1株植物乳杆菌的分离鉴定、生物学特性和安全性研究

doi:10.16431/j.cnki.1671-7236.2025.07.009

Abstract

Abstract: 【Objective】 This study aimed to isolate and screen a duck-origin probiotic strain,investigate its biological characteristics,and select candidate strains for meat duck production,providing reference for developing novel probiotic preparations. 【Method】 Bacterial strain was isolated and purified from duck cecal contents.Identification was performed through calcium dissolution zone assay,Gram staining,biochemical tests,and 16S rDNA sequencing.Growth characteristics,acid tolerance,bile salt tolerance,free amino acid production,and organic acid content were determined.Antimicrobial efficacy against Escherichia coli,Riemerella anatipestifer,Salmonella spp.,Staphylococcus aureus,Streptococcus suis,and Avibacterium paragallinarum were evaluated through inhibition rate and apoptosis rate assays.Scanning electron microscopy characterized antibacterial morphology.Duck-derived Lactiplantibacillus plantarum extracellular proteins were isolated by ultrafiltration,with antibacterial components analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS).Safety assessment was conducted in meat ducks. 【Result】 A Gram-positive,non-spore-forming bacillus was isolated.and it was determined to be Lactobacillus genus through biochemical identification tests.Then,it was confirmed to be Lactiplantibacillus plantarum through 16S rDNA detection.Compared with Lactiplantibacillus plantarum from pickle (pickle-LP),this Lactiplantibacillus plantarum from duck (duck-LP) could quickly enter the logarithmic phase and plateau of growth,and had a better acid and bile salt resistance.It could produce more free amino acids,lactic acid,and acetic acid (P＜0.05).It had a significant inhibitory effect on Escherichia coli and Avibacterium paragallinarum at a 1∶1 addition ratio (P＜0.05).At a 1∶4 addition ratio,there was a significant inhibitory effect on the growth of Escherichia coli,Riemerella anatipestifer,and Salmonella spp.(P＜0.05).At a 1∶9 addition ratio,there was a significant inhibitory effect on Escherichia coli and Riemerella anatipestifer (P＜0.05).At a 1∶4 addition ratio,the apoptosis rates of Escherichia coli,Riemerella anatipestifer, Streptococcus suis,and Avibacterium paragallinarum were significantly increased (P＜0.05). Through LC-MS/MS,a total of 10 proteins were detected,including two antimicrobial components:Peptidoglycan-binding protein and acetate kinase.The safety test on meat ducks showed that the bacterium was safe and had no toxic side effects on meat ducks. 【Conclusion】 This experiment isolated and screened a strain of duck-LP,which had excellent growth characteristics,acid/bile tolerance,amino acid/organic acid production,and broad-spectrum antimicrobial activity.Its antibacterial mechanism might be through synergistic effects of antimicrobial proteins and organic acids,disrupting cell wall/membrane integrity.The strain showed potential as a safe probiotic feed additive for meat ducks.

Key words: Lactiplantibacillus plantarum; isolation and identification; biological characteristics; safety; meat ducks

CLC Number:

S852.61

DAI Lin, HUAI Zhifang, WANG Baowei, ZHANG Ming'ai, FAN Wenlei, ZHANG Jing, WANG Binghan, QI Guofeng. Isolation, Identification and Biological Properties and Safety of A Lactiplantibacillus plantarum Strain[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3070-3083.

