, DNA KW - Swine KW - Swine Diseases KW - Tuberculosis AB -

Mycobacterium avium represents a health concern for both humans and pigs. The characterisation of its subspecies is an important step improving the understanding of the epidemiology and the control of this pathogen. Ninety-two human M. avium strains were selected for a retrospective study. Subspecies determination by rpoB sequencing and IS1245/IS901 analysis showed that 98.9% of Belgian human M. avium strains belong to the subspecies hominissuis (MAH). Some of these MAH strains present particular IS1245/IS901 profiles (absence of IS1245 and false IS901 detection provoked by the presence of ISMav6). In addition, 54 MAH strains isolated from submandibular lymph nodes of Belgian pigs with lymphadenitis were included in this study. Genotyping of human and porcine isolates was performed using multispacer sequence typing (MST). In total, 49 different MST types were identified among pig (n = 11) and human (n = 43) MA isolates, with only five shared by both hosts. Among these MST types, 34 were newly identified. Our findings demonstrate the extensive genetic diversity among MAH isolates. Some genotypes were more prevalent in human or pigs but no correlation was observed between MST type and place of residence or the farm of origin for human and porcine isolates respectively, suggesting an environmental source of infection.

VL - 21 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26835872?dopt=Abstract M3 - 10.2807/1560-7917.ES.2016.21.3.30111 ER - TY - Generic T1 - Infections mycobactériennes chez l'homme et l'animal Y1 - 2016 A1 - Vanessa Mathys A1 - David Fretin KW - ET KW - health KW - INFECTION KW - infections JF - One Health Seminar PB - NA CY - NA CP - WIV-ISP U1 - 37199 U2 - 25/05/2016 ER - TY - JOUR T1 - Seroprevalence of brucellosis in patients with prolonged fever in Bangladesh. JF - J Infect Dev Ctries Y1 - 2016 A1 - Rahman, Akm Anisur A1 - Berkvens, Dirk A1 - Saegerman, Claude A1 - David Fretin A1 - Muhammad Noor A1 - Hossain, Akram A1 - Abatih, Emmanuel KW - ADOLESCENT KW - Adult KW - Aged KW - Aged, 80 and over KW - Animal Husbandry KW - Animals KW - Bangladesh KW - Brucella abortus KW - Brucellosis KW - Cattle KW - Child KW - Child, Preschool KW - DNA, Bacterial KW - Female KW - Fever KW - Humans KW - Immunoassay KW - Male KW - middle aged KW - Real-Time Polymerase Chain Reaction KW - Risk Factors KW - Seroepidemiologic Studies KW - Young adult AB -

INTRODUCTION: This study describes the seroprevalence of human brucellosis among pyretic patients and detection of Brucella abortus DNA from seropositive pyretic patients using real-time polymerase chain reaction (rtPCR) for the first time in Bangladesh.

METHODOLOGY: Blood samples were collected from 300 pyretic patients from October 2007 to May 2008 and subjected to three serological tests: Rose-Bengal plate test (RBT), standard tube agglutination test (STAT), and indirect enzyme-linked immunosorbent assay (iELISA). Risk factors were identified by multivariate Firth's logistic regression analysis. Brucella genus (BCSP31) and species-specific (IS711) rtPCR were applied to six human sera samples.

RESULTS: The seroprevalence of brucellosis among pyretic patients was estimated to be 2.0% (95% confidence interval [CI]: 0.74-4.30). The odds of brucellosis seropositivity were 8.9 (95% CI: 1.26-63.0) times higher in pyretic patients who handled goats than those who handled only cattle, whereas the odds of brucellosis seropositivity were 9.7 (95% CI: 1.28-73.68) times higher in pyretic patients who had backache compared to those without backache. B. abortus DNA was amplified from all six human sera that tested positive by RBT, STAT, and iELISA. As the agreement between the tests was very strong, RBT is recommended as a screening test for the diagnosis of human brucellosis in Bangladesh because it is easier to use, cheaper, and faster.

CONCLUSIONS: Brucellosis among pyretic patients is common, and B. abortus is responsible for brucellosis in such patients. Pyretic patients who handle goats and those with backaches should be screened for brucellosis.

VL - 10 CP - 9 U1 - http://www.ncbi.nlm.nih.gov/pubmed/27694726?dopt=Abstract M3 - 10.3855/jidc.6844 ER - TY - RPRT T1 - Zoönosen en vectoroverdraagbare ziekten. Samenvattend jaaroverzicht 2015 Y1 - 2016 A1 - Javiera Rebolledo A1 - Tinne Lernout A1 - Amber Litzroth A1 - Dominique Van Beckhoven A1 - Bernard Brochier A1 - Delaere, B A1 - David Fretin A1 - Hing,M. A1 - Jacobs,J.A. A1 - B Kabamba Mukadi A1 - Marcella Mori A1 - Patteet,S. A1 - Saegeman,V. A1 - Vanessa Suin A1 - Truyens,C. A1 - Vanrompay, D A1 - Van Esbroeck, Marjan A1 - Steven Van Gucht A1 - P Wattiau KW - 2015 KW - Surveillance KW - vectoroverdraagbare ziekten KW - zoonosen PB - WIV-ISP CY - Brussel, België ER - TY - RPRT T1 - Zoonoses et maladies à transmission vectorielle. Surveillance épidémiologique en Belgique - Synthèse annuelle 2015 Y1 - 2016 A1 - Javiera Rebolledo A1 - Tinne Lernout A1 - Amber Litzroth A1 - Dominique Van Beckhoven A1 - Bernard Brochier A1 - Delaere,B. A1 - David Fretin A1 - Hing,M. A1 - B Kabamba Mukadi A1 - Marcella Mori A1 - Patteet,S. A1 - Saegeman,V. A1 - Vanessa Suin A1 - Truyens,C. A1 - Vanrompay, D A1 - Van Esbroeck, Marjan A1 - Steven Van Gucht A1 - P Wattiau KW - 2015 KW - Belgique KW - maladies à transmission vectorielle KW - Surveillance KW - Zoonoses PB - WIV-ISP CY - Bruxelles, Belgique ER - TY - JOUR T1 - About three cases of ulceroglandular tularemia, is this the re-emergence of Francisella tularensis in Belgium? JF - Acta Clin Belg Y1 - 2015 A1 - Dupont, E A1 - Van Eeckhoudt, S A1 - Thissen, X A1 - Ausselet, N A1 - David Fretin A1 - Stefanescu, I A1 - Glupczynski, Y A1 - Delaere, B KW - Adult KW - Animals KW - Belgium KW - Francisella tularensis KW - Humans KW - Male KW - Skin Ulcer KW - Tularemia KW - Zoonoses AB -

Tularemia is a zoonosis caused by Francisella tularensis that can be transmitted by several ways to human being and cause different clinical manifestations. We report three clinical cases of tularemia with ulceroglandular presentation in young males acquired during outdoor activities in Southern Belgium. Confirmation of the diagnosis was established by serology. Only three cases of tularemia have been reported in Belgium between 1950 and 2012 by the National Reference Laboratory CODA-CERVA (Ref Lab CODA-CERVA) but re-emergence of tularemia is established in several European countries and F. tularensis is also well known to be present in animal reservoirs and vectors in Belgium. The diagnosis of tularemia has to be considered in case of suggestive clinical presentation associated with epidemiological risk factors.

