籽鹅肠道中非解乳糖链球菌的分离鉴定及益生特性研究

doi:10.16431/j.cnki.1671-7236.2024.10.038

Abstract

Abstract: 【Objective】 The aim of this study was to isolate and screen potential probiotic strain from the intestinal tract of Zi geese to improve their health status，and analyze its probiotic properties. 【Method】 The contents of different intestinal tract of healthy Zi geese were collected，diluted in gradient and coated on (CaCO₃-MRS) solid medium containing calcium carbonate. After anaerobiccultivation，the isolates were identified through morphology，biochemical tests，16S rRNA gene PCR amplification，respectively. The probiotic properties of the isolates were screened by acid resistance and antibacterial tests. The antibacterial ability，antibiotic sensitivity，acid and bile salt tolerance，gastric and intestinal fluid tolerance were evaluated，and the growth curve and acid production curve were measured. 【Result】 4 suspected lactic acid bacteria strains were isolated from the intestinal tract of healthy Zi geese，named RS-1，RS-2，RS-3 and RS-4，of which RS-3 had good probiotic properties. The colony morphology of RS-3 in CaCO₃-MRS medium was smooth，light yellow and round colony with obvious calcium-soluble ring，Gram-positive Bacillus. The results of biochemical tests showed that the isolates could ferment maltose，cellobiose，sucrose，raffinose，inulin and lactose，as well as mannitol，sorbitol and salicin. The results of esculin test and 1% sodium hippurate test were positive，and the isolates were initially determined to be lactic acid bacteria. 16S rRNA gene sequence analysis showed that 4 isolates were all Streptococcus alactolyticus，and the tolerance of RS-3 was better than that of the other three strains. The fermentation broth of RS-3 had a good inhibitory effect on Staphylococcus aureus. The diameter of inhibitory zone was more than 10 mm，and it was resistant to kanamycin，amicacin，roxithromycin，sulfadiazine，lincomycin，polycolistin B and norfloxacin. Moreover，RS-3 had tolerance to acid，bile salts artificial gastric and intestinal fluids. The mice grew well after administration of RS-3. 【Conclusion】 4 isolates enterogenic Streptococcus alactolyticus of Zi geese were isolated in this experiment，of which RS-3 had good antibacterial and probiotic properties with acid and bile salt tolerance，and could be used as a reliable strain source for the subsequent development of probiotic preparations for geese.

Key words: Zi geese; Streptococcus alactolyticus; intestinal tract; probiotics; microecology

CLC Number:

S852.61⁺1

ZHANG Hong, TIAN Qiufeng, ZOU Yue, YIN Junyi, BAI Changsheng, HUANG Yuxiang, WANG Zhiqiang, MA Zhigang, HUO Mingdong, DONGJiaqiang, YANG Haotian, YANG Kun, MIAO Yan, CHEN Zhifeng. Isolation，Identification and Probiotic Characteristics of Streptococcus alactolyticus from Intestinal Tract of Zi Geese[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4578-4586.

