Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 5: 3372 - 3381

DOI: https://doi.org/10.12418/CJAN2023.313

RESEARCH PAPER

Isolation and Identification of Lactobacillus salivarius ZJBF005 from Alopex lagopus and Its Probiotic Function Evaluation in Vitro

  • MA Cuiliu , 1 ,
  • CAI Xiheng 1 ,
  • LIU Chaonan 2 ,
  • YUAN Weitao 1 ,
  • ZHAO Dehui 1, 3 ,
  • LIU Hanlu , 4, *
Expand
  • 1 Jilin Engineering Research Center of Microorganisms Feed for Special Wild Economic Animals, Innovation Engineering Centerof Feeding and Comprehensive Utilization for Special WildEconomic Animals, Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, Changchun 130112, China
  • 2 Laboratory Animal Center of Soochow University, Suzhou 215000, China
  • 3 Hebei Key Laboratory of Specialty Animal Germplasm Resources Exploration and Innovation, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China
  • 4 Chifeng University, Chifeng 024000, China
*associate professor, E-mail:

Received date: 2022-11-15

  Online published: 2023-05-11

Fold

Abstract

The objective of this study was to isolate and characterize lactic acid bacteria with probiotic potential in Alopex lagopus’s gut, and evaluate its probiotic function evaluation in vitro. The isolated strain identified by physiological, biochemical and 16S rRNA identification, measured its growth curve, acid production and acid resistance curves, bile salt tolerance and survival rate in artificial gastric juice and artificial intestinal juice were measured, the antibacterial activity and antibiotic sensitivity of the bacteria were detected by oxford cup agar diffusion test and disk diffusion test, respectively. Finally, twenty female Kunming mice were randomly divided into two groups, the control group was given normal saline intragastrically, and the experimental group was given Lactobacillus salivarius (1×109 CFU/mL), the experiment lasted for 7 days. The results showed that the strain was named Lactobacillus salivarius ZJBF005 according to 16S rRNA and physiological and biochemical identification results, it had a strong acid-producing ability and growth rapidly in the MRS medium; it could maintain good growth in the culture-medium with pH 4 to 7, while couldn’t grow in the culture-medium less than pH 2; the survival rate after incubation in artificial gastric juice and artificial intestinal juice for 3 h were 40.96% and 86.70%, respectively; it could grow in 0.2% bile salt environment; it could inhibit the growth of Salmonella and Staphylococcus aureus; it was not sensitive to antibiotics such as tetracycline, gentamicin, vancomycin, fosfomycin, azithromycin, trimethoprim and metronidazole, etc; the body weight change of mice after intragastric administration was normal, no death, and no pathological change in autopsy. In conclusion, the Lactobacillus salivarius ZJBF005 in healthy Alopex lagopus’s gut has pretty probiotic properties, it can be preliminarily determined as probiotic candidates.

Key words： Alopex lagopus; Lactobacillus salivarius; isolation; identification

Cite this article

MA Cuiliu , CAI Xiheng , LIU Chaonan , YUAN Weitao , ZHAO Dehui , LIU Hanlu . 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 . DOI: 10.12418/CJAN2023.313

益生菌是一种活的微生物,当给予足够的剂量时,会对宿主的健康产生良好的影响[1]。在人类中用于预防和治疗疾病、传染性和非传染性疾病,并在家禽、家畜、水产养殖中有广泛的应用[2]。对于性能良好的益生菌来说,其在口腔溶菌酶、胃pH和肠胆汁等酸性环境中的生存能力是一个重要因素[3]。益生菌的抑菌活性、抗炎活性和增强上皮细胞免疫屏障功能也是优异益生菌所要具备的关键因素[4]。
北极狐为犬科动物,目前对其肠道微生物的研究较少。前期研究表明,北极狐肠道存在复杂的微生物区系[5],在北极狐营养物质消化利用、维护机体健康和免疫功能方面发挥重要作用[6]。本项目组已在健康北极狐肠道中分离获得多种益生性能良好的益生菌[7⇓-9]。来自特定宿主的益生菌能更好地适应该宿主的环境。因此,本试验旨在从健康北极狐肠道中分离和鉴定具有益生菌潜能的乳酸菌,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提供理论基础。

1 材料与方法

1.1 试验材料

1.1.1 样品采集

2021年11月20日于长白山野生生物资源重点野外科学观测试验站采集24只冬毛期健康北极狐的肠道内容物。

1.1.2 菌株和细胞

唾液乳杆菌(Lactobacillus salivarius)ZJBF 005、大肠杆菌(Escherichia coli)ATCC 25922、鼠伤寒沙门氏菌(Salmonella typhimurium)ATCC 14028和金黄色葡萄球菌(Staphylococcus aureus)均由中国农业科学院特产研究所特种动物营养与饲养团队分离、鉴定和保存。

