Jane Beal

Probiotics, Prebiotics and Immunomodulation of Gut Mucosal Defences: Homeostasis and Immunopathology

Probiotics are beneficial microbes that confer a realistic health benefit on the host, which in combination with prebiotics, (indigestible dietary fibre/carbohydrate), also confer a health benefit on the host via products resulting from anaerobic fermentation. There is a growing body of evidence documenting the immune-modulatory ability of probiotic bacteria, it is therefore reasonable to suggest that this is potentiated via a combination of prebiotics and probiotics as a symbiotic mix. The need for probiotic formulations has been appreciated for the health benefits in “topping up your good bacteria” or indeed in an attempt to normalise the dysbiotic microbiota associated with immunopathology. This review will focus on the immunomodulatory role of probiotics and prebiotics on the cells, molecules and immune responses in the gut mucosae, from epithelial barrier to priming of adaptive responses by antigen presenting cells: immune fate decision—tolerance or activation? Modulation of normal homeostatic mechanisms, coupled with findings from probiotic and prebiotic delivery in pathological studies, will highlight the role for these xenobiotics in dysbiosis associated with immunopathology in the context of inflammatory bowel disease, colorectal cancer and hypersensitivity.

Effect on weaner pig performance and diet microbiology of feeding a liquid diet acidified to pH 4 with either lactic acid or through fermentation with Pediococcus acidilactici

The effect of feeding newly weaned pigs acidified liquid diets was investigated. The control diet was acidified to about pH 4 with lactic acid (LA). A second diet of the same formulation was acidified to about pH 4 by fermentation with Pediococcus acidilactici (PA). Forty-eight weaner pigs (weight 7 kg±1 kg, age 24±4 days) were allocated to the two dietary treatments according to a randomised block design and fed ad libitum for 28 days. Food intake, daily gain and water intake were recorded, and a microbial assessment of the liquid diet was conducted. Reducing pH<4.0 in either of the liquid diets was effective in eliminating coliform bacteria. There were no significant differences in any of the performance parameters measured. The average daily liveweight gain overall was 474 and 496±17.8 g d−1 for PA and LA, respectively, with a feed conversion ratio overall of 1.15 and 1.11±0.025 for PA and LA, respectively. Fermentation of liquid diets for newly weaned piglets could provide a more cost effective means of acidifying diets than the use of organic acids. Reducing the pH of the liquid diet to 4.00 by fermentation with Pediococcus acidilactici was a cost effective method of eliminating enteropathogens and spoilage organisms from the diet. © 1999 Society of Chemical Industry

The effect of temperature on the growth and persistence of Salmonella in fermented liquid pig feed

Two studies were conducted to investigate the effect of temperature on the fate of Salmonella enterica subsp. enterica serovar typhimurium DT104:30 in fermented liquid pig feed. These were (1) by co-inoculation of feed with S. typhimurium DT104:30 and Pediococcus pentosaceus, as the fermenting organism, and (2) by fermenting feed for 48, 72 or 96 h prior to inoculation with S. typhimurium DT104:30. In co-inoculated feed incubated at 20 degrees C, S. typhimurium DT104:30 persisted for at least 72 h. In contrast, in feed incubated at 30 degrees C, no S. typhimurium DT104:30 were detectable 48 h after inoculation. In prefermented feed, S. typhimurium DT104:30 died four to five times faster in feed maintained at 30 degrees C (D(value) 34-45 min) compared with feed maintained at 20 degrees C (D(value) 137-250 min). This was not entirely due to differences in lactic acid concentration as feed fermented for 72 or 96 h at 20 degrees C and feed fermented for 48 h at 30 degrees C contained similar concentrations of lactic acid (160-170 mM). Low numbers of S. typhimurium DT104:30 were still detectable in fermented feed 24 h after inoculation at 20 degrees C. In contrast, none were detectable 6-7 h after inoculation at 30 degrees C. The results of these studies indicate that it would be advisable for pig producers to control the temperature of liquid feed tanks to reduce the risk of Salmonella contamination.

Bacterial fermentation in the gastrointestinal tract of non-ruminants: Influence of fermented feeds and fermentable carbohydrates

The search for alternatives to in-feed antibiotics in animal nutrition has highlighted the role dietary modulation can play in improving gut health. Current antibiotic replacement strategies have involved the use of microbes beneficial to health (probiotics) or fermentable carbohydrates (prebiotics) or both (synbiotics). The present review recognises the contribution of fermented feeds and fermentable carbohydrates in improving the gut environment in non-ruminants. It proposes the screening of probiotic bacteria for the production of fermented feeds and supplementation of these feeds with fermentable carbohydrates prior to feeding animals. It is suggested that the term ‘fermbiotics’ should be used to describe this intervention strategy.

