Biomedical subjects

F Van Immerseel

Publications and source records attributed to F Van Immerseel.

17 recordsLinked to original sources

PubMed2006

The fibronectin binding protein ShdA is not a prerequisite for long term faecal shedding of Salmonella typhimurium in pigs.

Porcine carcasses contaminated with Salmonella typhimurium pose significant public health problems. Prolonged faecal shedding of Salmonella in pigs contributes to the contamination level of carcasses. Although the mechanism of prolonged faecal shedding is not yet clarified, the CS54 Island, and more specifically the shdA gene encoding a fibronectin binding autotransporter protein, was identified as an important locus for intestinal colonization and persistence of Salmonella typhimurium in mice. The aim of this study was to assess the contribution of ShdA in faecal shedding of Salmonella typhimurium in pigs. Pigs were orally inoculated with a Salmonella typhimurium wild type field strain or its isogenic shdA mutant strain. For the first few days after inoculation, the shdA mutant strain was excreted more, the diarrhoea was more pronounced and higher numbers of internal organs were infected. No effect on long-term shedding was found. In a porcine ileal loop model, the wild type strain and shdA mutant strain did not show any differences in the induction of neutrophil influx into the intestinal wall and lumen. In conclusion, we have shown that a Salmonella typhimurium deletion mutant in shdA is more virulent during the first days after inoculation and is not significantly impaired in persistence or prolonged shedding in pigs.

Animals↗

PubMed2006

The use of organic acids to combat Salmonella in poultry: a mechanistic explanation of the efficacy.

Salmonella is a human pathogen that is commonly found in poultry products. It is possible to decrease chicken carcass and egg contaminations by adding organic acids to the feed or drinking water at appropriate times. Medium-chain fatty acids are more antibacterial against Salmonella than short-chain fatty acids. The antibacterial effect of these acids is species specific. Bacteria that are unable to decrease intracellular pH accumulate organic acid anions in accordance with the pH gradient across their cell membranes. The short-chain fatty acid butyrate specifically down-regulates expression of invasion genes in Salmonella spp. at low doses. Also medium-chain fatty acids and propionate decrease the ability of Salmonella spp. to invade epithelial cells, in contrast to acetic acid. Because not all bacteria are affected in a similar fashion by organic acids, it may be possible to use probiotic and prebiotic bacteria to achieve beneficial effects. If diets can be designed to stimulate organic acid production in the caecum, it may be possible to control Salmonella spp. via even easier and more cost-effective measures, compared with addition of acids to feed or drinking water.

Animals↗

PubMed2006

Butyrate specifically down-regulates salmonella pathogenicity island 1 gene expression.

Invasion of intestinal epithelial cells by Salmonella enterica is decreased after exposure to butyric acid. To understand the molecular mechanisms of this phenomenon, a comparative transcriptomic analysis of Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium grown in medium supplemented with butyrate was performed. We found that butyrate down-regulated the expression of 19 genes common to both serovars by a factor of twofold or more, and 17 of these genes localized to the Salmonella pathogenicity island 1 (SPI1). These included the SPI1 regulatory genes hilD and invF. Of the remaining two genes, ampH has 91% homology to an Escherichia coli penicillin-binding protein and sopE2 encodes a type III-secreted effector protein associated with invasion but located at a separate site on the chromosome from SPI1.

Animals↗

PubMed2005

Role of SPI-1 in the interactions of Salmonella Typhimurium with porcine macrophages.

Salmonella Pathogenicity Island 1 (SPI-1) genes are indispensable for virulence of Salmonella Typhimurium in mice after oral challenge. These genes mediate invasion in intestinal epithelial cells and induce cell death in murine macrophages. The role of SPI-1 in the pathogenesis of Salmonella Typhimurium infections in food producing animals is not known. It was the aim of the present study to characterize the interactions of a porcine Salmonella Typhimurium field strain and its isogenic mutants in the SPI-1 genes hilA, sipA and sipB with porcine macrophages. SPI-1 was found to be important in the invasion of porcine pulmonary alveolar macrophages (PAM) and the induction of the formation of spacious phagosomes. Both early and delayed cytotoxicity were seen in PAM, but only the early cytotoxicity was SPI-1 dependent. Exposure of PAM to Salmonella Typhimurium induced the production of reactive oxygen species (ROS) and interleukin-8, but no differences were noticed between the induction mediated by the wild type strain and its SPI-1 mutant strains. In conclusion, invasion of porcine macrophages and the induction of early, but not delayed, cytotoxicity by Salmonella Typhimurium is SPI-1 dependent. SPI-1 dependent invasion, however, is not a prerequisite to induce a pro-inflammatory response.

