Potential Role of Probiotics in Mechanism of Intestinal Immunity

Probiotics are nonpathogenic bacteria exert a constructive influence on health or physiology of the host. Effect of probiotics in the intestinal defense against variety of diseases is well known. The probiotics are involved in the mechanism of intestinal defense, support as antagonist against pathogens, improve intestinal epithelial layer and boost the innate as well as adaptive immunity. However these responses are also exerted by intestinal components. The intestinal components as well as probiotics play a reciprocal role to enhance the immune response of the individual. The possibilities of mechanism of action include the stimulation of epithelial cells, activation of dendritic cells via toll-like receptors (TLRs), conversely produce cytokines. These observations reviewed together advocate that specific immunomodulatory properties of probiotic bacteria should be focusing on mechanism of action via antigen presenting cells (APC)

Effect of Orally Administered Enterococcus faecium EF1 on Intestinal Cytokines and Chemokines Production of Suckling Piglets

The objective of this study was to determine the effect of orally administered Enterococcus faecium EF1 on intestinal cytokines and chemokines production in piglets. Twenty-four newborn piglets were randomly divided into two groups. The treatment group (T1), orally administered sterilized (110 ºC for 30 min) skim milk 10% (2 ml/piglet/day) with addition of viable E. faecium EF1 (5~6×108 cfu/ml) on 1st, 3rd and 5th day after birth. The control group (T0), were fed the same volume of sterilized skim milk without addition of probiotics. Feeding trial was conducted for 25 days of suckling age. At the end of trail six piglets were randomly selected from each group to collect the samples of jejunum and ileum mucosa to observe the cytokines and chemokines production. The results showed that concentrations of IL-10 and TGF-β1 significantly increased in T1 group. Whereas, production of IL-1β, IL-6, IL-12, IFN-γ and IL-8 decreased in T1 compared to T0. Levels of TNF-α were increased in jejunal mucosa, while decreased in ileal mucosa comparatively in T1 group. Our findings revealed that oral administration of E. faecium EF1 induced a strong anti-inflammatory response in the small intestine. These immunomodulatory effects of this bacterium might contribute to maintenance of immune homeostasis in the intestine of piglets

Application of Probiotic (Bacillus subtilis) to Enhance Immunity, Antioxidation, Digestive Enzymes Activity and Hematological Profile of Shaoxing Duck

The study was designed to evaluate the effects of probiotics (Bacillus subtilis) to enhance immunity, antioxidation, digestive enzymes activity and hematological profile of Shaoxing duck. A population of 200 laying ducks (160 days old) was divided into two groups each further divided in five replications. The control (G1) were fed on basal diet and (G2) with B. subtilis 1×108 cfu/kg in addition of basal diet for thirty five days. The results showed that, ducks were treated with probiotics (B. subtilis), their serum IL-2 increased and IL-10 decreased (P<0.05). The concentrations of IgG, IgA and sIgA were observed significantly higher in (G2) as compared to (G1). Treatment group (G2), showed significantly improvement in (SOD), T-AOC and ASAFR activity in serum and liver. However, digestive enzymes amylase and trypsin activity also improved (P<0.05) in (G2). The blood chemistry analysis showed significant decrease in FT3 and no other significant change observed in hematological profile as compared to (G1). In conclusion, application of B. subtilis (1×108 cfu/kg) may be beneficial to improve antioxidation response, supportive in innate immunity and digestibility of fowls (Shaoxing duck)

Establishment and characterization of pygmy killer whale (Feresa attenuata) dermal fibroblast cell line.