VL - 70 CP - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25847026?dopt=Abstract M3 - 10.1179/2295333715Y.0000000022 ER - TY - Generic T1 - Analysis of the virulence and immunogenicity of porcine and human M. avium ssp. hominissuis isolates in an experimental mouse model. Y1 - 2015 A1 - Nicolas Bruffaerts A1 - Christelle Vluggen A1 - L Duytschaever A1 - J. Denoel A1 - Wattiez,R. A1 - J.J. Letesson A1 - Vanessa Mathys A1 - Virginie Roupie A1 - David Fretin A1 - Saegerman,C. A1 - K Huygen ED - Research Center Borstel KW - an KW - analysi KW - analysis KW - European KW - Human KW - M KW - MODEL KW - ON KW - symposium KW - virulence JF - European Symposium on Non-tuberculous Mycobacteria CP - Research Center Borstel U1 - 37032 U2 - 24/06/2015;27/06/2015 ER - TY - JOUR T1 - First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania. JF - BMC Vet Res Y1 - 2015 A1 - Mathew, C A1 - Stokstad, M A1 - Johansen, T B A1 - Klevar, S A1 - Mdegela, R H A1 - Mwamengele, G A1 - Michel, P A1 - Escobar, L A1 - David Fretin A1 - Godfroid, J KW - Animals KW - Brucella abortus KW - Brucellosis, Bovine KW - Cattle KW - Genotype KW - Seroepidemiologic Studies KW - Tanzania AB -

BACKGROUND: Brucellosis is a disease of worldwide public health and economic importance. Successful control is based on knowledge of epidemiology and strains present in an area. In developing countries, most investigations are based on serological assays. This study aimed at investigating a dairy herd experiencing abortions in order to establish within-herd seroprevalence to Brucella spp., identify, characterize Brucella strains by Multiple Loci Variable Number of Tandem Repeats Analysis (MLVA-VNTR) and investigate possible spillover to other species.

RESULTS: The within-herd seroprevalence in cattle (n = 200) was 48 % (95 % CI 41-55), using an indirect ELISA, while the Rose Bengal Test (RBT) yielded lower prevalence (21.5 %; 95 % CI 16-27). Two sheep (n = 35) and one goat (n = 50) were seropositive using ELISA while none of the dogs (n = 6) was positive with the RBT. Three Brucella were isolated from an aborted fetus and associated membranes. Real time PCR (IS711), Bruce-ladder and classical biotyping classified the isolates as B. abortus biovar 3. MLVA-VNTR revealed two different but closely related genotypes. The isolates showed unique profiles, providing the first genotypic data from Tanzania. These genotypes were not related to B. abortus biovar 3 reference strain Tulya originally isolated from a human patient in Uganda in 1958, unlike the genotypes isolated and characterized recently in Kenya. High within-herd prevalence, isolation of the pathogen and abortion confirm that B. abortus is circulating in this herd with cattle as reservoir hosts. A low seroprevalence in sheep and goats suggests a spillover of B. abortus from cattle to small ruminants in the herd.

CONCLUSIONS: This is the first isolation and characterization of B. abortus biovar 3 from a dairy cow with abortion in Tanzania. The origin of the Tanzanian genotypes remain elusive, although they seem to be related to genotypes found in Europe, Turkey and China but not related to B. abortus biovar 3 reference strain or genotypes from Kenya. Importantly, replacement heifers are commonly sourced from large farms like this to smallholder farmers, which poses risk of spread of bacteria to other herds. B. abortus is a significant zoonotic risk and animal health problem in this production system, therefore further studies on humans is recommended.

VL - 11 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26195218?dopt=Abstract M3 - 10.1186/s12917-015-0476-8 ER - TY - JOUR T1 - Outbreak of leptospirosis during a scout camp in the Luxembourg Belgian province, Belgium, summer 2012. JF - Epidemiol Infect Y1 - 2015 A1 - Marcella Mori A1 - M Van Esbroeck A1 - Depoorter, S A1 - Decaluwe, W A1 - Vandecasteele, S J A1 - David Fretin A1 - Reynders, M KW - ADOLESCENT KW - Animals KW - Anorexia KW - Arvicolinae KW - Belgium KW - Child KW - Conjunctivitis KW - Cough KW - Creatinine KW - Disease Outbreaks KW - Disease Reservoirs KW - Headache KW - Humans KW - Leptospirosis KW - Male KW - Myalgia KW - Proteinuria KW - Recreation KW - rivers KW - Vomiting AB -

An outbreak of leptospirosis occurred in the South of Belgium, during August 2012, in teenagers who participated in two consecutive adventure scout camps near the Semois river. Among the symptomatic patient population (ten scouts), clinical manifestations included headache (70%), myalgia (50%), fever (50%), bilateral conjunctival injection (50%), general malaise (30%), vomiting (20%), anorexia (20%) and cough (20%). Some of the cases presented elevated blood creatinine (40%), or proteinuria (30%). Three patients were confirmed by serology and one by polymerase chain reaction. Potential risk factors included direct contact with a muskrat and indirect contact with potentially contaminated environments including the river water. Prospective environmental investigation carried out near the river banks 2 weeks after the outbreak identified Ondatra zibethicus (muskrat) as one Leptospira sp. reservoir.

VL - 143 CP - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25311398?dopt=Abstract M3 - 10.1017/S0950268814002763 ER - TY - JOUR T1 - Serologic screening for 13 infectious agents in roe deer (Capreolus capreolus) in Flanders. JF - Infect Ecol Epidemiol Y1 - 2015 A1 - Tavernier, Paul A1 - Sys, Stanislas U A1 - Kris De Clercq A1 - Ilse De Leeuw A1 - Ann Brigitte Cay A1 - Miet De Baere A1 - Nick De Regge A1 - David Fretin A1 - Virginie Roupie A1 - Govaerts, Marc A1 - Heyman, Paul A1 - Vanrompay, Daisy A1 - Yin, Lizi A1 - Kalmar, Isabelle A1 - Vanessa Suin A1 - Bernard Brochier A1 - Alexandre Dobly A1 - Stéphane De Craeye A1 - Sophie Roelandt A1 - Goossens, Els A1 - S. Roels AB -

INTRODUCTION: In order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in Flanders, we conducted a serologic screening in 12 hunting areas.

MATERIALS AND METHODS: Roe deer sera collected between 2008 and 2013 (n=190) were examined for antibodies against 13 infectious agents, using indirect enzyme-linked immunosorbent assay, virus neutralisation, immunofluorescence, or microagglutination test, depending on the agent.

