Characterization of Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51 as novel dog probiotics with innate immune enhancing properties원문보기
Probiotics that are able to provide beneficial effects on animal health have become important ingredients of dog foods. This study was conducted to characterize the probiotic potentials of two strains, Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51, that were derived from feces of...
Probiotics that are able to provide beneficial effects on animal health have become important ingredients of dog foods. This study was conducted to characterize the probiotic potentials of two strains, Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51, that were derived from feces of healthy dogs and evaluated based on tolerance to low pH and bile acid, antimicrobial activities, enzyme profiles, sensitivity to antibiotics, and innate immune enhancing potentials. Both strains showed survival of more than 90% at pH 3 and 0.2% bile acid and exhibited broad antimicrobial activities against indicator bacteria. Moreover, both strains showed high sensitivity to antibiotics, except vancomycin, metronidazole, and gentamicin. The alkaline phosphatase was negligible (score 0), whereas they showed strong beta galactosidase activity (score range 5 or 3, respectively). The phagocytosis and oxidative burst activities of canine granulocytes were significantly enhanced in response to both strains. These results show that both strains have the capability to act as probiotics and the potential for application as ingredients in dog foods.
Probiotics that are able to provide beneficial effects on animal health have become important ingredients of dog foods. This study was conducted to characterize the probiotic potentials of two strains, Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51, that were derived from feces of healthy dogs and evaluated based on tolerance to low pH and bile acid, antimicrobial activities, enzyme profiles, sensitivity to antibiotics, and innate immune enhancing potentials. Both strains showed survival of more than 90% at pH 3 and 0.2% bile acid and exhibited broad antimicrobial activities against indicator bacteria. Moreover, both strains showed high sensitivity to antibiotics, except vancomycin, metronidazole, and gentamicin. The alkaline phosphatase was negligible (score 0), whereas they showed strong beta galactosidase activity (score range 5 or 3, respectively). The phagocytosis and oxidative burst activities of canine granulocytes were significantly enhanced in response to both strains. These results show that both strains have the capability to act as probiotics and the potential for application as ingredients in dog foods.
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문제 정의
The removal of intestinal commensals by oral administration of antibiotics increases the incidence and severity of diseases in these diverse systems, and supply of sufficient number of beneficial bacteria recovers the host from diseases [8]. These studies have highlighted the importance of probiotics that function as live beneficial bacteria in the host in maintaining health of dogs. To develop probiotics for dogs, we selected two strains of LABs, and their probiotic potentials were investigated.
제안 방법
Such effects are usually dependent on strains with different glycolipid and protein composition in their cell walls and dif-ferent CpG content of their DNA [22]. In this study, we evaluated the in vitro immunomodulatory effects of selected strains by using erythrocyte-deprived whole blood cells from dogs rather than peripheral blood mononuclear cells (PMBC). The reason we chose to use whole blood cells rather than PBMC is based on the fact that the immune cells in the blood should be exposed to live bacteria or molecules derived from the bacteria in the presence of other blood cells.
An aliquot of the cell suspension was inoculated into each well of the kit, and after addition of the reagents (Zym A and Zym B), the kit was incubated for 4 h at 37℃ under anaerobic condition. The evaluation of enzyme activity was carried out depending on the color intensity ranged from 0 to 5, which was determined by using a color code supplied by the manufacturers.
1 mL of 10-fold serially diluted samples with PBS were inoculated onto de Man-Rogosa-Sharpe (MRS; Merck, Germany) agar plate and incubated for 36 h at 37℃ anaerobically using a GasPak (Becton, Dickinson and Company, USA). The identification of resulting colonies was determined by sequencing 16S ribosomal RNA PCR product with the primer set 357F-926Rb (357F; CCTACGGGAGGCAG CAG, 926Rb; CCGTCAATTYMTT TRAGT). As a reference strain, we used Lactococcus lactis isolated from a commercial probiotic product (Activia; Pul-muon, Korea).
After washing the cells three times with PBS containing 3% FBS, the cells were analyzed within 30 min using a flow cytometer. The mean green fluorescence intensity of the granulocytes that were obtained by gating based on the forward and side scatter plot was analyzed to evaluate the oxidative burst activity.
