Lotte Bjerrum

The cereal type in feed influences gut wall morphology and intestinal immune cell infiltration in broiler chickens

In broiler chickens, a diet where the major cereal types are wheat, rye and/or barley has a lower digestibility compared with a diet in which maize is the major cereal type. In the present study, the effects of two different dietary cereal types, maize v. wheat/rye, on host factors (inflammation and gut integrity) and gut microbiota composition were studied. In addition, the effects of low-dose Zn-bacitracin supplementation were examined. Broilers given a wheat/rye-based diet showed more villus fusion, a thinner tunica muscularis, more T-lymphocyte infiltration, higher amount of immune cell aggregates in the mucosa, more and larger goblet cells and more apoptosis of epithelial cells in the mucosa than those given a maize-based diet. Adding Zn-bacitracin generally reversed these alterations. The microbiota composition was analysed by the use of terminal-restriction fragment length polymorphism, showing changes in the microbiota composition depending on the cereal type used in the diets. The effect of the change of cereal type on the gut microbiota composition was larger than that of Zn-bacitracin supplementation. In conclusion, a wheat/rye-based diet evoked mucosal damage, an alteration in the composition of the microbiota and an inflammatory bowel type of condition.

The influence of the cage system and colonisation of Salmonella Enteritidis on the microbial gut flora of laying hens studied by T-RFLP and 454 pyrosequencing

BackgroundIn the EU conventional cages for laying hens are forbidden beginning in January 2012, however concerns about a higher transmission rate of Salmonella in alternative cages systems have been raised. The extent to which cage systems may affect the intestinal microbiota of laying hens is not known, and different microbiota may demonstrate different resistance towards colonization with Salmonella. To investigate this, ileal and caecal samples from two experimental studies where laying hens were inoculated with Salmonella Enteritidis and housed in different systems (conventional cage, furnished cage or aviary), were compared using Terminal Restriction Fragment Length Polymorphism (T-RFLP). The distribution of genera in the microbiota in caecum was furthermore described by next generation sequencing of 16S rDNA libraries.ResultsHens in the same cage type developed similar T-RFLP fingerprints of the ileal and caecal microbiota, and these could be separated from layers in the other cages types. No significant difference in the fingerprint profiles could be observed between Salmonella positive and negative samples from same cage. By deep sequencing of 16S rDNA libraries from caecum, 197 different Operational Taxonomic Units (OTU) were identified, and 195 and 196 OTU respectively, were found in hens in aviary and furnished cages, but only 178 OTU of these were recovered from conventional cages. The ratio between the dominating phyla or families and genera in the microbiota remained fairly constant throughout the study. Faecalibacterium and Butyricimonas were the most prevalent genera found in the caecal microbiota of layers irrespective of the cage type.ConclusionsHens confined in the same cage group tend to develop similar microbiota in their ileum and caecum possibly due to isolation, while differences in the microbiota between cages may be caused by environmental or individual bird factors. Although the cages type had influence on composition of the microbiota in the layers by promoting higher diversity in furnished and aviary systems, we did not observe differences in colonization and excretion pattern of Salmonella from these groups. We suggest, that differences in group size and exposure to a more faecally contaminated environment in the alternative systems may explain the observed differences in diversity of the caecal microbiota.

Impact of salinomycin on the intestinal microflora of broiler chickens.

BackgroundThe ionophoric coccidiostat salinomycin is widely used in chicken feed. In the near future the use of ionophore coccidiostats may be banned as has been the case for other antimicrobial growth promoters. This study was conducted to examine the effect of salinomycin on Campylobacter jejuni infection and on the composition of the caecal microflora in broiler chickens.MethodsAn experimental infection study was carried out in isolators and the intestinal microflora was analyzed using quantitative cultivation, denaturant gradient gel electrophoresis (DGGE), cloning and sequencing.ResultsWe found no effect of salinomycin on C. jejuni but salinomycin significantly affected the composition of the microflora. In addition, salinomycin significantly reduced the prevalence of Clostridium perfringens and we observed a significant increase (62%) in the mean body weight of salinomycin treated chickens compared to un-treated controls.ConclusionTermination of the use of ionophore coccidiostats will not affect food safety related to campylobacter, but will increase the risk of necrotic enteritis in the broilers.