References

[1] 尹珺伊，刘秋瑾，田秋丰，等. 一株干酪乳杆菌的分离鉴定及生物学特性研究[J]. 中国畜牧兽医，2021，48(2)：516-524.YIN J Y，LIU Q J，TIAN Q F，et al. Isolation，identification and biological characteristics of a strain of Lactobacillus casei [J]. China Animal Husbandry & Veterinary Medicine，2021，48(2): 516-524. (in Chinese)
[2] 付金蕾，高丽，黄雯静，等. 鹅盲肠乳酸菌的筛选及抑菌能力分析[J]. 黑龙江畜牧兽医，2021，23:73-77.FU J L，GAO L，HUANG W J，et al. Screening of cecal lactic acid bacteria in goose and analysis of its antibacterial ability[J]. Heilongjiang Animal Science and Veterinary Medicine，2021，23:73-77.(in Chinese)
[3] 张名爱，王宝维. 鹅源高活力纤维素分解菌的分离鉴定[J]. 中国家禽，2012，34(5):12-15.ZHANG M A，WANG B W. Isolation and identification of a high active cellulose-decomposing strain from geese[J]. China Poultry，2012，34(5):12-15. (in Chinese)
[4] 张桂山，徐晶，娄玉杰，等. 鹅肠道纤维分解菌的分离筛选及酶活力研究[J]. 吉林农业大学学报，2009，31(2):195-199.ZHANG G S，XU J，LOU Y J,et al. Isolation and enzyme activity determination of cellulose-decomposing bacteria from goose intestinal tract[J]. Journal of Jilin Agricultural University，2009，31(2):195-199.(in Chinese)
[5] FUOCHI V，FURNERI P M. Applications of probiotics and their potential health benefits[J]. International Journal of Molecular Sciences，2023，24(21):15915.
[6] SINGH R P，SHADAN A，MA Y. Biotechnological applications of probiotics: A multifarious weapon to disease and metabolic abnormality[J]. Probiotics and Antimicrobial Proteins，2022，14(6):1184-1210.
[7] GOODOORY V C，FORD A C. Antibiotics and probiotics for irritable bowel syndrome[J]. Drugs，2023，83(8):687-699.
[8] CLSI. Performance standards for antimicrobial susceptibility testing.Twenty-seven informational supplement[S]. CLSI，2017.
[9] FARROW J A E，KRUZE J，PHILIPS B A，et al. Taxonomic studies on Streptococcus bovis and Streptococcus equinus: Description of Streptococcus alactolyticus sp. nov. and Streptococcus saccharolyticus sp. nov[J]. Systematic and Applied Microbiology，1984，5(4):467-482.
[10] BAELE M，DEVRIESE L A，BUTAYE P，et al. Composition of enterococcal and streptococcal flora from pigeon intestines[J]. Journal of Applied Microbiology，2002，92(2):348-351.
[11] ALMEIDA P，RAILSBACK J，GLEASON J B. A rare case of Streptococcus alactolyticus infective endocarditis complicated by septic emboli and mycotic left middle cerebral artery aneurysm[J]. Case Reports in Infectious Disease，2016，2016:9081352.
[12] RINKINEN M L，KOORT J M，OUWEHAND A C，et al. Streptococcus alactolyticus is the dominating culturable lactic acid bacterium species in canine jejunum and feces of four fistulated dogs[J]. FEMS Microbiology Letters，2004，230(1):35-39.
[13] 韩佳临，杨星月，聂毅桐，等. 南宁地区种猪源非解乳糖链球菌的分离鉴定及其益生性研究[J]. 家畜生态学报，2022，43(3):66-71.HAN J L，YANG X Y，NEI Y T，et al. Isolation，identification and probiotic charateristics of Streptococcus alactolyticus from breeding pigs in Nanning[J]. Journal of Domestic Animal Ecology，2022，43(3):66-71. (in Chinese)
[14] 李琳琳，王雅婷，杨欣，等. 猪源乳酸菌的分离及其生物学性能研究[J].西北农林科技大学学报(自然科学版)，2016，44(2):1-7.LI L L，WANG Y T，YANG X，et al. Isolation and characterization of lactic acid bacteria from swine[J]. Journal of Northwest A&F University (Natural Science Edition)，2016，44(2):1-7. (in Chinese)
[15] 李建喜. 中兽药生物转化技术研究[D]. 北京：中国农业科学院，2012.LI J X. Study on biotransformation of traditional chinese veterinary medicine[D]. Beijing: Chinese Academy of Agricultural Sciences，2012. (in Chinese)
