Papers by Freddy Haesebrouck
BMC Veterinary Research, 2014
Background: Bovine enterotoxemia is a major cause of mortality in veal calves. Predominantly veal... more Background: Bovine enterotoxemia is a major cause of mortality in veal calves. Predominantly veal calves of beef cattle breeds are affected and losses due to enterotoxemia may account for up to 20% of total mortality. Clostridium perfringens type A is considered to be the causative agent. Recently, alpha toxin and perfringolysin O have been proposed to play an essential role in the development of disease. However, other potential virulence factors also may play a role in the pathogenesis of bovine enterotoxemia. The aim of this study was to evaluate whether strains originating from bovine enterotoxemia cases were superior in in vitro production of virulence factors (alpha toxin, perfringolysin O, mucinase, collagenase) that are potentially involved in enterotoxemia. To approach this, a collection of strains originating from enterotoxemia cases was compared to bovine strains isolated from healthy animals and to strains isolated from other animal species.
Clostridium perfringens type A has been shown to be the causative agent of a wide variety of ente... more Clostridium perfringens type A has been shown to be the causative agent of a wide variety of enteric diseases in humans and animals, including bovine enterotoxemia. Enterotoxemia is a sudden death syndrome with necro hemorrhagic lesions in the small intestine, which mainly affects suckling calves and veal calves [1]. Predominantly veal calves of beef cattle breeds are affected, and losses due to enterotoxemia may be responsible for up to 20% of total mortality [2, 3]. Recently, alpha toxin has been proposed as an essential factor for induction of enterotoxemia in veal calves, which introduces the possibility to use it as a vaccine [4]. The use of only the main toxin instead of the whole arsenal of extracellular toxins and enzymes eliminates irrelevant or even immunosuppressive components and therefore may induce a stronger, protective immune response. In this study, we have compared a commercial multivalent vaccine with native alpha toxin, formalin inactivated alpha toxin and recomb...
Journal of Applied Microbiology, 2004
Avian Pathology, 2004
The incidence of Clostridium perfringens-associated necrotic enteritis in poultry has increased i... more The incidence of Clostridium perfringens-associated necrotic enteritis in poultry has increased in countries that stopped using antibiotic growth promoters. Necrotic enteritis and the subclinical form of C. perfringens infection in poultry are caused by C. perfringens type A, producing the alpha toxin, and to a lesser extent type C, producing both alpha toxin and beta toxin. Some strains of C. perfringens type A produce an enterotoxin at the moment of sporulation and are responsible for foodborne disease in humans. The mechanisms of colonization of the avian small intestinal tract and the factors involved in toxin production are largely unknown. It is generally accepted, however, that predisposing factors are required for these bacteria to colonize and cause disease in poultry. The best known predisposing factor is mucosal damage, caused by coccidiosis. Diets with high levels of indigestible, water-soluble non-starch polysaccharides, known to increase the viscosity of the intestinal contents, also predispose to necrotic enteritis. Standardized models are being developed for the reproduction of colonization of poultry by C. perfringens and the C. perfringens-associated necrotic enteritis. One such model is a combined infection with Eimeria species and C. perfringens. Few tools and strategies are available for prevention and control of C. perfringens in poultry. Vaccination against the pathogen and the use of probiotic and prebiotic products has been suggested, but are not available for practical use in the field at the present time. The most cost-effective control will probably be achieved by balancing the composition of the feed.
Avian Pathology, 2004
The incidence of Clostridium perfringens-associated necrotic enteritis in poultry has increased i... more The incidence of Clostridium perfringens-associated necrotic enteritis in poultry has increased in countries that stopped using antibiotic growth promoters. Necrotic enteritis and the subclinical form of C. perfringens infection in poultry are caused by C. perfringens type A, producing the alpha toxin, and to a lesser extent type C, producing both alpha toxin and beta toxin. Some strains of C. perfringens type A produce an enterotoxin at the moment of sporulation and are responsible for foodborne disease in humans. The mechanisms of colonization of the avian small intestinal tract and the factors involved in toxin production are largely unknown. It is generally accepted, however, that predisposing factors are required for these bacteria to colonize and cause disease in poultry. The best known predisposing factor is mucosal damage, caused by coccidiosis. Diets with high levels of indigestible, water-soluble non-starch polysaccharides, known to increase the viscosity of the intestinal contents, also predispose to necrotic enteritis. Standardized models are being developed for the reproduction of colonization of poultry by C. perfringens and the C. perfringens-associated necrotic enteritis. One such model is a combined infection with Eimeria species and C. perfringens. Few tools and strategies are available for prevention and control of C. perfringens in poultry. Vaccination against the pathogen and the use of probiotic and prebiotic products has been suggested, but are not available for practical use in the field at the present time. The most cost-effective control will probably be achieved by balancing the composition of the feed.