1.1.3 试验动物

21日龄无特定病原体(SPF)级雌性昆明小鼠20只,体重(22±2) g,由辽宁长生生物技术股份有限公司提供,动物生产许可证号:SCXK(辽)2020-0001。

1.2 试验方法

1.2.1 菌株的分离培养

采集北极狐空肠肠段内容物1 mL装于5 mL离心管,低温条件下运输回实验室。将离心管置于生物安全柜内操作,离心管内加入适量磷酸盐缓冲液(PBS)进行振荡混匀,吸取100μL混合液进行梯度稀释,随即取10-5、10-6、10-7稀释液100 μL分别涂于MRS平板,置于恒温培养箱。挑取单菌落在MRS平板中反复平板划线以获得纯化菌株,筛选和保存革兰氏阳性菌株,对纯化后的菌株进行冻干保存。

1.2.2 菌落形态及染色

取冻干菌粉于MRS液体试管中,37 ℃恒温培养48 h后传代,蘸取少量菌液采用平板划线法培养单菌落,观察菌落颜色和形态,采用革兰氏染色法进行染色后,显微镜观察。

1.2.3 16S rRNA基因序列测定

将待测菌株培养24 h后提取该细菌基因组DNA。以细菌基因组DNA作为目的基因进行PCR扩增。PCR反应体系见表1。PCR产物经1.2%琼脂糖凝胶电泳检测,菌株的保守序列约为1 500 bp,将片段长度正确的PCR产物送至天一辉远生物科技有限公司进行测序。从GenBank中获取的基因序列在NCBI网站进行BLAST比对,通过MEGA进行比对建树。
表1 PCR反应体系

Table 1 PCR reaction system

项目Items 体积Volume/μL 温度Temperature/℃ 时间Time
上游引物Forward primer 1 95 5 min
下游引物Reverse primer 1 95 30 s
Pre mix Taq 15 55 30 s
DNA 3 72 1.5 min
双蒸水ddH2O 10 72 10 min
合计Total 30

1.2.4 生理生化试验

对分离得到的乳酸菌进行生理生化特性鉴定,具体步骤参照《乳酸细菌分类鉴定及实验方法》[10]。将菌株接种培养24 h后,进行吲哚试验、过氧化氢酶试验、V-P试验、糖发酵试验等。

1.2.5 温度试验

于MRS液体培养基中培养菌株24 h后,以2.0%的接种量进行传代,分别于20、25、30、37、40和45 ℃恒温厌氧箱内培养,取其在培养0、1、2、3、4、5、6、8、10、12、14、24、36、48 h时的菌悬液测定600 nm处吸光度(OD600)值,每组设置3个平行。最终结果以每组平均值为标准绘制统计图。

1.2.6 产酸、耐酸性能试验

将培养24 h的菌悬液以2.0%的接种量接种于MRS液体培养基中,37 ℃恒温厌氧培养。分别于0、2、4、6、8、10、12、14、24 h利用pH计测定培养液pH,根据测定结果绘制产酸速率曲线。
将培养24 h的菌悬液以2.0%的接种量分别接种于不同初始pH(1、2、3、4、5、6、7)的MRS液体培养基中,37 ℃恒温厌氧培养,取其在培养0、3、6、12 h时的菌悬液测定OD600值,每组设置3个平行。最终结果以每组平均值为标准绘制统计图。

1.2.7 人工胃液和人工肠液耐受性试验

取培养24 h的菌悬液1 mL离心弃上清并加入生理盐水重悬,向重悬后的菌悬液中分别加入1 mL经滤膜过滤除菌的人工胃液和人工肠液,37 ℃恒温厌氧培养,取其在培养0和3 h时的菌悬液各100 μL,对其梯度稀释后涂布,用平板菌落计数法计算活菌数,以0 h为对照,计算存活率,测定菌株在胃液中的耐受性。人工胃液和人工肠液配制方法参照2020版《中华人民共和国药典》[11]。存活率计算公式如下:
存活率(%)=100×lgNt/lgN0。
式中:N0为0 h活菌数;Nt为3.0 h活菌数。

1.2.8 胆盐耐受性试验

将培养24 h的菌悬液以2.0%的接种量接种于胆盐浓度分别为0、0.1%、0.2%、0.3%、0.4%的MRS液体培养基中,37 ℃恒温厌氧培养,取其在培养0、3、6 h时的菌悬液测定OD600值,每组设置3个平行。最终结果以平均值为标准绘制统计图。