Antimicrobial resistance in Clostridium perfringens isolates from broilers in Belgium

The antimicrobial susceptibility of Clostridium perfringens strains isolated from Belgian broilers between May and September 2007 was investigated. All 39 tested isolates were sensitive to enrofloxacin, erythromycin, tylosin, florfenicol and bacitracin. Twenty-six (66%) and 24 (61%) out of the 39 tested isolates showed acquired resistance to tetracycline and lincomycin, respectively. The C. perfringens isolates were also screened by PCR for the presence of the resistance genes tet(K), tet(L), tet(M), tetB(P), tet(O), tet(W), lnu(A) and lnu(B). In 22/26 tetracycline resistant strains and 7/24 lincomycin resistant strains, resistance could be attributed to one or more of these genes. An extended frequency distribution range of MICs was seen for ampicillin. These data are consistent with data derived from studies carried out in 1980 and in 2004, indicating that no changes in antimicrobial resistance patterns have taken place during time in C. perfringens isolates from broilers in Belgium.

Co-Infection Dynamics of a Major Food-Borne Zoonotic Pathogen in Chicken

A major bottleneck in understanding zoonotic pathogens has been the analysis of pathogen co-infection dynamics. We have addressed this challenge using a novel direct sequencing approach for pathogen quantification in mixed infections. The major zoonotic food-borne pathogen Campylobacter jejuni, with an important reservoir in the gastrointestinal (GI) tract of chickens, was used as a model. We investigated the co-colonisation dynamics of seven C. jejuni strains in a chicken GI infection trial. The seven strains were isolated from an epidemiological study showing multiple strain infections at the farm level. We analysed time-series data, following the Campylobacter colonisation, as well as the dominant background flora of chickens. Data were collected from the infection at day 16 until the last sampling point at day 36. Chickens with two different background floras were studied, mature (treated with Broilact, which is a product consisting of bacteria from the intestinal flora of healthy hens) and spontaneous. The two treatments resulted in completely different background floras, yet similar Campylobacter colonisation patterns were detected in both groups. This suggests that it is the chicken host and not the background flora that is important in determining the Campylobacter colonisation pattern. Our results showed that mainly two of the seven C. jejuni strains dominated the Campylobacter flora in the chickens, with a shift of the dominating strain during the infection period. We propose a model in which multiple C. jejuni strains can colonise a single host, with the dominant strains being replaced as a consequence of strain-specific immune responses. This model represents a new understanding of C. jejuni epidemiology, with future implications for the development of novel intervention strategies.

Experimental infections with rifampicin-resistant Clostridium perfringens strains in broiler chickens using isolator facilities

Experimental infection studies were carried out on the ability of three Clostridium perfringens type A rifampicin-resistant strains to colonize the intestinal tract of broiler chickens kept in isolators from 1-day-old. Various doses of C. perfringens were given orally at 22 days, 9 days or at 1 day old. At 22 days none of the strains, given in doses of approximately 1010 colony-forming units, caused mortality or clinical necrotic enteritis. None was able to colonize the intestine permanently and all were eliminated within 9 days. One strain given to groups of 9-day-old birds was recovered only from those receiving high doses, but for no longer than 13 days. In chicks infected at 1-day-old there was transient colonization up to 15 days, and the most persistent colonization was in a group given a fresh broth culture of unwashed cells, including extracellular products. Test strains were rapidly replaced by naturally occurring strains of C. perfringens in all groups but they persisted for considerably longer in chickens inoculated at 1-day-old or at 9 days than those at 22 days, indicating a possible resistance to colonization with increasing age. The findings emphasize the difficulties of establishing a reproducible model for infection with C. perfringens in broiler chickens.

Reduction of Escherichia coli, Salmonella Enteritidis and Campylobacter jejuni in poultry manure by rearing of Musca domestica fly larvae

A major barrier for using animal waste as substrate for production of insects for feed or food is the concern for safety of the end products. In this study we investigated how rearing of fly larvae of Musca domestica in poultry manure influenced the counts of three pathogenic test strains (Escherichia coli, Salmonella Enteritidis and Campylobacter jejuni) and investigated whether these were transferred from manure to larvae, pupae or adults flies. We monitored quantitative microbiological changes over a study period of seven days. We showed that the decay in the numbers of E. coli, S. Enteritidis, and C. jejuni was faster in manure with rearing of fly larvae than in manure without larvae; an 8 log10 reduction of all three test bacteria was observed within four days in manure with larvae; compared to manure without larvae where a 1 to 2 log10 was observed. We found no sign of propagation of the pathogens in the larvae gut. All test strains were detectable in low numbers at day 4 in the larvae gut but only ...