华南虎源乳酸菌的分离筛选及体外益生特性

Isolation and Screening of Lactic Acid Bacteria from South China Tiger and Its Probiotic Characteristics in Vitro

乳酸菌对动物肠道健康具有非常重要的作用。为了筛选出具有益生特性的优良菌株,对健康华南虎(Panthera tigris amoyensis)粪便中的乳酸菌进行分离培养、纯化及鉴定,并通过研究菌株对大肠埃希氏杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)、沙门氏菌(Salmonella sp.)和迟钝爱德华氏菌(Edwardsiella tarda)的抑菌性及菌株的耐酸、耐胆盐、溶血性和对24种药敏纸片的耐药性,对菌株生物学特性进行评价。结果显示:共分离得到78株乳酸菌,其中15株具有较好的抑菌性能,对以上4种致病菌的抑菌圈均大于15 mm,含6株罗伊氏乳杆菌(Lactobacillus reuteri,22YY08、22DC20、22XX19、22Hui09、22LL14和22XX03)、3株卷曲乳杆菌(L. crispatus,22HH02、22YY13和22YY09)、2株动物乳杆菌(Ligilactobacillus animalis,22Hui15、22LL07)、1株戊糖片球菌(Pediococcus pentosaceus,22DC18)、1株黏膜乳杆菌(Lactobacillus mucosae,22DC10)、1株约氏乳杆菌(L. johnsonii,22DC09)和1株格氏乳球菌(Lactococcus garvieae,GaL110)。通过进一步试验筛选出8株乳酸菌,在pH=2.5和胆盐浓度为0.1%时表现出较好的耐酸、耐胆盐性能,均不产生溶血性,但其中3株罗伊氏乳杆菌22Hui09、22YY08和22LL14对多种药物耐药,其他菌株对多种药物敏感。本试验筛选获得的3株罗伊氏乳杆菌(22DC20、22XX03和22XX19)、1株戊糖片球菌(22DC18)和1株格氏乳球菌(GaL110)有希望作为具有益生特性的菌株应用到华南虎益生菌制剂的开发。

Lactic acid bacteria(LAB)plays an important role in the intestinal health of animals.In order to screen out ex⁃cellently properties LAB strains,in this study,LAB from the feces of healthy south China tigers(Panthera tigris amoyensis)were isolated,cultured,purified and identified.The biological characteristics of these strains were evaluated by studying its bacteriostatic properties against Escherichia coli,Staphylococcus aureus,Salmonella sp.and Edwardsiella tarda,acid re⁃sistance and bile salts resistance,hemolytic activity and drug against resistance test of 24 kinds of drugs.The results showed that a total of 78 strains of LAB were isolated,among which 15 strains had good antibacterial performance and the antibacterial circles were greater than 15 mm against the four pathogenic bacteria mentioned above,including six strains(22YY08,22DC20,22XX19,22Hui09,22LL14 and 22XX03)of Lactobacillus reuteri,three strains(22HH02,22YY13 and 22YY09)of L.crispatus,two strains(22Hui15 and 22LL07)of Ligilactobacillus animalis,one Pediococcus pentosaceus(22DC18),one Lactobacillus mucosae(22DC10),one L.johnsonii(22DC09),and one Lactococcus garvieae(GaL110).Further,eight LAB strains were screened out and showed good tolerance at pH 2.5 and bile salt concentration of 0.1%,and had no hemolytic activity.Among them,three strains of Lactobacillus reuteri(22Hui09,22YY08 and 22LL14)were re⁃sistant to multiple drugs while others were susceptible.In conclusion,three strains of L.reuteri(22DC20,22XX03 and 22XX19),Pediococcus pentosaceus 22DC18,and Lactococcus garvieae GaL110 obtained in this study are promising can⁃didates for the development of probiotic preparations in south China tiger.