Effect of dietary supplementation of immunobiotic lactiplantibacillus plantarum n14 fermented rakkyo (Allium chinense) pickled juice on the immunocompetence and production performance of pigs

Rakkyo (Allium chinense), is a Japanese leek that is primarily used to make a popular sweet or sour pickled dish. Lactic acid bacteria are often involved in the preparation steps of fermented pickles, which helps in the effective preservation of the natural bioactive compounds of fruits and vegetable, and thereby exert several health benefits including immunomodulation and growth per-formance. This work aimed to evaluate the in vivo effects of adding Lactiplantibacillus plantarum N14 fermented rakkyo pickled juice as feed supplement on the immunocompetence and production performance of pigs. We first analyzed the nutritional composition, which revealed that the proportion of protein, lipid, and water-soluble fiber content were estimated as of 4%, 5%, and 5% in rakkyo residual liquid or juice, while 22%, 15% and 14%, respectively, were estimated in rakkyo residual powder. For the in vivo feeding trials, three groups of pigs were treated either with 5%, 20%, or 40% mixture (v/v) of fermented rakkyo pickled juice and the grinded residual liquid supplemented in the drinking water in addition to standard feed. The results of the feeding trials showed that the administration of a juice mixture of 5% or 20% (fermented pickled juice and residual liquid) had a similar trend of effects in improving the complement activity, phagocytic activity and leucocytes counts in the peripheral blood when compared to pigs fed with 40% mixture or untreated controls. Those changes were related to an improved resistance to enteric infections. Moreover, animals receiving a mixture of fermented pickled juice and fermented rakkyo residues had a higher growth rate and carcass quality than controls. The results suggested that the use of 5% mixture of fermented rakkyo pickled juice and the residual liquid through drinking water could be a cost-effective approach to promote the immune-health and production performance of pigs. This approach would contribute not only to the sustainable management of food wastes but also to the application of a value-added feed supplement for the promotion of animal health and production.Fil: Islam, Md Aminul. Tohoku University; Japón. Bangladesh Agricultural University; BangladeshFil: Hashiguchi, Kenji. No especifíca;Fil: Humayun Kober, A.K.M.. Tohoku University; Japón. Chittagong Veterinary and Animal Sciences University; BangladeshFil: Morie, Kyoko. Tohoku University; JapónFil: Zhou, Binghui. Tohoku University; JapónFil: Tomokiyo, Mikado. Tohoku University; JapónFil: Shimazu, Tomoyuki. Miyagi University; JapónFil: Aso, Hisashi. Tohoku University; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Suda, Yoshihito. Miyagi University; JapónFil: Kitazawa, Haruki. Tohoku University; Japó

The toll-like receptor family protein RP105/MD1 complex is involved in the immunoregulatory effect of exopolysaccharides from Lactobacillus plantarum N14

