An in-depth investigation of the microbiota and its virulence factors associated with severe udder cleft dermatitis lesions.

Udder cleft dermatitis (UCD) is a skin condition affecting the anterior parts of the udder in dairy cattle. In the present study, we aimed to shed light on the microbiota in severe UCD lesions versus healthy udder skin by putting forward a taxonomic and functional profile based on a virulence factor analysis. Through shotgun metagenomic sequencing, we found a high proportion of bacteria in addition to a low abundance of archaea. A distinct clustering of healthy udder skin versus UCD lesion samples was shown by applying principal component analysis and (sparse) partial least squares analysis on the metagenomic data. Proteobacteria, Bacillota, and Actinomycetota were among the most abundant phyla in healthy udder skin samples. In UCD samples, Bacteroidota was the most abundant phylum. At genus level, Bifidobacterium spp. had the highest relative abundance in healthy skin samples, whereas Porphyromonas spp. and Corynebacterium spp. had the highest relative abundance in UCD samples. In the differential abundance analysis, Porphyromonas spp. and Bacteroides spp. were significantly differentially abundant in UCD samples, whereas Bifidobacterium spp., Staphylococcus sp. AntiMn-1, and Staphylococcus equorum were more commonly found in healthy samples. Moreover, the abundance of several treponeme phylotypes was significantly higher in lesion samples. The streptococcal cysteine protease speB was among the most abundant virulence factors present in severe UCD lesions, while a plethora of virulence factors such as the antitoxin relB were downregulated, possibly contributing to creating the ideal wound climate for the dysbiotic community. Network analysis showed healthy lesion samples had a large network ofpositive, correlations between the abundances of beneficial species such as Aerococcus urinaeequi and Bifidobacterium angulatum, indicating that the healthy skin microbiome forms an active protective bacterial network, which is disrupted in case of UCD. In UCD samples, a smaller microbial network mainly consisting of positive correlations between the abundances of Bacteroides fragilis and anaerobic Bacteroidota was exposed. Moreover, a high correlation between the taxonomic data and virulence factors was revealed, concurrently with 2 separate networks of microbes and virulence factors. One network, matching with the taxonomic findings in the healthy udder skin samples, showcased a community of harmless or beneficial bacteria, such as Bifidobacterium spp. and Butyrivibrio proteoclasticus, associated with hcnB, hcnC, relB, glyoxalase, and cupin 2. The other network, corresponding to UCD samples, consisted of pathogenic or facultative pathogenic and mainly anaerobic bacteria such as Treponema spp., Mycoplasmopsis spp., and bovine gammaherpesvirus 4, that correlated with virulence factors SpvB, fhaB, and haemagglutination activity domain-associated factor. Our results point toward a dysbiotic community with a notable decrease in diversity and evenness, with a loss of normal skin inhabitants and innocuous or useful species making way for predominantly anaerobic, facultative pathogens. The shift in the abundance of virulence factors such as fhaB and SpvB could play a role in the manifestation of a local micro-environment favorable to the microbiome associated with udder skin lesions. Lastly, the presence of specific networks between microbial species, and between microbes and virulence factors was shown.

Severe udder cleft dermatitis lesion transcriptomics points to an impaired skin barrier, defective wound repair and a dysregulated inflammatory response as key elements in the pathogenesis.

This study is the first to investigate the transcriptomic changes occurring in severe udder cleft dermatitis lesions (UCD) in Holstein-Friesian cows. An examination of the gene expression levels in natural UCD lesions and healthy udder skin through RNA Seq-Technology provided a deeper insight into the inflammatory pathways associated with this disease. A clear distinction between the gene expression patterns of UCD lesions and healthy skin was shown in the principal component analysis. Genes coding for inflammatory molecules were upregulated such as the chemokines C-X-C motif ligand 2 (CXCL2), 5 (CXCL5) and 8 (CXCL8), and C-C motif ligand 11 (CCL11). Moreover, the genes coding for the multifunctional molecules ADAM12 and SLPI were amongst the highest upregulated ones, whereas the most downregulated genes included the ones coding for keratins and keratin-associated molecules. Predominantly inflammatory pathways such as the chemokine signaling, cytokine receptor interaction and IL-17 signaling pathway were significantly upregulated in the pathway analysis. These results point towards a fulminant, dysregulated inflammatory response concomitant with a disruption of the skin barrier integrity and a hampered wound repair mechanism in severe UCD lesions.

