几种商用益生芽孢杆菌特性的比较研究

doi:10.16431/j.cnki.1671-7236.2018.05.015

摘要/Abstract

摘要：

为比较几种商用益生芽孢的菌株特性，为芽孢杆菌的选育提供理论基础，本试验收集整理了15种药用（Y1~Y6）和饲用益生芽孢杆菌（S7~S15）产品，对其芽孢杆菌进行分离鉴定，分别从产淀粉酶、纤维素酶和蛋白酶等酶活特性，抗菌活性，耐酸和耐0.3%胆盐稳定性，抗生素敏感性，耗氧及生长速率测试等方面对芽孢杆菌特性进行了比较研究。结果显示，分离的15株芽孢杆菌均能产蛋白酶、纤维素酶、淀粉酶，但不同菌株产酶能力差异较大，S9菌株表现出较高的产蛋白酶、纤维素酶、淀粉酶活力；所有分离的芽孢杆菌均能对金黄色葡萄球菌及藤黄微球菌产生显著抑制活性，但只有7株芽孢杆菌（Y2、Y6、S7、S9、S10、S11、S12）可对大肠杆菌产生抑制作用，尤其是S9菌株综合抗菌能力最强。S9、Y2和S8的营养体细胞具有较高的耐酸能力，S9、Y2菌株在0.3%胆盐环境中存活率超过90%。15株芽孢杆菌中有3株饲用来源菌株（S10、S12、S13）表现出对红霉素的耐受，S10菌株对四环素耐受。此外，S9菌株具有较快的耗氧速率和生长速率。上述结果表明，S9菌株（解淀粉芽孢杆菌）的益生特性更为优良，本试验结果可为芽孢杆菌益生菌的选育及活性评价提供参考依据。

关键词: 芽孢杆菌; 益生菌; 特性比较

Abstract:

The probiotic characteristics of commercial Bacillus products were compared using screening techniques in this study.Fifteen samples of medicinal (Y1-Y6) and feed sources (S7-S15) commercial Bacillus probiotics were collected to isolate and identify the Bacillus spp.,and their probiotic characteristics were compared including enzymic properties of amylase,cellulase and protease,antimicrobial activities,tolerance tests to acid and 0.3% bile salts, antibiotics sensitivity,oxygen-consumptions rate and growth rate.The results showed that all the 15 strains could produce amylase,cellulase and protease with different enzyme capacity,and that produced by S9 strain was the highest.All strains could inhibit the strains of Staphylococcus aureus and Micrococcus luteus.Only 7 strains (Y2,Y6,S7,S9,S10,S11 and S12) showed antimicrobial activity against E.coli.And the S9 strain showed the highest antimicrobial activity against all the three indicator bacteria.The vegetative cells of S9,Y2 and S8 had a strong tolerance to low acid environment and the survival rates of S9 and Y2 strains in 0.3% bile salt environment were more than 90%.Three strains (S10,S12 and S13) were resistant to erythrocin and S10 strain was resistant to tetracycline.The S9 strain grew faster with higher rate of oxygen consumption.In conclusion,S9 strain identified as Bacillus amyloliquefaciens showed better beneficial characteristics in all the 15 commercial probiotic Bacillus strains,and the study results could provide references for the selection and evaluation of probiotics of Bacillus.

Key words: Bacillus; probiotics; characterization comparison

中图分类号:

S816.7

杨坤明, 任航, 苏雅婷, 郭小华. 几种商用益生芽孢杆菌特性的比较研究[J]. 《中国畜牧兽医》, 2018, 45(5): 1241-1250.

YANG Kunming, REN Hang, SU Yating, GUO Xiaohua. Comparative Evaluation of Probiotic Characterization of Commercial Bacillus Products[J]. , 2018, 45(5): 1241-1250.

