Interference of LPS H. pylori with IL-33-Driven Regeneration of Caviae porcellus Primary Gastric Epithelial Cells and Fibroblasts

Background: Lipopolysaccharide (LPS) of Helicobacter pylori (Hp) bacteria causes disintegration of gastric tissue cells in vitro. It has been suggested that interleukin (IL)-33 is involved in healing gastric injury. Aim: To elucidate whether Hp LPS affects regeneration of gastric barrier initiated by IL-33. Methods: Primary gastric epithelial cells or fibroblasts from Caviae porcellus were transfected with siRNA IL-33. Such cells, not exposed or treated with LPS Hp, were sub-cultured in the medium with or without exogenous IL-33. Then cell migration was assessed in conjunction with oxidative stress and apoptosis, activation of extracellular signal-regulated kinase (Erk), production of collagen I and soluble ST2 (IL-33 decoy). Results: Control cells not treated with LPS Hp migrated in the presence of IL-33. The pro-regenerative activity of IL-33 was related to stimulation of cells to collagen I production. Wound healing by cells exposed to LPS Hp was inhibited even in the presence of IL-33. This could be due to increased oxidative stress and apoptosis in conjunction with Erk activation, sST2 elevation and modulation of collagen I production. Conclusions: The recovery of gastric barrier cells during Hp infection potentially can be affected due to downregulation of pro-regenerative activity of IL-33 by LPS Hp.

Using Nutritional Strategies to Shape the Gastro-Intestinal Tracts of Suckling and Weaned Piglets

This is a comprehensive review on the use of nutritional strategies to shape the functioning of the gastro-intestinal tract in suckling and weaned piglets. The progressive development of a piglet’s gut and the associated microbiota and immune system offers a unique window of opportunity for supporting gut health through dietary modulation. This is particularly relevant for large litters, for which sow colostrum and milk are insufficient. The authors have therefore proposed the use of supplemental milk and creep feed with a dual purpose. In addition to providing nutrients to piglets, supplemental milk can also serve as a gut modulator in early life by incorporating functional ingredients with potential long-term benefits. To prepare piglets for weaning, it is important to stimulate the intake of solid feed before weaning, in addition to stimulating the number of piglets eating. The use of functional ingredients in creep feed and a transition diet around the time of weaning helps to habituate piglets to solid feed in general, while also preparing the gut for the digestion and fermentation of specific ingredients. In the first days after weaning (i.e., the acute phase), it is important to maintain high levels of feed intake and focus on nutritional strategies that support good gastric (barrier) function and that avoid overloading the impaired digestion and fermentation capacity of the piglets. In the subsequent maturation phase, the ratio of lysine to energy can be increased gradually in order to stimulate piglet growth. This is because the digestive and fermentation capacity of the piglets is more mature at this stage, thus allowing the inclusion of more fermentable fibres. Taken together, the nutritional strategies addressed in this review provide a structured approach to preparing piglets for success during weaning and the period that follows. The implementation of this approach and the insights to be developed through future research can help to achieve some of the most important goals in pig production: reducing piglet mortality, morbidity and antimicrobial use.

Anti-Helicobacter activity of medicinal plants and probiotics as alternatives for Helicobacter pylori treatment: Review article

Helicobacter pylori, a gram-negative bacterium, known factor for chronic active gastritis, stomach and peptic ulceration, which may progress to gastric cancer. It is also associated with other non-gastric diseases such as stroke, diabetes mellitus and alzheimer’s disease. Although conventional treatment achieved a great advancements in controlling H.pylori infection nowadays it is not effective thus it’s intended to find some other alternative sources that may be used alone or in combination with antibiotics to eradicate the infection. Recently published literature of natural sources such as plant -derived bioactive compounds and probiotics are studied for their reduced side effects and for being safe and inexpensive. However, the mechanism of action by which these herbs and probiotics exert their medicinal properties in H.pylori treatment is still not fully clear. In this review, we highlight the potential antibacterial mechanisms of some traditionally used bioactive compounds and their possible role on H.pylori colonization. On the other hand, we focused on the possible inhibitory role of probiotics in the eradication of H. pylori infection through the release of organic acids and their role in the stabilization of the gastric barrier function in order to decrease the mucosal inflammation, modulate H.pylori colonization and enhance compliance in infected patients.