Salmonella enterica serovar Enteritidis is the serovar most frequently isolated from chicken eggs... more Salmonella enterica serovar Enteritidis is the serovar most frequently isolated from chicken eggs. Colonization of the upper oviduct of hens is believed to play an important role in egg contamination. The interaction of S. enteritidis with gland epithelial cells of the isthmus and the magnum was, therefore, studied in vitro and in vivo. In the first experiment, S. enteritidis bacteria were added to confluent monolayers of primary cultures of chicken tubular epithelial cells of the isthmus (ICTEC) or magnum (MCTEC). Intracellular bacteria in ICTEC and MCTEC were confirmed by a gentamicin protection assay. Internalization in the glandular cells was corroborated by confocal scanning microscopy. Although S. enteritidis was able to invade and proliferate intracellularly during 24 h in the cell culture of both segments, this was significantly more so in the ICTEC. In a second experiment, an in vivo loop model was developed for investiga-(Abbreviation Key: BGA = brilliant green agar; HBSS = Hanks' buffered salt solution; ICTEC = chicken tubular epithelial cells of the isthmus; MCTEC = chicken tubular epithelial cells of the magnum; MEM = minimum essential medium; TBS = Tris-buffered saline. by guest on September 7, 2014
Short-chain fatty acids (SCFA) are widely used as feed additives in poultry for the control of pa... more Short-chain fatty acids (SCFA) are widely used as feed additives in poultry for the control of pathogenic bacteria, such as Salmonella enteritidis. Recently, a new range of products was developed in which SCFA are encapsulated in mineral carriers, resulting in a slow release during the transport of these carriers through the intestinal tract. To test the efficacy of this type of products against early colonization after Salmonella infection in poultry, a challenge experiment with S. enteritidis was performed. Five groups of 20 chickens were given feed with no supplement or feed supplemented with acetic acid (0.24%), formic acid (0.22%), or propionic acid (0.27%) as film-coated microbeads or butyric acid (0.15%) (
Veterinary microbiology, Jan 12, 2009
In this study, the accuracy of two phenotypic tests, API Staph ID 32 and Staph-Zym, was determine... more In this study, the accuracy of two phenotypic tests, API Staph ID 32 and Staph-Zym, was determined for identification of coagulase-negative staphylococci (CNS) from bovine milk samples in comparison with identification based on DNA-sequencing. A total of 172 CNS isolated from bovine milk were classified into 17 species. The most frequently isolated species based on rpoB sequencing were Staphylococcus chromogenes and Staphylococcus epidermidis, followed by Staphylococcus xylosus, Staphylococcus warneri and Staphylococcus equorum (37, 13, 9, 8 and 6% of isolates, respectively). The API Staph ID 32 correctly identified 41% of the CNS isolates. Best agreement with rpoB sequence based species identification was found for S. epidermidis, Staphylococcus hyicus and S. xylosus (100, 89 and 87%, respectively). The positive predictive value was 89, 100 and 52%, respectively. Poor sensitivity was observed for 3 of the 5 most frequently found species, S. chromogenes (37%), Staphylococcus warneri...
Veterinary Microbiology, 2008
24 Virulence genes regulated by the SsrA/B system are indispensable for systemic disease in 25 BA... more 24 Virulence genes regulated by the SsrA/B system are indispensable for systemic disease in 25 BALB/c mice. The role of this regulating system in the pathogenesis of Salmonella 26 40 Key words 41 Salmonella Typhimurium -pig -macrophage -ssrA/B 42
Veterinary Research, 2012
Although aspergillosis is one of the most common diseases in captive birds, the pathogenesis of a... more Although aspergillosis is one of the most common diseases in captive birds, the pathogenesis of avian aspergillosis is poorly known. We studied the role of avian respiratory macrophages as a first line of defense against avian aspergillosis. The phagocytic and killing capacities of avian respiratory macrophages were evaluated using pigeon respiratory macrophages that were inoculated with Aspergillus fumigatus conidia. On average, 25% of macrophage-associated conidia were phagocytosed after one hour. Sixteen percents of these cell-associated conidia were killed after 4 h and conidial germination was inhibited in more than 95% of the conidia. A. fumigatus conidia were shown to be cytotoxic to the macrophages. Intracellularly germinating conidia were located free in the cytoplasm of necrotic cells, as shown using transmission electron microscopy. These results suggest that avian respiratory macrophages may prevent early establishment of infection, unless the number of A. fumigatus conidia exceeds the macrophage killing capacity, leading to intracellular germination and colonization of the respiratory tract.