References

[1] SALIM H M D,HUQUE K S,KAMARUDDIN K M,et al.Global restriction of using antibiotic growth promoters and alternative strategies in poultry production[J].Science Progress,2018,101(1):52-75.
[2] WANG B,ZHOU Y,MAO Y,et al.Dietary supplementation with Lactobacillus plantarum ameliorates compromise of growth performance by modulating short-chain fatty acids and intestinal dysbiosis in broilers under clostridium perfringens challenge[J].Frontiers in Nutrition,2021,8:1-11.
[3] 马晨.植物乳杆菌P-8对肠道菌群的调控及其机制[J].科学通报,2019,64(3):298-306.MA C.The regulation mechanism of Lactobacillus plantarum P-8 on intestinal microflora[J].Chinese Science Bulletin,2019,64(3):298-306.(in Chinese)
[4] 安柯颖,高文文,李蕾,等.植物乳杆菌对北京鸭脂质代谢和抗氧化功能的影响[J].动物营养学报,2022,34(12):7790-7800.AN K Y,GAO W W,LI L,et al.Effect of Lactobacillus plantarum on lipid metabolism and antioxidant function of Pekin ducks[J].Chinese Journal of Animal Nutrition,2022,34(12):7790-7800.(in Chinese)
[5] AN K,GAO W,LI P,et al.Dietary Lactobacillus plantarum improves the growth performance and intestinal health of Pekin ducks[J].Poultry Science,2022,101(6):101844.
[6] KAKTCHAM P M,TEMGOUA J B,ZAMBOU F N,et al.In vitro evaluation of the probiotic and safety properties of bacteriocinogenic and non-bacteriocinogenic lactic acid bacteria from the intestines of Nile tilapia and common carp for their use as probiotics in aquaculture[J].Probiotics and Antimicrobial Proteins,2018,10(1):98-109.
[7] 兰恬静.鸭疫里默氏菌对大环内酯-林可酰胺类耐药性及耐药机制的研究[D].成都：四川农业大学,2023.LAN T J.Resistance mechanism of Riemerella anatipestificus to macrolide and lincomamides[D].Chengdu: Sichuan Agricultural University,2023.(in Chinese)
[8] WU Y,WANG Y,YANG H,et al.Resident bacteria contribute to opportunistic infections of the respiratory tract[J].PLoS Pathogens,2021,17(3):e1009436.
[9] 万欣,江鹏,何苗,等.高效液相色谱法同时测定菌液中5种有机酸的含量[J].现代化工,2023,43(4):227-230.WAN X,JIANG P,HE M,et al.Determination of five kinds of organic acids in bacterial liquid by high performance liquid chromatography[J].Modern Chemical Industry,2023,43(4):227-230.(in Chinese)
[10] XU Z,WANG J,ZHENG W,et al.Antibacterial activity and mechanism of a novel bacteriocin produced by Lactiplantibacillus plantarum against Escherichia coli and Staphylococcus aureus[J].International Journal of Food Science & Technology,2023,58(1):181-193.
[11] LE LAY C,COTON E,LE BLAY G,et al.Identification and quantification of antifungal compounds produced by lactic acid bacteria and propionibacteria[J].International Journal of Food Microbiology,2016,239:79-85.
[12] 蔡红英,李道捷,孟昆,等.植物乳杆菌的生理功能及其在家禽中的应用研究进展[J].动物营养学报,2023,35(6):3410-3421.CAI H Y,LI D J,MENG K,et al.Research progress on physiological functions of Lactobacillus plantarum and its application in poultry[J].Chinese Science Bulletin,2023,35(6):3410-3421.(in Chinese)
[13] 张在,郑瑞程,方庆励,等.广西猪源益生菌的分离鉴定与生物学特性研究[J].中国畜牧兽医,2020,47(6):1921-1933.ZHANG Z,ZHENG R C,FANG Q L，et al.Isolation,identification and biological characteristics of probiotics from swine origin in Guangxi[J].China Animal Husbandry & Veterinary Medicine,2020,47(6):1921-1933.(in Chinese)
