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Biomedical subjects

Richard Ducatelle

Publications and source records attributed to Richard Ducatelle.

At least 37 records · Page 2Linked to original sources

Prevalence of Helicobacter pullorum among patients with gastrointestinal disease and clinically healthy persons.

Feces from 531 patients with gastroenteritis and from 100 clinically healthy individuals were tested for Helicobacter pullorum by use of PCR. Samples positive by PCR were qualified for isolation. H. pullorum DNA was demonstrated to be present in feces from 4.3% of patients with gastrointestinal disease but also in feces from 4.0% of clinically healthy persons. One strain was isolated from one patient with gastrointestinal disease.

DNA, Bacterial↗

Development of a gut perfusion model as an alternative to the use of live fish.

An alternative fish model with the principal aim of studying the interaction between fish pathogens and the intestinal tissue was developed. The preparation consisted of an excised gut tractus from rainbow trout (Oncorhynchus mykiss), perfused through cannulation of the aorta intestinalis ventralis with filtered and heparinized Cortland+dextran 1% as the perfusion fluid. This perfusion fluid was delivered by means of a drip. The perfused gut tractus was suspended in a circular bath filled with Ringer solution, which was aerated and kept at a constant temperature of 12 degrees C. Unperfused gut placed in Ringer solution at the same temperature served as the negative control. Perfusion was effective in maintaining the gut in a healthy condition for at least 60 min with only slight oedema and sloughing of the epithelium. Conversely, the unperfused gut revealed excessive tissue degeneration and severe necrosis.

Animals↗

Protection of laying hens against Salmonella Enteritidis by immunization with type 1 fimbriae.

Eighteen chickens were immunized subcutaneously with purified type 1 fimbriae from Salmonella enterica serotype Enteritidis at 18 and 21 weeks of age. Evidence of IgG and IgA responses was found in the eggs and in the sera of the immunized hens. Three weeks later, immunized and non-immunized chickens (n=18) were challenged intravenously with 2x10(7) live Salmonella enterica serotype Enteritidis. There was no significant difference in the numbers of eggs laid by immunized and non-immunized birds. The percentage of Salmonella contaminated eggs was significantly higher in the non-immunized group than in the immunized group due to a higher percentage of contamination of the externally disinfected egg shells. There were no statistical differences in the percentages of contaminated yolks and egg whites between control and immunized birds. No differences in the number of colonizing bacteria could be found in the spleen nor in the liver between the immunized and the control groups throughout the experiment. Salmonella was cleared from the ovary of the immunized birds in the second week p.i., in contrast to the control birds where Salmonella was isolated till the third week after infection. Oviducts were significantly more infected in the control group than in the immunized group. Salmonella was cleared from the oviducts at 3 weeks p.i. in the immunized hens but not in the control hens. In conclusion, we demonstrated that the immunization of laying hens with type 1 fimbriae reduced the number of contaminated eggs and reduced the colonization of the reproductive organs.

Animals↗

Efficacy of vaccines against bacterial diseases in swine: what can we expect?

This paper discusses what can be expected with regard to efficacy of antibacterial vaccines used in swine, based on the present knowledge of pathogen-host interactions. First, vaccination against bacteria that mainly cause disease by production of exotoxins is considered. Vaccines containing the inactivated toxin or a non-toxic but antigenic recombinant protein derived from the exotoxin can be expected to provide protection against disease. The degree of protection induced by such vaccines varies, however, depending amongst other things on the pathogenesis of the disease. Vaccination against clostridial infections, Actinobacillus pleuropneumoniae infections, progressive atrophic rhinitis and enterotoxigenic Escherichia coli, is considered. The second part of the article deals with vaccination against extracellular bacteria. Protection against these bacteria is generally mediated by antibodies against their surface antigens and certain secreted antigens, but cellular immunity may also play a role. Efficacy of vaccines against swine erysipelas, Streptococcus suis infections, Mycoplasma hyopneumoniae infections and swine dysentery is discussed. Finally, vaccination against facultatively intracellular bacteria is considered. For protection against these bacteria cell-mediated immunity plays an important role, but antibodies may also be involved. It is generally accepted that live-attenuated vaccines are more suitable for induction of cell-mediated immunity than inactivated vaccines, although this also depends on the adjuvant used in the vaccine. As an example, vaccination against Salmonella enterica serotype Typhimurium is discussed.