A field study on correlations between macroscopic gut health scoring, histological measurements and performance parameters in broilers.

Optimal intestinal health is a prerequisite for sustainable poultry production. Intestinal health can be evaluated by scoring macroscopic intestinal abnormalities and by histological measurements. The aim of this study was to evaluate correlations between macroscopic gut appearance scoring (GAS), coccidiosis lesion scoring, histological measurements and performance parameters in broilers under field conditions. Therefore, a cross-sectional study was performed on 50 broiler farms where birds were sampled at 28 days of age. The GAS was determined by scoring the absence or presence of 10 macroscopically visible parameters of the gastrointestinal tract, yielding a GAS between 0 and 10, with 0 meaning no gut abnormalities. On individual bird level, when birds had a GAS score of 6 or higher, significantly shorter villi were found in the duodenum. Also, CD3+ T-lymphocyte area percentage in the duodenal mucosa was significantly negatively correlated to villus length. On farm level, the average feed conversion ratio (FCR) was 1.59 ± 0.04 [1.52-1.73]. However, no correlations were found between the GAS at the age of 28 days and the prevalence of coccidiosis, nor did coccidiosis lesion scoring correlate with the FCR. Also, a higher GAS could not be associated with a higher FCR. In conclusion, on all farms a certain degree of macroscopic visible gut and coccidiosis lesions were present in birds of 28 days of age, while this did not correlate with the FCR. This suggests that broilers are able to cope with a certain level of gut damage before it influences the overall performance.

Dietary supplementation of micro-encapsulated sodium butyrate in healthy horses: effect on gut histology and immunohistochemistry parameters.

BACKGROUND: As colic and intestinal disorders are a major concern in horses, the aim of the present study was to investigate the effect of dietary supplementation of butyrate, known to have a diverse array of beneficial effects on intestinal health. The effect of micro-encapsulated sodium butyrate supplementation on gut histology and immunohistochemistry parameters was studied in 14 healthy warmblood horses destined for slaughter in two separate periods. Horses were fed a low fiber - high starch diet, designed to induce subsequent starch overflow in the large intestine, aiming to create a mild challenge for large intestinal health. Treatment included supplementation with either micro-encapsulated sodium butyrate (Excential Butycoat®, Orffa, Werkendam, the Netherlands) or placebo (containing only coating material). The horses were fed for 20 consecutive days at a dosage of 0.4 g/kg BW (body weight). At day 21, the horses were slaughtered and intestinal samples were collected for determination of gut pH, villus length, crypt depth and area % of CD3+ and CD20+ cells. RESULTS: Horses on the butyrate supplemented diet had significantly reduced crypt depths in the right dorsal colon compared to placebo-fed horses (P < 0.001). However, a treatment x period interaction (P = 0.002) was discovered regarding this parameter, which could not be explained by the authors. Further investigation into the number of KI67+ cells in the RDC crypts did not reveal any significant differences between treatments (P = 0.650), indicating that the reduction in crypt depth in butyrate-fed horses could not be explained by a significant difference in cellular proliferation. Intestinal pH, villus length and expression of intestinal CD3+ and CD20+ cells were not significantly affected by treatment at any intestinal level. CONCLUSIONS: Our data indicate that supplementation of micro-encapsulated sodium butyrate to the equine diet did not influence gut histology (with the exception of a decrease found in the crypts of the RDC) or immunohistochemistry parameters in healthy horses. Further research is warranted to investigate the impact of butyrate supplementation in horses with intestinal disease.

Spotlight on avian pathology: untangling contradictory disease descriptions of necrotic enteritis and necro-haemorrhagic enteritis in broilers.

Necrotic enteritis (NE) is one of the most detrimental infectious diseases in the modern poultry industry, characterized by necrosis in the small intestine. It is commonly accepted that NetB-producing C. perfringens type G strains are responsible for the disease. However, based on both macroscopic and histopathological observations, two distinct types of NE are observed. To date, both a haemorrhagic form of NE and the type G-associated non-haemorrhagic disease entity are commonly referred to as NE and the results from scientific research are interchangeably used, without distinguishing between the disease entities. Therefore, we propose to rename the haemorrhagic disease entity to necro-haemorrhagic enteritis.

Antimicrobial susceptibility pattern of Helicobacter suis isolates from pigs and macaques.