1.2.9 抑菌试验

将培养24 h的菌悬液离心取上清液备用,活化大肠杆菌、沙门氏菌和金黄色葡萄球菌并制备菌悬液,分别取200 μL均匀涂布于LB平板上,上清液通过牛津杯琼脂扩散法进行抑菌试验,测定抑菌圈。大于牛津杯直径(8 mm)1 mm的抑菌圈直径视为具有抑菌性,小于等于1 mm的抑菌圈直径视为无抑菌性。

1.2.10 耐药性试验

取培养24 h的菌悬液200 μL均匀涂布于MRS平板上,室温放置3~5 min,利用纸片扩散法将抗菌药物纸片贴于已接种细菌的琼脂表面,测定菌株对青霉素G、氯霉素、头孢孟多、克林霉素、环丙沙星、四环素、庆大霉素、万古霉素、磷霉素、阿奇霉素、甲氧苄啶和甲硝唑12种抗生素的耐药性。大于药敏片直径(6 mm)1 mm的抑菌圈直径视为具有抑菌性,小于等于1 mm的抑菌圈直径视为无抑菌性。

1.2.11 动物安全性试验

将20只体重(22±2) g的昆明系雌性小鼠随机分为2组,每组10只。将菌液浓度调整为1×109 CFU/mL备用,试验组小鼠每只一次性腹腔注射0.3 mL菌液,对照组小鼠腹腔注射等量的生理盐水,留察7 d,记录各组小鼠每日的体重变化、死亡情况,7 d后对每组小鼠进行剖检,对比2组间小鼠内脏器官变化。

1.3 数据分析

数据通过SPSS 26.0软件中t检验进行分析,结果以“平均值±标准差”表示,采用Graph Pad Prism 8.0.1软件作图,P<0.05为差异显著。

2 结果与分析

2.1 菌株形态学分析

在MRS培养基中分离出多株乳酸菌,将其中一株菌株命名为ZJBF005,进行后续分析与鉴定。菌落呈乳白色、圆形、不透明状,大而平坦,表面光滑,边缘整齐(图1)。菌株ZJBF005为直或微弯杆状、两端钝圆、单个或成对存在、无芽孢的革兰氏染色阳性细菌。
图1 菌株ZJBF005的菌落形态(左)及细胞形态(右,1 000×)

Fig.1 Colony morphology (left) and cell morphology (right, 1 000×) of strain ZJBF005

2.2 菌株同源比对分析

基于16S rRNA基因序列比对结果,以酒类酒球菌(Oenococcus oeni)JCM 6125 (AB022924.1)为外枝构建菌株ZJBF005的Neighbor-Joining系统发育树(图2)。由16S rRNA基因测序BLAST结果得知,菌株ZJBF005与唾液乳杆菌之间的成对序列相似度高达99.52%,因此确定菌株ZJBF005为唾液乳杆菌。
图2 菌株ZJBF005系统发育树

Lactobacillus aviarius subsp. araffinosus DSM 20653 (AYYZ01000003):鸟乳杆菌不解棉籽糖亚种DSM 20653 (AYYZ01000003);Lactobacillus aviarius subsp. aviarius DSM 20655 (AYZA01000007):鸟乳杆菌鸟亚种DSM 20655 (AYZA01000007);Lactobacillus hayakitensis JCM 14209(BAML0100063):早来乳杆菌JCM 14209(BAML0100063);Lactobacillus salivarius BCRC 14759(CP024067):唾液乳杆菌BCRC 14759(CP024067)。

Fig.2 Evolutionary tree of strain ZJBF005

2.3 菌株生理生化特性

对菌株ZJBF005进行生理生化特性鉴定,检测结果见表2,菌株ZJBF005可利用单糖(甘露糖、山梨糖、半乳糖、木糖、果糖、鼠李糖、葡萄糖等)、二糖(纤维二糖、麦芽糖、蔗糖、乳糖等)和多糖(棉子糖等)产酸。硫化氢试验、葡萄糖产气试验、淀粉水解试验、硝酸盐试验、过氧化氢酶试验和吲哚试验等结果均为阴性,V-P试验、甲基红试验和明胶液化试验结果呈阳性,此外菌株ZJBF005具有运动性。试验结果参照《伯杰细菌鉴定手册》[12]进行比对发现,菌株ZJBF005生理生化特性与唾液乳杆菌相符。
表2 菌株ZJBF005的生理生化特性