Animals↗

Patulin produced by an Aspergillus clavatus isolated from feed containing malting residues associated with a lethal neurotoxicosis in cattle.

A severe neurotoxicosis, comprising tremors, ataxia, paresis, recumbency and death, occurred simultaneously among several herds of beef cattle in the region of Flanders (Belgium). After a first multi-toxin screening of some suspected diet elements, verruculogen was detected in a sample of a common feed ingredient. However, when the first animal necropsies revealed serious nervous lesions, including neuronal degeneration of the central nervous system and axonal degeneration in the peripheral nervous system, further investigations focused on fungal isolation. As expected from the pathological lesions, Aspergillus clavatus was found to be the dominant fungal species in a sample of compacted fodder, containing malting residues, consumed by all the affected herds. The isolated fungus appeared to produce patulin in culture medium. Traces of patulin were also detected in the fodder. These findings and their possible role in the intoxication are discussed.

Animal Feed↗

Clostridium perfringens in poultry: an emerging threat for animal and public health.

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.

Animal Feed↗

Effect of type 1 fimbriae of Salmonella enterica serotype Enteritidis on bacteraemia and reproductive tract infection in laying hens.

Research on the role of type 1 fimbriae of Salmonella enterica serotype Enteritidis in poultry to date has been focused on the intestinal phase of the infection. This study aimed to investigate the role of type 1 fimbriae in a systemic infection by intravenously inoculating chickens with a fimD mutant or its parent strain. The fimD mutant was present in the blood for 3 weeks after infection, while the wild type parent strain was cleared within the first 3 days. The fimD mutant was isolated at least as much as the parent strain from the liver and spleen for up to 3 weeks after inoculation. The wild type strain was cleared from the caeca in the second week, while the fimD mutant was isolated from the caeca for up to 3 weeks after infection. The ovaries were more heavily infected by the fimD mutant than by the wild-type strain. In the first and second week after inoculation, the oviducts were more frequently infected by the mutant strain. The eggs of birds infected with the fimD mutant were less frequently contaminated with Salmonella. The shells of the eggs were more frequently contaminated by the wild type strain than with the mutant strain. Thus, the absence of type 1 fimbriae prolongs bacteraemia, modifies reproductive tract infection and reduces egg shell contamination by Salmonella enterica serovar Enteritidis.

Animals↗

Assessment of virulence of pigeon isolates of Salmonella enterica subsp. enterica serovar typhimurium variant copenhagen for humans.

Salmonella enterica serovar Typhimurium variant Copenhagen was isolated from 5 of 152 (3.3%) feral pigeons from the city of Ghent (Belgium) and from 26 pooled fecal samples from 114 pigeon lofts (22.8%). These isolates belonged to phage type (PT) 99. Seven of the pigeon isolates were further compared in vitro to five human variant Copenhagen isolates, 2 isolates of PT 208, 1 isolate each of PT 120 and U302, and a nontypeable isolate. No differences in invasiveness in human intestinal epithelial Caco-2 cells were found. The human strains, however, were able to multiply significantly more inside human THP-1 macrophages than the pigeon strains. After inoculation of mice with a pigeon PT 99 strain, high numbers of Salmonella bacteria were shed with the feces, the internal organs were heavily colonized, and the animals showed severe clinical symptoms resulting in death. In conclusion, the less-pronounced ability of the pigeon variant Copenhagen strains to multiply inside human macrophages than human strains as well as the lack of human PT 99 isolates during 2002, despite the relatively high frequency of this PT in the pigeon population, suggest these strains to be of low virulence to humans. However, the high virulence for mice of the tested strain implies that rodents may act as reservoirs.

Animals↗

Cats as a risk for transmission of antimicrobial drug-resistant Salmonella.

To determine whether cats were a risk for transmission of Salmonella to humans, we evaluated the excretion of Salmonella by pet cats. Rectal-swab specimens were taken from 278 healthy house cats, from 58 cats that died of disease, and from 35 group-housed cats. Group-housed cats were kept in one room with three cat trays and a common water and feed tray. Eighteen (51.4%) of 35 group-housed cats, 5 (8.6%) of 58 diseased cats (5/58), and 1 (0.36%) of 278 healthy house cats excreted Salmonella. Salmonella isolates were of serotypes Typhimurium, Enteritidis, Bovismorbificans and 4:i:-. Acquired antimicrobial resistance was found in serotype Typhimurium (resistance to ampicillin, chloramphenicol, and tetracycline; to ampicillin; and to chloramphenicol) and 4:i:- strains (resistance to ampicillin, chloramphenicol, sulfonamides, trimethoprim, and sulfamethoxazole/trimethoprim). Cats that excrete Salmonella can pose a public health hazard to people who are highly susceptible to Salmonella, such as children, the elderly, and immunocompromised persons.