Helicobacter suis is a fastidious, Gram negative bacterium that colonizes the stomach of pigs and non-human primates. It has also been associated with gastric disease in humans. A combined agar and broth dilution method was used to analyze the activity of 15 antimicrobial agents against 20 and 15 H. suis isolates obtained from pigs and macaques, respectively. After 48 h microaerobic incubation, minimal inhibitory concentrations (MICs) were determined by software-assisted calculation of bacterial growth as determined by quantitative real-time PCR. A monomodal distribution of MICs was seen for ß-lactam antibiotics, macrolides, gentamicin, neomycin, doxycycline, metronidazole, and rifampicin. Presence of a bimodal distribution of MICs indicated that 2 porcine isolates did not belong to the wild type population (WTP) for fluoroquinolones. This was also the case for 1 porcine isolate for tetracycline, 1 porcine and 2 primate isolates for lincomycin, and 1 primate isolate for spectinomycin. Single nucleotide polymorphisms (SNPs) were present in the gyrA gene of the isolates not belonging to the WTP for fluoroquinolones and in ribosomal protein encoding genes of the isolates not belonging to the WTP for tetracycline and spectinomycin. MICs of ampicillin, tetracycline and doxycycline were higher for porcine H. suis isolates compared to primate isolates and in these porcine isolates SNPs were detected in genes encoding penicillin binding and ribosomal proteins. This study indicates that acquired resistance occasionally occurs in H. suis isolates and that zoonotically important porcine isolates may be intrinsically less susceptible to ß-lactam antibiotics and tetracyclines than primate isolates.

Rapid growth predisposes broilers to necrotic enteritis.

Over the past 50 years, intentional genetic selection within the broiler industry has led to major improvements in both body weight gain (BWG) and feed conversion efficiency. Next to its economic advantages, enhancing BWG can increase the risk of metabolic and skeletal disorders. The aim of this study was to examine whether higher BWG is a predisposing factor for broiler necrotic enteritis. In this study, 300 broilers were challenged with Clostridium perfringens using a well-established, previously described challenge model. It was found that birds with higher body weight (BW) and BWG before challenge were predisposed to develop more severe necrotic enteritis lesions. After challenge, the average BWG of the birds developing mild to severe lesions dropped significantly, negatively affecting bird welfare and performance. These results show a significant interplay between BWG and the development of necrotic enteritis lesions. This raises the question whether there is a limit to broiler performance with respect to maintaining intestinal health, and whether decreasing BWG (at certain stages of the growth cycle) can be part of a plan to prevent intestinal pathology. RESEARCH HIGHLIGHTS Higher body weight is a predisposing factor to necrotic enteritis in broilers.

The Salmonella Enteritidis TolC outer membrane channel is essential for egg white survival.

Salmonella Enteritidis has developed the potential to contaminate eggs by surviving in the antimicrobial environment of the hen's egg white. This has led to a worldwide pandemic of foodborne salmonellosis infections in humans due to the consumption of contaminated eggs and egg-derived products. The molecular mechanisms of Salmonella Enteritidis egg white survival are not fully clear. Using in vivo expression technology and promoter-reporter fusions we showed that the promoter of the tolC gene, encoding the TolC outer membrane channel that is used by multidrug efflux pumps to export harmful molecules and to secrete bacterial products, is activated by egg white at the chicken body temperature. Using a Salmonella Enteritidis tolC deletion mutant we showed that TolC has an important role in egg white survival. Chromatographic separation techniques and subsequent testing of antimicrobial activities of separated egg white fractions led to the identification of ovotransferrin as the egg white antimicrobial factor which is capable of inhibiting growth of a tolC deletion strain but not the wild type strain. We provide evidence that TolC protects Salmonella Enteritidis against ovotransferrin-mediated growth inhibition in egg white.

Heat stress as a predisposing factor for necrotic enteritis in broiler chicks.