RESULTS AND DISCUSSION: High numbers of seropositives were found for Anaplasma phagocytophilum (45.8%), Toxoplasma gondii (43.2%) and Schmallenberg virus (27.9%), the latter with a distinct temporal distribution pattern following the outbreak in domestic ruminants. Lower antibody prevalence was found for Chlamydia abortus (6.7%), tick-borne encephalitis virus (5.1%), Neospora caninum (4.8%), and Mycobacterium avium subsp paratuberculosis (4.1%). The lowest prevalences were found for Leptospira (1.7%), bovine viral diarrhoea virus 1 (1.3%), and Coxiella burnetii (1.2%). No antibodies were found against Brucella sp., bovine herpesvirus 1, and bluetongue virus. A significant difference in seroprevalence between ages (higher in adults >1 year) was found for N. caninum. Four doubtful reacting sera accounted for a significant difference in seroprevalence between sexes for C. abortus (higher in females).

CONCLUSIONS: Despite the more intensive landscape use in Flanders, the results are consistent with other European studies. Apart from maintaining C. abortus and MAP, roe deer do not seem to play an important role in the epidemiology of the examined zoonotic and domestic animal pathogens. Nevertheless, their meaning as sentinels should not be neglected in the absence of other wild cervid species.

VL - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26609692?dopt=Abstract M3 - 10.3402/iee.v5.29862 ER - TY - RPRT T1 - Zoönosen en Vector-Overdraagbare Ziekten. Epidemiologische surveillance in België, 2013 en 2014 Y1 - 2015 A1 - Javiera Rebolledo A1 - Tinne Lernout A1 - Amber Litzroth A1 - Dominique Van Beckhoven A1 - Bernard Brochier A1 - Delaere,B. A1 - David Fretin A1 - Heuninckx,W. A1 - Hing,M. A1 - Jacobs, J. A1 - B Kabamba Mukadi A1 - Maes,P. A1 - Marcella Mori A1 - Patteet,S. A1 - Sophie Quoilin A1 - Saegeman,V. A1 - Vanessa Suin A1 - Truyens,C. A1 - Vanrompay, D A1 - M. Van Esbroeck A1 - Steven Van Gucht A1 - P Wattiau KW - 2013 KW - 2014 KW - jaarrapport KW - overdraagbare ziekten KW - Vector KW - zoonosen PB - WIV-ISP CY - Brussel, België ER - TY - RPRT T1 - Zoonoses et maladies à transmission vectorielle. Surveillance épidémiologique en Belgique, 2013 et 2014 Y1 - 2015 A1 - Javiera Rebolledo A1 - Tinne Lernout A1 - Amber Litzroth A1 - Dominique Van Beckhoven A1 - Bernard Brochier A1 - Delaere,B. A1 - David Fretin A1 - Heuninckx,W. A1 - Hing,M. A1 - Jacobs,J.A. A1 - B Kabamba Mukadi A1 - Maes,P. A1 - Marcella Mori A1 - Patteet,S. A1 - Sophie Quoilin A1 - Saegeman,V. A1 - Vanessa Suin A1 - Truyens,C. A1 - Vanrompay, D A1 - Van Esbroeck, Marjan A1 - Steven Van Gucht A1 - P Wattiau KW - 2013 KW - 2014 KW - maladies à transmission vectorielle KW - Surveillance KW - Zoonoses PB - WIV-ISP CY - Bruxelles, Belgique ER - TY - JOUR T1 - Prevalence and molecular typing of Coxiella burnetii in bulk tank milk in Belgian dairy goats, 2009-2013. JF - Vet Microbiol Y1 - 2014 A1 - Samira Boarbi A1 - Marcella Mori A1 - Rousset, Elodie A1 - Sidi-Boumedine, Karim A1 - Van Esbroeck, Marjan A1 - David Fretin KW - Animals KW - Belgium KW - Coxiella burnetii KW - Genetic Variation KW - Genotype KW - Goat Diseases KW - Goats KW - Humans KW - milk KW - Molecular Typing KW - prevalence KW - Q Fever KW - Vaccination AB -

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.

VL - 170 CP - 1-2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/24598136?dopt=Abstract M3 - 10.1016/j.vetmic.2014.01.025 ER - TY - JOUR T1 - Bayesian estimation of the true prevalence, sensitivity and specificity of the Rose Bengal and indirect ELISA tests in the diagnosis of bovine brucellosis. JF - Vet J Y1 - 2013 A1 - Sanogo, Moussa A1 - Thys, Eric A1 - Achi, Yaba L A1 - David Fretin A1 - Michel, Patrick A1 - Abatih, Emmanuel A1 - Berkvens, Dirk A1 - Saegerman, Claude KW - Animals KW - Bayes Theorem KW - Brucella abortus KW - Brucellosis, Bovine KW - Cattle KW - Cote d'Ivoire KW - Enzyme-Linked Immunosorbent Assay KW - Rose Bengal KW - Sensitivity and Specificity KW - Serologic Tests KW - Serotyping AB -

Serology is the most convenient method for detecting brucellosis but the efficient use of such tests in disease control requires evaluation of diagnostic performance and discriminative ability. The objective of this study was to assess the performance of the Rose Bengal test (RBT) and an indirect ELISA (iELISA) in diagnosing brucellosis in 995 serum samples collected from cattle in the Ivory Coast between 2005 and 2009. A Bayesian approach was used to evaluate the two tests by estimating their sensitivities and specificities. The correlation-adjusted sensitivity of the iELISA was estimated to be 96.1% (credibility interval [CrI], 92.7-99.8), whereas that of the RBT was 54.9% (CrI, 23.5-95.1). High correlation-adjusted specificities were found for both tests (95.0%; [CrI, 91.1-99.6] for the iELISA and 97.7%; [CrI, 95.3-99.4] for the RBT, respectively). The true prevalence of brucellosis was estimated from the serum samples to be 4.6% (95%; [CrI, 0.6-9.5]). The level of agreement between the two tests was evaluated using indices of agreement (n=995). Good agreement was found for negative results (96.6%; confidence interval [CI], 95.7-97.4), a finding supported by an estimated significant correlation of 0.37 (95%; CI, 0.01-0.73) within the sera testing negative. Agreement was lower for sera testing positive (52.2% CI: 41.9-62.5). The findings highlight the importance of using these two tests in combination as part of any brucellosis control programme.