The pygmy killer whale (Feresa attenuata) (PKW) is a tropical and subtropical marine mammal commonly found in the Atlantic, Indian and Pacific oceans. Since the PKWs live in offshore protected territories, they are rarely seen onshore. Hence, PKW are one of the most poorly understood oceanic species of odontocetes. The dermal tissue comes primarily from stranding events that occur along the coast of the Shantou, Guangdong, China. The sampled tissues were immediately processed and attached on collagen-coated 6-well tissue culture plate. The complete medium (DMEM and Ham's F12, fetal bovine serum, antibiotic and essential amino acids) was added to the culture plates. The primary culture (PKW-LWH) cells were verified as fibroblast by vimentin and karyotype analyses, which revealed 42 autosomes and two sex chromosomes X and Y. Following transfection of PKW-LWH cells with a plasmid encoding, the SV40 large T-antigens and the transfected cells were isolated and expanded. Using RT-PCR, western blot, immunofluorescence analysis and SV40 large T-antigen stability was confirmed. The cell proliferation rate of the fibroblast cells, PKW-LWHT was faster than the primary cells PKW-LWH with the doubling time 68.9h and 14.4h, respectively. In this study, we established PKW dermal fibroblast cell line for the first time, providing a unique opportunity for in vitro studies on the effects of environmental pollutants and pathogens that could be determined in PKW and/or Cetaceans

Induction of Probiotic Strain Enterococcus faecium EF1 on the Production of Cytokines, Superoxide Anion and Prostaglandin E2 in a Macrophage Cell Line

Immunological effects of probiotic strain Enterococcus faecium EF1 was evaluated through the production of cytokines, superoxide anion (O2.-) and prostaglandin E2 (PGE2) in murine macrophage cell line RAW 264.7. In addition, the responses were contrasted with a pathogenic coliform, Escherichia coli strain K88. After 12-h co-incubation of RAW 264.7 with E. faecium EF1 or E. coli K88, the culture supernatants were collected for further analysis of cytokines (TNF-α, IFN-γ, IL-1β, IL-6, IL-12, IL-8 and IL-10) and inflammatory mediators including superoxide anion (O2.-) and PGE2. The results showed that, E. faecium EF1 induced an increased (P<0.01) release of TNF-α, IFN-γ, IL-6, IL-10 and O2.- in RAW 264.7 cells. However, levels of cytokines induced by E. faecium EF1 were far lower (P<0.01) than those observed following co-culture with E. coli K88. In addition, when RAW 264.7 cells were first treated with E. faecium EF1 and then infected with E. coli K88, the production of IL-12 and O2.- in response to E. coli K88 were significantly suppressed, indicating that E. faecium EF1 may limit the ability of macrophages to induce excessive inflammation even to potent inflammatory bacteria. Our current findings concluded that, E. faecium EF1 was capable of triggering a moderate innate inflammatory response on direct contact with murine macrophage cell line RAW264.7

Proanthocyanidins Alleviates AflatoxinB1-Induced Oxidative Stress and Apoptosis through Mitochondrial Pathway in the Bursa of Fabricius of Broilers

Aflatoxin B1 (AFB1) is a serious threat to the poultry industry. Proanthocyanidins (PCs) demonstrates a broad range of biological, pharmacological, therapeutic, and chemoprotective properties. The aim of this study was to investigate the ameliorative effects of PCs against AFB1-induced histopathology, oxidative stress, and apoptosis via the mitochondrial pathway in the bursa of Fabricius (BF) of broilers. One hundred forty-four one-day old Cobb chicks were randomly assigned into four treatment groups of six replicates (6 birds each replicate) for 28 days. Groups were fed on the following four diets; (1) Basal diet without addition of PCs or AFB1 (Control); (2) basal diet supplemented with 1 mg/kg AFB1 from contaminated corn (AFB1); (3) basal diet supplemented with 250 mg/kg PCs (PCs); and (4) basal diet supplemented with 1 mg/kg AFB1 + 250 mg/kg PCs (AFB1+ PCs). The present study results showed that antioxidant enzymes activities of total superoxide dismutase (T-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione S-transferase (GST) in AFB1 treated group were (p &lt; 0.05) decreased, whereas malondialdehyde (MDA) contents were significantly increased in comparison with the control group. Furthermore, we found that dietary PCs treatment ameliorated AFB1-induced oxidative stress in the BF through inhibiting the accumulation of MDA content and enhancing the antioxidant enzymes activities (T-SOD, CAT, GSH-Px, and GST). Similarly, PCs markedly enhanced messenger RNA (mRNA) expression of antioxidant genes (SOD, CAT, GPx1, and GST) in comparison with AFB1 group. Moreover, histological results showed that PCs alleviated AFB1-induced apoptotic cells in the BF of broilers. In addition, both mRNA and protein expression results manifested that mitochondrial-apoptosis-associated genes (Bax, caspase-9, caspase-3, and p53 and cytochrome c) showed up-regulation, while (Bcl-2) showed down-regulation in AFB1 fed group. The supplementation of PCs to AFB1 diet significantly reversed the mRNA and protein expression of these apoptosis-associated genes, as compared to the AFB1 group. Our results demonstrated that PCs ameliorated AFB1-induced oxidative stress by modulating the antioxidant defense system and apoptosis in the BF through mitochondrial pathway in broilers