대상 데이터
Fecal samples were obtained from 5 healthy female beagle dogs (age, 1 to 3 years; body weight, 12 to 15 kg). One gram of feces from healthy dogs was suspended in 10 mL phosphate-buffered saline (PBS), and 0.
이론/모형
The remaining pellets were extensively washed with PBS, and finally resuspended in PBS. The protein content in the solution was determined by the Bradford assay [2]. All reagents in this assay were purchased from Sigma-Aldrich.
성능/효과
In conclusion, this study shows that L. reuteri BCLR 42 and L. plantarum BCLP 51, which were isolated from dogs, can function as probiotics for dogs. Several studies have reported potential properties of L.
lactis. The MFIs by L. reuteri BCLR 42 in the form of live, inactivated and cell wall were lower than those by L. lactis, metabolites from L. reuteri BCLR 42 generated significantly higher MFI than L. lactis.
참고문헌 (42)
Adams CA. The probiotic paradox: live and dead cells are biological response modifiers. Nutr Res Rev 2010, 23, 37-46.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976, 72, 248-254.
Brink M, Todorov SD, Martin JH, Senekal M, Dicks LM. The effect of prebiotics on production of antimicrobial compounds, resistance to growth at low pH and in the presence of bile, and adhesion of probiotic cells to intestinal mucus. J Appl Microbiol 2006, 100, 813-820.
Bron PA, Tomita S, Mercenier A, Kleerebezem M. Cell surface-associated compounds of probiotic lactobacilli sustain the strain-specificity dogma. Curr Opin Microbiol 2013, 16, 262-269.
Camacho AI, Souza-Reboucas J, Irache JM, Gamazo C. Towards a non-living vaccine against Shigella flexneri: from the inactivation procedure to protection studies. Methods 2013, 60, 264-268.
Cuello-Garcia CA, Brozek JL, Fiocchi A, Pawankar R, Yepes-Nunez JJ, Terracciano L, Gandhi S, Agarwal A, Zhang Y, Schunemann HJ. Probiotics for the prevention of allergy: a systematic review and meta-analysis of randomized controlled trials. J Allergy Clin Immunol 2015, 136, 952-961.
Di Pierro F, Bertuccioli A, Marini E, Ivaldi L. A pilot trial on subjects with lactose and/or oligosaccharides intolerance treated with a fixed mixture of pure and enteric-coated $\alpha$ - and $\beta$ -galactosidase. Clin Exp Gastroenterol 2015, 8, 95-100.
Gilliland SE, Staley TE, Bush LJ. Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct. J Dairy Sci 1984, 67, 3045-3051.
Grazul H, Kanda LL, Gondek D. Impact of probiotic supplements on microbiome diversity following antibiotic treatment of mice. Gut Microbes 2016, 7, 101-114.
Handwerger S, Pucci MJ, Volk KJ, Liu J, Lee MS. Vancomycin-resistant Leuconostoc mesenteroides and Lactobacillus casei synthesize cytoplasmic peptidoglycan precursors that terminate in lactate. J Bacteriol 1994, 176, 260-264.
Helander IM, Mattila-Sandholm T. Permeability barrier of the Gram-negative bacterial outer membrane with special reference to nisin. Int J Food Microbiol 2000, 60, 153-161.
Holst BS, Gustavsson MH, Lilliehook I, Morrison D, Johannisson A. Leucocyte phagocytosis during the luteal phase in bitches. Vet Immunol Immunopathol 2013, 153, 77-82.
Kapila R, Sebastian R, Varma D VP, Sharma R, Kapasiya M, Salingati V, Kapila S, Dang AK. Comparison of innate immune activation after prolonged feeding of milk fermented with three species of Lactobacilli. Microbiol Immunol 2013, 57, 778-784.
Klaenhammer TR, Azcarate-Peril MA, Altermann E, Barrangou R. Influence of the dairy environment on gene expression and substrate utilization in lactic acid bacteria. J Nutr 2007, 137 (3 Suppl 2), 748S-750S.
