Exploring probiotic potential and antimicrobial properties of lactic acid bacteria from cow's milk

Jaqueline Milagres de Almeida , Juliana Takahashi Maffei , Clarice Gebara , Cintia Minafra , Bruno Toledo-Silva , Mônica Correia Gonçalves , Helio Langoni , André Thaler Neto , Fernando Nogueira Souza , Nathalia Cristina Cirone Silva
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Abstract

One of the most important foods in the world is milk, due to its nutrients and composition. Milk also contains lactic acid bacteria (LABs), which can produce antimicrobial compounds and organic acids; an essential factor in developing different methods to control other bacteria. Another critical factor is that several species of LABs are considered potential probiotics, offering several health benefits. This study aimed to isolate, identify LABs, characterize the antimicrobial susceptibility profile, and evaluate the inhibition potential against Staphylococcus aureus (ATCC 25923), Listeria monocytogenes (ATCC 7644), Escherichia coli (ATCC 25922) and Salmonella Typhimurium (ATCC 14028); as well as verifying the acidification capacity, survival of simulated in vitro gastrointestinal conditions and identification of virulence genes and bacteriocin-producing genes. The study was carried out with LABs previously isolated from milk of cows with clinical mastitis, low and high (Somatic Cells Count) in herds from all regions of Brazil (South, Southeast, Central-West, North, and Northeast). One hundred and ten strains were selected for identification using the MALDI-TOF technique. The genera with the most significant spectrum of action were Lactococcus, Lactobacillus, and Pediococcus in the antimicrobial action test against pathogens, concluding that Lactobacillus showed resistance to several antibiotics. The analysis of antimicrobial action made it possible to verify the potential production of antimicrobial compounds by LABs against the primary pathogens of food origin. The most significant reduction in pH could be observed from 24 h to 48 h, ranging from 6.8 (0 h) to 4.74 (48 h) for Lactobacillus and Lactococcus. All isolates survived tolerance tests at pH=3.0 and bile salts (0.5 %), with survival rates ranging from 34.2 to 69.9 % at pH = 3.0 and above 82 % at 0.5 % bile salts. Lacotococcus and Lactobacillus isolates presented the plnEF and plnNC8 genes. This study contributes to the search for alternatives in the treatment of bacterial infections with a reduction in the use of antibiotics, which, due to the increase in bacterial resistance, is causing difficulties in treating diseases caused by these microorganisms. pH is an important parameter when it comes to food safety and conservation. Tolerance to acid stress and bile salts are important properties in evaluating probiotic potential and the ability to maintain high counts under these conditions. The presence of the plnEF and plnNC8 genes suggests bacteriocin production.

探索牛奶中乳酸菌的益生潜力和抗菌特性
牛奶是世界上最重要的食品之一,因为它富含营养和成分。牛奶中还含有乳酸菌(LABs),它们能产生抗菌化合物和有机酸;这是开发不同方法来控制其他细菌的一个重要因素。另一个关键因素是,有几种 LABs 被认为是潜在的益生菌,可提供多种健康益处。本研究旨在分离、鉴定 LABs,确定其抗菌敏感性特征,评估其对金黄色葡萄球菌(ATCC 25923)、单核细胞增生李斯特菌(ATCC 7644)、大肠杆菌(ATCC 25922)和鼠伤寒沙门氏菌(ATCC 14028)的抑制潜力;以及验证酸化能力、模拟体外胃肠道条件下的存活率,并鉴定毒力基因和细菌素产生基因。这项研究使用的是以前从巴西所有地区(南部、东南部、中西部、北部和东北部)患有临床乳腺炎、低体细胞数和高体细胞数的奶牛乳汁中分离出来的 LABs。通过 MALDI-TOF 技术,选出了 110 株菌株进行鉴定。在对病原体的抗菌作用试验中,作用谱最明显的菌属是乳球菌属、乳酸杆菌属和木薯球菌属。通过对抗菌作用的分析,可以验证 LABs 生产抗菌化合物的潜力,以对付食品来源的主要病原体。从 24 小时到 48 小时,乳酸菌和乳球菌的 pH 值明显下降,从 6.8(0 小时)到 4.74(48 小时)不等。在 pH=3.0 和胆盐(0.5%)条件下,所有分离菌都能通过耐受性测试,在 pH=3.0 条件下存活率为 34.2% 至 69.9%,在胆盐(0.5%)条件下存活率超过 82%。漆球菌和乳酸杆菌分离物含有 plnEF 和 plnNC8 基因。这项研究有助于寻找治疗细菌感染的替代品,减少抗生素的使用,由于细菌抗药性的增加,治疗由这些微生物引起的疾病变得困难。 pH 值是食品安全和保存方面的一个重要参数。对酸压力和胆盐的耐受性是评估益生菌潜力和在这些条件下保持高数量的能力的重要特性。plnEF 和 plnNC8 基因的存在表明可以产生细菌素。
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