Heat stress is a physical environmental stressor, which can affect performance, health and welfare of poultry. The present study investigates the effect of cyclic acute heat stress as a predisposing factor for necrotic enteritis in broiler chicks in an experimental challenge model. Two hundred and forty broiler chicks were randomly allocated to four treatment groups, as follows: group A served as negative control (25°C), group B birds were subjected to cyclic acute heat stress (35°C), group C birds were challenged and group D birds were both subjected to heat stress and challenged. From each bird, the intestine, gizzard and liver were collected and scored for gross lesions. The intestinal digesta was collected for pH and viscosity determination. One caecum from each bird was taken for microbiological analysis. The statistical analysis and evaluation of data revealed that the heat stress in challenged birds showed a relative trend to increase the severity and the incidence of necrotic enteritis lesions, although it was not considered as statistically significant (Ρ = 0.077). Additionally, the heat stress induced necrotic enteritis outbreak in unchallenged birds; the challenge of birds as well as its combination with the heat stress affected significantly (Ρ ≤ 0.05) the pH and viscosity of intestinal digesta and the caecal Clostridium perfringens counts. The study provides evidence that cyclic acute heat stress is an environmental stressor, which can significantly affect necrotic enteritis and gut health, and thus should be taken into account in warmer areas of the world where poultry farming becomes a major industry.

The role of infectious agents in the development of porcine gastric ulceration.

Ulceration of the non-glandular part of the stomach is a common disease entity of pigs worldwide, with prevalences of up to 93%. It may result in decreased daily weight gain, decreased feed intake and sudden death, thus leading to significant economic losses, as well as animal welfare issues. The aetiology is multifactorial. Diet particle size, management and infectious agents have been hypothesised to be involved. The exact mechanism behind porcine gastric ulceration is, however, not completely clear. The aim of this article is to provide an overview of the role of infectious agents in the development of porcine gastric ulceration. Results of recent studies indicate that Helicobacter suis infection plays an important role in gastric ulceration, probably by affecting gastric acid secretion through alteration of the number and/or function of parietal, D and G cells. In a gastric environment altered by H. suis, higher numbers of Fusobacterium gastrosuis are present and this novel pathogen has a potential role in the development of porcine gastric ulceration. Inoculation of pigs with Lactobacillus sp., Bacillus sp. or Helicobacter pylori-like bacteria in combination with a carbohydrate-rich diet may induce gastric lesions. It has been hypothesised that production of short chain fatty acids by some of these bacteria might be involved in the pathogenesis of porcine gastric ulceration, but the lack of taxonomic characterisation hampers the interpretation of these studies. Severe infectious diseases have also been associated with gastric lesions, probably due to interruption in feed intake and/or histamine release. Other agents, including fungi and parasites such as Hyostrongylus rubidus and Ascaris suum, have occasionally been associated with gastric lesions in pigs.

Presence of Helicobacter and Campylobacter species in faecal samples from zoo mammals.

Helicobacter and Campylobacter species (spp.) colonize the gastrointestinal tract of various domesticated animals. Apart from their pathogenic significance in animals, several species are of zoonotic importance as well. For most non-domesticated animal spp., however, little is known on the presence and importance of these agents. Therefore, we investigated the presence of Helicobacter and Campylobacter spp. in marine and terrestrial zoo mammals. Faecal samples of various marine and terrestrial zoo mammals were collected from 6 different zoos in Belgium. These samples were tested for the presence of Helicobacter and Campylobacter spp. by isolation and direct demonstration of DNA using genus-specific PCRs, followed by sequencing of the obtained amplicons. Helicobacter spp. were detected in 12 and Campylobacter spp. in 5 of the 22 faecal samples from marine mammals. In 4 of 5 dolphins, H. cetorum was demonstrated, a well-known pathogen associated with gastritis and gastric ulceration in marine mammals. C. insulaenigrae was isolated from 4 of 6 sea lions and from 1 of 11 seals. To our knowledge, this is the first description of the presence of C. insulaenigrae on the European mainland. Helicobacter spp. were detected in 5 and Campylobacter spp. (mainly C. jejuni subsp. jejuni and C. coli) in 9 of the 26 faecal samples from terrestrial mammals. Potential novel enterohepatic Helicobacter spp. were detected in both marine and terrestrial zoo mammals. For the first time, the presence of several known and unknown Campylobacter and Helicobacter spp. was demonstrated in the gastrointestinal tract of various marine and terrestrial zoo mammals. Further investigation is needed on the pathogenic and zoonotic importance of these species.

Scoring biosecurity in European conventional broiler production.