参考文献

[1] ZHANG W,ZHU Y H,ZHOU D,et al.Oral administration of a select mixture of Bacillus probiotics affects the gut microbiota and goblet cell function following Escherichia coli challenge in newly weaned pigs of genotype muc4 that are supposed to be enterotoxigenic E.coli F4ab/ac receptor negative[J].Applied and Environmental Microbiology,2017,83(3):e02747.
[2] HONG H A,HUANG J M,KHANEJA R,et al.The safety of Bacillus subtilis and Bacillus indicus as food probiotics[J].Journal of Applied Microbiology,2008,105(2):510-520.
[3] HONG H A,DUC L H,CUTTING S M.The use of bacterial spore formers as probiotics[J].FEMS Mcrobiol, 2005,29(4):813-815.
[4] ALEXOPOULOS C,GEORGOULAKIS I E,TZIVARA A,et al.Field evaluation of the effect of a probiotic-containing Bacillus licheniformis and Bacillus subtilis spores on the health status,performance,and carcass quality of grower and finisher pigs[J].Journal of Veterinary Medicine Series A-Physiology Pathology Clinical Medicine,2004,51(6):306-312.
[5] EFSA PANEL ON ADDITIVES AND PRODUCTS OF SUBSTANCES USED IN ANIMAL FEED (FEEDAP).Scientific opinion on the safety and efficacy of Calsporin^® (Bacillus subtilis) as a feed additive for piglets on request from the European Commission[J].EFSA Journal,2010,8(1):1426-1436.
[6] WILLIAMS L D,BURDOCK G A,JIMENEZ G,et al.Literature review on the safety of Toyocerin, a non-toxigenic and non-pathogenic Bacillus cereus var.toyoi preparation[J].Regulatory Toxicology and Pharmacolog, 2009,55(2):236-246.
[7] MANHAR A K,BASHIR Y,SAIKIA D,et al.Cellulolytic potential of probiotic Bacillus subtilis AMS6 isolated from traditional fermented soybean (Churpi):An in vitro study with regards to application as an animal feed additive[J].Microbiological Research,2016,186-187:62-70.
[8] AZIMIRAD M,ALEBOUYEH M,NAJI T.Inhibition of lipopolysaccharide-induced interleukin 8 in human adenocarcinoma cell line ht-29 by spore probiotics:B.coagulans and B.subtilis (natto)[J].Probiotics & Antimicrobial Proteins,2016:9(1):56-63.
[9] HEO J,KIM S K,PARK K S,et al.A double-blind,randomized,active drug comparative,parallel-group, multi-center clinical study to evaluate the safety and efficacy of probiotics (Bacillus licheniformis, Zhengchangsheng^® capsule) in patients with diarr-hea[J].Intestinal Research,2014,12(3):236-244.
[10] CASULA G,CUTTING S M.Bacillus probiotics:Spore germination in the gastrointestinal tract[J].Applied and Environmental Microbiology,2002,68(5):2344-2352.
[11] 陶荣霞.17株芽孢杆菌产酶特性及益生特性的研究[D].武汉:华中农业大学,2012. TAO R X.Study on characteristics of enzyme production and probiotics by 17Bacillus strains[D].Wuhan:Huazhong Agricultural University,2012.(in Chinese)
[12] GUO X H,LI D F,LU W Q,et al.Screening of Bacillus strains as potential probiotics and subsequent confirmation of the in vivo effectiveness of Bacillus subtilis MA139 in pigs[J].Antonie van Leeuwenhoek,2006,90(2):139-146.
[13] 任平,阮祥稳,赵文娟,等.产酶益生芽孢杆菌的筛选[J].江苏农业科学,2014,42(10):362-365. REN P,RUAN X W,ZHAO W J,et al.Screening on enzyme production of Bacillus probiotics[J].Jiangsu Agricultural Sciences,2014,42(10):362-365.(in Chinese)
[14] 刘清源,赵献军.两株益生菌对16种抗菌药敏感性测定[J].动物医学进展,2002,22(4):73-74. LIU Q Y,ZHAO X J.The determination of the sensitivity of the two probiotics to the 16 kinds of antibacterial[J].Progress in Veterinary Medicine,2002,22(4):73-74.(in Chinese)
[15] BARBOSA T M,SERRA C R,WOODWARD M J,et al.Screening for Bacillus isolates in the broiler gastrointestinal tract[J]. Applied and Environmental Microbiology,2005,71(2):968-978.
[16] DUC LE H,HONG H A,BARBOSA T,et al.Characterization of Bacillus probiotics available for human use[J]. Applied and Environmental Microbiology,2004,70(4):2161-2171.
[17] HOA N T,BACCIGALUPI L,HUXHAM A,et al.Characterization of Bacillus species used for oral bacteriotherapy and bacterioprophylaxis of gastrointestinal disorders[J]. Applied and Environmental Microbiology,2000,66(12):5241-5247.
[18] JADAMUS A,VAHJEN W,SIMON O.Growth behaviour of a spore forming probiotic strain in the gastrointestinal tract of broiler chicken and piglets[J].Archives of Animal Nutrition,2001,54(1):1-17.
[19] 李雅丽,秦艳,周绪霞,等.6株芽孢杆菌的生物学特性比较研究[J].中国畜牧兽医,2011,38(4):62-66. LI Y L,QIN Y,ZHOU X X,et al.Comparative study on biological characteristics of six Bacillus strains[J].China Animal Husbandry & Veterinary Medicine,2011,38(4):62-66.(in Chinese)
[20] 李冠楠,夏雪娟.蚕用益生芽孢杆菌SWL-19的筛选鉴定及其对肠道菌群多样性的影响[J].中国农业科学,2015, 48(9):1845-1853. LI G N,XIA X J.Screening and identification of silkworm probiotic Bacillus SWL-19 and its effect on intestinal microflora diversity[J].Scientia Agricultura Sinaca,2015,48(9):1845-1853.(in Chinese)
[21] 张娟,杨彩梅.解淀粉芽孢杆菌及其作为益生菌的应用[J].动物营养学报,2014,26(4):863-867. ZHAN J,YANG C M.Bacillus amylolytius and its application as a probiotic[J].Chinese Journal of Animal Nutrition,2014,26(4):863-867.(in Chinese)
[22] HYRONIMUS B,MARREC C L E,SASSI A H,et al.Acid and bile tolerance of spore-forming lactic acid bacteria[J].International Journal of Food Microbiology,2000,61(2-3):193-197.
[23] ELSHAGHABEE F M F,ROKANA N,GULHANE R D,et al.Bacillus as potential probiotics:Status,concern, and future perspectives[J].Frontiers in Microbiology,2017,8:1490-1497.