Biomedical subjects

F Haesebrouck

Publications and source records attributed to F Haesebrouck.

At least 91 records · Page 5Linked to original sources

PubMed2001

Prevalence and mechanism of resistance against macrolides and lincosamides in Streptococcus suis isolates.

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 five other antimicrobial agents were determined. All strains were susceptible to penicillin. Ninety-nine percent of the isolates were susceptible to enrofloxacin and tiamulin. Eighty-five percent of the strains were resistant to doxycycline. A 540bp fragment of the ermB genes of eight S. suis strains was sequenced and compared with ermB genes of five S. pneumoniae and five 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. This study demonstrates that resistance against macrolides, lincosamides and streptogramin B is widespread in S. suis and mediated by ribosome methylation, encoded by the ermB gene.

Animals↗

PubMed2001

Detection of Actinobacillus pleuropneumoniae in cultures from nasal and tonsillar swabs of pigs by a PCR assay based on the nucleotide sequence of a dsbE-like gene.

A PCR assay for the detection of Actinobacillus pleuropneumoniae was developed based on the amplification of a dsbE-like gene. All of 157 field isolates of A. pleuropneumoniae reacted in the PCR by the amplification of a 342bp product. No reaction was observed with related bacterial species or other bacterial species isolated from pigs, except for A. lignieresii. The lower detection limit of the PCR was 10(2) CFU per PCR test tube and was not affected by the addition of 10(6) CFU Escherichia coli. The PCR was evaluated on mixed bacterial cultures from nasal and tonsillar swabs as well as suspensions of nasal conchae and tonsils obtained from specific pathogen-free (SPF) pigs, experimentally infected pigs, and pigs from farrow-to-finish herds. The results of the new PCR were compared with a PCR based on the detection of the omlA gene coding for an outer membrane protein, with a commercially available PCR (Adiavet APP, Adiagène, Saint-Brieuc, France), and with conventional culturing. No positive reactions were observed with any of the PCR methods in samples of SPF animals. In samples of the other animals, no or low significant differences between nasal swabs and suspensions as well as tonsillar swabs and suspensions were observed in any method. In general, more positive results were obtained from tonsillar samples in comparison to nasal samples. Interassay sensitivity and specificity values were assessed for each test by pair wise comparisons between assays. The agreement between tests was evaluated by calculating Cohen's kappa coefficient. From these analyses the three PCR assays showed a good agreement. The dsbE-based PCR proved to be highly sensitive (95 and 93%) and specific (82 and 74%) in comparison to the omlA-based PCR and the commercially available PCR, respectively. It was concluded that the dsbE-like gene-based PCR is a reliable diagnostic assay for demonstration of A. pleuropneumoniae. Furthermore, it was demonstrated that tonsillar swabs can be used for the detection of the pathogen in healthy carrier animals.

Actinobacillus Infections↗

PubMed2001

Sequence analysis of a RAPD band differentiating high and low virulence Staphylococcus aureus strains from rabbits.

RAPD typing revealed the presence of a nucleotide band in typical high virulence rabbit Staphylococcus aureus strains which was absent in low virulence strains and in an atypical high virulence strain. The nucleotide sequence of this band was determined. Primers within this sequence were developed and PCR products of eight typical high virulence, one atypical high virulence and nine low virulence rabbit S. aureus strains were sequenced. All low virulence strains and the atypical high virulence strain revealed a constant difference with the typical high virulence strains for nucleotide 377 of the 1055bp sequence. The eight typical high virulence strains possessed a guanine base on this site, while the other strains tested showed an adenine base. These findings support the hypothesis on the clonal origin of typical high virulence rabbit S. aureus strains. After comparison with databases, two open reading frames (ORF) were identified within the sequence, which appeared to encode two structural ribosomal proteins. The single nucleotide mutation does not affect the amino acid sequence of the protein it encodes for.

Animals↗

PubMed2001

Secreted antigens as virulence-associated markers in Staphylococcus aureus strains from rabbits.