Animals↗

PubMed2005

Virulence-associated traits in avian Escherichia coli: comparison between isolates from colibacillosis-affected and clinically healthy layer flocks.

Colibacillosis appears to be of increasing importance in layer flocks. The aim of this study was to determine characteristics of avian pathogenic Escherichia coli associated with the occurrence of colibacillosis outbreaks at flock level. Forty E. coli strains originating from layers from healthy flocks ('control isolates'), consisting of 25 caecal and 15 extra-intestinal isolates, were compared with 40 strains isolated from layers originating from colibacillosis-affected flocks ('outbreak isolates'), consisting of 20 caecal and 20 extra-intestinal isolates. The examined characteristics were adhesins, invasivity in T84 cell culture, serum resistance, iron uptake, colicin production, and toxinogenicity. The following traits were significantly more often detected in the outbreak isolates than in the control isolates: tsh, iss, iucA, iutA, irp2, fyuA, iroC, cvaC, colicin and colicin V production. A comparison of the extra-intestinal outbreak isolates and the caecal control isolates yielded the same results as when the caecal isolates, extra-intestinal isolates and total number of isolates of the outbreak and the control group were compared. When comparing the caecal and extra-intestinal isolates within the control and within the outbreak group, no significant differences were detected. The O78 and O2 groups showed significant differences with other O-types and NT strains for prevalence of most of the same characteristics. The combination of type 1 fimbriae, tsh, serum resistance, iss, traT, iucA, fyuA, iroC and colicin or colicin V production was significantly more often present in extra-intestinal outbreak isolates than in extra-intestinal control isolates. Only the combination of serum resistance, fyuA and colicin production was present in all outbreak isolates, with a significantly lower prevalence in the control isolates. None of the characteristics or combinations examined were exclusive to the outbreak isolates.

Adhesins, Escherichia coli↗

PubMed2005

Survival of Salmonella serovar Typhimurium inside porcine monocytes is associated with complement binding and suppression of the production of reactive oxygen species.

The development of the carrier state in swine after infection with Salmonella serovar Typhimurium (S. Typhimurium) has not been elucidated yet. Possibly, phagocytes like macrophages play a crucial role. It was the aim of the present study to characterize the interaction of a S. Typhimurium strain and its hilA and ssrA mutants with porcine peripheral blood monocytes (PBM). Exposure of porcine PBM to S. Typhimurium induced the production of reactive oxygen species (ROS), requiring bacterial protein synthesis. The numbers of intracellular bacteria sharply decreased over a period of 3h. Monocytes obtained from different pigs differed markedly in their ROS production and in their ability to kill the bacteria. Interestingly, high ROS production did not coincide with increased intracellular killing. Using diphenylene iodonium inhibition of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, it was shown that bacterial killing was ROS-dependent only within 1h post inoculation, but was ROS-independent from 1h post inoculation onwards. This might be explained by the finding that metabolically active Salmonella bacteria were capable of suppressing the respiratory burst activity in a SPI-1- and SPI-2-independent manner without causing measurable cell damage. Opsonization with complement did not alter the ROS production. Nevertheless, it increased intracellular survival of the bacteria. In conclusion, survival of S. Typhimurium inside porcine PBM is promoted by suppression of respiratory burst activity and complement binding.

Animals↗

PubMed2005

Vaccination and early protection against non-host-specific Salmonella serotypes in poultry: exploitation of innate immunity and microbial activity.