The radioprotective 105 (RP105)/MD1 complex is a member of the Toll-like receptor (TLR) family. It was reported that RP105/MD1 cooperates with the lipopolysaccharide (LPS) receptor TLR4/MD2 complex and plays a crucial role in the response of immune cells to LPS. This work evaluated whether RP105, TLR4 or TLR2 were involved in the immunoregulatory capacities of Lactobacillus plantarum N14 (LP14) or its exopolysaccharides (EPS). EPS from LP14 were fractionated into neutral (NPS) and acidic (APS) EPS by anion exchange chromatography. Experiments with transfectant HEKRP105/MD1 and HEKTLR2 cells demonstrated that LP14 strongly activated NF-B via RP105 and TLR2. When we studied the capacity of APS to activate NF-B pathway in HEKRP105/MD1 and HEKTLR4 cells; we observed that APS strongly stimulated both transfectant cells. Our results also showed that LP14 and APS were able to decrease the production of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) in porcine intestinal epithelial (PIE) cells in response to enterotoxigenic Escherichia coli (ETEC) challenge. In order to confirm the role of TLR2, TLR4 and RP105 in the immunoregulatory effect of APS from LP14, we used small interfering RNA (siRNA) to knockdown these receptors in PIE cells. The capacity of LP14 and APS to modulate pro-inflammatory cytokine expression was significantly reduced in PIERP105−/− cells. It was also shown that LP14 and APS were capable of upregulating negative regulators of the TLR signaling in PIE cells. This work describes for the first time that a Lactobacillus strain and its EPS reduce inflammation in intestinal epithelial cells in a RP105/MD1-dependend manner.Fil: Murofushi, Yo. Tohoku University; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina. Tohoku University; JapónFil: Morie, Kyoko. Tohoku University; JapónFil: Kanmani, Paulraj. Tohoku University; JapónFil: Tohno, Masanori. National Agricultural Research Organization. National Institute of Livestock and Grassland Science; JapónFil: Shimazu, Tomoyuki. Tohoku University; JapónFil: Aso, Hisashi. Tohoku University; JapónFil: Suda, Yoshihito. Miyagi University; JapónFil: Hashiguchi, Kenji. Momoya. Research Center; JapónFil: Saito, Tadao. Tohoku University; JapónFil: Kitazawa, Haruk. Tohoku University; Japó

Toll-like receptor-2-activating bifidobacteria strains differentially regulate inflammatory cytokines in the porcine intestinal epithelial cell culture system: Finding new anti-inflammatory immunobiotics

A total of 23 strains of bifidobacteria taxonomically belonging to five species were tested for their potent immunomodulatory effect using a combination of two methods: the NF-κB-reporter assay using a toll-like receptor 2-expressing transfectant (HEK pTLR2 system) and the mitogenic assay using porcine Peyer's patches immunocompetent cells. Among the four preselected strains from different immunomodulatory groups, Bifidobacterium breve MCC-117 was able to efficiently modulate the inflammatory response triggered by enterotoxigenic Escherichia coli (ETEC) in a porcine intestinal epithelial (PIE) cell line. Moreover, using PIE cells and swine Peyer's patches immunocompetent cell co-culture system, we demonstrated that the immunoregulatory effect of B. breve MCC-117 was related to the capacity of the strain to influence PIE and immune cell interactions, leading to the stimulation of regulatory T cells. The results suggested that bifidobacteria that express high activity in both the HEK pTLR2 and the mitogenic assays may behave like potential anti-inflammatory strains. The combination of the HEK pTLR2 system, the evaluation of mitogenic activity and PIE cells will be of value for the development of new immunologically functional foods and feeds that could prevent inflammatory intestinal disorders. Although our findings should be proven in appropriate experiments in vivo, the results of the present work provide a scientific rationale for the use of B. breve MCC-117 to prevent ETEC-induced intestinal inflammation.Fil: Fujie, Hitomi. Tohoku University; JapónFil: Villena, Julio Cesar. Tohoku University; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Tohno, Masanori. National Agricultural Research Organization. National Institute of Livestock and Grassland Science. Functional Feed Research Team; JapónFil: Morie, Kyoko. Tohoku University; JapónFil: Shimazu, Tomoyuki. Tohoku University; JapónFil: Aso, Hisashi. Tohoku University; JapónFil: Suda, Yoshihito. Miyagi University; JapónFil: Shimosato, Takeshi. Shinshu University; JapónFil: Iwabuchi, Noriyuki. Morinaga Milk Industry Co. Ltd. Food Science and Technology Institute; JapónFil: Xiao, Jin-Zhong. Morinaga Milk Industry Co. Ltd. Food Science and Technology Institute; JapónFil: Yaeshima, Tomoko. Morinaga Milk Industry Co. Ltd. Food Science and Technology Institute; JapónFil: Iwatsuki, Keiji. Morinaga Milk Industry Co. Ltd. Food Science and Technology Institute; JapónFil: Saito, Tadao. Tohoku University; JapónFil: Numasaki, Muneo. Josai University; JapónFil: Kitazawa, Haruki. Tohoku University; Japó