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

Abstract

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

CLC Number:

S811.6

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.

References

[1] 李玉梅.东北地区梅花鹿四种疫病的血清学调查[D].长春:吉林农业大学,2005. LI Y M.Serological examination of sika deer on four loemias in Dongbei district[D].Changchun:Jilin Agricultural University,2005.(in Chinese)
[2] DAVIES J,DAVIES D.Origins and evolution of antibiotic resistance[J].Microbiology and Molecular Biology Reviews,2010,74(3):417-433.
[3] 刘政.酵母菌微生态制剂制备工艺研究及应用[D].呼和浩特:内蒙古农业大学,2017. LIU Z.Studies and application on the processing technology of the probiotic with yeast[D].Hohhot:Inner Mongolia Agricultural University,2017.(in Chinese)
[4] 刘程.复合益生菌对泌乳中期奶牛生产性能和瘤胃菌群的影响[D].杭州:浙江大学,2020. LIU C.Effect of compound probiotics on milk performance and rumen flora of mid-lactation dairy cows[D].Hangzhou:Zhejiang University,2020.(in Chinese)
[5] 王鸿泽,康坤,李彪,等.活酵母在反刍动物生产中的应用及作用机制[J].中国饲料,2023,7:167-172. WANG H Z,KANG K,LI B,et al.The application and mechanism of action of live yeast in ruminant production[J].China Feed,2023,7:167-172. (in Chinese)
[6] AKRAMIENE D,KONDROTAS A,DIDZIAPETRIENE J,et al.Effects of beta-glucans on the immune system[J].Medicina (Kaunas),2007,43(8):597-606.
[7] 陈玉洁.酸马奶源酵母菌代谢物对致病性大肠杆菌的抑菌作用机理研究[D].呼和浩特:内蒙古农业大学,2015. CHEN Y J.Study of antibacterial mechanism of yeasts metabolites from koumiss on pathogenic Escherichia coli[D].Hohhot:Inner Mongolia Agricultural University,2015.(in Chinese)
[8] 卢奇成,吴妍妍,张文举.布拉迪酵母菌的生物学功能、作用机制及其应用[J].中国畜牧兽医,2020,47(8):2472-2480. LU Q C,WU Y Y,ZHANG W J.Biological function,mechanism of action and application of Saccharomyces boulardii[J].China Animal Husbandry & Veterinary Medicine,2020,47(8):2472-2480.(in Chinese)
[9] 张惠,魏宗友,王栋麟,等.活性酵母对湖羊生长性能、血液指标的影响[J].饲料工业,2024,45(6):57-61. ZHANG H,WEI Z Y,WANG D L,et al.Effects of active yeast on growth performance and blood indexes of Hu sheep[J].Feed Industry,2024,45(6):57-61.(in Chinese)
[10] 杨佳嘉,黄顺红.酿酒酵母及其培养物对动物生产和经济效益的影响[J].饲料研究,2021,44(12):156-159. YANG J J,HUANG S H.Effect of Saccharomyces cerevisiae and its culture on animal production and economic benefits[J].Feed Research,2021,44(12):156-159.(in Chinese)
[11] 桂林生,陈渲,周力,等.日粮中添加酵母培养物对青海藏羊生长性能和瘤胃微生物多样性的影响[J].四川农业大学学报,2023,41(5):912-917. GUI L S,CHEN X,ZHOU L,et al.Effects of adding yeast cultures to diets on growth performance and rumen microbial diversity of Qinghai Tibetan sheep[J].Journal of Sichuan Agricultural University,2023,41(5):912-917.(in Chinese)
[12] AUNG M,OHTSUKA H,IZUMI K.Short communication:Effect of yeast cell wall supplementation on peripheral leukocyte populations and mRNA expression of cytokines in lactating dairy cows[J].Journal of Dairy Science,2020,103(6):5634-5640.
