从泰国鸡粪中筛选和评价潜在益生菌用乳酸菌。
Selection and evaluation of lactic acid bacteria from chicken feces in Thailand as potential probiotics.
机构信息
Department of Veterinary Microbiology, Faculty of Veterinary Science., Chulalongkorn University, Bangkok, Thailand.
College of Public Health Sciences, Chulalongkorn University, Bangkok, Thailand.
出版信息
PeerJ. 2023 Dec 14;11:e16637. doi: 10.7717/peerj.16637. eCollection 2023.
BACKGROUND
Lactic acid bacteria (LAB) are widely used as probiotics in poultry production due to their resilience to low pH and high bile salt concentrations, as well as their beneficial effects on growth performance and antagonistic activity against enteric pathogens. However, the efficacy of probiotics depends on strain selection and their ability to colonize the host's intestine. This study aimed to select, identify, and evaluate LAB strains isolated from chicken feces in Thailand for potential use as probiotics in the chicken industry.
METHODS
LAB strains were isolated from 58 pooled fresh fecal samples collected from chicken farms in various regions of Thailand, including commercial and backyard farms. Gram-positive rods or cocci with catalase-negative characteristics from colonies showing a clear zone on MRS agar supplemented with 0.5% CaCO were identified using MALDI-TOF mass spectrometry. The LAB isolates were evaluated for acid (pH 2.5 and pH 4.5) and bile salt (0.3% and 0.7%) tolerance. Additionally, their cell surface properties, resistance to phenol, antimicrobial activity, hemolytic activity, and presence of antimicrobial resistance genes were determined.
RESULTS
A total of 91 LAB isolates belonging to the , and genera were obtained from chicken feces samples. Backyard farm feces exhibited a greater LAB diversity compared to commercial chickens. Five strains, including BF12 and BF9, BF14, BYF20, and BYF26, were selected based on their high tolerance to acid, bile salts, and phenol. BF12 and BF14 demonstrated strong adhesion ability. The five LAB isolates exhibited significant cell-cell interactions (auto-aggregation) and co-aggregation with . All five LAB isolates showed varying degrees of antimicrobial activity against strains, with BYF20 displaying the highest activity. None of the LAB isolates exhibited beta-hemolytic activity. Whole genome analysis showed that BF12 contained , and , whereas strains BF9 and BF14 carried , and .
CONCLUSION
The selected LAB isolates exhibited basic probiotic characteristics, although some limitations were observed in terms of adhesion ability and the presence of antibiotic resistance genes, requiring further investigation into their genetic location. Future studies will focus on developing a probiotic prototype encapsulation for application in the chicken industry, followed by evaluations of probiotic efficacy.
背景
由于耐低 pH 值和高胆盐浓度，以及对生长性能的有益影响和对肠道病原体的拮抗活性，乳酸菌（LAB）广泛用作家禽生产中的益生菌。然而，益生菌的功效取决于菌株的选择及其在宿主肠道中定植的能力。本研究旨在从泰国鸡粪便中分离、鉴定和评估 LAB 菌株，以评估其作为家禽益生菌的潜力。
方法
从泰国不同地区（包括商业和后院农场）的鸡场采集的 58 个新鲜粪便样本中分离 LAB 菌株。使用 MALDI-TOF 质谱法，从在添加 0.5% CaCO 的 MRS 琼脂上显示清晰区域的菌落中鉴定出革兰氏阳性杆菌或球菌，其特征为过氧化氢酶阴性。评估 LAB 分离株对酸（pH 2.5 和 pH 4.5）和胆盐（0.3% 和 0.7%）的耐受性。此外，还测定了其细胞表面特性、对苯酚的抗性、抗菌活性、溶血活性以及抗菌药物耐药基因的存在。
结果
从鸡粪便样本中获得了 91 株属于 、 和 属的 LAB 分离株。与商业鸡相比，后院鸡的粪便显示出更高的 LAB 多样性。根据对酸、胆盐和苯酚的高耐受性，从鸡粪便中分离到了 5 株菌株，包括 BF12 和 BF9、BF14、BYF20 和 BYF26。BF12 和 BF14 表现出很强的粘附能力。这五个 LAB 分离株表现出很强的细胞间相互作用（自聚集）和与 的共聚集。所有五个 LAB 分离株对 菌株均表现出不同程度的抗菌活性，其中 BYF20 表现出最高的活性。没有一个 LAB 分离株表现出β溶血活性。全基因组分析表明 BF12 含有 、 和 ，而 BF9 和 BF14 菌株则携带 、 和 。
结论
所选的 LAB 分离株表现出基本的益生菌特性，但在粘附能力和抗生素耐药基因的存在方面存在一些局限性，需要进一步研究其遗传位置。未来的研究将集中于开发一种益生菌原型包封物，用于家禽行业，并随后评估益生菌的功效。
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