Western blot analysis was performed from the culture supernatant of 59 rabbit Staphylococcus aureus strains, classified as high and low virulence strains according to their epidemiological behaviour in commercial rabbitries, bio-, phage- and RAPD-type. Fourteen extracellular antigen bands (A-N) were recognised using sera of rabbits immunised with washed, viable high virulence S. aureus bacteria. Eleven of these bands were found in high virulence as well as in low virulence strains. The band A, approximately 78 kDa, was not seen in any of the 27 high virulence strains, except for one strain which was also typical in other aspects, was detected in all, but one of the low virulence strains. The M and N bands with molecular masses of approximately 29 and 27 kDa, respectively, were recognised in all high virulence strains except for the atypical strain, but in none of the low virulence strains. This indicates that the latter two antigens may be virulence-associated markers for S. aureus strains from rabbits.

Animal Diseases↗

PubMed2001

Induction of the respiratory burst in turtle peritoneal macrophages by Salmonella muenchen.

Peritoneal macrophages were collected from juvenile turtles 72h after intraperitoneal inoculation with a 3% Sephadex suspension. The macrophages were assayed for their chemiluminescent (CL) properties, reflecting their respiratory burst activity, after stimulation with Zymosan A, phorbol 12-myristate 13-acetate (PMA), N-formyl-methionyl-leucyl-phenylalanine (fMLP), and calcium ionophore A23187. Except for fMLP, all triggering agents induced a marked CL response. Luminol was used as the chemiluminescent probe. When comparing CL responses in temperatures ranging from 15 to 35 degrees C, lower assay temperatures induced lower and slower CL responses. Stimulation with viable Salmonella muenchen resulted in a distinct response. Bacteria, inactivated by means of heat or acetone, induced a faster and stronger oxidative burst. Opsonization of either viable or heat-inactivated S. muenchen with non-inactivated anti-S. muenchen serum, prepared in turtles, induced faster and higher CL responses. On the other hand, opsonization of acetone-inactivated S. muenchen caused CL responses to be slower and weaker. S. muenchen, opsonized with heat-inactivated turtle anti S. muenchen serum, induced higher responses than non-opsonized bacteria, but slower and weaker responses than bacteria opsonized with native turtle antiserum. No response was recorded after stimulation with LPS and the supernatant of heat-inactivated bacteria.

Acetone↗

PubMed2001

Lactobacillus agilis is an important component of the pigeon crop flora.

AIMS: To examine the presence of lactic acid bacteria (LAB: enterococci, streptococci and lactobacilli) in the pigeon crop. METHODS AND RESULTS: The crops of 10 pigeons were sampled and inoculated on agar plates for isolation of streptococci, enterococci and lactobacilli. The isolates were identified using tDNA-PCR. Lactobacillus agilis, a species described in 1981 from municipal sewage, was the dominant component in eight of these pigeon crop sacs. A Lactobacillus species related to L. fermentum and L. mucosae but probably not belonging to one of these species was isolated from five birds. Three pigeons carried Enterococcus cecorum. Minor species found were E. columbae, E. faecalis, E. hirae, L. johnsonii, L. salivarius, and Streptococcus gallolyticus. A description is given of the phenotypic characteristics of the L. agilis pigeon strains. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: L. agilis is found to be the main component of the LAB flora in the pigeon crop.

Animals↗

PubMed2001

The gram-positive tonsillar and nasal flora of piglets before and after weaning.

AIMS: To investigate gram-positive nasal and tonsillar microbial flora of piglets before and after weaning. METHODS AND RESULTS: The nasal and tonsillar gram-positive bacterial flora of 20 non-weaned piglets (2 weeks of age) and 20 weaned piglets (6 weeks of age), obtained from four different piggeries, was quantified by culture and identified by tDNA-PCR. The most widely occurring species from nasal conchae before as well as after weaning in the different piglets investigated were Streptococcus suis and Rothia nasimurium. After weaning a wide variety of Lactobacillus species appeared but in low numbers. In the tonsils, Strep. suis, Strep. dysgalactiae, S. hyicus, S. aureus, Arcanobacterium pyogenes and Actinomyces hyovaginalis were the species isolated from the largest number of pigs before and after weaning. S. aureus and most lactobacilli became more prevalent after weaning. Bacteria not known to be associated with pigs found in the present study included R. nasimurium, Strep. gallolyticus, Pediococcus pentosaceus and some Lactobacillus species. CONCLUSIONS: Over 30 different gram-positive bacterial species may occur in nasal conchae and tonsils of unweaned piglets at 2 weeks of age and of 6-week-old weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that weaning is associated with changes in prevalence of only a small minority of the highly diversified bacterial flora of the nares and tonsils of pigs.