Animals↗

Adhesion of high and low virulence Flavobacterium psychrophilum strains to isolated gill arches of rainbow trout Oncorhynchus mykiss.

The ability of Flavobacterium psychrophilum to adhere to the gill tissue of rainbow trout Oncorhynchus mykiss was evaluated. A gill perfusion model was adopted, offering a number of advantages compared to other in vitro as well as in vivo models. A comparison between the adhesion capacity of a high and low virulence F. psychrophilum strain was made. Experiments were additionally carried out to assess the influence of water quality (organic material, nitrite) and temperature on the adhesion process of the bacterial cells. The high virulence strain attached more readily to the gill tissue than did the low virulence strain. Moreover, the adherence of the high virulence strain of F. psychrophilum was influenced by a number of factors. These were immersion of the gill arches in water to which organic material or nitrite were added, and elevated temperature. The former 2 increased the adhesion ability, while the latter had a negative influence on the adherence process.

Animals↗

Adhesion of Salmonella enterica serotype Enteritidis isolates to chicken isthmal glandular secretions.

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.

Animals↗

Host adaptation of pigeon isolates of Salmonella enterica subsp. enterica serovar Typhimurium variant Copenhagen phage type 99 is associated with enhanced macrophage cytotoxicity.

Phage type 99 of Salmonella enterica subsp. enterica serovar Typhimurium variant Copenhagen strains isolated from pigeons were examined for the presence of genotypic and phenotypic characteristics. The pulsed-field gel electrophoresis patterns obtained with XbaI and BlnI from 38 pigeon strains were compared with those obtained from 89 porcine, poultry, and human strains of variant Copenhagen. Identical patterns with XbaI and four closely related patterns with BlnI were obtained with the pigeon strains, whereas 16 XbaI patterns were found with the other strains. The XbaI patterns of the pigeon strains showed a low genetic similarity to the patterns of the porcine, poultry, and human strains and invariably showed a low-molecular-weight band that was absent in the majority of the other strains. The virulence genes shdA, spvR, pefA, sopE, and spvB were uniformly present in six pigeon isolates representing the genetic diversity found with BlnI. These six pigeon-derived strains were highly cytotoxic for pigeon macrophages compared to three porcine strains. After experimental infection of pigeons with a pigeon strain, clinical symptoms, fecal shedding, and colonization of internal organs were more pronounced than those after infection with a porcine strain. These data suggest that the phage type 99 strains used in this study are highly adapted to pigeons and should be classified as a host-restricted lineage of the serovar Typhimurium.

Adaptation, Physiological↗

Evaluation of serology, bacteriological isolation and polymerase chain reaction for the detection of pigs carrying Actinobacillus pleuropneumoniae in the upper respiratory tract after experimental infection.

Pigs, asymptomatically infected with Actinobacillus pleuropneumoniae in their upper respiratory tract, can transmit the infection. Detection of such animals is indispensable to prevent the intake of the disease in a herd. This study was conducted to evaluate bacteriology, polymerase chain reaction (PCR) and serology for the detection of subclinically infected pigs. Pigs were inoculated onto the tonsils with an A. pleuropneumoniae serotype 9 strain (n=12, group 1) or phosphate buffered saline solution (PBSS) (n=5, group 2). To prevent infection of the lungs, pigs of group 1 were treated three times with sodium ceftiofur as an aerosol. A third group (n=5) was inoculated intranasally with the same strain. All animals were euthanized 30 days post-inoculation (dpi). In pigs of group 1, clinical signs were not observed. A small lung lesion was found in only one pig and A. pleuropneumoniae was isolated from this lesion. The bacterium was not isolated from the lungs of animals that did not develop lung lesions. A. pleuropneumoniae was demonstrated in tonsils of 9/12 animals using bacteriological isolation, whereas it was demonstrated in mixed bacterial cultures from tonsils of all 12 animals by PCR. In non-infected animals (group 2), clinical signs were not observed and A. pleuropneumoniae was not demonstrated in any sample. All intranasally infected animals (group 3) developed disease signs and lung lesions. High antibody titers against ApxI, ApxII and heat-stable antigens were detected in animals that developed lung lesions. Antibody titers against these antigens were low or absent in all other pigs. It was concluded that pigs carrying A. pleuropneumoniae in the upper respiratory tract generally do not show measurable antibodies in serum. Therefore, sensitive methods for the detection of the etiological agent such as PCR are required to identify carrier animals, while serological methods are not suitable.