[16] 李亚菲，张秀凤，张灵璇，等. 应对细菌耐药策略研究进展[J]. 中国畜牧兽医，2023，50(11): 4568-4576.LI Y F，ZHANG X F，ZHANG L X，et al. Research progress on strategies to control bacterial resistance: A review[J]. China Animal Husbandry & Veterinary Medicine，2023，50(11): 4568-4576. (in Chinese)
[17] 耿爽，耿春银，杨濛，等. 两种无抗制剂对育肥猪生猪性能及肠道健康的影响[J]. 中国畜牧兽医，2021，48(4):1240-1250.GENG S，GENG C Y，YANG M，et al. The influence of two different kinds of antibiotic-freeaddictive on growth[J]. China Animal Husbandry & Veterinary Medicine，2021，48(4):1240-1250. (in Chinese)
[18] SHI N，LI N，DUAN X，et al. Interaction between the gut microbiome and mucosal immune system[J]. Military Medical Research，2017，4:14.
[19] JUNG A，CHEN L R，SUYEMOTO M M，et al. A review of Enterococcus cecorum infection in poultry[J]. Avian Diseases，2018，62(3):261-271.
[20] WILLIAMS N T. Probiotics[J]. American Journal of Health-System Pharmacy，2010，67(6):449-458.
[21] SHOKRYZADAN P，FASELEH J M，LIANG J B，et al. Probiotics: From isolation to application[J]. Journal of the American College of Nutrtion，2017，36(8):666-676.
[22] RAMOS C L，THORSEN L，SCHWAN R F，et al. Strain-specific probiotics properties of Lactobacillus fermentum，Lactobacillus plantarum and Lactobacillus brevis isolates from Brazilian food products[J]. Food Microbiology，2013，36(1):22-29.
[23] ROSIYANTI A S，ADRIANI L，RAMADHAN R F，et al. Effect of lactic acid and pH of probiotic yogurt on peak production of laying hens[J]. Pakistan Journal of Biological Sciences，2023，26(10):529-533.
[24] KUSADA H，MORINAGA K，TAMAKI H. Identification of bile salt hydrolase and bile salt resistance in a probiotic bacterium Lactobacillus gasseri JCM1131T[J]. Microorganisms，2021，9(5):1011.
[25] HASSANZADAZAR H，EHSANI A，MARDANI K，et al. Investigation of antibacterial，acid and bile tolerance properties of Lactobacilli isolated from Koozeh cheese[J]. Veterinary Research Forum，2012，3(3):181-185.
[26] TURPIN W，HUMBLOT C，TOMAS M，et al. Lactobacilli as multifaceted probiotics with poorly disclosed molecular mechanisms[J]. International Journal of Food Microbiology，2010，143(3):87-102.
[27] TRAN C，HORYANTO D，STANLEY D，et al. Antimicrobial properties of Bacillus probiotics as animal growth promoters[J]. Antibiotics-Basel，2023，12(9):407.
[28] GOODMAN C，KEATING G，GEORGOUSOPOULOU E，et al. Probiotics for the prevention of antibiotic-associated diarrhoea: A systematic review and meta-analysis[J]. BMJ Open，2021，11(8): e043054.
[29] LONE A，MOTTAWEA W，MEHDI Y，et al. Bacteriocinogenic probiotics as an integrated alternative to antibiotics in chicken production—Why and how?[J]. Critical Reviews in Food Science and Nutrition，2022，62(31)，8744-8760.
[30] YU Q，XU C，WANG M，et al. The preventive and therapeutic effects of probiotics on mastitis: A systematic review and Meta-analysis[J]. PLoS One，2022，17(9):e0274467.
[31] KICECKA A，MACURA B，SZCZEPANIK M. Modulation of allergic contact dermatitis via gut microbiota modified by diet，vitamins，probiotics，prebiotics，and antibiotics[J]. Pharmacological Reports，2023，75(2):236-248.
[32] DOBSON A，COTTER P D，ROSS R P，et al. Bacteriocin production: A probiotic trait? [J]. Applied and Environmental Microbiology，2012，78(1):1-6.
[33] SZAJEWSKA H，KOŁODZIEJ M. Systematic review with Meta-analysis: Lactobacillus rhamnosus GG in the prevention of antibiotic-associated diarrhoea in children and adults[J]. Alimentary Pharmacology & Therapeutics，2015，42(10):1149-1157.
[34] LOW C X，TAN L T，AB MUTALIB N S，et al. Unveiling the impact of antibiotics and alternative methods for animal husbandry: A review[J]. Antibiotics-Basel，2021，10(5):578.