Good biosecurity procedures are crucial for healthy animal production. The aim of this study was to quantify the level of biosecurity on conventional broiler farms in Europe, following a standardized procedure, thereby trying to identify factors that are amenable to improvement. The current study used a risk-based weighted scoring system (biocheck.ugent ®) to assess the level of biosecurity on 399 conventional broiler farms in 5 EU member states. The scoring system consisted of 2 main categories, namely external and internal biosecurity, which had 8 and 3 subcategories, respectively. Biosecurity was quantified by converting the answers to 97 questions into a score from 0 to 100. The minimum score, "0," represents total absence of any biosecurity measure on the broiler farm, whereas the maximum score, "100," means full application of all investigated biosecurity measures. A possible correlation between biosecurity and farm characteristics was investigated by multivariate linear regression analysis. The participating broiler farms scored better for internal biosecurity (mean score of 76.6) than for external biosecurity (mean 68.4). There was variation between the mean biosecurity scores for the different member states, ranging from 59.8 to 78.0 for external biosecurity and from 63.0 to 85.6 for internal biosecurity. Within the category of external biosecurity, the subcategory related to "infrastructure and vectors" had the highest mean score (82.4), while the subcategory with the lowest score related to biosecurity procedures for "visitors and staff" (mean 51.5). Within the category of internal biosecurity, the subcategory "disease management" had the highest mean score (65.8). In the multivariate regression model a significant negative correlation was found between internal biosecurity and the number of employees and farm size. These findings indicate that there is a lot of variation for external and internal biosecurity on the participating broiler farms, suggesting that improvements are possible. Since the subcategory "visitors and staff" scored the lowest, better education of broiler farmers and their staff may help to improve overall biosecurity on broiler farms in Europe.

Short-chain arabinoxylans prepared from enzymatically treated wheat grain exert prebiotic effects during the broiler starter period.

Carbohydrate-degrading multi-enzyme preparations (MEP) are used to improve broiler performances. Their mode of action is complex and not fully understood. In this study, we compared the effect of water-soluble fractions isolated at the pilot scale from wheat grain incubated with (WE) and without (WC) MEP. The fractions were incorporated in a wheat-based diet (0.1% w/w) to feed Ross PM3 broilers and compared with a non-supplemented control group (NC). The body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) until d 14 were determined. At d 14, ileal and cecal contents and tissue samples were collected from euthanized animals. The intestinal contents were used to measure the short-chain fatty acids (SCFA) concentration using gas chromatography and to determine the abundance and composition of microbiota using 16S sequencing. Villi length of ileal samples was measured, while L-cell and T-cell densities were determined using immuno-histochemistry. The MEP treatment increased the amount of water-soluble arabinoxylans (AX) and reduced their molecular weight while retaining their polymer behavior. The WE fraction significantly (P < 0.05) increased FI by 13.8% and BWG by 14.7% during the first wk post hatch when compared to NC. No significant effect on FCR was recorded during the trial. The WE increased the abundance of Enterococcus durans and Candidatus arthromitus in the ileum and of bacteria within the Lachnospiraceae and Ruminococcaceae families, containing abundant butyrate-producing bacteria, in the ceca. It also increased the concentration of SCFA in the ceca, decreased the T-lymphocyte infiltration in the intestinal mucosa, and increased the glucagon-like-peptide-2 (GLP-2)-producing L-cell density in the ileal epithelium compared with WC and NC. No significant effects were observed on villi length. These results showed that AX present in the WE fraction altered the microbiota composition towards butyrate producers in the ceca. Butyrate may be responsible for the reduction of inflammation, as suggested by the decrease in T-lymphocyte infiltration, which may explain the higher feed intake leading to improved animal growth.

The choroid plexus epithelium as a novel player in the stomach-brain axis during Helicobacter infection.

Several studies suggest a link between shifts in gut microbiota and neurological disorders. Recently, we reported a high prevalence of Helicobacter suis (H. suis) in patients with Parkinson's disease. Here, we evaluated the effect of gastric H. suis infection on the brain in mice. One month of infection with H. suis resulted in increased brain inflammation, reflected in activation of microglia and cognitive decline. Additionally, we detected choroid plexus inflammation and disruption of the epithelial blood-cerebrospinal fluid (CSF) barrier upon H. suis infection, while the endothelial blood-brain barrier (BBB) remained functional. These changes were accompanied by leakage of the gastrointestinal barrier and low-grade systemic inflammation, suggesting that H. suis-evoked gastrointestinal permeability and subsequent peripheral inflammation induces changes in brain homeostasis via changes in blood-CSF barrier integrity. In conclusion, this study shows for the first time that H. suis infection induces inflammation in the brain associated with cognitive decline and that the choroid plexus is a novel player in the stomach-brain axis.