Saccharomyces boulardii and Bacillus subtilis B10 modulate TLRs and cytokines expression patterns in jejunum and ileum of broilers.

The present study was designed to evaluate the effects of Saccharomyces boulardii (Sb) and Bacillus subtilis B10 (Bs) on intestinal epithelial Toll like receptors (TLR), and Cytokine expression response to understand the intestinal epithelial innate immune mechanism in broilers. A total of 300 birds (Sanhuang broilers) were allotted into three groups (n = 100) and each divided into five replications (n = 20). Control group (Ctr) birds were fed basal diet, broilers in experimental groups received (1×108cfu/kg feed) Sb and Bs respectively in addition to basal diet for 72 days. The result showed significant increase in mRNA expression level of TLR2, TLR4 and TLR15. Down streaming MyD88, TRAF6, TAB2 and NF-κB mRNA level noted higher, in the jejunum and ileum as compared to control group. Meanwhile, IL-6, TNFα, IL-10, TGF-β expression levels showed high expression in the jejunum of Sb and Bs groups. IL-10 expression level increased in the ileum and IL-6, TNFα, IL-10 and TGF-β expression levels increased in the jejunum of Sb group. Levels of IL-1 β, IL-17, and IL-4, increased merely in Sb group. Ileal cytokines IL-1β, IL-17 and IL-4concentration were noted higher in Sb group, and IL-1β, and IL-4 levels were up-regulated in Bs group. The results indicated that the INF-γ and IL-8 level decreased in Sb and BS groups. Serum IgA and sIgA level increased in both treatment groups. Our findings illustrated that S. boulardii and B. subtilis B10 may have a role to induce mucosal immunity by activating the TLRs and cytokines expressions in broilers

Establishment and characterization of pygmy killer whale (<i>Feresa attenuata</i>) dermal fibroblast cell line

<div><p>The pygmy killer whale (<i>Feresa attenuata</i>) (PKW) is a tropical and subtropical marine mammal commonly found in the Atlantic, Indian and Pacific oceans. Since the PKWs live in offshore protected territories, they are rarely seen onshore. Hence, PKW are one of the most poorly understood oceanic species of odontocetes. The dermal tissue comes primarily from stranding events that occur along the coast of the Shantou, Guangdong, China. The sampled tissues were immediately processed and attached on collagen-coated 6-well tissue culture plate. The complete medium (DMEM and Ham’s F12, fetal bovine serum, antibiotic and essential amino acids) was added to the culture plates. The primary culture (PKW-LWH) cells were verified as fibroblast by vimentin and karyotype analyses, which revealed 42 autosomes and two sex chromosomes X and Y. Following transfection of PKW-LWH cells with a plasmid encoding, the SV40 large T-antigens and the transfected cells were isolated and expanded. Using RT-PCR, western blot, immunofluorescence analysis and SV40 large T-antigen stability was confirmed. The cell proliferation rate of the fibroblast cells, PKW-LWHT was faster than the primary cells PKW-LWH with the doubling time 68.9h and 14.4h, respectively. In this study, we established PKW dermal fibroblast cell line for the first time, providing a unique opportunity for <i>in vitro</i> studies on the effects of environmental pollutants and pathogens that could be determined in PKW and/or Cetaceans.</p></div