Isolation，Identification and Probiotic Characteristics of Streptococcus alactolyticus from Intestinal Tract of Zi Geese

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	TANG Jun, HE Li, WANG Penghui, HE Xiaolong, YI Huanming, CHENG Xiao, REN Chunhuan, CHEN Jiahong, WANG Qiangjun, ZHANG Zijun. Research Progress on the Role of Intestinal Microbes in Healthy Production of Ruminants [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1466-1479.
[2]	ZHAO Xiaonan, HU Youjun, CHENG Huangzuo, WEN Wei, LU Xinfang, HE Zishi, CHEN Jiemei, E Xiaodi, HE Qiantai, ZOU Tiande. Effects of Different Types of Benzoic Acid on Growth Performance, Serum Biochemical Indices and Intestinal Flora of Weaned Piglets [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(3): 1031-1040.
[3]	JIANG Shanyi, ZHOU Bijun, WEN Ming, WANG Kaigong. Prevention of Traditional Chinese Medicine with Probiotics on Eimeria tenella [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2092-2102.
[4]	GUO Hongwei, CAI Yongchao, FAN Chunguang, HAO Kaixuan, CHANG Juan, WANG Ping, LIU Chaoqi, YIN Qingqiang. Effect of Compound Biological Detoxifier on Aflatoxin B₁ Degradation Rate Under Artificial Gastrointestinal Fluid Condition [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1395-1403.
[5]	CHEN Shaojun, LI Gang, NAI Zida, LIU Di, JIANG Xinpeng. Research and Application of Gene-edited Probiotics in Animal Intestinal Health [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1016-1024.
[6]	ZHANG Mingliang, ZHANG Hailong, MA Shipeng, WANG Cailian, LANG Xia. Effects of Zanthoxylum bungeanum Essential Oil on Production Performance and Gastrointestinal Tissue Structure of Small-tail Han Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 556-563.
[7]	WANG Wei, LI Chunzhu, WANG Shasha, SANG Rui, GE Bingjie, LI Chunting, ZHAO Xin, LIU Xinman, YU Minghong, ZHANG Xuemei. Study on the Protective Mechanism of Traditional Chinese Medicine Combined with Probiotics on Diarrhea Caused by E.coli in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 779-788.
[8]	LI Chunting, WANG Shasha, ZHAO Xin, WANG Wei, GE Bingjie, ZHANG Xuemei. Protective Effect of Traditional Chinese Medicine Combined with Probiotics on E.coli Diarrheain Mice [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3226-3234.
[9]	ZHANG Yu, MA Xi, ZHAO Jinbiao, LU Lin. Optimization of Fermentation Conditions and Determination of Antibacterial Activity of Bacillus licheniformis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2812-2819.
[10]	ZHANG Lihuan, JIA Hao, ZHANG Ruonan, WANG Yanfei, LIU Xuan. Effects of Compound Probiotics on the Intestinal Microflora of Broilers [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 488-500.
[11]	WANG Zhihua, JIA Junpeng, RADEBE Stoffel Matjeke, YU Qinghua. Study on the Growth Performance and Intestinal Flora of Beagle Puppies Regulated by Probiotics [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 510-520.
[12]	LI Yang, ZHU Siming, ZHANG Jianling, ZHENG Yue, CHEN Yingqiang, LUO Dizhou, YU Ting, ZHANG Yanbin, WANG Zhilin. Optimization of Fermentation Process of Oregano Powder by Response Surface Method and Study on Its Antibacterial Ability [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4795-4806.
[13]	ZHAO Dehui, ZHANG Haihua, WANG Jinming, MA Cuiliu, LIU Hanlu. Research Progress on Maintain Intestinal Health and Antiviral Effect of Probiotics [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4252-4260.
[14]	BU Ying, ZHENG Nan, WANG Jiaqi, ZHAO Shengguo. Research Progress on Immunoglobulin Regulating Gastrointestinal Health and Microbial Function in Animals [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3800-3808.
[15]	LIU Shi, LIU Jing, CHEN Zhuang, YU Ting, BEI Jinlong, WANG Lei, LI Zhengmin, WANG Zhilin. Effects of Compound Probiotics Added to Drinking Water on Growth Performance and Intestinal Microflora in Lingnan Yellow Feather Broilers [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 98-108.