Strategies to augment non-immune system based defence mechanisms against gastrointestinal diseases in pigs

AbstractOur study addresses the first two weeks of the weaning period of piglets during which stressful physiological and environmental conditions experienced by the animals can promote the proliferation of pathogens in the digestive tract. The aim of the study was to identify new feeding strategies that result in boosting the gastrointestinal tract (GIT) microbiota of piglets and improve growth performance, reducing the negative impact of weaning. In order to identify a new synbiotic combination, 12 new putative probiotic strains of Bifidobacterium spp. and three non-digestible oligosaccharides [NDO] were screened in newly weaned piglets. The ability to increase the level of autochthonous bifidobacteria and improve growth performance were assessed. Bifidobacteria strains with a similar ability to develop in the hindgut showed a different effect on piglet performance depending on the dose in which they were provided. Our data support the idea that the presence of fructo-oligosaccharides would stimulate the occurrence of bifidobacteria in the caecum. It was shown that dietary intake of nitrate can generate salivary nitrite, which in turn is acidified in the stomach and could have antimicrobial activity against swallowed pathogens. The efficacy of the resulting synbiotic formula was improved by adding nitrate as antimicrobial. To enhance probiotic survival during gastric transit, a novel technology of microencapsulation was developed and applied to bacteria. The final synbiotic, containing the strain RA 18 of Bifidobacterium animalis subsp. lactis [1011cfu/day], the prebiotic Actilight® [4% of the diet], and nitrate [150mg KNO3/kg feed/day] was tested in organic weaned piglets reared under field conditions. Results show that the strain Ra 18 had a probiotic effect in organic weaned piglets, as it colonized and remained detectable in faecal samples until two weeks after addition. The use of our synbiotic formula improved weight gain, feed efficiency and health status of the weaned piglets

Lyophilized alginate-based microspheres containing Lactobacillus fermentum D12, an exopolysaccharides producer, contribute to the strain\u2019s functionality in vitro

Lactobacillus (Limosilactobacillus) fermentum D12 is an exopolysaccharide (EPS) producing strain whose genome contains a putative eps operon. Whole-genome analysis of D12 was performed to disclose the essential genes correlated with activation of precursor molecules, elongation and export of the polysaccharide chain, and regulation of EPS synthesis. These included the genes required for EPS biosynthesis such as epsA, B, C, D and E, also gt, wzx, and wzy and those involved in the activation of the precursor molecules galE, galT and galU. Both the biosynthesis and export mechanism of EPS were proposed based on functional annotation. When grown on MRS broth with an additional 2% w/v glucose, L. fermentum D12 secreted up to 200\ua0mg/L of a mixture of EPSs, whose porous structure was visualized by scanning electron microscopy (SEM). Structural information obtained by 1HNMR spectroscopy together with composition and linkage analyses, suggested the presence of at least two different EPSs, a branched heteropolysaccharide containing t-Glcp and 2,6-linked Galf, and glycogen. Since recent reports showed that polysaccharides facilitate the probiotic-host interactions, we at first sought to evaluate the functional potential of L. fermentum D12. Strain D12 survived simulated gastrointestinal tract (GIT) conditions, exhibited antibacterial activity against enteropathogenic bacteria, adhered to Caco-2 cells in vitro, and as such showed potential for in vivo functionality. The EPS crude extract positively influenced D12 strain capacity to survive during freeze-drying and to adhere to extracellular matrix (ECM) proteins but did not interfere Caco-2 and mucin adherence when added at concentrations of 0.2, 0.5, and 1.0\ua0mg/mL. Since the viable bacterial count of free D12 cells was 3 logarithmic units lower after the exposure to simulated GIT conditions than the initial count, the bacterial cells had been loaded into alginate for viability improvement. Microspheres of D12 cells, which were previously analyzed at SEM, significantly influenced their survival during freeze-drying and in simulated GIT conditions. Furthermore, the addition of the prebiotic substrates mannitol and lactulose improved the viability of L. fermentum D12 in freeze-dried alginate microspheres during 1-year storage at 4\ua0\ub0C compared to the control. [Figure not available: see fulltext.

Building an all-hazards agricultural emergency response system to maintain business continuity and promote the sustainable supply of food and agricultural products

The response to an agricultural emergency that threatens to destroy crops or animals requires a rapid, coordinated state-level response from the outset. An authority should be established at the local level to initiate and enforce food embargoes, quarantine livestock or poultry premises, depopulate affected or potentially affected animals, and provide indemnity, when appropriate, for those depopulated animals or destroyed products. Depending on the scale of the threat, industry needs, state resources, and response capacity, the authority for these activities currently resides with the state and is supported by federal agencies. However, an all-encompassing all-hazards agricultural emergency response system can be constructed through collaborations with agricultural industry, state responders, and federal agencies. The formed response should include development of permitting guidance for controlled harvest and movement of unaffected crops, animals, and animal products. The ultimate goal is to effectively manage the emergency yet maintain agricultural business continuity

Flow Cytometric Assessment of Viability of Lactic Acid Bacteria

The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA binding probes propidium iodide (PI) and TOTO-1 were tested for live/dead discrimination using a Lactococcus, a Streptococcus, three Lactobacillus, two Leuconostoc, an Enterococcus, and a Pediococcus species. Plate count experiments were performed to validate the results of the FCM assays. The results showed that cFDA was an accurate stain for live cells; in exponential-phase cultures almost all cells were labeled, while 70°C heat-killed cultures were left unstained. PI did not give clear live/dead discrimination for some of the species. TOTO-1, on the other hand, gave clear discrimination between live and dead cells. The combination of cFDA and TOTO-1 gave the best results. Well-separated subpopulations of live and dead cells could be detected with FCM. Cell sorting of the subpopulations and subsequent plating on agar medium provided direct evidence that cFDA labels the culturable subpopulation and that TOTO-1 labels the nonculturable subpopulation. Applied to cultures exposed to deconjugated bile salts or to acid, cFDA and TOTO-1 proved to be accurate indicators of culturability. Our experiments with lactic acid bacteria demonstrated that the combination of cFDA and TOTO-1 makes an excellent live/dead assay with versatile applications