[13] LI Z,FAN Y,BAI H,et al.Live yeast supplementation altered the bacterial community’s composition and function in rumen and hindgut and alleviated the detrimental effects of heat stress on dairy cows[J].Journal of Animal Science,2023,101:skac410.
[14] 高晓莎.瘤胃源酵母菌的分离筛选及其对绵羊-亚急性瘤胃酸中毒的防治效果[D].太谷:山西农业大学,2020. GAO X S.Isolation and screening of rumen-derived yeast and its application in prevention and treatment its application in prevention and treatment effect of sheep SARA[D].Taigu:Shanxi Agricultural University,2020.(in Chinese)
[15] KUMAR S,PATTANAIK A K,SHARMA S,et al.Comparative assessment of canine-origin Lactobacillus johnsonii CPN23 and dairy-origin Lactobacillus acidophillus NCDC 15 for nutrient digestibility,faecal fermentative metabolites and selected gut health indices in dogs[J].Journal of Nutritional Science,2017,6:e38.
[16] 全海志.犬源乳酸菌的分离鉴定及生化特性研究[D].延吉:延边大学,2003. QUAN H Z.The isolation,identification and research of biological property of Lactobacillus from the canine[D].Yanji:Yanbian University,2003.(in Chinese)
[17] 路福平,李玉.微生物学实验技术[M].2版.北京:中国轻工业出版社,2020. LU F P,LI Y.Experimental Techniques of Microbiology[M].2nd edition.Beijing:China Light Industry Press,2020.(in Chinese)
[18] 胡瑞卿.酵母菌的特征与鉴定手册[M].青岛:中国海洋大学出版社,1991. HU R Q.Yeasts Characteristics and Identification[M].Qingdao:China Ocean University Press,1991.(in Chinese)
[19] ELGHANDOUR M M Y,KHUSRO A,ADEGBEYE M J,et al.Dynamic role of single-celled fungi in ruminal microbial ecology and activities[J].Journal of Applied Microbiology,2020,128(4):950-965.
[20] AMIN A B,MAO S.Influence of yeast on rumen fermentation,growth performance and quality of products in ruminants:A review[J].Animal Nutrition,2021,7(1):31-41.
[21] 白楠.改性酵母葡聚糖的制备及其作为免疫增强剂在两种甲壳动物上的应用[D].青岛:中国海洋大学,2013. BAI N.Modified yeast glucan preparation and its application as immunostimulant for two crustaceans[D].Qingdao:Ocean University of China,2013.(in Chinese)
[22] GRZE AŚG KOWIAK Ł,ENDO A,BEASLEY S,et al.Microbiota and probiotics in canine and feline welfare[J].Anaerobe,2015,34:14-23.
[23] SUNTARA C,CHERDTHONG A,WANAPAT M,et al.Isolation and characterization of yeasts from rumen fluids for potential use as additives in ruminant feeding[J].Veterinary Sciences,2021,8(3):52.
[24] SORO A B,OLIVEIRA M,O’DONNELL C P,et al.Ultrasound assisted modulation of yeast growth and inactivation kinetics[J].Ultrasonics Sonochemistry,2021,80:105819.
[25] 廖娟,李嘉宇,黄杰惠,等.耐低温酵母的分离鉴定及对低温酿造米酒品质的影响[J].食品工业科技,2024,45(7):159-166. LIAO J,LI J Y,HUANG J H,et al.Isolation and identification of cold adaptive yeast and its influence on the quality of low temperature brewing rice wine[J].Science and Technology of Food Industry,2024,45(7):159-166.(in Chinese)