VL - 195 CP - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22831991?dopt=Abstract M3 - 10.1016/j.tvjl.2012.06.007 ER - TY - JOUR T1 - Bayesian estimation of true prevalence, sensitivity and specificity of indirect ELISA, Rose Bengal Test and Slow Agglutination Test for the diagnosis of brucellosis in sheep and goats in Bangladesh. JF - Prev Vet Med Y1 - 2013 A1 - Rahman, A K M Anisur A1 - Saegerman, Claude A1 - Berkvens, Dirk A1 - David Fretin A1 - Gani, Md Osman A1 - Ershaduzzaman, Md A1 - Ahmed Muzahed Uddin A1 - Emmanuel Abatih KW - Agglutination Tests KW - Animals KW - Antibodies, Bacterial KW - Bangladesh KW - Bayes Theorem KW - Brucella abortus KW - Brucella melitensis KW - Brucellosis KW - Enzyme-Linked Immunosorbent Assay KW - Goat Diseases KW - Goats KW - Immunoglobulins KW - prevalence KW - Rose Bengal KW - Sensitivity and Specificity KW - Serologic Tests KW - Sheep KW - Sheep Diseases AB -

The true prevalence of brucellosis and diagnostic test characteristics of three conditionally dependent serological tests were estimated using the Bayesian approach in goats and sheep populations of Bangladesh. Serum samples from a random selection of 636 goats and 1044 sheep were tested in parallel by indirect ELISA (iELISA), Rose Bengal Test (RBT) and Slow Agglutination Test (SAT). The true prevalence of brucellosis in goats and sheep were estimated as 1% (95% credibility interval (CrI): 0.7-1.8) and 1.2% (95% CrI: 0.6-2.2) respectively. The sensitivity of iELISA was 92.9% in goats and 92.0% in sheep with corresponding specificities of 96.5% and 99.5% respectively. The sensitivity and specificity estimates of RBT were 80.2% and 99.6% in goats and 82.8% and 98.3% in sheep. The sensitivity and specificity of SAT were 57.1% and 99.3% in goats and 72.0% and 98.6% in sheep. In this study, three conditionally dependent serological tests for the diagnosis of small ruminant brucellosis in Bangladesh were validated. Considerable conditional dependence between IELISA and RBT and between RBT and SAT was observed among sheep. The influence of the priors on the model fit and estimated parameter values was checked using sensitivity analysis. In multiple test validation, conditional dependence should not be ignored when the tests are in fact conditionally dependent.

VL - 110 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23276401?dopt=Abstract M3 - 10.1016/j.prevetmed.2012.11.029 ER - TY - JOUR T1 - Importance of identification and typing of Brucellae from West African cattle: a review. JF - Vet Microbiol Y1 - 2013 A1 - Sanogo, Moussa A1 - Abatih, Emmanuel A1 - Thys, Eric A1 - David Fretin A1 - Berkvens, Dirk A1 - Saegerman, Claude KW - Africa, Western KW - Animals KW - Bacterial Typing Techniques KW - Brucella KW - Brucellosis KW - Cattle KW - Cattle Diseases KW - Host-Parasite Interactions KW - prevalence AB -

Bovine brucellosis is an endemic infectious disease which can impact cattle productivity and welfare negatively, as well as human health. Sufficient knowledge on its epidemiology, particularly on species and biotypes of Brucella at national and/or regional scale are important to set up and implement efficient control measures against brucellosis in a "One health" perspective. The main objective of this review was to investigate available literature on strains of Brucella in order to provide a state of art-knowledge on species and biovars reported in cattle from West Africa. A literature search was conducted to identify relevant data on species and biovars of Brucella in cattle from Western African countries. This search included studies presenting bacteriological and/or molecular results of identification and typing, relied on international classification methods with no time limit and no language restrictions. Studies reporting results of identification at genus level only were not considered for this review. This review revealed that Brucella abortus was the most prevalent species in cattle from West Africa, in line with host preference for Brucellae. So far, biovars 1, 2, 3, 4, 6 and intermediate biovar 3/6 of B. abortus were reported in cattle in the region. Among these strains, biovars 3, recently identified in The Gambia and Ivory Coast, was the most commonly isolated. Brucella melitensis and/or B. suis have not been mentioned yet in cattle in this part of Africa. The public health significance of prevailing strains is discussed and a regional collaborative control program of brucellosis is suggested.

VL - 164 CP - 3-4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23499188?dopt=Abstract M3 - 10.1016/j.vetmic.2013.02.009 ER - TY - JOUR T1 - In vitro and in vivo infectious potential of coxiella burnetii: a study on Belgian livestock isolates. JF - PLoS One Y1 - 2013 A1 - Marcella Mori A1 - Samira Boarbi A1 - Michel, Patrick A1 - Bakinahe, Raïssa A1 - Rits, Katleen A1 - P Wattiau A1 - David Fretin KW - Animals KW - Cattle KW - Cell Line KW - Cell Line, Tumor KW - Coxiella burnetii KW - Female KW - Genotype KW - Goats KW - HeLa Cells KW - Humans KW - Livestock KW - mice KW - Mice, Inbred BALB C KW - Netherlands KW - Q Fever AB -

Q-fever is a zoonosis caused by the gram-negative obligate intracellular pathogen Coxiella burnetii. Since its discovery, and particularly following the recent outbreaks in the Netherlands, C. burnetii appeared as a clear public health concern. In the present study, the infectious potential displayed by goat and cattle isolates of C. burnetii was compared to a reference strain (Nine Mile) using both in vitro (human HeLa and bovine macrophage cells) and in vivo (BALB/c mice) models. The isolates had distant genomic profiles with one--the goat isolate--being identical to the predominant strain circulating in the Netherlands during the 2007-2010 outbreaks. Infective doses were established with ethidium monoazide-PCR for the first time here applied to C. burnetii. This method allowed for the preparation of reproducible and characterized inocula thanks to its capacity to discriminate between live and dead cells. Globally, the proliferative capacity of the Nine Mile strain in cell lines and mice was higher compared to the newly isolated field strains. In vitro, the bovine C. burnetii isolate multiplied faster in a bovine macrophage cell line, an observation tentatively explained by the preferential specificity of this strain for allogeneic host cells. In the BALB/c mouse model, however, the goat and bovine isolates multiplied at about the same rate indicating no peculiar hypervirulent behavior in this animal model.

VL - 8 CP - 6 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23840751?dopt=Abstract M3 - 10.1371/journal.pone.0067622 ER - TY - JOUR T1 - Seroprevalence and potential risk factors for Brucella spp. infection in traditional cattle, sheep and goats reared in urban, periurban and rural areas of Niger. JF - PLoS One Y1 - 2013 A1 - Boukary, Abdou Razac A1 - Saegerman, Claude A1 - Abatih, Emmanuel A1 - David Fretin A1 - Rianatou Bada Alambédji A1 - De Deken, Reginald A1 - Halimatou Adamou Harouna A1 - Yenikoye, Alhassane A1 - Thys, Eric KW - Animal Husbandry KW - Animals KW - Antibodies, Bacterial KW - Brucella KW - Brucellosis KW - Cattle KW - Cattle Diseases KW - cross-sectional studies KW - Goat Diseases KW - Goats KW - Humans KW - Niger KW - Risk Factors KW - Rural Population KW - Seroepidemiologic Studies KW - Sheep KW - Sheep Diseases KW - Urban Population AB -

INTRODUCTION: In Niamey, Niger, interactions within the interface between animals, humans and the environment induce a potential risk of brucellosis transmission between animals and from animals to humans. Currently, little is known about the transmission of Brucella in this context.