[14] 尹杨燕,潘艳,李小宁,等.1株植物乳杆菌的生物学特性及体外抑菌活性研究[J].中国畜牧兽医,2023,50(2):666-673.YIN Y Y,PAN Y,LI X N,et al.Study on biological characteristics and antibacterial activity in vitro of a Lactobacillus plantarum strain[J].China Animal Husbandry & Veterinary Medicine,2023,50(2):666-673.(in Chinese)
[15] 夏超笃,湛穗璋,艾琴,等.一株解淀粉芽孢杆菌的益生特性及抑菌性研究[J].中国畜牧兽医,2020,47(2):425-432.XIA C D,ZHAN S Z,AI Q,et al.Study on the probiotic characteristics and antibacterial activity of a strain Bacillus amyloliquefaciens[J].China Animal Husbandry & Veterinary Medicine,2020,47(2):425-432.(in Chinese)
[16] DEC M,PUCHALSKI A,URBAN-CHMIEL R,et al.Screening of Lactobacillus strains of domestic goose origin against bacterial poultry pathogens for use as probiotics[J].Poultry Science,2014,93(10):2464-2472.
[17] REUBEN R C,ROY P C,SARKAR S L,et al.Isolation,characterization,and assessment of lactic acid bacteria toward their selection as poultry probiotics[J].BMC Microbiology,2019,19(1):253.
[18] IBEGBULEM C O,IGWE C U,OKWU G N,et al.Total amino acid profiles of heat-processed fresh Elaeis guineensis and Raphia hookeri wines[J].Food Chemistry,2013,138(2):1616-1620.
[19] SUSHIDA H,ISHIBASHI N,ZENDO T,et al.Evaluation of leader peptides that affect the secretory ability of a multiple bacteriocin transporter,EnkT[J].Journal of Bioscience and Bioengineering,2020,130(4):377-384.
[20] GAO Y,LIU Y,SUN M,et al.Physiological function analysis of Lactobacillus plantarum Y44 based on genotypic and phenotypic characteristics[J].Journal of Dairy Science,2020,103(7):5916-5930.
[21] WEI M,WANG S,GU P,et al.Comparison of physicochemical indexes,amino acids,phenolic compounds and volatile compounds in bog bilberry juice fermented by Lactobacillus plantarum under different pH conditions[J].Journal of Food Science and Technology,2018,55(6):2240-2250.
[22] 章检明,张兰威,易华西,等.乳酸菌Ⅱ类细菌素生物合成及其代谢调控策略[J].中国酿造,2019,38(2):1-6.ZHANG J M,ZHANG L W,YI H X,et al.Biosynthesis and strategies of metabolism regulation for class Ⅱ bacteriocins produced by lactic acid bacteria[J].China Brewing,2019,38(2):1-6.(in Chinese)
[23] THOMPSON H O,ÖNNING G,HOLMGREN K,et al.Fermentation of cauliflower and white beans with Lactobacillus plantarum—Impact on levels of riboflavin,folate,vitamin B₁₂,and amino acid composition[J].Plant Foods for Human Nutrition,2020,75(2):236-242.
[24] YOO D,BAGON B B,VALERIANO V D V,et al.Complete genome analysis of Lactobacillus fermentum SK152 from kimchi reveals genes associated with its antimicrobial activity[J].FEMS Microbiology Letters,2017,364(18):fnx185.
[25] PARLINDUNGAN E,MAY B K,JONES O A H.Metabolic insights into the effects of nutrient stress on Lactobacillus plantarum B21[J].Frontiers in Molecular Biosciences,2019,6:1-10.
[26] JANKOWSKA E,CHWIALKOWSKA J,STODOLNY M,et al.Volatile fatty acids production during mixed culture fermentation—— The impact of substrate complexity and pH[J].Chemical Engineering Journal,2017,326:901-910.
[27] 曾美端,洪艺萍,蔡玉凤,等.产酸丙酸杆菌FS1171产丙酸发酵动力学研究[J].微生物学杂志,2022,42(6):32-38.ZENG M D,HONG Y P,CAI Y F,et al.Fermentation kinetics of Propionibacterium acidogenes FS1171 producing propanoic acid[J].Journal of Microbiology,2022,42(6):32-38.(in Chinese)