Animals↗

PubMed2001

Effect of endobronchial challenge with Actinobacillus pleuropneumoniae serotype 9 of pigs vaccinated with a vaccine containing Apx toxins and transferrin-binding proteins.

The efficacy of a subunit vaccine containing the Apx toxins of Actinobacillus pleuropneumoniae and transferrin-binding proteins was determined. Ten pigs were vaccinated twice with the vaccine. Eight control animals were injected twice with a saline solution. Three weeks after the second vaccination, all pigs were endobronchially inoculated with 10(6.5) colony-forming units (CFU) of an A. pleuropneumoniae serotype 9 strain. In the vaccine group, none of the pigs died after inoculation. Only one pig of the control group survived challenge. Surviving pigs were killed at 7 days after challenge. The mean percentage of affected lung tissue was 64% in the control group and 17% in the vaccine group. Actinobacillus pleuropneumoniae was isolated from the lungs of all animals. The mean bacterial titres of the caudal lung lobes were 5.0 x 10(8) CFU/g in the control group and 3.0 x 10(6) CFU/g in the vaccine group. It was concluded that the vaccine induced partial protection against severe challenge.

Actinobacillus Infections↗

PubMed2001

Pathogenicity of Actinobacillus minor, Actinobacillus indolicus and Actinobacillus porcinus strains for gnotobiotic piglets.

The purpose of the study was to evaluate the clinical significance of Actinobacillus minor, Actinobacillus porcinus and Actinobacillus indolicus strains in gnotobiotic piglets. Twenty-two 6-h-old Caesarean-delivered and colostrum-deprived piglets were intranasally and orally inoculated with 2 x 10(6) colony-forming units of an A. minor (group 2; n = 9), A. indolicus (group 3; n = 5), or A. porcinus (group 4; n = 8) strain. Six other piglets were inoculated in the same way with phosphate-buffered saline solution and used as controls (group 1). All pigs were observed for clinical signs and rectal temperatures were taken until euthanasia 7 days after inoculation. At necropsy, conchae, tonsils, lungs, brains, liver, spleen and kidneys were macroscopically examined for lesions and samples were taken for bacteriology. None of the pigs developed fever. Mild ataxia was observed in one pig from group 3 for 2 days. Clinical signs were not observed in the other animals. In none of the animals were macroscopic lesions detected at necropsy. NAD-dependent Pasteurellaceae were not isolated from control animals (group 1). The A. minor, A. indolicus and A. porcinus strains were isolated from the tonsils of one, two and one pigs, respectively. Actinobacillus porcinus was isolated from the brains of the pig with central nervous symptoms and from the conchae of another pig. The inoculation strains were not demonstrated in the other samples. It was concluded that, using these inoculation routes and dose, the A. minor, A. indolicus and A. porcinus strains had low capacity to colonize the upper respiratory tract of gnotobiotic piglets and demonstrated low or no pathogenicity in such animals.

Actinobacillus↗

PubMed2001

Enterococcus villorum sp. nov., an enteroadherent bacterium associated with diarrhoea in piglets.

The taxonomic positions of five enteroadherent bacterial pig isolates, showing phenotypic characteristics most similar to those of Enterococcus durans and Enterococcus hirae, were investigated in a polyphasic study that included 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determinations, whole-cell protein fingerprinting, D11344-primed PCR typing and an extensive examination of phenotypic properties. The results demonstrated that the organisms represent a new species in the Enterococcus faecium species group, for which the name Enterococcus villorum sp. nov. is proposed. The type strain is LMG 12287T (= CCM 4887T).

Animals↗

PubMed2001

Enterococcus haemoperoxidus sp. nov. and Enterococcus moraviensis sp. nov., isolated from water.