RESULTS: 5,192 animals from 681 herds were included in the study. Serum samples and hygroma fluids were collected. A household survey enabled to identify the risk factors for transmission of brucellosis. The true adjusted herd-level prevalence of brucellosis ranged between 11.2% and 17.2% and the true adjusted animal-population level prevalence was 1.3% (95% CI: 0.9-1.8%) based on indirect ELISA test for Brucella antibodies. Animals aged of 1-4 years were found to be more susceptible than animals less than 1 year old (Odds ratio [OR] of 2.7; 95% CI: 1.43-5.28). For cattle, the odds of brucellosis seropositivity were higher in rural compared to the periurban areas (OR of 2.8; 95% CI: 1.48-5.17) whereas for small ruminants the risk of seropositivity appeared to be higher in urban compared to periurban areas (OR of 5.5; 95% CI: 1.48-20.38). At herd level, the risk of transmission was increased by transhumance (OR of 5.4; 95% CI: 2.84-10.41), the occurrence of abortions (OR of 3.0; 95% CI: 1.40-6.41), and for herds having more than 50 animals (OR of 11.0; 95% CI: 3.75-32.46). Brucella abortus biovar 3 was isolated from the hygromas.

CONCLUSION: brucellosis in Niger is a serious problem among cattle especially in the rural areas around Niamey and among sheep in the urban areas of Niamey. The seroprevalence varies across strata and animal species with important risk factors including herd size, abortion and transhumance at herd level and age at animal population level. For effective control of brucellosis, an integrated approach seems appropriate involving all stakeholders working in public and animal health.

VL - 8 CP - 12 U1 - http://www.ncbi.nlm.nih.gov/pubmed/24358261?dopt=Abstract M3 - 10.1371/journal.pone.0083175 ER - TY - JOUR T1 - Unexpected Brucella suis biovar 2 Infection in a dairy cow, Belgium. JF - Emerg Infect Dis Y1 - 2013 A1 - David Fretin A1 - Marcella Mori A1 - Czaplicki, Guy A1 - Quinet, Christian A1 - Maquet, Benoît A1 - Godfroid, Jacques A1 - Saegerman, Claude KW - Animals KW - Belgium KW - Brucella suis KW - Brucellosis KW - Cattle KW - Cattle Diseases KW - Disease Outbreaks KW - Serotyping VL - 19 CP - 12 U1 - http://www.ncbi.nlm.nih.gov/pubmed/24274041?dopt=Abstract M3 - 10.3201/eid1912.130506 ER - TY - JOUR T1 - B. anthracis in a wool-processing factory: seroprevalence and occupational risk. JF - Epidemiol Infect Y1 - 2012 A1 - Kissling, E A1 - P Wattiau A1 - Bernard China A1 - Poncin, M A1 - David Fretin A1 - Pirenne, Y A1 - Hanquet, G KW - Adult KW - Animals KW - Anthrax KW - Antibodies, Bacterial KW - Bacillus anthracis KW - Belgium KW - Female KW - Goats KW - Humans KW - Male KW - middle aged KW - Occupational Exposure KW - Risk Assessment KW - Seroepidemiologic Studies KW - wool AB -

In a Belgian wool-processing factory, living anthrax spores were found in raw goat hair and air dust, but confirmed anthrax cases had never been reported. Anthrax vaccines are not licensed nor recommended in Belgium. We conducted a B. anthracis seroprevalence study to investigate risk factors associated with positive serology and advise on protective measures. Overall 12·1% (8/66) employees were seropositive; 30% of persons processing raw goat hair and 20% of persons sorting raw goat hair were seropositive compared to 3% in less exposed jobs [adjusted prevalence ratio (aPR) 44·4, P=0·001; aPR 14·5, P=0·016, respectively). The number of masks used per day was protective (aPR 0·3, P=0·015). Results suggest a dose-response association for those processing raw goat hair. Host-related factors probably played a role as antibody response varied from person to person within an exposure group. Workers exposed to raw goat hair should be offered higher protection against anthrax and have access to anthrax vaccines.

VL - 140 CP - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21835070?dopt=Abstract M3 - 10.1017/S0950268811001488 ER - TY - JOUR T1 - Risk factors associated with brucellosis seropositivity among cattle in the central savannah-forest area of Ivory Coast. JF - Prev Vet Med Y1 - 2012 A1 - Sanogo, Moussa A1 - Abatih, Emmanuel A1 - Thys, Eric A1 - David Fretin A1 - Berkvens, Dirk A1 - Saegerman, Claude KW - Animals KW - Antibodies, Bacterial KW - Brucella KW - Brucellosis, Bovine KW - Cattle KW - Cote d'Ivoire KW - Enzyme-Linked Immunosorbent Assay KW - Female KW - Logistic Models KW - Male KW - Seroepidemiologic Studies AB -

Serological results obtained from 907 serum samples collected from unvaccinated cattle of at least 6 months of age in the savannah-forest region of Ivory Coast were used to investigate risk factors associated with bovine brucellosis seropositivity. Serum samples were tested using the Rose Bengal test (RBT) and indirect enzyme linked immunosorbent assay (iELISA). Using a parallel interpretation, RBT and iELISA results showed that 10.3% (95% confidence interval (CI): 8.4, 12.4) of the cattle had antibodies against Brucella in our study area. The logistic regression analysis indicated that brucellosis seropositivity was associated with age and herd size. Cattle above 5 years of age were found to have a higher chance of being seropositive (odd ratio (OR)=2.8; 95% CI: 1.3, 6.4) compared to cattle under 3 years of age. Similarly, the odd of brucellosis seropositivity for herds with more than 100 cattle was 3.3 (95% CI: 1.2, 8.9) times higher compared to those with less than 50 cattle.

VL - 107 CP - 1-2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22704904?dopt=Abstract M3 - 10.1016/j.prevetmed.2012.05.010 ER - TY - JOUR T1 - Brucellosis at the animal/ecosystem/human interface at the beginning of the 21st century. JF - Prev Vet Med Y1 - 2011 A1 - Godfroid, J A1 - Scholz, H C A1 - Barbier, T A1 - Nicolas, C A1 - P Wattiau A1 - David Fretin A1 - Whatmore, A M A1 - Cloeckaert, A A1 - Blasco, J M A1 - Moriyon, I A1 - Saegerman, C A1 - Muma, J B A1 - Al Dahouk, S A1 - Neubauer, H A1 - Letesson, J-J KW - Animals KW - Brucella KW - Brucellosis KW - Female KW - Humans KW - Pregnancy KW - Zoonoses AB -

Following the recent discovery of new Brucella strains from different animal species and from the environment, ten Brucella species are nowadays included in the genus Brucella. Although the intracellular trafficking of Brucella is well described, the strategies developed by Brucella to survive and multiply in phagocytic and non-phagocytic cells, particularly to access nutriments during its intracellular journey, are still largely unknown. Metabolism and virulence of Brucella are now considered to be two sides of the same coin. Mechanisms presiding to the colonization of the pregnant uterus in different animal species are not known. Vaccination is the cornerstone of control programs in livestock and although the S19, RB51 (both in cattle) and Rev 1 (in sheep and goats) vaccines have been successfully used worldwide, they have drawbacks and thus the ideal brucellosis vaccine is still very much awaited. There is no vaccine available for pigs and wildlife. Animal brucellosis control strategies differ in the developed and the developing world. Most emphasis is put on eradication and on risk analysis to avoid the re-introduction of Brucella in the developed world. Information related to the prevalence of brucellosis is still scarce in the developing world and control programs are rarely implemented. Since there is no vaccine available for humans, prevention of human brucellosis relies on its control in the animal reservoir. Brucella is also considered to be an agent to be used in bio- and agroterrorism attacks. At the animal/ecosystem/human interface it is critical to reduce opportunities for Brucella to jump host species as already seen in livestock, wildlife and humans. This task is a challenge for the future in terms of veterinary public health, as for wildlife and ecosystem managers and will need a "One Health" approach to be successful.