Helicobacter suis induces changes in gastric inflammation and acid secretion markers in pigs of different ages.

Gastric mRNA expression of markers for acid secretion and inflammation and presence of gastric ulceration was studied in naturally Helicobacter suis-infected and non-infected 2-3 months old, 6-8 months old and adult pigs. In H. suis-infected 2-3 months old pigs, IL-8 and IL-1ß transcript levels were upregulated in the pyloric gland zone, indicating an innate immune response. A similar response was demonstrated in the fundic gland zone of adult pigs, potentially due to a shift of H. suis colonization from the pyloric to the fundic gland zone. A Treg response in combination with decreased expressions of IL-8, IL-17A and IFN-γ was indicated to be present in the H. suis-infected 6-8 months old pigs, which may have contributed to persistence of H. suis. In H. suis-infected adult pigs, a Treg response accompanied by a Th17 response was indicated, which may have played a role in the decreased number of H. suis bacteria in the stomach of this age group. The decreased G-cell mass and upregulated expression of somatostatin indicated decreased acid secretion in H. suis-infected 6-8 months old pigs. In H. suis-infected adult pigs, upregulation of most markers for gastric acid secretion and increased G-cell mass was detected. Presence of severe hyperkeratosis and erosions in the non-glandular part of the stomach were mainly seen in the H. suis-positive groups. These results show that H. suis infection affects the expression of markers for acid secretion and inflammation and indicate that these effects differ depending on the infection phase.

A Deep Nasopharyngeal Swab Versus Nonendoscopic Bronchoalveolar Lavage for Isolation of Bacterial Pathogens from Preweaned Calves With Respiratory Disease.

BACKGROUND: Nonendoscopic bronchoalveolar lavage (BAL) is a practical alternative for a deep nasopharyngeal swab (DNS) to sample the airways of a large number of calves in a short period of time. The extent of commensal overgrowth and agreement of BAL with DNS culture results in preweaned calves are unknown. OBJECTIVES: To compare commensal overgrowth and bacterial culture results between DNS and BAL samples. ANIMALS: A total of 183 preweaned calves (144 with bovine respiratory disease and 39 healthy animals). METHODS: Cross-sectional study. Deep nasopharyngeal swab and BAL samples were taken from each calf and cultured to detect Pasteurellaceae and Mycoplasma bovis. Agreement and associations between culture results of DNS and BAL samples were determined by kappa statistics and logistic regression. RESULTS: Bronchoalveolar lavage samples were less often polymicrobial, more frequently negative and yielded more pure cultures compared to DNS, leading to a clinically interpretable culture result in 79.2% of the cases compared to only in 31.2% of the DNS samples. Isolation rates were lower in healthy animals, but not different between DNS and BAL samples. Only Histophilus somni was more likely to be isolated from BAL samples. In clinical cases, a polymicrobial DNS culture result did not increase the probability of a polymicrobial BAL result by ≥30%, nor did it influence the probability of a negative culture. A significant herd effect was noted for all observed relationships. CONCLUSIONS AND CLINICAL RELEVANCE: Nonendoscopic BAL samples are far less overgrown by bacteria compared to DNS samples under the conditions of this study, facilitating clinical interpretation and resulting in a higher return on investment in bacteriologic culturing.

Reduced particle size wheat bran is butyrogenic and lowers Salmonella colonization, when added to poultry feed.

Feed additives, including prebiotics, are commonly used alternatives to antimicrobial growth promoters to improve gut health and performance in broilers. Wheat bran is a highly concentrated source of (in)soluble fiber which is partly degraded by the gut microbiota. The aim of the present study was to investigate the potential of wheat bran as such to reduce colonization of the cecum and shedding of Salmonella bacteria in vivo. Also, the effect of particle size was evaluated. Bran with an average reduced particle size of 280µm decreased levels of cecal Salmonella colonization and shedding shortly after infection when compared to control groups and groups receiving bran with larger particle sizes. In vitro fermentation experiments revealed that bran with smaller particle size was fermented more efficiently, with a significantly higher production of butyric and propionic acid, compared to the control fermentation and fermentation of a larger fraction. Fermentation products derived from bran with an average particle size of 280µm downregulated the expression of hilA, an important invasion-related gene of Salmonella. This downregulation was reflected in an actual lowered invasive potential when Salmonella bacteria were pretreated with the fermentation products derived from the smaller bran fraction. These data suggest that wheat bran with reduced particle size can be a suitable feed additive to help control Salmonella infections in broilers. The mechanism of action most probably relies on a more efficient fermentation of this bran fraction and the consequent increased production of short chain fatty acids (SCFA). Among these SCFA, butyric and propionic acid are known to reduce the invasion potential of Salmonella bacteria.