A polyphasic taxonomic approach was used to study atypical enterococci isolated from surface waters. All strains were characterized by physiological and biochemical tests as well as by genotyping. The results of biochemical tests and tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all studied strains uniformly into two groups. Because these groups were clearly separated from all enterococcal species described to date, 16S rDNA sequence analysis, DNA base composition analysis and DNA-DNA hybridization of representative strains were done to elucidate the taxonomic position of the analysed groups. On the basis of the results obtained, the names Enterococcus haemoperoxidus (type strain CCM 4851T = LMG 19487T) and Enterococcus moraviensis (type strain CCM 4856T = LMG 19486T) are proposed for the two hitherto undescribed species. The type strains and reference cultures have been deposited in the Czech Collection of Microorganisms (CCM), Masaryk University, Brno, Czech Republic, and in the BCCM/LMG Culture Collection, Ghent University, Belgium.

Base Composition↗

PubMed2001

Globicatella sulfidifaciens sp. nov., isolated from purulent infections in domestic animals.

DNA-DNA hybridization experiments and comparative 165 rDNA sequence analysis revealed that six isolates from purulent joint and lung infections in calves, from a lung lesion in a sheep, and from a joint infection of a pig represented a novel species belonging to the genus Globicatella. Whole-cell protein electrophoresis and biochemical activity testing revealed that the isolates formed a homogeneous group differing from Globicatella sanguinis, the only species of this genus described to date. These animal isolates were classified as Globicatella sulfidifaciens sp. nov. with LMG 188441 (= CCUG 44365T), isolated from the lung of a calf, as the type strain. A detailed description of its phenotypic characteristics is presented. Hydrogen sulphide production was found to be a very useful diagnostic feature.

Animals↗

PubMed2001

Differences in antibiotic resistance patterns of Enterococcus faecalis and Enterococcus faecium strains isolated from farm and pet animals.

The prevalence of acquired resistance in 146 Enterococcus faecium and 166 Enterococcus faecalis strains from farm and pet animals, isolated in 1998 and 1999 in Belgium, against antibiotics used for growth promotion and for therapy was determined. Acquired resistance against flavomycin and monensin, two antibiotics used solely for growth promotion, was not detected. Avoparcin (glycopeptide) resistance was found sporadically in E. faecium only. Avilamycin resistance was almost exclusively seen in strains from farm animals. Resistance rates were higher in E. faecium strains from broiler chickens than in strains from other animal groups with tylosin and virginiamycin and in E. faecalis as well as in E. faecium strains with narasin and bacitracin. Resistance against ampicillin was mainly found among E. faecium strains from pets and was absent in E. faecalis. Tetracycline resistance occurred most often in strains from farm animals, while enrofloxacin resistance, only found in E. faecalis, occurred equally among strains from all origins. Resistance against gentamicin was very rare in broiler strains, whereas resistance rates were high in strains from other origins. It can be concluded that resistance against antibiotics used solely for growth promotion was more prevalent in E. faecium strains than in E. faecalis strains. With few exceptions, resistance against the different categories of antibiotics was more prevalent in strains from farm animals than in those from pets.

Animals↗

PubMed2001

Evaluation of a group-specific 16S ribosomal DNA-based PCR for detection of Helicobacter bizzozeronii, Helicobacter felis, and Helicobacter salomonis in fresh and paraffin-embedded gastric biopsy specimens.

A new specific and sensitive 16S ribosomal DNA-based PCR assay was developed. The assay targets a 78-bp DNA fragment unique to Helicobacter bizzozeronii, Helicobacter felis, and Helicobacter salomonis and can be used with freshly frozen and formalin-fixed paraffin-embedded gastric biopsy specimens.

Animals↗

PubMed2001

Application and evaluation of the interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) for identification of Streptococcus species.

The discriminatory power, speed, and interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) combined with capillary electrophoresis was evaluated for the identification of streptococci. This method was carried out in three different laboratories under highly standardized conditions for 54 strains belonging to 18 different species. It was concluded that interlaboratory reproducibility of tDNA fingerprints produced by means of capillary electrophoresis was sufficiently high to permit the exchange between different laboratories and the construction of common libraries which can be consulted for comparison with fingerprints obtained independently in separate laboratories. In a second step, 17 other species were included in the study and examined in one of the participating laboratories. All Streptococcus species studied, except S. mitis, S. oralis, S. parasanguinis, S. pneumoniae, S. thermophilus, and S. vestibularis, showed distinguishable tDNA fingerprints. A database of well-characterized strains was constructed to enable computer-aided identification of unknown streptococcal isolates.

