RESUMO
Cow mastitis, which significantly lowers milk quality, is mainly caused by pathogenic bacteria such as E. coli. Previous studies have suggested that lactic acid bacteria can have antagonistic effects on pathogenic bacteria that cause mastitis. In the current study, we evaluated the in vitro and in vivo alleviative effects of L. plantarum KLDS 1.0344 in mastitis treatment. In vitro antibacterial experiments were performed using bovine mammary epithelial cell (bMEC), followed by in vivo studies involving mastitis mouse models. In vitro results indicate that lactic acid was the primary substance inhibiting the E. coli pathogen. Meanwhile, treatment with L. plantarum KLDS 1.0344 can reduce cytokines' mRNA expression levels in the inflammatory response of bMEC induced by LPS. In vivo, the use of this strain reduced the secretion of inflammatory factors IL-6, IL-1ß, and TNF-α, and decreased the activity of myeloperoxidase (MPO), and inhibited the secretion of p-p65 and p-IκBα. These results indicate that L. plantarum KLDS 1.0344 pretreatment can reduce the expression of inflammatory factors by inhibiting the activation of NF-κB signaling pathway, thus exerting prevent the occurrence of inflammation in vivo. Our findings show that L. plantarum KLDS 1.0344 has excellent properties as an alternative to antibiotics and can be developed into lactic acid bacteria preparation to prevent mastitis disease.
Assuntos
Escherichia coli/imunologia , Lactobacillus plantarum/imunologia , Glândulas Mamárias Animais/imunologia , Mastite/imunologia , Animais , Antibiose/imunologia , Bovinos , Células Cultivadas , Citocinas/imunologia , Citocinas/metabolismo , Células Epiteliais/imunologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Escherichia coli/metabolismo , Escherichia coli/fisiologia , Feminino , Inflamação/imunologia , Inflamação/metabolismo , Lactobacillus plantarum/fisiologia , Lipopolissacarídeos , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Animais/microbiologia , Mastite/induzido quimicamente , Mastite/microbiologia , Camundongos Endogâmicos BALB C , NF-kappa B/imunologia , NF-kappa B/metabolismo , Transdução de Sinais/imunologiaRESUMO
Macrophages are professional phagocytic cells that play a critical role in initiating immune responses by presenting antigen and phagocytic clearance. The macrophages can be targeted for immunomodulation by beneficial microbes, such as probiotics. The aim of this study is to investigate the protective effect of Saccharomyces boulardii against Clostridium perfringens infection in avian macrophage cell line HD11. In this study, HD11 macrophages were prestimulated with S. boulardii for 6 h and then infected with C. perfringens for 3 h. Results showed that S. boulardii enhanced phagocytosis and bactericidal capacity against C. perfringens by HD11 cells. The S. boulardii effectively promoted the mRNA expression of CD80, CD83, and CD197 cell-surface molecules in C. perfringens-infected HD11 cells. Moreover, we found that prestimulation with S. boulardii reduced the mRNA expression of CD40, toll-like receptor [TLR] 4, and TLR15 induced by C. perfringens and thereby downregulated the mRNA expression of myeloid differentiation primary response 88, TNF receptor associated factor 6, nuclear factor kappa-B p65 subunit, and c-Jun N-terminal kinase genes in HD11 cells. The upregulation of cytokines (interleukin [IL]-6, tumor necrosis factor alpha, and IL-10) and inducible nitric oxide synthase mRNA expression in C. perfringens-infected HD11 cells were noticeably inhibited by S. boulardii pretreatment. Conclusively, these results might provide a new insight into the role of S. boulardii in regulating avian immune defense against C. perfringens invasion and immune escape.