Differences in extracellular matrix proteins between Friesian horses with aortic rupture, unaffected Friesians and Warmblood horses.

BACKGROUND: Unlike in Warmblood horses, aortic rupture is quite common in Friesian horses, in which a hereditary trait is suspected. The aortic connective tissue in affected Friesians shows histological changes such as medial necrosis, elastic fibre fragmentation, mucoid material accumulation and fibrosis with aberrant collagen morphology. However, ultrastructural examination of the collagen fibres of the mid-thoracic aorta has been inconclusive in further elucidating the pathogenesis of the disease. OBJECTIVES: To assess several extracellular matrix (ECM) components biochemically in order to explore a possible underlying breed-related systemic ECM defect in Friesians with aortic rupture. STUDY DESIGN: Cadaver study. METHODS: Tissues from affected Friesians (n = 18), unaffected Friesians (n = 10) and Warmblood horses (n = 30) were compared. Samples were taken from the thoracic aorta at the level of the rupture site, from two locations caudal to the rupture and from the deep digital flexor tendon. Total collagen content, post-translational modifications of collagen formation including lysine hydroxylation, and hydroxylysylpyridinoline (HP), lysylpyridinoline (LP) and pyrrole cross-links were analysed. Additionally, elastin cross-links, glycosaminoglycan content and matrix metalloproteinase (MMP) activity were assessed. RESULTS: Significantly increased MMP activity and increased LP and HP cross-linking, lysine hydroxylation and elastin cross-linking were found at the site of rupture in affected Friesians. These changes may reflect processes involved in healing and aneurysm formation. Unaffected Friesians had less lysine hydroxylation and pyrrole cross-linking within the tendons compared with Warmblood horses. No differences in the matrix of the aorta were found between normal Warmbloods and Friesian horses. MAIN LIMITATIONS: Small sample size. CONCLUSIONS: The differences in collagen parameters in tendon tissue may reflect differences in connective tissue metabolism between Friesians and Warmblood horses.

Detection, isolation and characterization of Fusobacterium gastrosuis sp. nov. colonizing the stomach of pigs.

Nine strains of a novel Fusobacterium sp. were isolated from the stomach of 6-8 months old and adult pigs. The isolates were obligately anaerobic, although they endured 2h exposure to air. Phylogenetic analysis based on 16S rRNA and gyrase B genes demonstrated that the isolates showed high sequence similarity with Fusobacterium mortiferum, Fusobacterium ulcerans, Fusobacterium varium, Fusobacterium russii and Fusobacterium necrogenes, but formed a distinct lineage in the genus Fusobacterium. Comparative analysis of the genome of the type strain of this novel Fusobacterium sp. confirmed that it is different from other recognized Fusobacterium spp. DNA-DNA hybridization, fingerprinting and genomic %GC determination further supported the conclusion that the isolates belong to a new, distinct species. The isolates were also distinguishable from these and other Fusobacterium spp. by phenotypical characterization. The strains produced indole and exhibited proline arylamidase and glutamic acid decarboxylase activity. They did not hydrolyse esculin, did not exhibit pyroglutamic acid arylamidase, valine arylamidase, α-galactosidase, ß-galactosidase, ß-galactosidase-6-phosphate or α-glucosidase activity nor produced acid from cellobiose, glucose, lactose, mannitol, mannose, maltose, raffinose, saccharose, salicin or trehalose. The major fatty acids were C16:0 and C18:1ω9c. The name Fusobacterium gastrosuis sp. nov. is proposed for the novel isolates with the type strain CDW1(T) (=DSM 101753(T)=LMG 29236(T)). We also demonstrated that Clostridium rectum and mortiferum Fusobacterium represent the same species, with nomenclatural priority for the latter.