A recent European Union Directive required member states to put monitoring and control programmes in place, of which vaccination is a central component. Live Salmonella vaccines generally confer better protection than killed vaccines, because the former stimulate both cell-mediated and humoral immunity. Administering Salmonella bacteria orally to newly hatched chickens results in extensive gut colonization and a strong adaptive immune stimulus but broiler chickens are immunologically immature. However, colonization exerts a variety of rapid (within 24 h) protective effects. These include specific colonization-inhibition (competitive exclusion) in which the protective bacteria exert a profound resistance to establishment and colonization by other related bacteria. This is thought to be primarily a metabolic attribute of the vaccinating bacteria but may also involve competition for attachment sites. The presence of large numbers of bacteria originating from a live Salmonella vaccine in the intestine can also induce infiltration of polymorphonuclear cells into the intestinal wall, which confers resistance to invasion and systemic spread by virulent Salmonella strains. This opens new perspectives for vaccine usage in broilers, layers and breeding poultry but also in other animals which show increased susceptibility to infection because of their young age or for other reasons, such as oral chemoprophylaxis or chemotherapy, where the lack of established normal gut flora is an issue. We recommend that all live vaccines considered for oral administration should be tested for their ability to induce the two protective effects described above. Further developments in live Salmonella vaccines are, however, currently hindered by fears associated with the use and release of live vaccines which may be genetically modified.

Animals↗

PubMed2005

Supplementation of coated butyric acid in the feed reduces colonization and shedding of Salmonella in poultry.

Short-chain fatty acids have been widely used as feed additives to control Salmonella in poultry. Data on the use of butyric acid in poultry are lacking. In this study, powder form and coated butyric acid were compared in their ability to reduce Salmonella colonization of ceca and internal organs shortly after infection of young chickens with Salmonella enteritidis. In the first trial, 4 groups of 25 specific pathogen free layer chickens were given feed either supplemented with powder form butyric acid, coated butyric acid, a combination of powder form and coated butyric acid (all groups received a total of 0.63 g of butyric acid/kg) or nonsupplemented feed. The specific pathogen free layer chickens were orally infected with 10(6) cfu of S. enteritidis. Coated butyric acid significantly decreased cecal colonization 3 d post-infection compared with control chickens, and powder form butyric acid had no effect. To study long-term shedding and colonization of Salmonella in broilers given coated butyric acid as feed additive (0.63 g of active product butyric acid/kg), 10 Ross broiler chickens were infected at d 5 with 10(5) cfu of S. enteritidis and housed together with 40 noninfected broilers. A control group received nonsupplemented feed. The group of broilers receiving coated butyric acid had a significantly lower number of broilers shedding Salmonella bacteria, but cecal colonization at slaughter age was equal for both groups. In conclusion, butyric acid decreases cecal colonization shortly after infection, decreases fecal shedding, and as a consequence, decreases environmental contamination by S. enteritidis-infected broilers. However, complete elimination can probably only be achieved with a combined approach using both hygienic measures and different protection measures, as the broilers still carried S. enteritidis bacteria in the ceca at slaughter age, although at enrichment level.

Aging↗

PubMed2004

Bacteria-host interactions of Salmonella Paratyphi B dT+ in poultry.

In recent years, a dramatic increase in incidence of the dextro-rotatory tartrate-positive variant (dT+) of Salmonella enterica subspecies enterica serovar Paratyphi B has been observed in poultry and poultry products. In the present study the interactions of this bacterium with the host were studied in vivo and in vitro in an attempt to explain the preferential association of this serotype with poultry. The ability of this organism to invade and multiply in chicken intestinal epithelial cells and the intracellular behaviour in chicken macrophages was studied in vitro using chicken cell lines. In vivo challenge experiments in specific pathogen-free chickens were carried out to determine the level of colonization of caeca and internal organs early after experimental infection. An in vivo trial with commercial broiler chickens, using a seeder model, was performed to determine whether S. Paratyphi B dT+ could persist and spread in broilers until slaughter. S. Paratyphi B dT+ invaded and multiplied in the chicken epithelial cell line and survived in a chicken macrophage cell line. The strain used colonized caeca and internal organs of chickens to a high extent 1 week after infection with a low-dose inoculum. Moreover, the strain was efficiently transmitted within a group of broilers and persisted until slaughter. It was concluded that S. Paratyphi B dT+ was well adapted to poultry and therefore it is suggested that specific control measures against this serotype should be considered.

Animals↗

PubMed2004

Microencapsulated short-chain fatty acids in feed modify colonization and invasion early after infection with Salmonella enteritidis in young chickens.