[28] FERNANDEZ B,VIMONT A,DESFOSSÉS-FOUCAULT É,et al.Antifungal activity of lactic and propionic acid bacteria and their potential as protective culture in cottage cheese[J].Food Control,2017,78:350-356.
[29] 刘颖颖,卢艳波,杨小雁,等.植物乳杆菌DY6的生理特性及代谢机制分析[J].食品与发酵工业,2022,48(18):11-19.LIU Y Y,LU Y B,YANG X Y,et al.Physiological characteristics of Lactobacillus plantarum DY6 and its metabolic mechanisms analysis[J].Food and Fermentation Industries,2022,48(18):11-19.(in Chinese)
[30] 孟怀旺,蓝洪,米自由,等.乳杆菌抗病机制研究进展[A].中国畜牧兽医学会禽病学分会第十五次学术研讨会论文集[C].2010.MENG H W,LAN H,MI Z Y,et al.Research progress on the disease resistance mechanism of Lactobacilli[A].The 15th Academic Seminar of the Poultry Disease Branch of the Chinese Society of Animal Husbandry and Veterinary Medicine[C].2010.(in Chinese)
[31] GONG Y,LI T,LI S,et al.Achieving high yield of lactic acid for antimicrobial characterization in cephalosporin-resistant Lactobacillus by the co-expression of the phosphofructokinase and glucokinase[J].Journal of Microbiology and Biotechnology,2016,26(6):1148-1161.
[32] 吴轶,贡汉生,刘文丽,等.一株产细菌素乳杆菌的鉴定及其细菌素编码基因的获得[J].食品科学,2015,36(11):110-113.WU Y,GONG H S,LIU W L,et al.Identification of a bacteriocin-producing Lactobacillus and acquisition of bacteriocin-encoding genes[J].Food Science,2015,36(11):110-113.(in Chinese)
[33] 王鸿蕉,蔡昀洁,岳明祥,等.植物乳杆菌的抑菌性能及其影响因素研究进展[J].食品研究与开发,2022,43(14):212-218.WANG H J,CAI Y J,YUE M X,et al.Antibacterial properties of Lactobacillus plantarum and their influencing factors[J].Food Research and Development,2022,43(14):212-218.(in Chinese)
[34] 杨广珠,张淑红,李滢,等.植物乳杆菌拮抗致病大肠杆菌机制与应用研究进展[J].现代食品科技,2023,39(12):310-319.YANG G Z,ZHANG S H,LI Y,et al.The progress on the mechanism and application of Lactiplantibacillus plantarum inhibiting pathogenic Escherichia coli[J].Modern Food Science and Technology,2023,39(12):310-319.(in Chinese)
[35] RUIZ M J,GARCÍA M D,CANALEJO L M M,et al.Antimicrobial activity of Lactiplantibacillus plantarum against Shiga toxin-producing Escherichia coli[J].Journal of Applied Microbiology,2023,134(9):lxad202.
[36] LI Z,GUO Q,LIN F,et al.Lactobacillus plantarum supernatant inhibits growth of Riemerella anatipestifer and mediates intestinal antimicrobial defense in Muscovy ducks[J].Poultry Science,2024,103(2):103216.
[37] 李忠琴,王泽楠,黄文树,等.仿刺参(Apostichopus japonicus)肠道源植物乳杆菌(Lactobacillus plantarum)HY21的生长及益生特性分析[J].微生物学报,2024,64(3):840-853.LI Z Q,WANG Z N,HUANG W S,et al.Growth and probiotic properties of Lactobacillus plantarum sp. HY21 from intestinal tract of Apostichopus japonicus[J].Acta Microbiologica Sinica,2024,64(3):840-853.(in Chinese)
[38] SONG X,LIN Z,YU C,et al.Effects of Lactobacillus plantarum on growth traits,slaughter performance,serum markers and intestinal bacterial community of Daheng broilers[J].Journal of Animal Physiology and Animal Nutrition,2022,106(3):575-585.
[39] TANG Q,YI H,HONG W,et al.Comparative effects of L.plantarum CGMCC 1258 and L.reuteri LR1 on growth performance,antioxidant function,and intestinal immunity in weaned pigs[J].Frontiers in Veterinary Science,2021,8:1-11.