Escherichia coli O157:H7 Stationary-Phase Acid Resistance and Assessment of Survival in a Model Vegetable Fermentation System

ABSTRACT Escherichia coli O157:H7 (STEC) acid resistance may aid the pathogen's ability to cross the human gastric barrier, which makes it an organism of concern in acidic foods. Our objective was to determine how STEC acid resistance may correlate with survival during vegetable fermentations. Seven E. coli O157:H7 strains were screened to assess acid resistance in simulated stomach acid at pH 2. The strains were separated into two groups that differed in acid resistance (P < 0.05), with three being acid sensitive and four acid resistant. The growth rates of these strains were measured in a Luria broth at pH values from 4.2 to 6.8. Two strains having similar growth kinetics, B201 (acid sensitive) and B241 (acid resistant), were selected for further analysis. B201 was found to be missing (compared with B241) two glutamic acid decarboxylase regulatory genes required for acid resistance, gadE and gadX. These strains were challenged in lactic acid (100 mM) solutions, including cucumber juice (CJ) media at pH 3.3. As expected, B201 was more acid sensitive than B241, and a filtered fermented CJ was more inhibitory than similarly acidified CJ. In competitive growth studies with Lactobacillus plantarum LA445 in CJ, B201 or B241 grew from approximately 104 to 108 CFU/mL within 24 h, but the STEC strains were below the limit of detection by 48 h. In all fermentations, L. plantarum reached 108 CFU/mL by 48 h. However, in three of four independent fermentation experiments, strain B201 survived longer than B241. This was possibly due to buffering in B241-LA445 fermentation brines that had increased lactic acid for a given pH compared with B201-LA445. These data indicate that stationary-phase acid resistance may not accurately predict STEC survival during vegetable fermentations. HIGHLIGHTS

Chlamydia Deficient in Plasmid-Encoded pGP3 Is Prevented from Spreading to Large Intestine

ABSTRACT The cryptic plasmid pCM is critical for chlamydial colonization in the gastrointestinal tract. Nevertheless, orally inoculated plasmid-free Chlamydia sp. was still able to colonize the gut. Surprisingly, orally inoculated Chlamydia sp. deficient in only plasmid-encoded pGP3 was no longer able to colonize the gut. A comparison of live organism recoveries from individual gastrointestinal tissues revealed that pGP3-deficient Chlamydia sp. survived significantly better than plasmid-free Chlamydia sp. in small intestinal tissues. However, the small intestinal pGP3-deficient Chlamydia sp. failed to reach the large intestine, explaining the lack of live pGP3-deficient Chlamydia sp. in rectal swabs following an oral inoculation. Interestingly, pGP3-deficient Chlamydia sp. was able to colonize the colon following an intracolon inoculation, suggesting that pGP3-deficient Chlamydia sp. might be prevented from spreading from the small intestine to the large intestine. This hypothesis is supported by the finding that following an intrajejunal inoculation that bypasses the gastric barrier, pGP3-deficient Chlamydia sp. still failed to reach the large intestine, although similarly inoculated plasmid-free Chlamydia sp. was able to do so. Interestingly, when both types of organisms were intrajejunally coinoculated into the same mouse small intestine, plasmid-free Chlamydia sp. was no longer able to spread to the large intestine, suggesting that pGP3-deficient Chlamydia sp. might be able to activate an intestinal resistance for regulating Chlamydia sp. spreading. Thus, the current study has not only provided evidence for reconciling a previously identified conflicting phenotype but also revealed a potential intestinal resistance to chlamydial spreading. Efforts are under way to further define the mechanism of the putative intestinal resistance.