Veterinary Microbiology, 2003
The ability of Salmonella enterica serotype Enteritidis isolates to adhere to immobilized secreti... more The ability of Salmonella enterica serotype Enteritidis isolates to adhere to immobilized secretions of the isthmus of the laying hen was determined in an ELISA-type assay. One-third of the 56 isolates tested in the logarithmic growth phase, adhered to the isthmal secretions. Using a binding assay of the isolates to thin paraffin sections of the oviduct, we demonstrated that the receptor of the adhesion was localized inside the tubular gland cells of the isthmus. The adhesion to immobilized isthmal secretions as well as to the paraffin sections was blocked by the addition of mannose. A fimD mutant of S. Enteritidis, lacking type 1 fimbriae, did not adhere, confirming that the adhesion was mediated by type 1 fimbriae. Mannosylated glycoproteins were demonstrated in the isthmus glandular cells using confocal laser scanning microscopy by FITC-labelled Lens culinaris lectins. It is hypothesized that the binding of S. Enteritidis to isthmal secretions could play a role in the contamination of eggs through incorporation of the bacteria in the shell membranes.
Veterinary Microbiology, 2001
Eighty-seven Streptococcus suis isolates recovered in 1999±2000 from diseased pigs, all from diff... more Eighty-seven Streptococcus suis isolates recovered in 1999±2000 from diseased pigs, all from different farms, were screened for resistance against macrolide and lincosamide antibiotics by the disk diffusion and agar dilution test and a PCR assay, amplifying the ermB gene and the mefA/E gene. Seventy-one percent of the isolates showed constitutive resistance to macrolide and lincosamide antibiotics (MLS B -phenotype). All these isolates were positive for the ermB gene in the PCR, but negative for the mefA/E gene. For all strains minimum inhibitory concentrations (MIC) against ®ve other antimicrobial agents were determined. All strains were susceptible to penicillin. Ninety-nine percent of the isolates were susceptible to enro¯oxacin and tiamulin. Eighty-®ve percent of the strains were resistant to doxycycline. A 540 bp fragment of the ermB genes of eight S. suis strains was sequenced and compared with ermB genes of ®ve S. pneumoniae and ®ve S. pyogenes strains of human origin. A 100% homology was found between these fragments in seven S. suis, one S. pneumoniae and three of the S. pyogenes isolates.
Veterinary Microbiology, 2010
Veterinary Microbiology, 2009
The Veterinary Journal, 2010
The objective of this study was to evaluate an automated blood culture system for the isolation o... more The objective of this study was to evaluate an automated blood culture system for the isolation of microorganisms from infected equine synovial fluid (SF). Samples were collected from 220 severely inflamed synovial joints and classified as either presumably infected (group A: n = 149) or not infected (group B: n = 71), based on a combination of clinical history, clinical signs and cytological analysis of the SF. Samples were inoculated into blood culture bottles and after incubation were subcultured onto agar media to confirm the results and to facilitate full bacterial identification. Microorganisms were isolated from 107 group A samples (71.8%) and from three group B samples (4.2%). Overall, the detection system identified 117 bottles as positive and 103 as negative, including nine instrument-false-positives and two instrument-false-negatives. The median time-to-detection for Gram-positive bacteria, Gram-negative bacteria, and for fungi was 14.3 (interquartile range [I.R.] 10.0) h, 8.8 (I.R. 12.8) h, and 72.0 (range 60.8-74.8) h, respectively. It was concluded that culture of infected SF using the automated system combines the advantages of enrichment in specialised medium with the rapid detection of bacterial growth.
PLoS ONE, 2010
Background: ''Helicobacter (H.) heilmannii'' type 1 is the most prevalent gastric non-H. pylori H... more Background: ''Helicobacter (H.) heilmannii'' type 1 is the most prevalent gastric non-H. pylori Helicobacter species in humans suffering from gastric disease. It has been shown to be identical to H. suis, a bacterium which is mainly associated with pigs. To obtain better insights into the long-term pathogenesis of infections with this micro-organism, experimental infections were carried out in different rodent models.
Developmental and Comparative Immunology, 2002
Cellular Microbiology, 2011
Helicobacter (H.) suis is the most prevalent non-H. pylori Helicobacter species colonizing the st... more Helicobacter (H.) suis is the most prevalent non-H. pylori Helicobacter species colonizing the stomach of humans suffering from gastric disease. In the present study, we aimed to unravel the mechanism used by H. suis to induce gastric epithelial cell damage. H. suis lysate induced mainly apoptotic death of human gastric epithelial cells. Inhibition of !-glutamyl transpeptidase (GGT) activity present in H. suis lysate and incubation of AGS cells with purified native and recombinant H. suis GGT showed that this enzyme was partly responsible for the observed apoptosis. Supplementation of H. suis or H. pylori GGT-treated cells with glutathione strongly enhanced the harmful effect of both enzymes and resulted in the induction of oncosis/necrosis, demonstrating that H. suis and H. pylori GGT-mediated degradation of glutathione and the resulting formation of glutathione degradation products play a direct and active role in the induction of gastric epithelial cell death. This was preceded by an increase of extracellular H 2 O 2 concentrations, generated in a cell-independent manner and causing lipid peroxidation. In conclusion, H. suis and H. pylori GGT-mediated generation of pro-oxidant glutathione degradation products brings on cell damage and causes apoptosis or necrosis, dependent on the amount of extracellular glutathione available as a GGT substrate.
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Papers by Freddy Haesebrouck