Isolation, Identification and Biological Properties and Safety of A Lactiplantibacillus plantarum Strain

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LU Chong, WANG Yuyan, FU Han, LI Tongyang, MIAO Ronghao, LU Yabin, LI Jianlong, LIU Jianhua, GUO Qingyong, MAI Zhanhai, KUANG Ling. Isolation and Identification of the Main Pathogenic Bacteria of Aborted Yili Foals and Their Pathogenicity [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3344-3358.
[2]	ZHAO Yilong, TANG Na, JING Changhua, XU Qingqing, YIN Xiusheng, WANG Haiming, SUN Jing, LIN Panpan, DONG Lin, LIU Jishan, CAO Rongfeng. Isolation,Identification and Biological Characteristics of a Strain of Lumpy Skin Disease Virus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2790-2799.
[3]	YANG Qinhong, YANG Jia, DUAN Wenjie, ZHANG Yongxian, ZHU Youshuai, YANG Shaohong, HE Cheng, YIN Hongbin, LI Suhua. Isolation and Identification of a High-temperature Tolerant Bacillus thermoamylovorans and Optimization of Its Enzyme Production Conditions [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2839-2850.
[4]	ZHOU Bowu, ZHAO Wen, ZHANG Yehuai, GU Zuhua, YANG Kai, XIONG Biao, JU Rongcheng, DENG Genfu, HUANG Wenfei, HU Tingjun, FENG Xiangren. Study on the Acute Toxicity and Sub-chronic Toxicity of Polygonum hydropiper Powder [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2915-2924.
[5]	WU Peng, YUAN Yang, YANG Yunyun, FENG Yi, WANG Yan, YANG Ying, CHEN Jiangfeng, JIANG Haibo, WEN Ming. Diagnosis and Pathogen Identification of a Case of Mixed Infection of Duck Plague Virus and Pasteurella multocida in Breeding Goose [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2253-2265.
[6]	WU Yue, SHI Xingya, ZHANG Shuai, ZHAO Yunhuan, GUO Liming, ZUO Yuzhu, FAN Jinghui. Isolation and Identification of Porcine Pseudorabies Virus and Genetic Evolution Analysis of gE and gC Genes [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2266-2277.
[7]	YU Kun, ZHAO Jie, MA Qin, SHI Yanhong, ZHANG Xiao, LIU Zihan, ZHANG Xinting, WANG Jianhua, LI Yufeng. Isolation,Identification,Drug Resistance and Pathogenicity Analysis of Salmonella Enteritidis from Commercial Meat Ducks [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2353-2363.
[8]	LONG Baoqin, WANG Huixiang, YU Linjin, HAERLEHA·Amantai, CHEN Haoran, XU Mengjiao, SHI Longxing, LI Youwen. Isolation,Identification and Biological Characteristics Analysis of Two Strains of Klebsiella pneumoniae from Quail [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2364-2378.
[9]	WU Jiaxin, SUN Yue, MAO Wei, LIU Shuying, YIN Kaiwen, ZHANG Zhidan, HAN Kaifan, ZHAO Hongxia. Isolation and Identification of Mannheimia haemolytica from Sheep Respiratory Tract and Its Pathogenicity and Drug Resistance [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2421-2431.
[10]	LI Yongjian, ZHANG Runze, WU You, ZHAO Pengfei, JIAO Longling, ZHOU Ming, QI Yayin, REN Jingjing. Isolation,Identification and Probiotic Characteristics Analysis of Bacillus subtilis from Sheep [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1590-1602.
[11]	CHEN Zuoxin, CHEN Yuxin, PAN Yanlin, HUANG Yunzhen, LI Linlin, DONG Jiawen, XIANG Yong, XU Zhihong, SUN Minhua, ZHANG Junqin, HUANG Shujian, LIAO Ming. Isolation,Identification and Pathogenicity of Two Strains of Cluster 3 Goose Tembusu Virus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1750-1762.
[12]	GUO Qing, WANG Hongli, FENG Yuxuan, ZHANG Yumeng, MA Qiyin, CHENG Jia, LIU Mingchao. Isolation and Identification of Chicken Probiotics and Analysis of Their Probiotic Properties [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1852-1861.
[13]	LIN Bingbing, ZHAO Hongzhe, GUAN Na, WU Rigumula, QI Gen, ZHANG Yang, WEN Yongjun, WANG Fengxue. Isolation,Identification and Drug Resistance Analysis of Clostridium perfringens from Cattle in Some Areas of Inner Mongolia [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1873-1883.
[14]	HE Yuxuan, CIRING Zhuoma, WANG Yu, LIU Huaizhi, YANG Jinpeng, WEI Mingbang, SHANG Peng. Isolation and Identification of Tibetan Pig-derived Enterococcus faecalis and Detection of Drug Resistance and Virulence Genes [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1895-1904.
[15]	LI Na, LIU Chongyang, ZHANG Jingjing, MALIYA Qiqige, ZHU Na, LU Bin, HAI Ying. Isolation,Identification and Drug Resistance Analysis of a Sheep-derived Strain of Clostridium perfringens Type D [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1352-1359.