epiglucan and Mastitis

Mastitis, caused by mammary pathogenic bacteria which are frequent implications of Escherichia coli, is an important disease affecting women and dairy animals worldwide. The β-glucan binding of dectin-1 can induce its own intracellular signaling and can mediate a variety of cellular responses. This work was to investigate the effect of β-glucan on the lipopolysaccharide (LPS)-induced inflammatory response and related innate immune signaling in primary rat mammary epithelial cells. Cells were treated with serum-free medium added with a DMSO solution containing β-glucans at concentrations of 0, 1, 5, 25 μmol/L for 12h, and then exposed to 10 μg/mL LPS for 40 min. Moreover, cells were pretreated with BAY 11-7082 to inhibit NF-κB and then successively exposed to 5 μmol/L β-glucan, 10 μg/mL LPS, 5 μmol/L β-glucan and 10 μg/mL LPS, according to the specific experimental design. Normal control cultures contained an equal volume of DMSO, which was collected at the same time. After incubating rat mammary epithelial cells for 40 min with 10 μg/mL LPS, TLR4, MyD88 and NF-κB expression all increased (P<0.05), as did the secretion of TNF-α and IL-1β (P<0.05), but IκB and β-casein expression both decreased (P<0.05). Treatment with different concentrations of β-glucan for 12h activated Dectin1/Syk, which subsequently suppressed TLR4, MyD88 and NF-κB expression and TNF-α and IL-1β secretion. However, it restored the IκB and β-casein expression that had been induced by the 40 min incubation with 10 μg/mL LPS. Pretreatment with BAY 11-7082 at 10 µmol/L for 2h partially prevented NF-κB induction by LPS, but the presence of β-glucan prevented this inactivation. BAY 11-7082 could not simultaneously inhibit LPS induction of TLR4, MyD88 and β-glucan activation of Dectin1/Syk in rat mammary epithelial cells. These findings demonstrated that β-glucan activation of Dectin1/Syk attenuated LPS induction of TLR4/MyD88/NF-κB and inhibited the LPS-induced inflammation factors in mammary epithelial cells, thereby providing a possibly protective effect of β-glucan in the prevention of LPS-induced dysfunction in mammary epithelial cells.

Topics: Animals; Anti-Inflammatory Agents; beta-Glucans; Cells, Cultured; Disease Models, Animal; Epithelial Cells; Female; Immunity, Innate; Interleukin-1beta; Lectins, C-Type; Lipopolysaccharides; Mammary Glands, Animal; Mastitis; Myeloid Differentiation Factor 88; NF-kappa B; Nitriles; Primary Cell Culture; Rats; Rats, Sprague-Dawley; Signal Transduction; Sulfones; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

A lipopolysaccharide (LPS)-induced mastitis model in rats was used to study the protective effect of β-glucan.. On gestation day 10, β-glucan was administered to rats daily by gavage at either 0 (control), 0.1 mg/kg BW, 1 mg/kg BW and 10 mg/kg BW until parturition. LPS or pyrogen-free physiological saline was inoculated into the mammary gland 72 h after parturition and the rats were euthanized at 12 h post-infection.. LPS increased dectin1 and toll-like receptor 4 (TLR4) mRNA and protein expression significantly in mammary tissues. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, N-acetyl-β-D-glucosaminidase (NAGase), inducible nitric oxide synthase (iNOs) in mammary tissues and serum, and myeloperoxidase (MPO) in mammary tissues were significantly increased 12 h after LPS infusion. β-glucan administration could enhance dectin1 mRNA and protein expression while downregulating the expression of TLR4. Level of TNF-α, IL-1β in mammary tissues and serum, MPO, NAGase and iNOs activity in mammary tissues was decreased, but the level of IL-2 in serum was increased by β-glucan.. The results indicate that β-glucan may protect mammary tissue against LPS-induced rat acute mastitis.

Topics: Animals; beta-Glucans; Female; Interleukin-1beta; Interleukin-2; Lectins, C-Type; Lipopolysaccharides; Male; Mammary Glands, Animal; Mastitis; Membrane Proteins; Nerve Tissue Proteins; Nitric Oxide Synthase Type II; Proteoglycans; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

Staphylococcus aureus vaccines based on bacterins surrounded by slime, surface polysaccharides coupled to protein carriers and polysaccharides embedded in liposomes administered together with non-biofilm bacterins confer protection against mastitis. However, it remains unknown whether protective antibodies are directed to slime-associated known exopolysaccharides and could be produced in the absence of bacterin immunizations. Here, a sheep mastitis vaccination study was carried out using bacterins, crude bacterial extracts or a purified exopolysaccharide from biofilm bacteria delivered in different vehicles. This polysaccharide reacted specifically with antibodies to poly-N-acetyl-beta-1,6-glucosamine (PNAG) and not with antibodies to other capsular antigens or bacterial components. Following intra-mammary challenge with biofilm-producing bacteria, antibody production against the polysaccharide, milk bacterial counts and mastitis lesions were determined. Bacterins from strong biofilm-producing bacteria triggered the highest production of antibodies to PNAG and conferred the highest protection against infection and mastitis, compared with weak biofilm-producing bacteria and non-cellular inocula. Thus, bacterins from strong biofilm bacteria, rather than purified polysaccharide, are proposed as a cost-efficient vaccination against S. aureus ruminant mastitis.

Topics: Animals; Antibody Formation; beta-Glucans; Biofilms; Female; Mammary Glands, Animal; Mastitis; Milk; Pregnancy; Sheep; Staphylococcal Infections; Staphylococcal Vaccines; Staphylococcus aureus; Treatment Outcome

The objective of this prospective study was to determine if Candida albicans is present in the milk of women suffering from symptoms of severe nipple and deep breast pain.. The symptomatic group included women who reported sore, inflamed, or traumatized nipples or intense stabbing or burning pain. The control group included breastfeeding women without symptoms. The skin of the nipple and areola were washed with detergent and thoroughly rinsed. Milk samples were analyzed for (1 --> 3)-beta-D-glucan and grown on Candida growth medium.. There was no significant difference in (1 --> 3)-beta-D-glucan levels between the control and symptomatic group. No Candida species were culturable either before or after the addition of iron to stimulate growth, with the exception of one patient. The addition of pure C. albicans to milk samples suggested that milk does not inhibit Candida growth.. These data suggest that C. albicans is not present in milk ducts and may not be associated with this syndrome.

Topics: Adult; beta-Glucans; Breast Feeding; Candida albicans; Candidiasis; Case-Control Studies; Female; Humans; Mastitis; Milk, Human; Nipples; Pain; Prospective Studies