DNA Fingerprinting↗

PubMed2000

Effect of Flavobacterium psychrophilum strains and their metabolites on the oxidative activity of rainbow trout oncorhynchus mykiss phagocytes.

The oxidative activity of rainbow trout phagocytes was studied using a chemiluminescence technique using 12 different Flavobacterium psychrophilum strains and their metabolites. Phagocytes were obtained from the head kidney of rainbow trout Oncorhynchus mykiss. The addition of viable F. psychrophilum or their metabolites to the phagocytes resulted in an immediate chemiluminescence response. The stimulating effects of both the F. psychrophilum and their metabolites on the phagocytes were found to be heat stable. No significant differences in stimulation capacity were found between the strains tested. To investigate the nature of the stimulating agent, both the bacteria and the supernatant were treated with either sodium metaperiodate or polymyxin B. Adding polymyxin B to the bacterial cells and supernatant did not change the chemiluminescence pattern, suggesting that the capacity of F. psychrophilum to stimulate the phagocytes probably is not due to lipopolysaccharides (LPS). However, following incubation of the bacteria and their metabolites with sodium metaperiodate, the capacity to stimulate phagocytes was significantly impaired. This suggests that a carbohydrate component most likely plays an important role in the ability of F. psychrophilum to stimulate phagocytes. Opsonisation of the bacteria with native trout serum or with rabbit anti-F. psychrophilum serum resulted in an additional chemiluminescence peak which was significantly higher than the first peak. This extra peak disappeared following heat treatment of the trout serum and the rabbit anti-F. psychrophilum serum, pointing towards the involvement of heat labile complement in opsonisation.

Animals↗

PubMed2000

Adhesion of Streptococcus gallolyticus strains to extracellular matrix proteins.

Fourteen pigeon Streptococcus gallolyticus strains of differing virulence, were tested for their ability to adhere to immobilised fibronectin, collagen types I, III and IV. Eight, 2 and 13 strains were able to bind fibronectin, collagen types III and IV, respectively. None of the strains adhered to collagen type I. Heat treatment, proteolytic digestion or periodate treatment reduced the binding of S. gallolyticus to fibronectin and collagen type IV, suggesting that surface receptors contain proteins and carbohydrates. Although binding to these extracellular matrix proteins can play a role in the pathogenesis of streptococcosis in pigeons, binding properties could not be related to virulence, indicating that other factors determine differences in virulence among pigeon S. gallolyticus strains. Adhesion to collagen type IV may account in part for the distribution pattern of the lesions observed in naturally and experimentally infected pigeons.

Animals↗

PubMed2000

Identification of virulence associated markers in the cell wall of pigeon Streptococcus gallolyticus strains.

The cell wall protein profiles of 56 isolates of Streptococcus gallolyticus of differing virulence for pigeons were compared by SDS-PAGE. Additionally, Western blot analysis was performed on the cell wall proteins of 14 strains using sera of pigeons, experimentally infected with A(+)T1 or A(-)T2 strains of S. gallolyticus. The profile of silver stained gels exhibited a complex array of 20-50 bands ranging from less than 6.5-210kDa. A band with molecular mass of 114kDa was only observed in isolates that belonged to the highly virulent A(+)T1, A(+)T2, A(+)T3 and A(-)T1 culture supernatant groups. A band with a slightly higher molecular mass (115kDa) as well as a 207kDa band were only detected in isolates that belonged to the moderately A(-)T3 or low A(-)T2 virulent culture supernatant groups. The 114 and 115kDa band were recognised by all homologous and heterologous pigeon sera used whereas the 207kDa band was only recognised by sera of pigeons infected with a A(-)T2 strain. These findings may indicate that the 114, 115 and 207kDa bands are useful as additional virulence associated markers for pigeon S. gallolyticus strains.

Animals↗