Actinobacillus Infections↗

Characterization of the in vitro adhesion of Actinobacillus pleuropneumoniae to swine alveolar epithelial cells.

Actinobacillus pleuropneumoniae biovar 1 serotypes 2, 5a, 9 and 10 strains were tested for their ability to adhere to alveolar epithelial cells in culture. For the serotypes 5a, 9 and 10 strains, optimal adherence was observed after growth of bacterial cells in a NAD-restricted medium (0.001% NAD). This condition was also associated with the expression of a 55 kDa outer membrane protein (OMP) and of fimbriae. For the serotype 2 strain, adherence and expression of fimbriae and a 55 kDa OMP was less influenced by the growth conditions. The N-terminal amino acid sequence of the 55 kDa OMP had no homology with any known sequence, suggesting that it is an as yet unknown protein. Adherence capabilities were significantly reduced following treatment of the bacteria with proteolytic enzymes or heat. These findings suggest that proteins are involved in adhesion. The hydrophobic bond-breaking agent tetramethylurea was unable to inhibit the adherence of A. pleuropneumoniae to alveolar epithelial cells. Treatment of the bacteria with sodium metaperiodate resulted in lower adhesion scores for the serotypes 2 and 9 strains but the inhibition of adhesion was clearly lower than after treatment with proteolytic enzymes. This indicates that, besides proteins, carbohydrates might also be involved in adhesion of A. pleuropneumoniae to alveolar epithelial cells. The finding that inhibition of adhesion was very high when bacteria were treated with a combination of sodium metaperiodate and pronase also suggests that more than one adhesin is involved.

Actinobacillus pleuropneumoniae↗

The effect of vaccination with a Salmonella enteritidis aroA mutant on early cellular responses in caecal lamina propria of newly-hatched chickens.

When newly hatched chicks are inoculated with a Salmonella strain, they induce a rapid onset of resistance to intestinal colonization by other Salmonella strains. The exact mechanism of this early colonization-inhibition is not known. To study host-related contributions to this phenomenon, the kinetics of immune cell infiltration in the caecal wall was analyzed during the first 10 days after vaccination of newly hatched chickens with a Salmonella enterica serovar Enteritidis aroA mutant, and infection 1 day later with a virulent S. enterica serovar Enteritidis strain. These data were correlated with bacterial colonization and clearance of the Salmonella Enteritidis challenge strain. Bacteriological data showed that vaccinated animals had a much lower number of challenge bacteria in their organs and caecal contents the first days post-challenge, relative to unvaccinated animals. Immunohistochemical analysis of the caecal lamina propria revealed that heterophils started infiltrating the caecal lamina propria from 12 h post-vaccination. Macrophages and T-lymphocytes started infiltrating from 20 h and B-lymphocytes from 24 h post-vaccination. These data imply that immune cells already colonized the caecal wall at the time of challenge in vaccinated animals. The presence and activity of these cells in the caecal wall shortly after administration of a Salmonella Enteritidis aroA mutant might contribute to the inhibition of colonization of a virulent Salmonella strain, subsequently administered.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗

Actinobacillus pleuropneumoniae infections in closed swine herds: infection patterns and serological profiles.