VL - 102 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21571380?dopt=Abstract M3 - 10.1016/j.prevetmed.2011.04.007 ER - TY - JOUR T1 - The first International Standard anti-Brucella melitensis Serum. JF - Rev Sci Tech Y1 - 2011 A1 - McGiven, J A1 - Taylor, A A1 - Duncombe, L A1 - Sayers, R A1 - Albert, D A1 - Banai, M A1 - Blasco, J M A1 - Elena, S A1 - David Fretin A1 - Garin-Bastuji, B A1 - Melzer, F A1 - Muñoz, P M A1 - Nielsen, K A1 - Nicola, A A1 - Scacchia, M A1 - Tittarelli, M A1 - I Travassos Dias A1 - Walravens, K A1 - Stack, J KW - Analysis of Variance KW - Animals KW - Antibodies, Bacterial KW - Brucella melitensis KW - Brucellosis KW - Complement Fixation Tests KW - Dose-Response Relationship, Drug KW - Enzyme-Linked Immunosorbent Assay KW - Female KW - Fluorescence Polarization Immunoassay KW - Goat Diseases KW - Goats KW - Immune Sera KW - Pregnancy KW - Reference Standards KW - Sheep KW - Sheep Diseases AB -

The World Organisation for Animal Health (OIE) requested an International Standard anti-Brucella melitensis Serum (ISaBmS) to standardise diagnostic tests and reagents for sheep and goats. The agreed criteria were the highest dilution (in negative serum) of the standard which must give a positive result and the lowest dilution (in negative serum) which must simultaneously give a negative result. The two dilutions for each assay were, respectively: indirect enzyme-linked immunosorbent assay (iELISA) 1/64 and 1/750, competitive ELISA (cELISA) 1/8 and 1/300, fluorescent polarisation assay (FPA) 1/16 and 1/200, Rose Bengal test (RBT) 1/16 and 1/200. The OIE International Standard Serum (OIEISS) will remain the primary standard for the RBT; the ISaBmS is an additional standard. It was impossible to set criteria for the complement fixation test, therefore the OIEISS will remain the primary standard. The ISaBmS can be used to standardise iELISA, cELISA and FPA to diagnose sheep and goat brucellosis. This standard should facilitate harmonisation of tests used for brucellosis surveillance and international trade in these species.

VL - 30 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22435193?dopt=Abstract ER - TY - JOUR T1 - Nucleotide polymorphism-based single-tube test for robust molecular identification of all currently described Brucella species. JF - Appl Environ Microbiol Y1 - 2011 A1 - P Wattiau A1 - Whatmore, Adrian M A1 - Van Hessche, Mieke A1 - Godfroid, Jacques A1 - David Fretin KW - Bacteriological Techniques KW - Brucella KW - DNA, Bacterial KW - Electrophoresis, Capillary KW - Genetic Markers KW - Ligase Chain Reaction KW - Polymorphism, Genetic AB -

Among the numerous molecular methods described during the last 20 years to identify Brucella, multiplexed amplification methods offer the cheapest and simplest technical solution for molecular identification. However, one disadvantage of such methods is their need to undergo technical revalidation each time a new marker is added to the system. Moreover, polymorphic markers cannot be assessed at the single-nucleotide level in these assays. Since new Brucella species are continuously being described, open methodologies able to accommodate new markers while preserving all other system parameters have an obvious advantage. We present a ligase chain reaction (LCR)-based method that simultaneously assesses multiple genetic markers at the single-nucleotide level. Most of the selected markers originate from a multilocus sequence typing (MLST) database that has been extensively validated on hundreds of different Brucella strains. When assayed on both reference and field strains, the method yields characteristic capillary electrophoresis profiles for each of the 10 Brucella species described to date and displays discriminatory potential below the species level for some. Since the LCR methodology is insensitive to interference resulting from the use of multiple oligonucleotides in a single mixture, the way is open for smooth future updates of the proposed system. Such updates are inevitable, given the pending description of new Brucella species.

VL - 77 CP - 18 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21803907?dopt=Abstract M3 - 10.1128/AEM.00767-11 ER - TY - JOUR T1 - Q fever in Woolsorters, Belgium. JF - Emerg Infect Dis Y1 - 2011 A1 - P Wattiau A1 - Boldisova, Eva A1 - Toman, Rudolf A1 - Van Esbroeck, Marjan A1 - Sophie Quoilin A1 - Hammadi, Samia A1 - Tissot-Dupont, Hervé A1 - Raoult, Didier A1 - Henkinbrant, Jean-Marie A1 - Van Hessche, Mieke A1 - David Fretin KW - Animals KW - Antibodies, Bacterial KW - Belgium KW - Cohort Studies KW - Communicable Diseases, Emerging KW - Coxiella burnetii KW - Goats KW - Humans KW - Occupational Diseases KW - Occupational Exposure KW - Q Fever KW - Seroepidemiologic Studies KW - Sheep, Domestic KW - wool VL - 17 CP - 12 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22172399?dopt=Abstract M3 - 10.3201/eid1712.101786 ER - TY - JOUR T1 - Brucella ceti infection in harbor porpoise (Phocoena phocoena). JF - Emerg Infect Dis Y1 - 2010 A1 - Jauniaux, Thierry P A1 - Brenez, Cecile A1 - David Fretin A1 - Godfroid, Jacques A1 - Haelters, Jan A1 - Jacques, Thierry A1 - Kerckhof, Francis A1 - Jan Mast A1 - Sarlet, Michael A1 - Coignoul, Freddy L KW - Animals KW - Belgium KW - Brucella KW - Brucellosis KW - Fatal Outcome KW - Female KW - Phocoena AB -

We describe Brucella sp. infection and associated lesions in a harbor porpoise (Phocoena phocoena) found on the coast of Belgium. The infection was diagnosed by immunohistochemistry, transmission electron microscopy, and bacteriology, and the organism was identified as B. ceti. The infection's location in the porpoise raises questions of abortion and zoonotic risks.

