Recovery of probiotic bacteria from the intestinal tract of broilers after noninvasive pre-hatch application

This paper reports the success of intestinal colonisation of chickens and foetuses by probiotics after different methods of pre-hatch application. Hatcheries not using in ovo injection of probiotics or wish to avoid the reduced hatchability associated with in ovo injections prefer using alternatives to in ovo technologies. Therefore, we used noninvasive pre-hatch application methods. This included the vertical transmission of probiotics from the mother hen to offspring, application of probiotic late in incubation and transmission of probiotics during hatch. Enterococcus faecium (NCIMB11181) and Lactobacillus animalis (DSM33570) were used as probiotics. Probiotics were applied either through drinking water for the mother hens, by dipping the eggs in a probiotic solution on days 16-18 of incubation or through drops/spray on the eggshell of the fertilised eggs. Similarly, intestinal colonisation of the probiotic in chickens was investigated either before hatch (pre-hatch) or immediately after hatch (post-hatch). Based on the performed experiments, it is concluded that E. faecium was vertically transmitted from the mother hen to the offspring, as E. faecium was recovered in 20 and 33% of the offspring pre- and post-hatch, respectively. When applied on the eggshell, the recovery of E. faecium before hatch depended on the application method and ranged from 0 to 9%. In contrast, L. animalis was not recovered before hatch. Moreover, when sampling post-hatch 100% of the chickens were colonised when E. faecium was used and 54% were colonised when L. animalis was used. Furthermore, spray application with E. faecium was the most successful application method as 9% of the foetuses were colonised pre-hatch and 100% became colonised post-hatch. Therefore, pre-hatch application by, for example, spray of probiotics on the eggshell can be used as an easy-to-use, noninvasive method for early life colonisation of chicken gut.

Campylobacter jejuni transmission and colonisation in broiler chickens is inhibited by Faecal Microbiota Transplantation

ABSTRACTBACKGROUNDCampylobacter jejuni, the most frequent cause of foodborne bacterial infection, is found on around 70% of retail chicken. As such there is a need for effective controls in chicken production. Microbial-based controls such as probiotics are attractive to the poultry industry, but of limited efficacy. Furthermore, as commercially-produced chickens have no maternal contact, their pioneer microbiome is likely to come from the hatchery environment. Early delivery of microbials that lead to a more ‘natural avian’ microbiome may, therefore, improve bird health and reduce susceptibility to C.jejuni colonisation.A faecal microbiota transplant (FMT) was used to transfer a mature cecal microbiome to newly-hatched broiler chicks and its effects on C.jejuni challenge assessed. We used both a seeder-bird infection model that mimics natural bird-to-bird infection alongside a direct-challenge model. We used a 16S rRNA gene sequencing-based approach to characterize the transplant material itself alongside changes to the chicken microbiome following FMT.RESULTSFMT changes the composition of the chicken intestinal microbiome. We observed little change in species richness following FMT compared to untreated samples, but there is an increase in phylogenetic diversity within those species. The most significant difference in the ceca is an increase in Lactobacilli, although not a major component of the transplant material, suggesting the FMT results in a change in the intestinal milieu as much as a direct change to the microbiome.Upon direct challenge, FMT resulted in lower initial intestinal colonisation with C.jejuni. More significantly, in a seeder-bird challenge of infection transmission, FMT reduced transmission and intestinal colonisation until common UK retail age of slaughter. In a repeat experiment, transmission was completely blocked following FMT treatment. Delayed FMT administration at 7 days of-age had limited effect on colonisation and transmission.CONCLUSIONSWe show that transfer of a whole mature microbiome to newly-hatched chicks reduces transmission and colonisation of C.jejuni. This indicates that modification of the broiler chick microbiome can reduce intestinal colonisation of C.jejuni to levels projected to lead to lower the human infection rate. We believe these findings offer a way to identify key taxa or consortia that are effective in reducing C.jejuni colonisation and improving broiler gut health.

Opposing effect of Lactobacillus on in vitro Klebsiella pneumoniae in biofilm and in an in vivo intestinal colonisation model

Beneficial bacteria represent potential sources of therapy, particularly in the battle against antibiotic-resistant pathogens. The Gram-negative bacillus Klebsiella pneumoniae is not only a paradigm of multi-resistant opportunistic pathogen, but it is also able to colonise the human intestine and displays a high capacity to form biofilm. In this study, the anti-biofilm activity of 140 neutralised Lactobacillus supernatants was assessed against K. pneumoniae. Among the 13 strains whose supernatant significantly impaired biofilm formation, Lactobacillus plantarum CIRM653 was selected because it was also able to impair K. pneumoniae preformed biofilm, independently of a bactericidal effect. Mixed K. pneumoniae/L. plantarum CIRM653 biofilms had reduced tridimensional structures associated with a significant decrease in K. pneumoniae biomass. Further investigation showed that L. plantarum CIRM653 supernatant induced transcriptional modifications of K. pneumoniae biofilm-related genes, including down-regulation of the quorum sensing-related lsr operons and over-expression of type 3 pili structure genes. Increased production of type 3 pili was validated by Western-blot, hemagglutination and adhesion assays. L. plantarum CIRM653 activity against K. pneumoniae was also assessed in a murine intestinal colonisation model: a constant faecal pathogen burden was observed, as against a gradual decrease in the control group. These results reveal that an in vitro a priori attracting anti-biofilm activity of Lactobacillus might be counterbalanced by an in vivo behaviour in a complex microbiota environment with potential deleterious dispersal of highly adherent K. pneumoniae cells, raising the question of the accuracy of in vitro assays in screening of beneficial microbes.