[26] WANG Y,LI Z,JIN W,et al.Isolation and characterization of ruminal yeast strain with probiotic potential and its effects on growth performance,nutrients digestibility,rumen fermentation and microbiota of Hu sheep[J].Journal of Fungi (Basel,Switzerland),2022,8(12):1260.
[27] 张一平,田雨晴,李秋凤,等.单宁提取物对西门塔尔育肥牛生长性能、营养物质表观消化率、血清生化和抗氧化指标的影响[J].动物营养学报,2024,36(3):1713-1723. ZHANG Y P,TIAN Y Q,LI Q F,et al.Effects of tannin extract on growth performance,nutrient apparent digestibility,serum biochemical and antioxidant indices of Simmental fattening cattle[J].Chinese Journal of Animal Nutrition,2024,36(3):1713-1723.(in Chinese)
[28] 姚志芳,冯宇哲,王磊,等.益生性酵母菌的筛选与鉴定[J].家畜生态学报,2021,42(5):38-44. YAO Z F,FENG Y Z,WANG L,et al.Screening and identification of probiotic yeast[J].Journal of Domestic Animal Ecology,2021,42(5):38-44.(in Chinese)
[29] HALDER S,YADAV K K,SARKAR R,et al.Alteration of Zeta potential and membrane permeability in bacteria:A study with cationic agents[J].Springerplus,2015,4:672.
[30] 钟舒红,彭红艳,李军,等.1株牛源屎肠球菌的分离鉴定及其益生特性[J].江苏农业科学,2024,52(2):198-204. ZHONG S H,PENG H Y,LI J,et al.Isolation,identification and probiotic characteristics of a bovine Enterococcus faecium[J].Jiangsu Agricultural Sciences,2024,52(2):198-204.(in Chinese)
[31] HU X Q,LIU Q,HU J P,et al.Identification and characterization of probiotic yeast isolated from digestive tract of ducks[J].Poultry Science,2018,97(8):2902-2908.
[32] 马翠柳,蔡熙姮,刘超楠,等.北极狐源唾液乳杆菌ZJBF005的分离鉴定与体外益生功能的评价[J].动物营养学报,2023,35(5):3372-3381. MA C L,CAI X H,LIU C N,et al.Isolation and identification of Lactobacillus salivarius ZJBF005 from Alopex lagopus and its probiotic function evaluation in vitro[J].Chinese Journal of Animal Nutrition,2023,35(5):3372-3381.(in Chinese)
[33] SELVIN J,MAITY D,SAJAYAN A,et al.Revealing antibiotic resistance in therapeutic and dietary probiotic supplements[J].Journal of Global Antimicrobial Resistance,2020,22:202-205.
[34] SHEN Y,BAI X,ZHANG Y,et al.Evaluation of the potential probiotic yeast characteristics with anti-MRSA abilities[J].Probiotics and Antimicrobial Proteins,2022,14(4):727-740.
[35] LIU C,ZHANG Z Y,DONG K,et al.Antibiotic resistance of probiotic strains of lactic acid bacteria isolated from marketed foods and drugs[J]. Biomedical and Environmental Sciences,2009,22(5):401-412.
[36] KAEWNOPPARAT S,DANGMANEE N,KAEWNOPPARAT N,et al. In vitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina of a healthy woman[J].Anaerobe,2013,22:6-13.
[37] 王卓,刘晓颖,李光玉,等.鹿源枯草芽孢杆菌对小鼠生长性能、细菌易位、血清生化指标和血液学指标的影响[J].特产研究,2019,41(4):47-52. WANG Z,LIU X Y,LI G Y,et al.Effects of the treatment of Bacillus subtilis isolated from sika deer on the growth performance,bacterial translocation,serum biochemical indices and hematological indices in mice[J].Special Wild Economic Animal and Plant Research,2019,41(4):47-52.(in Chinese)