Lammers KM, Brigidi P, Vitali B, Gionchetti P, Rizzello F, Caramelli E, Matteuzzi D, Campieri M. Immunomodulatory effects of probiotic bacteria DNA: IL-1 and IL-10 response in human peripheral blood mononuclear cells. FEMS Immunol Med Microbiol 2003, 38, 165-172.
Lee Y, Lee TS. Enhancement in ex vivo phagocytic capacity of peritoneal leukocytes in mice by oral delivery of various lactic-acid-producing bacteria. Curr Microbiol 2005, 50, 24-27.
Maeda K, Sakonju I, Kanda A, Suzuki T, Kakuta T, Shimamura S, Okano S, Takase K. Priming effects of lipopolysaccharide and inflammatory cytokines on canine granulocytes. J Vet Med Sci 2010, 72, 55-60.
Matsumoto S, Hara T, Hori T, Mitsuyama K, Nagaoka M, Tomiyasu N, Suzuki A, Sata M. Probiotic Lactobacillus-induced improvement in murine chronic inflammatory bowel disease is associated with the down-regulation of pro-inflammatory cytokines in lamina propria mononuclear cells. Clin Exp Immunol 2005, 140, 417-426.
McCoy S, Gilliland SE. Isolation and characterization of Lactobacillus species having potential for use as probiotic cultures for dogs. J Food Sci 2007, 72, M94-97.
Moretti J, Blander JM. Insights into phagocytosis-coupled activation of pattern recognition receptors and inflammasomes. Curr Opin Immunol 2014, 26, 100-110.
Nilsson M, Weineisen M, Andersson T, Truedsson L, Sjobring U. Critical role for complement receptor 3 (CD11b/CD18), but not for Fc receptors, in killing of Streptococcus pyogenes by neutrophils in human immune serum. Eur J Immunol 2005, 35, 1472-1481.
Oh C, Lee K, Cheong Y, Lee SW, Park SY, Song CS, Choi IS, Lee JB. Comparison of the oral microbiomes of canines and their owners using next-generation sequencing. PLoS One 2015, 10, e0131468.
Paturi G, Phillips M, Kailasapathy K. Effect of probiotic strains Lactobacillus acidophilus LAFTI L10 and Lactobacillus paracasei LAFTI L26 on systemic immune functions and bacterial translocation in mice. J Food Prot 2008, 71, 796-801.
Perelmuter K, Fraga M, Zunino P. In vitro activity of potential probiotic Lactobacillus murinus isolated from the dog. J Appl Microbiol 2008, 104, 1718-1725.
Rabiu BA, Jay AJ, Gibson GR, Rastall RA. Synthesis and fermentation properties of novel galacto-oligosaccharides by $\beta$ -galactosidases from Bifidobacterium species. Appl Environ Microbiol 2001, 67, 2526-2530.
Saez-Lara MJ, Gomez-Llorente C, Plaza-Diaz J, Gil A. The role of probiotic lactic acid bacteria and bifidobacteria in the prevention and treatment of inflammatory bowel disease and other related diseases: a systematic review of randomized human clinical trials. Biomed Res Int 2015. Epub ahead of print. doi: 10.1155/2015/505878.
Sanchez de Medina F, Martinez-Augustin O, Gonzalez R, Ballester I, Nieto A, Galvez J, Zarzuelo A. Induction of alkaline phosphatase in the inflamed intestine: a novel pharmacological target for inflammatory bowel disease. Biochem Pharmacol 2004, 68, 2317-2326.
Schmitz S, Henrich M, Neiger R, Werling D, Allenspach K. Comparison of $TNF{\alpha}$ responses induced by Toll-like receptor ligands and probiotics Enterococcus faecium in whole blood and peripheral blood mononuclear cells of healthy dogs. Vet Immunol Immunopathol 2013, 153, 170-174.
Stoyanovski S, Gacovski Z, Antonova-Nikolova S, Kirilov N, Ivanova I, Tenev T, Hadjinesheva V. API ZYM enzymatic profile of lactic acid bacteria isolated from traditional Bulgarian meat product "Lukanka". Bulg J Agric Sci 2013, 19 (Suppl 2), 86-89.
Strompfova V, Laukova A. Isolation and characterization of faecal bifidobacteria and lactobacilli isolated from dogs and primates. Anaerobe 2014, 29, 108-112.
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