Table 2 Physiological and biochemical characteristics of strain ZJBF005

项目
Items		 结果
Results		 项目
Items		 结果
Results
甘露糖Mannose + 阿拉伯糖Pectinose -
山梨糖Sorbitan + 硫化氢试验Hydrogen sulfide test -
半乳糖Galactose + 葡萄糖产气试验Glucose gas production test -
木糖Xylose + 淀粉水解试验Starch hydrolysis test -
果糖Fructose + 硝酸盐试验Nitrate test -
鼠李糖Rhamnose + 过氧化氢试验Hydrogen peroxide test -
葡萄糖Glucose + 吲哚试验Indole test -
纤维二糖Cellobiose + V-P试验V-P test -
麦芽糖Maltose + 甲基红试验Methyl red test +
蔗糖Sucrose + 明胶液化试验Gelatin liquidized test +
乳糖Lactose + 运动性Motility +
棉子糖Raffinose + 甘露醇Mannitol +
七叶苷Aesculin + 山梨醇Sorbitol -

+:阳性 positive;-:阴性 negative。

2.4 菌株适宜生长温度

对在20、25、30、35、37、40和45 ℃环境下生长的唾液乳杆菌ZJBF005进行OD600值测定,结果如图3所示。唾液乳杆菌ZJBF005在20~45 ℃条件下皆可生长,适宜生长温度为37 ℃,其最大OD600值可达1.7。接种2 h后进入对数生长期,培养至25 h后进入生长平台期。
图3 唾液乳杆菌ZJBF005不同温度下的生长曲线

Fig.3 Growth curves at different temperatures of Lactobacillus salivarius ZJBF005

2.5 菌株产酸及耐酸特性

由图4可知,唾液乳杆菌ZJBF005在MRS培养液中迅速产酸,6 h内pH从5.97下降至3.28,此阶段对应该菌株的生长对数期;6~24 h内培养液pH略微降低,直至稳定在3.14。唾液乳杆菌ZJBF005的菌体数量随着pH的升高而增加,在pH为1和2的培养液中无法生长;在pH为3的培养液中生长缓慢;当pH提高为4或者更高时,菌体生长速度明显加快,菌体在pH为7的培养液中生长最佳。
图4 唾液乳杆菌ZJBF005的产酸(左)及耐酸曲线(右)

Fig.4 Curves of acid producing (left) and acid resisting (right) of Lactobacillus salivarius ZJBF005

2.6 菌株对人工胃液和人工肠液的耐受特性

由表3可知,经3 h人工胃液培养后,唾液乳杆菌ZJBF005活菌数从8.39 lg(CFU/mL)降至3.44 lg(CFU/mL),存活率为(40.96±0.21)%。经3 h人工肠液培养后,唾液乳杆菌ZJBF005活菌数从8.23 lg(CFU/mL)降至7.14 lg(CFU/mL),存活率为(86.70±0.14)%。
表3 唾液乳杆菌ZJBF005在人工胃液和人工肠液中的存活率

Table 3 Survival rate of Lactobacillus salivarius ZJBF005 in artificial gastric juice and artificial gastric juice

项目
Items		 人工胃液
Artificial gastric juice		 人工肠液
Artificial intestinal juice
耐受时间Tolerance time/h 0 3 0 3
活菌数Viable count/[lg(CFU/mL)] 8.39 3.44 8.23 7.14
存活率Survival rate/% 40.96±0.21 86.70±0.14

2.7 菌株对胆盐的耐受特性

由图5可知,对唾液乳杆菌ZJBF005进行胆盐耐受性试验,在无胆盐培养基中菌体正常生长;在含有0.1%和0.2%的胆盐条件下培养至3 h时生长较为良好,随时间的推移菌体在0.2%的胆盐培养基中生长受到抑制,而在含有0.1%的胆盐环境下还可以缓慢生长;当胆盐浓度超过0.3%时,菌体几乎不能生长。
图5 唾液乳杆菌ZJBF005的胆盐耐受性

Fig.5 Bile salt resistance of Lactobacillus salivarius ZJBF005

2.8 菌株对常见致病菌的抑制作用

由表4可知,通过牛津杯法抑菌试验发现,除大肠杆菌外,唾液乳杆菌ZJBF005对沙门氏菌和金黄色葡萄球菌均具有生长抑制作用。唾液乳杆菌ZJBF005对沙门氏菌和金黄色葡萄球菌的抑制程度相同,抑菌圈直径分别为(14±1) mm和(13±1) mm。
表4 唾液乳杆菌ZJBF005抑菌试验结果

Table 4 Results of bacteriostatic test of Lactobacillus salivarius ZJBF005

项目Items 抑菌程度Bacteriostatic level
大肠杆菌Escherichia coli -
沙门氏菌Salmonella +
金黄色葡萄球菌Staphylococcus aureus +