Isolation and Identification of Candida rugosa DZ018 from Deer and Its Probiotic Properties in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	SUN Huimin, FAN Bingfeng, ZHANG Xulin, SHAO Jing, LIU Lixiang, XU Baozeng. Transcriptomic Analysis of Blood in Early Pregnancy at Different Time Points in Sika Deer [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3892-3908.
[2]	XU Fei, WEN Guilan, GONG Xinyong, WEN Ming, CHEN Bofan. Isolation and Identification of a Multidrug-resistant Strain of Enterococcus faecalis from Dog and Analysis of Its Pathogenicity [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(8): 3615-3624.
[3]	LIU Wenkai, WANG Ping, JIN Yinghong, XUE Jing, LI Yue, LIANG Qianqian, LI Xiaozhuo, HAN Xiangshu, ZHENG Qiming, JIANG Song, XIA Jun. Isolation,Identification and Genetic Evolution Analysis of a Strain of Bovine Coronavirus [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 3100-3108.
[4]	ZHAO Yiduo, ZHANG Kang, WANG Lei, LU Xiaorong, ZHANG Jingyan, CHENG Jia, GUO Zhiting, XU Guowei, HUANG Liping, NIU Yuhui, MA Ting, FENG Xiaowei, EGIDE, LIU Mingchao, LI Jianxi. Isolation,Identification and Pathogen Characteristics Evaluation of a Strain of Pasteurella multocida D:L3:L5 from Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 3128-3141.
[5]	CHEN Yixing, ZHANG Meilin, LI Shikai, YANG Zhican, LI Wanjia, CHEN Jidang, ZHU Wanjun, ZHANG Jipei. Isolation,Identification and Drug Resistance Analysis of Riemerella anatipestifer from Some Areas in Guangdong [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 3185-3194.
[6]	YU Fuxin, WU Siyu, YU Jieshi, ZHANG Xiaoai, YU Ting, DONG Bo, LI Jianbo, WANG Lei, WEI Wenkang. Isolation,Identification,Whole Genome Sequencing and Genetic Evolution Analysis of Goose Astrovirus in Some Areas of Guangdong Province [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(6): 2633-2643.
[7]	WU Yuxing, ZOU Weili, CHENG Zixin, MO Yupeng, ZHANG Lingyuan, JI Wantong, CHEN Siyu, ZHENG Haodong, MA Runwen, LI Xun, LI Maoning, WANG Xiaoye. Isolation, Identification and Pathogenicity Analysis of a Strain of Swine-derived Clostridium perfringens Type A [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(6): 2697-2706.
[8]	LIU Dingyu, LIU Baoling, HE Zhenwen, LUO Qin, QIAO Changhong, CHEN Xiangyu, WANG Xiaohu, XIANG Hua, WANG Gang, TAN Chen, CAI Rujian. Isolation,Identification and Pathogenicity Analysis of a Strain of Streptococcus suis Causing Meningitis in Piglets [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2143-2153.
[9]	MA Ziwei, MA Xuejun, MA Yanan, SUN Yawei, OMAR Yakup, SHAOBAI Xinyue, YANG Zuobin, WANG Xuanyi, WANG Mengjiao, ZHONG Qi, MA Xuelian, YAO Gang. Isolation,Identification and Drug Resistance Analysis of Clostridium perfringens in Calves in a Large-scale Angus Beef Cattle Farm in Kashgar,Xinjiang [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2188-2197.
[10]	DUAN Xingxun, LIU Lu, GAO Shiwen, MA Yuhui, CAI Tao, LIU Laizhen, GU Weifang, ZHAO Hongqiong. Isolation and Identification,Drug Resistance and Pathogenicity Analysis of A Rhodococcus equi Strain from Horse [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2210-2218.
[11]	HAN Hui, GUO Yajing, YAN Guangzhi, CHEN Shengnan, LIU Mingjie, MO Meilian, HUANG Liangzong. Isolation,Identification and Genetic Evolution Analysis of a Eurasian Avian-like Swine Influenza Virus [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1642-1650.
[12]	LIU Chunhong, LYU Zhihang, ZHANG Yuqian, LIAN Chunyang, ZHENG Xucan, YAN Changyan, ZHANG Xuelian. Isolation and Identification of Chicken-derived Salmonella Senftenberg and Detection of Drug Resistance and Virulence Genes [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1773-1783.
[13]	LI Yuqi, ZHU Zhihao, ZHENG Xin, XU Guizong, CAI Sisi, ZHOU Xianjing, JIN Yuxin, LIU Lu, XU Jingru, CHEN Fangting, XUE Mengru, KANG Yali, XUE Yuxi, CHEN Tengteng, LIN Xipan, LIN Kaixiong, TAN Congling, ZHANG Min, FAN Kewei. Isolation,Identification and Pathogenicity of Providencia alcalifaciens from South China Tiger [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(3): 1320-1328.
[14]	LIU Huanghao, HUANG Yanhua, LUO Yuhang, REN Tongwei, QIN Yifeng, WEI Zuzhang, OUYANG Kang, CHEN Ying, XIE Jiang, LI Fengmei, CHEN Jicheng, WANG Xiaoling, PAN Yan, HUANG Weijian. Isolation and Identification of Bovine Enterovirus Types E and F in Guangxi [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5489-5498.
[15]	MENG Fanxing, ZENG Jiangyong, QIN Haofeng, ZHAO Congming, YUAN Zhenjie, WANG Dongjing, GESANG Zhuoga, ZHUO Ma, WU Qingxia, MA Hongcai. Isolation and Identification of Staphylococcus chromogenes from Yak Milk and Analysis of Virulence and Drug Resistance [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5539-5548.