VL - 16 CP - 12 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21122233?dopt=Abstract M3 - 10.3201/eid1612.101008 ER - TY - JOUR T1 - Phenotypic and genotypic characterisation of Brucella strains isolated from cattle in the Gambia. JF - Vet Rec Y1 - 2010 A1 - Bankole, A A A1 - Saegerman, C A1 - Berkvens, D A1 - David Fretin A1 - Geerts, S A1 - Ieven, G A1 - Walravens, K KW - Animals KW - Bacterial Typing Techniques KW - Brucella KW - Brucella abortus KW - Brucellosis, Bovine KW - Cattle KW - Enzyme-Linked Immunosorbent Assay KW - Female KW - Gambia KW - Genotype KW - Phenotype KW - Rose Bengal AB -

Thirty-five serum samples and six hygroma fluid samples were collected from sexually mature cattle in one herd with clinical signs of brucellosis (abortion and hygromas) in the Western Region of the Gambia in order to isolate and characterise Brucella species. Information on the sex, age, number of calvings, number of abortions, presence of hygromas, and presence of orchitis was also collected for each animal sampled. Twenty-six (74 per cent) of the serum samples were positive in the rose bengal test and 29 (83 per cent) were positive by indirect ELISA. Three isolates of Brucella, biotyped as Brucella abortus biovar 3, were cultured from six hygroma fluid samples. The multiple locus variable number tandem repeat analysis assay clustered the isolates as B abortus with the same profile for the three isolates, suggesting a common origin of contamination.

VL - 166 CP - 24 U1 - http://www.ncbi.nlm.nih.gov/pubmed/20543166?dopt=Abstract M3 - 10.1136/vr.b4862 ER - TY - JOUR T1 - Immunologic response of unvaccinated workers exposed to anthrax, Belgium. JF - Emerg Infect Dis Y1 - 2009 A1 - P Wattiau A1 - Govaerts, Marc A1 - Frangoulidis, Dimitrios A1 - David Fretin A1 - Kissling, Esther A1 - Van Hessche, Mieke A1 - Bernard China A1 - Poncin, Martine A1 - Pirenne, Yvo A1 - Hanquet, Germaine KW - Anthrax KW - Anthrax Vaccines KW - Antibodies, Bacterial KW - Bacillus anthracis KW - Belgium KW - Blotting, Western KW - Cell Proliferation KW - Enzyme-Linked Immunosorbent Assay KW - Humans KW - Inhalation Exposure KW - Occupational Exposure KW - T-Lymphocytes AB -

To determine immunologic reactivity to Bacillus anthrax antigens, we conducted serologic testing of workers in a factory that performed scouring of wool and goat hair. Of 66 workers, approximately 10% had circulating antibodies or T lymphocytes that reacted with anthrax protective antigen. Individual immunity varied from undetectable to high.

VL - 15 CP - 10 U1 - http://www.ncbi.nlm.nih.gov/pubmed/19861061?dopt=Abstract M3 - 10.3201/eid1510.081717 ER - TY - JOUR T1 - Brucella suis identification and biovar typing by real-time PCR. JF - Vet Microbiol Y1 - 2008 A1 - David Fretin A1 - Whatmore, Adrian M A1 - Sascha Al Dahouk A1 - Neubauer, Heinrich A1 - Garin-Bastuji, Bruno A1 - Albert, David A1 - Van Hessche, Mieke A1 - Ménart, Marie A1 - Godfroid, Jacques A1 - Walravens, Karl A1 - P Wattiau KW - Brucella suis KW - Genetic Markers KW - polymerase chain reaction KW - Polymorphism, Genetic AB -

Fast and accurate identification of Brucella suis at the biovar level is an important issue for public health laboratories because some of the biovars that infect suidae (boars and pigs) are pathogenic for humans while others are not. Since classical biovar typing methods are often time-consuming, hard to standardize and require high-level biosafety containment, methodological improvements are desirable. This article describes new single nucleotide polymorphism (SNP) signatures for the rapid identification and biovar characterization of B. suis. These SNPs were included together with previously described ones in real-time PCR assays applicable to low-biosafety conditions. Allelic profiles unique for each B. suis biovar were defined and the most relevant signatures were determined on a collection of 137 field strains of worldwide origin characterized previously. Biovars assigned with both present and classical methods were globally consistent except for some biovar 3 field strains which matched the allelic profile of biovar 1.

VL - 131 CP - 3-4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/18499359?dopt=Abstract M3 - 10.1016/j.vetmic.2008.04.003 ER - TY - JOUR T1 - Occurrence and genetic diversity of Bacillus anthracis strains isolated in an active wool-cleaning factory. JF - Appl Environ Microbiol Y1 - 2008 A1 - P Wattiau A1 - Klee, Silke R A1 - David Fretin A1 - Van Hessche, Mieke A1 - Ménart, Marie A1 - Franz, Tatjana A1 - Camille Chasseur A1 - Butaye, Patrick A1 - Hein Imberechts KW - Animals KW - Bacillus anthracis KW - Bacterial Typing Techniques KW - Culture Media KW - Dust KW - Fresh Water KW - Genetic Variation KW - Goats KW - Hair KW - Industrial Waste KW - Industry KW - Minisatellite Repeats KW - Sheep KW - wool AB -

Culturable microorganisms from various samples taken at an active factory performing wool and goat hair cleaning were isolated and analyzed. Bacillus anthracis was found in air filter dust, wastewater, and goat hairs, where it accounted for approximately 1% of the total counts of viable bacteria. Consistent with the countries of origin of the processed material (South Caucasian and Middle Eastern), all B. anthracis isolates belonged to the same phylogenetic cluster, as determined by variable-number tandem repeat (VNTR) typing at eight loci. Within this cluster, five closely related VNTR subtypes could be identified, of which two were previously unreported. Additional diversity was observed when more sensitive genetic markers were assayed, demonstrating the multifocal nature of goat hair contamination. Goat hair originating from areas where anthrax is endemic remains a material with high biological risk for modern woolworkers.

VL - 74 CP - 13 U1 - http://www.ncbi.nlm.nih.gov/pubmed/18487406?dopt=Abstract M3 - 10.1128/AEM.00417-08 ER - TY - JOUR T1 - Isolation of Morganella morganii from a domestic rabbit with bronchopneumonia. JF - Vet Rec Y1 - 2007 A1 - S. Roels A1 - P Wattiau A1 - David Fretin A1 - Butaye, P A1 - Vanopdenbosch, E KW - Animals KW - Bronchopneumonia KW - Enterobacteriaceae Infections KW - Female KW - Lung KW - Male KW - Morganella morganii KW - Rabbits VL - 161 CP - 15 U1 - http://www.ncbi.nlm.nih.gov/pubmed/17938414?dopt=Abstract ER - TY - JOUR T1 - Real,time PCR typing of single nucleotide polymorphism in DNA containing inverted repeats. JF - Biotechniques Y1 - 2006 A1 - P Wattiau A1 - David Fretin KW - Dna KW - polymerase chain reaction KW - Polymorphism, Single Nucleotide KW - Repetitive Sequences, Nucleic Acid KW - Sequence Analysis, DNA VL - 41 CP - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/17140109?dopt=Abstract ER - TY - JOUR T1 - From the discovery of the Malta fever's agent to the discovery of a marine mammal reservoir, brucellosis has continuously been a re-emerging zoonosis. JF - Vet Res Y1 - 2005 A1 - Godfroid, Jacques A1 - Cloeckaert, Axel A1 - Liautard, Jean-Pierre A1 - Kohler, Stephan A1 - David Fretin A1 - Walravens, Karl A1 - Garin-Bastuji, Bruno A1 - Letesson, Jean-Jacques KW - Animals KW - Animals, Wild KW - Bioterrorism KW - Brucella KW - Brucellosis KW - Cetacea KW - Communicable Diseases, Emerging KW - Disease Reservoirs KW - Humans KW - Pinnipedia KW - prevalence KW - Zoonoses AB -