Probiotic bacterial strains differentially modulate macrophage cytokine production in a strain-dependent and cell subset-specific manner

Gut mucosal macrophages play a pivotal role in driving mucosal immune responses, resulting in either activation of inflammatory immune responses to pathogenic challenge or tolerance to beneficial luminal contents such as food and commensal bacteria. Macrophage responses elicited are dependent on tissue environment and the resulting cell subset, where homeostatic macrophages resemble the M2 macrophage subset and inflammatory macrophages resemble M1s. Probiotics can modulate macrophage function with outcome dependent on subset present. Using a THP-1 monocyte cell line-derived model of CD14high/low M1 and M2 macrophages, the aim of this study was to investigate the immunomodulatory effects of a panel of heat-killed probiotic bacteria and their secreted proteins on the subset-specific inflammatory marker profile of TNFα, IL-6 and NFκB. M1 and M2 cells were generated by differentiation of monocyte stable transfectants for high and low CD14 expression with phorbol 12-myristate 13-acetate and vitamin D3, respectively, where the resulting CD14lo M2 and CD14hi M1s mimicked homeostatic and inflammatory mucosal macrophages. Subsets were stimulated by enteropathic lipopolysaccharides in the presence or absence of heat-killed (HK) or secreted proteins (SP) from a panel of probiotic bacteria. Regulation of cytokine expression was measured by ELISA and NFκB activity by reporter assay. HK probiotics suppress CD14lo and augment CD14hi M1 and M2 production of TNFα whereas SPs augmented CD14hi M1 TNFα and were generally suppressive in the other subtypes. M2 macrophage IL-6 production was suppressed by both HK and SPs and differentially regulated in CD14lo and CD14hi M1s. NFκB activation failed to parallel the regulatory profiles for TNFα and IL-6 which is suggestive of probiotic bacteria exerting their regulatory effects on these cytokines in an NFκB-independent manner. In conclusion, HK and SP probiotics differentially regulate macrophage cytokines and NFκB activation in a subset-dependent manner and suggest a cautionary approach to probiotic treatment of mucosal inflammation.

The effect of copper on the death rate of Salmonella typhimurium DT104:30 in food substrates acidified with organic acids

Aims:  The objective of this study was to investigate the effect of copper ions on the survival of Salmonella typhimurium DT104:30 in acidified liquid food substrates.

Heat-killed probiotic bacteria differentially regulate colonic epithelial cell production of human β-defensin-2: dependence on inflammatory cytokines

The inducible antimicrobial peptide human β-defensin-2 (hBD-2) stimulated by pro-inflammatory cytokines and bacterial products is essential to antipathogen responses of gut epithelial cells. Commensal and probiotic bacteria can augment such mucosal defences. Probiotic use in the treatment of inflammatory bowel disease, however, may have adverse effects, boosting inflammatory responses. The aim of this investigation was to determine the effect of selected probiotic strains on hBD-2 production by epithelial cells induced by pathologically relevant pro-inflammatory cytokines and the role of cytokine modulators in controlling hBD-2. Caco-2 colonic intestinal epithelial cells were pre-incubated with heat-killed probiotics, i.e. Lactobacillus casei strain Shirota (LcS) or Lactobacillus fermentum strain MS15 (LF), followed by stimulation of hBD-2 by interleukin (IL)-1β and tumour necrosis factor alpha (TNF-α) in the absence or presence of exogenous IL-10 or anti-IL-10 neutralising antibody. Cytokines and hBD-2 mRNA and protein were analysed by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. LcS augmented IL-1β-induced hBD-2, whereas LF enhanced TNF-α- and suppressed IL-1β-induced hBD-2. LF enhanced TNF-α-induced TNF-α and suppressed IL-10, whereas augmented IL-1β-induced IL-10. LcS upregulated IL-1β-induced TNF-α mRNA and suppressed IL-10. Endogenous IL-10 differentially regulated hBD-2; neutralisation of IL-10 augmented TNF-α- and suppressed IL-1β-induced hBD-2. Exogenous IL-10, however, suppressed both TNF-α- and IL-1β-induced hBD-2; LcS partially rescued suppression in TNF-α- and IL-1β-stimulation, whereas LF further suppressed IL-1β-induced hBD-2. It can be concluded that probiotic strains differentially regulate hBD-2 mRNA expression and protein secretion, modulation being dictated by inflammatory stimulus and resulting cytokine environment.

Evaluation of the microbial community, acidity and proximate composition of akamu, a fermented maize food

BACKGROUND Akamu is a lactic acid-fermented cereal-based food that constitutes a major infant complementary food in most West African countries. The identities of LAB populations from DGGE analysis and conventionally isolated LAB and yeasts from traditionally fermented akamu were confirmed by PCR sequencing analysis. The relationships between pH, acidity and lactic acid levels and proximate composition of the akamu samples were investigated. RESULTS The LAB communities in the akamu samples comprised mainly Lactobacillus species, including Lb. fermentum, Lb. plantarum, Lb. delbrueckii ssp. bulgaricus and Lb. helveticus, as well as Lactococcus lactis ssp. cremoris. Identified yeasts were Candida tropicalis, Candida albicans, Clavispora lusitaniae and Saccharomyces paradoxus. Low pH (3.22-3.95) was accompanied by high lactic acid concentrations (43.10-84.29 mmol kg⁻¹). Protein (31.88-74.32 g kg⁻¹) and lipid (17.74-36.83 g kg⁻¹ contents were negatively correlated with carbohydrate content (897.48-926.20 g kg⁻¹, of which ≤1 g kg⁻¹ was sugars). Ash was either not detected or present only in trace amounts (≤4 g kg⁻¹). Energy levels ranged from 17.29 to 18.37 kJ g⁻¹. CONCLUSION The akamu samples were predominantly starchy foods and had pH < 4.0 owing to the activities of fermentative LAB.