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%) as spray-cooled microcapsules. The 5 groups were challenged with 5 x 10(3) cfu S. enteritidis at d 5 and 6 posthatch, and samples of ceca, liver, and spleen were taken at d 8 and analyzed for the number of colony-forming units of Salmonella per gram of tissue. Feed supplementation with acetic acid, and to a lesser extent formic acid, resulted in an increase of colonization of ceca and internal organs. Birds receiving propionic acid-coated microbeads as feed supplement were colonized with Salmonella to the same extent as controls. Butyric acid-impregnated microbeads in the feed, however, resulted in a significant decrease of colonization by S. enteritidis in the ceca but not in liver and spleen.

Animal Feed↗

PubMed2004

The impact of prebiotics and salmonellosis on apparent nutrient digestibility and Salmonella typhimurium var. Copenhagen excretion in adult pigeons (Columba livia domestica).

The effects of lactose or fructo-oligosaccharide (FOS) supplementation on the excretion of salmonellae, apparent digestibilities and excreta consistency were studied. Thirty-two male pigeons (Columba livia domestica) were randomly divided into 4 equal groups: 3 of 4 groups were orally infected with 10(9) Salmonella Typhimurium var. Copenhagen, after being offered a drinking water supplement of 2% FOS, 2% lactose, or no supplement, respectively, for 2 wk. Pigeons in the fourth group were not challenged with S. Typhimurium and remained unsupplemented. Initially, FOS increased water intake, resulting in more watery excreta. After infection, supplementation showed no major effects on S. Typhimurium excretion, nitrogen retention, or apparent nutrient digestibilities, although lactose--and to a lesser extent FOS--improved apparent fiber digestibility during recovery from the S. Typhimurium infection. The excreta consistency of all pigeons returned to normal when recovering from the Salmonella infection. In this trial, neither FOS nor lactose was successful in tempering the negative aspects of Salmonella infection in pigeons. Nevertheless, it should be stated that future investigations should clarify the importance of duration and level of prebiotic supplementation and infection level.

Animal Feed↗

PubMed2004

Intermittent long-term shedding and induction of carrier birds after infection of chickens early posthatch with a low or high dose of Salmonella enteritidis.

Poultry are very likely to become infected with Salmonella in the early posthatch period, due to environmental contamination. The purpose of this study was to evaluate the effect of infection dose on the risk of persistent infection in laying hens. In this study, young layer chicks were orally infected with a low (10(2) at 1 d posthatch) or a high dose (10(9) cfu at 1 wk posthatch) of Salmonella Enteritidis. The pattern and duration of fecal shedding was studied for 18 wk. All chickens shed Salmonella early after infection and shed Salmonella intermittently during the whole study period. There were more positive birds in the high-dose group than in the low-dose group in the first weeks following Salmonella exposure. From 10 wk postinfection onwards, however, birds that were orally infected with the low dose of Salmonella Enteritidis shed more Salmonella than the birds that received the high dose. At 18 wk of age, there was no difference in cecal colonization between the treatment groups. It can be concluded that infection of newly hatched chicks with a low dose of Salmonella Enteritidis can lead to persistent infection until onset of lay, hereby excreting Salmonella bacteria intermittently.

Animals↗

PubMed2004

Tubular glands of the isthmus are the predominant colonization site of Salmonella enteritidis in the upper oviduct of laying hens.

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 investigation of the invasiveness of S. enteritidis in the oviduct of laying hens. Loops in the isthmus and magnum of laying hens were made under anaesthesia. S. enteritidis was inoculated into each loop. After 1 h, tubular gland cells were isolated from the infected tissue under gentamicin. S. enteritidis invasiveness was measured as the ratio of isolated bacteria per isolated tubular gland cell. This ratio was higher (P < 0.01) in the isthmus (1.3 x 10(-3)) than in the magnum (5.3 x 10(-5)). In a third experiment, laying hens were intravenously infected with 5 x 10(7) cfu S. enteritidis bacteria. The number of intracellular bacteria was counted in the tubular gland cells of the isthmus and magnum after isolation under gentamicin. The ratio of isolated bacteria per isolated tubular gland cell was again significantly higher in the isthmus as compared with in the magnum. In all 3 assays, the tubular gland cells of the isthmus were more heavily invaded than those of the magnum.