Brucellosis is not a sustainable disease in humans. The source of human infection always resides in domestic or wild animal reservoirs. The routes of infection are multiple: food-borne, occupational or recreational, linked to travel and even to bioterrorism. New Brucella strains or species may emerge and existing Brucella species adapt to changing social, cultural, travel and agricultural environment. Brucella melitensis is the most important zoonotic agent, followed by Brucella abortus and Brucella suis. This correlates with the fact that worldwide, the control of bovine brucellosis (due to B. abortus) has been achieved to a greater extent than the control of sheep and goat brucellosis (due to B. melitensis), these latter species being the most important domestic animals in many developing countries. The long duration and high cost of treatment of human brucellosis reduces the efficacy of the therapy. There is no human vaccine for brucellosis and the occurrence of brucellosis is directly linked to the status of animal brucellosis in a region. In this context, the Word Health Organization has defined the development of a human vaccine, besides the implementation of control and eradication programs in animals, as a high priority. The pathogenicity for humans of B. suis biovars 1, 3 and 4 is well established, whereas B. suis biovar 2 seems to be less pathogenic. Indeed, although hunters and pig farmers have repeatably experienced infectious contact with B. suis biovar 2 (found in wild boar and outdoor-rearing pigs in Europe), isolation of B. suis biovar 2 from human samples have only been seldom reported. Marine mammal brucellosis, due to two new proposed Brucella species i.e. B. cetaceae and B. pinnipediae, represents a new zoonotic threat but the pathogenicity for humans of the different Brucella species found in cetaceans and pinnipeds still has to be clearly established.

VL - 36 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/15845228?dopt=Abstract M3 - 10.1051/vetres:2005003 ER - TY - JOUR T1 - Fun stories about Brucella: the "furtive nasty bug". JF - Vet Microbiol Y1 - 2002 A1 - Letesson, J-J A1 - Lestrate, P A1 - Delrue, R-M A1 - Danese, I A1 - Bellefontaine, F A1 - David Fretin A1 - Taminiau, B A1 - Tibor, A A1 - Dricot, A A1 - Deschamps, C A1 - Haine, V A1 - Leonard, S A1 - Laurent, T A1 - Mertens, P A1 - Vandenhaute, J A1 - De Bolle, X KW - Animals KW - Brucella KW - Brucellosis KW - Cell Communication KW - Cell Cycle KW - Flagella KW - Humans KW - Luminescent Measurements KW - Vibrio KW - Zoonoses AB -

Although Brucella is responsible for one of the major worldwide zoonosis, our understanding of its pathogenesis remains in its infancy. In this paper, we summarize some of the research in progress in our laboratory that we think could contribute to a better understanding of the Brucella molecular virulence mechanisms and their regulation.

VL - 90 CP - 1-4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/12414152?dopt=Abstract ER - TY - Generic T1 - Belgische H3N1 Epizoötie Y1 - 0 A1 - Mieke Steensels A1 - Philippe Lagacé-Wiens A1 - Thierry van den Berg A1 - Steven Van Borm A1 - David Fretin A1 - Virginie Roupie A1 - Fabienne Rauw A1 - Bénédicte Lambrecht KW - 2019 KW - Avian Influenza KW - H3N1 AB -

Vanaf begin april (week 14) tot op 24.07.2019 (week 29), werden 82 bedrijven positief H3-AI gedetecteerd. Bij de getroffen bedrijven bevinden zich zowel verschillende types van bedrijven als verschillende species, verspreid in de tijd. De getroffen bedrijven bevinden zich voornamelijk in een beperkte geografische regio in het noordwesten van het land.

De karakterisatie van het virus wijst op een typisch laag pathogeen virus dat niet meldingsplichtig is volgens de EU-richtlijnen. De genetische informatie wijst op een oorsprong vanuit in wilde vogel circulerende AI virussen, met een aanpassing aan pluimvee ten gevolge van circulatie. In vivo experimenten in jonge SPF leggers bevestigden dit laag pathogeen pathotype. Bij leggers in productie zijn de klinische verschijnselen in het veld problematisch. Een infectie van AI-vrije conventionele leggers uit het veld toonden een eerder beperkte sterfte van 2/6 dieren één week na infectie, zonder sterfte bij de 6 contactdieren, en dit tijdens een 21 dagen studieduur.

ER - TY - Generic T1 - Epizootie H3N1 en Belgique Y1 - 0 A1 - Mieke Steensels A1 - Philippe Lagacé-Wiens A1 - Thierry van den Berg A1 - Steven Van Borm A1 - David Fretin A1 - Virginie Roupie A1 - Fabienne Rauw A1 - Bénédicte Lambrecht AB -

Depuis début avril (semaine 14) jusqu’au 4.07.2019 (semaine 29), 82 exploitations ont été détectées positive H3. Le secteur a été fortement impacté, tant les différents types d’élevages qu’une variété d’espèces ont été touchés dans cette période.  La majorité des élevages infectés étaient localisés dans une région limitée au nord-ouest du pays.

La caractérisation du virus a identificié un influenza aviaire faiblement pathogène, non notifiable conformémént aux directives de l’EU. Les informations génétiques indiquent une origine de virus de l'IA circulant chez les oiseaux sauvages, avec une adaptation à la volaille due à la circulation. Des expériences in vivo sur de jeunes poules pondeuses SPF ont confirmé ce pathotype faiblement pathogène. Par contre, les signes cliniques sur le terrain étaient surtout problématiques pour des pondeuses en production. Une infection des poules pondeuses conventionnelles terrain, confirmé négatif d'IA, a démontré une mortalité plutôt limitée de 2/6 animaux une semaine après l'infection, sans mortalité chez les 6 animaux de contact, et ceci pendant une période d'étude de 21 jours.

ER - TY - RPRT T1 - La résistance antimicrobienne chez les E. coli BLSE et Commensales, Campylobacter spp., Salmonella spp., Staphylococcus aureus résistants à la méthicilline (MRSA) et Enterococcus faecalis et faecium isolés des populations d’animaux producteurs d’aliments Y1 - 0 A1 - Cristina Garcia-Graells A1 - François Bricteux A1 - Inge Van Damme A1 - Cécile Boland A1 - Carole Kowalewicz A1 - Koenraad Van Hoorde A1 - David Fretin A1 - Katelijne Dierick ER -