The Cryptic Plasmid Improves Chlamydia Fitness in Different Regions of the Gastrointestinal Tract

ABSTRACT The cryptic plasmid is important for chlamydial colonization in the gastrointestinal tract. We used a combination of intragastric, intrajejunal, and intracolon inoculations to reveal the impact of the plasmid on chlamydial colonization in distinct regions of gastrointestinal tract. Following an intragastric inoculation, the plasmid significantly improved chlamydial colonization. At the tissue level, plasmid-positive Chlamydia produced infectious progenies throughout gastrointestinal tract. However, to our surprise, plasmid-deficient Chlamydia failed to produce infectious progenies in small intestine, although infectious progenies were eventually detected in large intestine, indicating a critical role of the plasmid in chlamydial differentiation into infectious particles in small intestine. The noninfectious status may represent persistent infection, since Chlamydia genomes proliferated in the same tissues. Following an intrajejunal inoculation that bypasses the gastric barrier, plasmid-deficient Chlamydia produced infectious progenies in small intestine but was 530-fold less infectious than plasmid-positive Chlamydia, suggesting that (i) the noninfectious status developed after intragastric inoculation might be induced by a combination of gastric and intestinal effectors and (ii) chlamydial colonization in small intestine was highly dependent on plasmid. Finally, following an intracolon inoculation, the dependence of chlamydial colonization on plasmid increased over time. Thus, we have demonstrated that the plasmid may be able to improve chlamydial fitness in different gut regions via different mechanisms, which has laid a foundation to further reveal the specific mechanisms.

The Plasmid-Encoded pGP3 PromotesChlamydiaEvasion of Acidic Barriers in Both Stomach and Vagina

ABSTRACTAlthoughChlamydia trachomatisis a human genital tract pathogen, chlamydial organisms have frequently been detected in both vaginal and rectal swab samples of animals and humans. The plasmid-encoded pGP3, a genital tract virulence factor, is essential forChlamydia muridarumto colonize the mouse gastrointestinal tract. However, intracolon inoculation to bypass the gastric barrier rescued the colonization ability of a pGP3-deficientC. muridarummutant, suggesting that pGP3 is required forC. muridarumto reach but not to colonize the large intestine. The pGP3-deficient mutant was rapidly cleared in the stomach and was 100-fold more susceptible to gastric killing. In mice genetically deficient in gastrin, a key regulator for gastric acid production, or pharmacologically treated with a proton pump inhibitor, the ability of pGP3-deficientC. muridarumto colonize the gastrointestinal tract was rescued. The pGP3-dependent resistance was further recapitulatedin vitrowith treatments with HCl, pepsin, or sarkosyl. In the genital tract, deficiency in pGP3 significantly reducedC. muridarumsurvival in the mouse vagina and increasedC. muridarumsusceptibility to vaginal killing by ∼8 times. The pGP3-deficientC. muridarumwas more susceptible to lactic acid killing, and the pGP3 deficiency also significantly increasedC. trachomatissusceptibility to lactic acid. The above-described observations together suggest thatChlamydiamay have acquired the plasmid-encoded pGP3 to overcome the gastric barrier during its adaptation to the gastrointestinal tract and the pGP3-dependent resistance may enable chlamydial evasion of the female lower genital tract barrier during sexual transmission.

Omics of bifidobacteria: research and insights into their health-promoting activities

Members of the genus Bifidobacterium include gut commensals that are particularly abundant among the microbial communities residing in the gut of healthy breast-fed infants, where their presence has been linked to many beneficial host effects. Next-generation DNA sequencing and comparative and functional genome methodologies have been shown to be particularly useful in exploring the diversity of this genus. These combined approaches have allowed the identification of genetic features related to bifidobacterial establishment in the gut, involving host–microbe as well as microbe–microbe interactions. Among these, proteinaceous structures, which protrude from the bacterial surface, i.e. pili or fimbriae, and exopolysaccharidic cell surface layers or capsules represent crucial features that assist in their colonization and persistence in the gut. As bifidobacteria are colonizers of the large intestine, they have to be able to cope with various sources of osmotic, oxidative, bile and acid stress during their transit across the gastric barrier and the small intestine. Bifidobacterial genomes thus encode various survival mechanisms, such as molecular chaperones and efflux pumps, to overcome such challenges. Bifidobacteria represent part of an anaerobic gut community, and feed on nondigestible carbohydrates through a specialized fermentative metabolic pathway, which in turn produces growth substrates for other members of the gut community. Conversely, bifidobacteria may also be dependent on other (bifido)bacteria to access host- and diet-derived glycans, and these complex co-operative interactions, based on resource sharing and cross-feeding strategies, represent powerful driving forces that shape gut microbiota composition.