抑菌程度用抑菌圈大小表示,+++、++和+分别表示抑菌圈大小为23~30 mm、16~23 mm和9~16 mm,-表示无抑菌圈。下表同。

The inhibition zone diameter shows the bacteriostatic level, +++, ++ and + represent the inhibition zone diameter 23 to 30 mm, 16 to 23 mm和9 to 16 mm, respectively, - was no inhibition zone. The same as below。

2.9 菌株的抗生素敏感性

由表5可知,利用纸片扩散法测定唾液乳杆菌ZJBF005对12种抗生素的耐药性,结果发现,唾液乳杆菌ZJBF005对青霉素G、氯霉素、头孢孟多、克林霉素、环丙沙星等抗生素敏感;而对四环素、庆大霉素、万古霉素、磷霉素、阿奇霉素、甲氧苄啶和甲硝唑等抗生素不敏感。
表5 唾液乳杆菌ZJBF005抗生素敏感试验结果

Table 5 Results of antibiotic susceptibility experiment of Lactobacillus salivarius ZJBF005

抗生素Antibiotics 敏感性Susceptibility 抗生素Antibiotics 敏感性Susceptibility
青霉素G Penicillin G 氯霉素Chloramphenicol
头孢孟多Cefmenador 克林霉素Clindamycin ++
环丙沙星Ciprofloxacin + 四环素Tetracycline -
庆大霉素Gentamicin - 万古霉素Vancomycin -
磷霉素Fosfomycin - 阿奇霉素Azithromycin -
甲氧苄啶Trimethoprim - 甲硝唑Metronidazole -

2.10 动物安全性试验

动物安全性试验期间,2组之间小鼠体重无显著差异,无生病或者死亡;剖检后观察其内脏均无出血现象,肝脏、脾脏均无眼观病变。这表明唾液乳杆菌ZJBF005对小鼠无毒副作用。

3 讨论

本研究从北极狐的肠道中分离出唾液乳杆菌ZJBF005,并进行一系列益生菌所必需的特性评价试验,目的是提供有关北极狐乳酸菌益生菌潜力的基本信息。大多数益生菌制剂经口服后到达胃肠道,因此菌株对胃肠液和胆盐的耐受性是评判益生菌功能的关键。动物胃黏膜分泌的酸有助于消化,是益生菌到达胃肠道末端的主要问题之一[13]。因此,pH被认为是微生物作为潜在益生菌的重要标准之一[14]。在大多数益生菌耐pH评价研究中,通常将pH设置为2~5[15],因为当食物被吞下时,胃的pH根据食物成分的不同在3~6变化[16]。许多研究表明,乳酸菌可以在低pH下存活[17]。本试验结果与这些公认的事实一致,唾液乳杆菌ZJBF005能抵抗低pH为3的环境并保持相当数量的活菌。在其他研究中也有类似的结果,认为唾液乳杆菌可以在低pH环境中大量存活[18]。然而,当pH降低时,唾液乳杆菌ZJBF005的活菌数显著降低。这与其他研究相似,有报道称当pH为2时,乳酸菌的存活数量显著减少[19]。乳酸菌在pH低于5的环境下生存时,其机制会发生改变,如细胞膜组成和细胞密度、中和过程、多种代谢途径、质子泵、蛋白质和DNA损伤修复等[20⇓⇓-23]。
胃蛋白酶和胰蛋白酶的抑菌作用是导致胃肠道内微生物死亡的两大因素。结果表明,唾液乳杆菌ZJBF005能在模拟胃液中存活,但随着时间的增加,存活率显著下降。抗胰蛋白酶的能力是选择益生菌的另一个重要标准。胰蛋白酶参与脂肪、蛋白质和碳水化合物的消化,并通过胰管在小肠中分泌[24]。唾液乳杆菌ZJBF005可以在模拟肠液中存活,具有一定的抗胰蛋白酶的能力。随着培养时间的增加,存活率明显下降,这表明胃蛋白酶和胰蛋白酶的抑菌作用更强。乳酸菌对人工胃液和人工肠液的耐受活性在许多其他研究中也有报道[25-26]。
同样,对胆盐的耐受性也是益生菌的重要特性之一[27]。大多数细菌对胆盐敏感,在胆盐存在的情况下无法生长,因为胆盐具有强大的抗菌活性,主要是通过触发细胞膜结构和损伤DNA[28]。当细菌接触到胆盐时,它们的细胞内稳态受到干扰,由于细胞膜分解,细菌内容物泄漏而导致细胞死亡[29]。但是乳酸菌能够通过胆汁盐水解酶来应对这种不利的条件,把胆盐解耦成氨基酸和胆固醇[30-31]。在本研究中,唾液乳杆菌ZJBF005也表现出这种能力,并能抵抗不同浓度的胆盐,在动物胃肠道中具有良好的生存潜力。唾液乳杆菌ZJBF005可在0.2%胆盐浓度下存活,而在0.3%胆盐浓度下细菌的生长明显受到抑制。另一项研究也表明,唾液乳杆菌具有良好的抗胆盐能力[32]。许多其他关于乳酸菌的研究也显示了它们在胆盐中的生存能力[33-34]。
抑制有害细菌生长是益生菌的一个显著特性。在本研究中,菌株对被测病原体表现出不同程度的抑制作用。唾液乳杆菌ZJBF005上清液对沙门氏菌和金黄色葡萄球菌的抑菌效果较为显著,而对大肠杆菌无抑制作用。实验室菌株对革兰氏阴性菌和革兰氏阳性菌均表现出广泛的拮抗活性,这与其他研究人员的发现一致。这些结果同样表明,益生菌的代谢产物有抑菌作用。乳酸菌可产生多种抗菌分子,如过氧化氢、细菌素、类细菌素化合物、细胞外有机酸等[35]。酸性微环境进一步增强了乳酸菌分泌这些化合物的能力[36]。
本研究中,唾液乳杆菌ZJBF005对万古霉素具有耐药性。万古霉素的耐药性被认为是固有的或自然的耐药菌株,在大多数情况下,它们的先天阻力来自于膜的不透水性[37]。因此,根据以往的研究[38],万古霉素的耐药性对乳酸菌菌株是不可传播的。对四环素和甲氧苄啶的耐药性与Kim等[39]的发现一致。对磷霉素有内源性耐药性,一些具有内在抗生素耐药性的益生菌株在抗生素治疗后可能有助于恢复肠道菌群[40]。这些有益微生物的抗生素耐药性,无论是内在的还是由于任何突变引起的,并不能说明菌株本身存在安全问题。