Many farrow-to-finish herds are endemically infected with Actinobacillus pleuropneumoniae. In order to control the disease efficiently, a better knowledge of the ages at which pigs become infected is necessary. Furthermore, no information is available concerning the influence of maternally derived antibodies on the colonization of the upper respiratory tract. Therefore, A. pleuropneumoniae infection patterns were studied in five farrow-to-finish pig herds (A-E) with a history of pleuropneumonia. A longitudinal study was carried out in herds A and B. In these herds, piglets from sows carrying A. pleuropneumoniae in their noses or tonsils were sampled. Nasal and tonsillar swabs as well as sera, were collected from these animals at the age of 4, 8, 12, 16 (herds A and B) and 23 weeks (herd B). At these ages other pigs from the same sows were euthanized. The lungs were macroscopically examined and samples from nose, tonsils and lungs were collected at necropsy. A cross-sectional study was performed in herds C-E. In these herds nasal and tonsillar swabs, as well as sera, were taken from 10 animals of 4, 8, 12 and 16 weeks of age. Lung, nasal and tonsillar samples were tested for the presence of A. pleuropneumoniae by routine bacteriology and PCR with mixed bacterial cultures. The sera were examined for the presence of Apx toxin neutralizing antibodies. In herd A, A. pleuropneumoniae serotype 2 and 10 strains were isolated, whereas serotype 2, 3, 5b and 8 strains were demonstrated in herd B. In most herds, A. pleuropneumoniae was detected in mixed bacterial cultures of tonsillar and/or nasal samples by PCR from the age of 4 weeks onwards. Colonization of the lungs and development of lung lesions was observed in 12- and 16-week-old animals of herd A and 23-week-old animals of herd B. In most herds, high antibody titres were detected in 4-week-old piglets. These titres decreased during the first 12 weeks of age, but thereafter, increased. It was concluded that PCR with mixed bacterial cultures from tonsillar swabs is a valuable tool for the detection of infected animals. It was also concluded that colonization of tonsils and nasal mucosae can occur in the presence of maternally derived antibodies. Infection of the upper respiratory tract without lung involvement did not result in development of Apx toxin neutralizing antibodies. Therefore, such serological assays cannot be used for the detection of subclinically infected animals.

Actinobacillus Infections↗

Possible role for insulin-like growth factor-I in the pathogenesis of cystic endometrial hyperplasia pyometra complex in the bitch.

Cystic endometrial hyperplasia (CEH) is an important pathologic condition in the canine uterus and recognized as a common cause of illness and death in this species. The underlying cause and pathogenic mechanism responsible for this condition remains incompletely understood. Aberrant sex steroid hormone receptor expression in the uterus of dogs with CEH has been documented but not explained. In the dog there is an exceptionally high, progestin induced production of insulin-like growth factor-I (IGF-I) which is now generally accepted to be one of the most important growth factors with a high mitogenic effect on the uterus. Therefore, in this study the immunohistochemical staining intensity for IGF-I was compared among the uteri of 25 adult female dogs that had developed CEH and 14 healthy dogs in comparable stages of the estrus cycle. Specific staining for IGF-I was found in the cytoplasm epithelial cells and in smooth muscle cells of endometrium and myometrium. A marked increase in specific staining intensity for IGF-I was found in the surface epithelium, glandular epithelium and in the stroma of the uteri of dogs with CEH. The increase in IGF-I specific staining intensity was most prominent in the superficial endometrial stroma. Based on the known role of IGF-I in endometrial proliferation, it was concluded from the present study that high concentrations of IGF-I located in and around the epithelial cells of the endometrium in dogs with CEH, could play an important role in the development of CEH.

Anestrus↗

Dynamics of immune cell infiltration in the caecal lamina propria of chickens after neonatal infection with a Salmonella enteritidis strain.

Dynamics of leucocyte infiltration and bacterial invasion in the caecal wall were studied after oral infection of 2-day-old chicks with Salmonella enterica ser. Enteritidis. Bacteria invaded the lamina propria of the caecal wall from 12h post-challenge onwards. Bacteriological examination of internal organs (liver, spleen) showed a peak in Salmonella bacteria at 3days post-infection, after which the number of bacteria decreased. Immunohistochemistry revealed macrophages and T-lymphocytes invading the caecal propria mucosae from 24h after challenge onwards, while B-lymphocytes came somewhat later, subsequently organising into follicular aggregates. An early increase in granulocytes was partly masked by the response to natural flora. While the B-lymphocyte and granulocyte populations were maintained, T-lymphocyte and macrophage populations were already reducing by 10days post-challenge. The infiltration of macrophages and T-lymphocytes in the caecal wall, followed by B-lymphocytes, is the result of an inflammatory response, caused by invading bacteria at this site. The structural maturation of the mucosa-associated lymphoid tissues is antigen driven, since B-cells organise in a follicular pattern.

Animals↗