鹿源藿香假丝酵母菌DZ018的分离鉴定及其体外益生特性研究

doi:10.16431/j.cnki.1671-7236.2025.01.035

摘要/Abstract

摘要： 【目的】从梅花鹿瘤胃液中分离和鉴定出具有益生菌潜能的酵母菌,并对其体外益生功能进行评价,为其在动物营养与饲料中应用以及鹿源益生菌制剂的研制提供参考。【方法】采用YPD固体培养基从健康雄性梅花鹿瘤胃液中分离出酵母菌菌株,对分离株进行形态学观察、生理生化和分子生物学鉴定,测定其生长性能及产酸性能,并绘制生长曲线;探究其在不同pH、不同浓度胆盐及人工胃液和人工肠液中的耐受性;采用纸片扩散法(K-B法)检测菌株的药物敏感性,并对菌株进行小鼠安全性试验。【结果】从梅花鹿瘤胃液中分离到1株优势酵母菌,并将其命名为DZ018,其菌落菌体形态特征、生理生化特性和ITS rRNA序列均与藿香假丝酵母菌(Candida rugosa)的特征相符。生长性能测定结果显示,分离株在YPD液体培养基中生长迅速,培养2 h时进入对数生长期,且具有一定的产酸能力。耐受性评价结果显示,分离株对低pH有较好的耐受性,在pH为2.5~6.5范围内均能正常生长,最适生长pH为4.5;对胆盐耐受性较强,在0.3%和1.0%胆盐中存活率均＞100%,在2.0%胆盐中存活率可达74.27%;在人工胃液和人工肠液中的存活率分别为31.64%和64.80%。药敏试验结果显示,分离株对氟胞嘧啶、氟康唑和酮康唑表现敏感,对两性霉素B耐药。小鼠安全性试验结果显示,分离株对小鼠生长性能和脏器系数均无不良影响。【结论】本研究从梅花鹿瘤胃液中分离获得1株具有一定益生潜力的藿香假丝酵母菌DZ018,其耐受性强、安全性好。试验结果为鹿源益生菌在鹿科动物生产中的应用及微生态制剂的开发提供参考和理论依据。

关键词: 藿香假丝酵母菌; 梅花鹿; 分离鉴定; 益生特性

Abstract: 【Objective】 Yeast with probiotic potential was isolated and identified from the rumen fluid of sika deer,and its probiotic functions in vitro were evaluated to provide a reference for its application in animal nutrition and feed and the development of deer derived probiotic preparations. 【Method】 The yeast strain was isolated from rumen fluid of healthy male sika deer by YPD solid medium.The isolate was identified by morphological observation,physiological,biochemical and molecular biology.The growth performance and acid production were measured,and the growth curve was drawn.The tolerance of the isolate in different pH,different concentration of bile salts,artificial gastric fluid and artificial intestinal fluid was investigated.The drug sensitivity of the strain was detected by Kirby-Bauer (K-B) method,and the safety of the strain was tested in mice. 【Result】 A dominant yeast strain was isolated from rumen fluid of sika deer and named DZ018.The morphological characteristics,physiological and biochemical characteristics and ITS rRNA sequence of the colony were consistent with those of Candida rugosa.The results of growth performance test showed that the isolate grew rapidly in YPD liquid medium,entered logarithmic growth stage after cultured for 2 h,and had a certain ability to produce acid.The results of tolerance evaluation showed that the isolate had good tolerance to low pH,and could grow normally in the range of pH 2.5-6.5,and the optimal growth pH was 4.5.The isolate had strong tolerance to bile salts,and the survival rate was ＞100% in 0.3% and 1.0% bile salts,and the survival rate reached 74.27% in 2.0% bile salt.The survival rate of artificial gastric fluid and intestinal fluid was 31.64% and 64.80%,respectively.The results of drug sensitivity test showed that the isolate was sensitive to fluorocytosine,fluconazole and ketoconazole,and resistant to amphotericin B.The results of safety test in mice showed that the isolate had no adverse effects on the growth performance and organ coefficient of mice. 【Conclusion】 In this study,a strain of Candida rugosa DZ018 with probiotic potential was isolated from rumen fluid of sika deer,with strong tolerance and good safety.The experimental results provided reference and theoretical basis for the application of deer probiotics in the production of deer animals and the development of microecological preparations.

Key words: Candida rugosa; sika deer; isolation and identification; probiotic properties

中图分类号:

S811.6

黄姚, 么乃全, 袁伟涛, 毛爱鹏, 张婷, 徐超. 鹿源藿香假丝酵母菌DZ018的分离鉴定及其体外益生特性研究[J]. 中国畜牧兽医, 2025, 52(1): 389-399.

HUANG Yao, YAO Naiquan, YUAN Weitao, MAO Aipeng, ZHANG Ting, XU Chao. Isolation and Identification of Candida rugosa DZ018 from Deer and Its Probiotic Properties in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 389-399.

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