4 结论

本研究从北极狐肠道中分离出一株产酸能力较强的唾液乳杆菌ZJBF005,在体外环境中表现出良好的益生菌特性和抗菌能力,而且对临床使用的多种抗生素不敏感,对动物无毒副作用。因此,唾液乳杆菌ZJBF005可作为动物益生菌制剂的候选菌株。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
SPANO G. Editorial:lactic acid bacteria (probiotics),fermented milk and health[J]. Food Bioscience, 2021, 41:100938.

DOI

[2]
RAHEEM A, LIANG L, ZHANG G Z, et al. Modulatory effects of probiotics during pathogenic infections with emphasis on immune regulation[J]. Frontiers in Immunology, 2021, 12:616713.

DOI

[3]
CHALAS R, JANCZAREK M, BACHANEK T, et al. Characteristics of oral probiotics—a review[J]. Current Issues in Pharmacy and Medical Sciences, 2016, 29(1):8-10.

DOI

[4]
KLIMKO A I, CHERDYNTSEVA T A, BRIOUKHANOV A L, et al. In vitro evaluation of probiotic potential of selected lactic acid bacteria strains[J]. Probiotics and Antimicrobial Proteins, 2020, 12(3):1139-1148.

DOI

[5]
LIU H L, LI Z P, SI H Z, et al. Comparative analysis of the gut microbiota of the blue fox (Alopex lagopus) and raccoon dog (Nyctereutes procyonoides)[J]. Archives of Microbiology, 2020, 202(1):135-142.

DOI PMID

[6]
陈双双. 蓝狐肠道菌群多样性分析及其体外培养方法比较研究[D].硕士学位论文. 北京: 中国农业科学院, 2019.

CHEN S S. Analysis of gut microbiota diversity in blue fox and comparative study on its enrichment methods in vitro[D].Master’s Thesis. Beijing: Chinese Academy of Agricultural Sciences, 2019. (in Chinese)

[7]
张如春, 崔焕忠, 蔡熙姮, 等. 狐源罗伊氏乳杆菌ZJF036益生特性研究[J]. 动物营养学报, 2020, 32(8):3819-3829.

DOI

ZHANG R C, CUI H Z, CAI X H, et al. Probiotic characteristics of Lactobacillus reuteri ZJF036 isolated from fox[J]. Chinese Journal of Animal Nutrition, 2020, 32(8):3819-3829. (in Chinese)

[8]
蔡熙姮, 朱翔宇, 张如春, 等. 蓝狐源嗜酸乳杆菌ZJBF003株的分离鉴定及其体外益生潜能的评价[J]. 中国兽医科学, 2020, 50(8):1001-1011.