Assuntos
Antibiose , Infecções por Clostridium , Clostridium perfringens , Doenças das Aves Domésticas , Saccharomyces boulardii , Animais , Antibiose/imunologia , Galinhas , Infecções por Clostridium/imunologia , Infecções por Clostridium/microbiologia , Infecções por Clostridium/veterinária , Inflamação/imunologia , Macrófagos/imunologia , Macrófagos/microbiologia , Fator 88 de Diferenciação Mieloide/imunologia , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/microbiologia , Saccharomyces boulardii/imunologia , Receptor 4 Toll-Like/imunologia , Receptores Toll-Like/imunologiaRESUMO
BACKGROUND AND OBJECTIVE: Although previous studies revealed the potential use of probiotics in the control of periodontitis, little is known about their interactions with gingival epithelial cells (GECs). Since GECs comprise the first defense in the subgingival microenvironment, the aim of this study was to evaluate the effect of probiotic lactobacilli and bifidobacteria strains on OBA-9 cells challenged with Porphyromonas gingivalis. METHODS: Immortalized human GECs (OBA-9) were challenged with live P. gingivalis (strains W83 and ATCC33277) and co-infected with one of 12 tested probiotic strains at a multiplicity of infection (MOI) of 1:1000 for 2 hours. Bacterial adhesion and invasion were determined by antibiotic exclusion analysis and CFU counting. OBA-9 viability was assessed by MTT assay, and levels of inflammatory mediators (TNF-α, IL-1ß, and CXCL8) in the supernatants were determined by ELISA. The expression of genes encoding Toll-like receptors (TLR2, TLR4) was evaluated by RT-qPCR. RESULTS: Both strains of P. gingivalis were able to adhere and invade OBA-9 cells, with significant loss in cell viability, increase in the levels of TNF-α and IL-1ß, and upregulation of TLR4. However, co-infection with probiotics attenuated these effects in P. gingivalis challenged GECs. Most probiotics maintained OBA-9 viability and reduced pathogens adhesion and invasion. Furthermore, probiotics were able to adhere to GECs, which was enhanced for most strains in the presence of P. gingivalis. The synthesis of IL-1ß and TNF-α by P. gingivalis in challenged GECs was reduced in co-culture with most of the tested probiotics, whereas the secretion of CXCL8 increased, and TLR4 was downregulated. CONCLUSION: Probiotics can alter the interaction of GECs with P. gingivalis by modulating the pathogen's ability to adhere and invade these cells, as well as by regulating the innate immune response. Such properties are strain-specific and may indicate the most efficient probiotics to control periodontitis.
Assuntos
Antibiose/imunologia , Bifidobacterium/fisiologia , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Gengiva/citologia , Gengiva/imunologia , Imunidade Inata , Lactobacillus/fisiologia , Periodontite/prevenção & controle , Periodontite/terapia , Porphyromonas gingivalis/imunologia , Porphyromonas gingivalis/patogenicidade , Probióticos , Células Cultivadas , Microambiente Celular/imunologia , Humanos , Interleucina-1beta/metabolismo , Interleucina-8/metabolismo , Periodontite/imunologia , Periodontite/microbiologia , Porphyromonas gingivalis/metabolismo , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Background: the solts of the southern part of Chile, that are isolated, cold, humid, poorly axygenated and with a low acitity, could contain new strains of antimicrobial producing Streptomyces strains with antimicrobial activity towards pathogenic bacteria and fungi. Material and methods: two hundred fifty eight soil and sediment samples were collected from 148 paces in Souther regions of Chile. They were cultured in Kuster-Williams growth media and the presence of Streptomyces was confirmed by microscopic examination and biochemical characterization. The antimicrobial activity against reference microorganisms. Results: Seventy seven percent of soils were positive and 542 wild strains of Streptomyces were isolates; of these, 266 had antimicrobial activity. Fifty three percent of isolates had activity against S aureus 43 percent against B subtilis and 0.7 percent against E coli. Most Streptomyces were active against more than one organism. When there was activity against single organisms, these were mostly eucariotic, such as C albicans and T mentagrophytes. Among clinical microorganisms, 29 percent of S aureus strains were inhibited, while P aeruginosa, Alternaria sp, P vulgaris and Y enterocolitica strains were not inhibited. The most frequent Streptomyces morphotypes were those showing pigmented colonies with flexuous and spiral shaped chains of arthopores. Conclusions: soils of the southern region Chile allow the grouth of abundant native strains of Streptomyces with a promising anticrobial activity