Animals↗

PubMed2004

Colonization of the chicken reproductive tract and egg contamination by Salmonella.

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Animals↗

PubMed2004

Medium-chain fatty acids decrease colonization and invasion through hilA suppression shortly after infection of chickens with Salmonella enterica serovar Enteritidis.

The most common source of Salmonella infections in humans is food of poultry origin. Salmonella enterica serovar Enteritidis has a particular affinity for the contamination of the egg supply. In this study, the medium-chain fatty acids (MCFA), caproic, caprylic, and capric acid, were evaluated for the control of Salmonella serovar Enteritidis in chickens. All MCFA were growth inhibiting at low concentrations in vitro, with caproic acid being the most potent. Contact of Salmonella serovar Enteritidis with low concentrations of MCFA decreased invasion in the intestinal epithelial cell line T84. By using transcriptional fusions between the promoter of the regulatory gene of the Salmonella pathogenicity island I, hilA, and luxCDABE genes, it was shown that all MCFA decreased the expression of hilA, a key regulator related to the invasive capacity of Salmonella. The addition of caproic acid (3 g/kg of feed) to the feed of chicks led to a significant decrease in the level of colonization of ceca and internal organs by Salmonella serovar Enteritidis at 3 days after infection of 5-day-old chicks. These results suggest that MCFA have a synergistic ability to suppress the expression of the genes required for invasion and to reduce the numbers of bacteria in vivo. Thus, MCFA are potentially useful products for reducing the level of colonization of chicks and could ultimately aid in the reduction of the number of contaminated eggs in the food supply.

Animals↗

PubMed2003

Invasion of Salmonella enteritidis in avian intestinal epithelial cells in vitro is influenced by short-chain fatty acids.

Fermentation reactions in the caeca of chickens, the predominant place for Salmonella colonization, result in high concentrations of short-chain fatty acids (SCFA). Thus Salmonella bacteria are in close contact with SCFA during their life cycle. A study was carried out to analyse the effects of SCFA on invasion of Salmonella enteritidis in an avian intestinal epithelial cell line. Preincubation of S. enteritidis for 4 h in growth media supplemented with various concentrations of propionate or butyrate resulted in decreased invasion compared to bacteria, preincubated in nonsupplemented media, and to bacteria, preincubated in media supplemented with formate or acetate. Incubation of the S. enteritidis bacteria in media supplemented with mixtures of SCFA mimicking the in vivo caecal concentrations resulted in increased invasion compared with butyrate-exposed bacteria, but equal invasion compared with nonexposed bacteria. Increasing the butyrate concentration in these mixtures did not modify invasion compared with the original mixtures.

Animals↗

PubMedDate not supplied

Interactions of Salmonella enterica subsp. enterica serovar Muenchen with intestinal explants of the turtle Trachemys scripta scripta.

Salmonella infections in reptiles, in contrast to those in birds and mammals, are limited to the intestinal tract. In this study, interactions of a strain of Salmonella enterica subsp. enterica serovar Muenchen (SEEM) with intestinal explants of the turtle Trachemys scripta scripta were examined by scanning electron microscopy (SEM). Adhesion and invasion in the chelonian intestinal explants at 30 degrees C and 37 degrees C were evaluated quantitatively. For purposes of comparison, the invasive capacity of SEEM in the continuous avian epithelial cell line DIV-1 at 30 degrees C and 37 degrees C was determined. Small numbers of M-like cells were found in the ileum of the turtles. The bacteria adhered mainly to the mucus of the intestinal explants. Only small numbers of salmonellae were associated with epithelial cells. Higher numbers of bacteria adhered at 30 degrees C than at 37 degrees C. Epithelial damage, embedding of bacteria in the epithelial surface and a ruffling-like process were noted only at 37 degrees C. Minimal numbers of salmonellae invaded the explants at 30 degrees C and 37 degrees C. Invasion of DIV-1 cells was greater at 37 degrees C than at 30 degrees C. The study suggested that the intestinal mucous layer provides an important site of colonization for salmonellae in the chelonian host and protects the underlying epithelial cells.

Animal Diseases↗