CAI X H, ZHU X Y, ZHANG R C, et al. Isolation and identification of Lactobacillus acidophilus ZJBF004 from Alopex lagopus and evaluation of its probiotic potential in vitro[J]. Chinese Veterinary Science, 2020, 50(8):1001-1011. (in Chinese)

[9]
蔡熙姮, 刘晗璐, 张如春, 等. 蓝狐源约氏乳杆菌菌株ZJBF004分离鉴定与体外益生潜能的评价[J]. 动物营养学报, 2020, 32(7):3313-3323.

DOI

CAI X H, LIU H L, ZHANG R C, et al. Isolation and identification of Lactobacillus johnsonii strain ZJBF004 from Alopex lagopus and its probiotic potential evaluation in vitro[J]. Chinese Journal of Animal Nutrition, 2020, 32(7):3313-3323. (in Chinese)

[10]
凌代文, 东秀珠. 乳酸细菌分类鉴定及实验方法[M]. 北京: 中国轻工业出版社,1999.

LING D W, DONG X Z. Taxonomic analysis and experimental method of Lactobacillus[M]. Beijing: China Light Industry Press,1999. (in Chinese)

[11]
国家药典委员会. 中华人民共和国药典[M]. 北京: 中国医药科技出版社, 2020.

Chinese Pharmacopoeia Commission. Chinese pharmacopoeia[M]. Beijing: China Medical Science Press, 2020. (in Chinese)

[12]
布坎南, 吉本斯. 伯杰细菌鉴定手册[M].中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组,译. 北京: 科学出版社,1984.

BUCHANAN R E, GIBBONS N E. Bergey’s manual of determinative bacteriology[M]. Translation team of Berger’s Handbook of Bacterial Identification,Institute of Microbiology,Chinese Academy of Sciences,translated. Beijing: Science Press,1984. (in Chinese)

[13]
GAO H X, MA L, SUN W X, et al. Impact of encapsulation of probiotics in oil-in-water high internal phase emulsions on their thermostability and gastrointestinal survival[J]. Food Hydrocolloids, 2022, 126:107478.

DOI

[14]
HORACKOVA S, VESELA K, KLOJDOVA I, et al. Bile salt hydrolase activity,growth characteristics and surface properties in Lactobacillus acidophilus[J]. European Food Research and Technology, 2020, 246(8):1627-1636.

DOI

[15]
TENEA G N, GONZALEZ G L, MORENO J L. Probiotic characteristics and antimicrobial potential of a native Bacillus subtilis strain Fa17.2 rescued from wild Bromelia sp. flowers[J]. Microorganisms, 2022, 10(5):860.

DOI

[16]
NADIA J, OLENSKYJ A G, SUBRAMANIAN P, et al. Influence of food macrostructure on the kinetics of acidification in the pig stomach after the consumption of rice- and wheat-based foods:implications for starch hydrolysis and starch emptying rate[J]. Food Chemistry, 2022, 394:133410.

DOI

[17]
PANICKER A S, ALI S A, ANAND S, et al. Evaluation of some in vitro probiotic properties of Lactobacillus fermentum strains[J]. Journal of Food Science and Technology, 2018, 55(7):2801-2807.

DOI

[18]
YANG Y, SONG X, XIONG Z Q, et al. Complete genome sequence of Lactobacillus salivarius AR809,a probiotic strain with oropharyngeal tract resistance and adhesion to the oral epithelial cells[J]. Current Microbiology, 2022, 79(9):280.

DOI

[19]
LIU J J, WANG Y P, LI A Y, et al. Probiotic potential and safety assessment of Lactobacillus isolated from yaks[J]. Microbial Pathogenesis, 2020, 145:104213.

DOI

[20]
DA CRUZ NIZER W S, INKOVSKIY V, OVERHAGE J. Surviving reactive chlorine stress:responses of Gram-negative bacteria to hypochlorous acid[J]. Microorganisms, 2020, 8(8):1220.

DOI

[21]
FOSTER J W. Low pH adaptation and the acid tolerance response of Salmonella typhimurium[J]. Critical Reviews in Microbiology, 1995, 21(4):215-237.

DOI

[22]
COTTER P D, HILL C. Surviving the acid test:responses of Gram-positive bacteria to low pH[J]. Microbiology and Molecular Biology Reviews, 2003, 67(3):429-453.

DOI

[23]
WANG C, CUI Y H, QU X J. Mechanisms and improvement of acid resistance in lactic acid bacteria[J]. Archives of Microbiology, 2018, 200(2):195-201.

DOI PMID

[24]
FERREIRA-LAZARTE A, MORENO F J, VILLAMIEL M. Bringing the digestibility of prebiotics into focus:update of carbohydrate digestion models[J]. Critical Reviews in Food Science and Nutrition, 2021, 61(19):3267-3278.

DOI

[25]
KIM K T, KIM J W, KIM S I, et al. Antioxidant and anti-inflammatory effect and probiotic properties of lactic acid bacteria isolated from canine and feline feces[J]. Microorganisms, 2021, 9(9):1971.

DOI

[26]
TOPÇU K C, KAYA M, KABAN G. Probiotic properties of lactic acid bacteria strains isolated from pastırma[J]. LWT, 2020, 134:110216.

DOI

[27]
SELLE K, KLAENHAMMER T R. Genomic and phenotypic evidence for probiotic influences of Lactobacillus gasseri on human health[J]. FEMS Microbiology Reviews, 2013, 37(6):915-935.

DOI

[28]
FALAH F, VASIEE A, BEHBAHANI B A, et al. Evaluation of adherence and anti-infective properties of probiotic Lactobacillus fermentum strain 4-17 against Escherichia coli causing urinary tract infection in humans[J]. Microbial Pathogenesis, 2019, 131:246-253.

DOI

[29]
RUIZ L, MARGOLLES A, SÁNCHEZ B. Bile resistance mechanisms in Lactobacillus and Bifidobacterium[J]. Frontiers in Microbiology, 2013, 4:396.

[30]
FOLEY M H, O’FLAHERTY S, BARRANGOU R, et al. Bile salt hydrolases:gatekeepers of bile acid metabolism and host-microbiome crosstalk in the gastrointestinal tract[J]. PLoS Pathogens, 2019, 15(3):e1007581.

DOI

[31]
GUO C F, ZHANG S, YUAN Y H, et al. Bile salt hydrolase and S-layer protein are the key factors affecting the hypocholesterolemic activity of Lactobacillus casei-fermented milk in hamsters[J]. Molecular Nutrition & Food Research, 2018, 62(24):e1800728.

[32]
LI C Y, WANG H N, HE R J, et al. Discovery and characterization of amentoflavone as a naturally occurring inhibitor against the bile salt hydrolase produced by Lactobacillus salivarius[J]. Food & Function, 2022, 13(6):3318-3328.

[33]
WANG G Q, YU H N, FENG X, et al. Specific bile salt hydrolase genes in Lactobacillus plantarum AR113 and relationship with bile salt resistance[J]. LWT, 2021, 145:111208.

DOI

[34]
HERNÁNDEZ-GÓMEZ J G, LÓPEZ-BONILLA A, TREJO-TAPIA G, et al. In vitro bile salt hydrolase (BSH) activity screening of different probiotic microorganisms[J]. Foods, 2021, 10(3):674.

DOI

[35]
AZIZI F, HABIBI NAJAFI M B, EDALATIAN DOVOM M R. The biodiversity of Lactobacillus spp. from Iranian raw milk Motal cheese and antibacterial evaluation based on bacteriocin-encoding genes[J]. AMB Express, 2017, 7(1):176.

DOI

[36]
YANG E, FAN L H, YAN J P, et al. Influence of culture media,pH and temperature on growth and bacteriocin production of bacteriocinogenic lactic acid bacteria[J]. AMB Express, 2018, 8(1):10.

DOI

[37]
STOGIOS P J, SAVCHENKO A. Molecular mechanisms of vancomycin resistance[J]. Protein Science, 2020, 29(3):654-669.

DOI PMID

[38]
URBAN-CHMIEL R, MAREK A, STEPIEÑ-PYŚNIAK D, et al. Antibiotic resistance in bacteria-a review[J]. Antibiotics, 2022, 11(8):1079.

DOI

[39]
KIM J Y, YOUNG J A, GUNTHER N W IV, et al. Inhibition of Salmonella by bacteriocin-producing lactic acid bacteria derived from U.S. kimchi and broiler chicken[J]. Journal of Food Safety, 2015, 35(1):1-12.

DOI

[40]
SHARMA P, TOMAR S K, GOSWAMI P, et al. Antibiotic resistance among commercially available probiotics[J]. Food Research International, 2014, 57:176-195.

DOI

Options
Outlines

/

〈 〉

京ICP备17050989号-2
Copyright © Chinese Journal of Animal Nutrition, All Rights Reserved.
Tel: 010－62817823　E-mail: cjan@vip.163.com
Powered by Beijing Magtech Co., Ltd.