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通过体外、体内和基因组研究揭示了嗜热链球菌 KLDS 3.1003 对一些食源性致病菌的抑制作用。

Suppressive effects of Streptococcus thermophilus KLDS 3.1003 on some foodborne pathogens revealed through in vitro, in vivo and genomic insights.

机构信息

Key Laboratory of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin 150030, China.

School of Biosciences and Veterinary Medicine, University of Camerino 60232, Camerino Marche - Floristic Research Centre of the Apennine Gran Sasso and Monti della Laga National Park, San Colombo, 67021 Barisciano, L'Aquila, Italy.

出版信息

Food Funct. 2020 Jul 22;11(7):6573-6587. doi: 10.1039/d0fo01218a.

Abstract

Foodborne diseases (FBDs) remain a persistent global challenge and recent research efforts suggest that lactic acid bacteria (LAB) strains can contribute towards their prevention and treatment. This study investigates the genetic properties of Streptococcus thermophilus KLDS 3.1003 as a potential probiotic and health-promoting LAB strain as well as its in vitro and in vivo activities against two foodborne pathogens. In vitro, its antimicrobial activities and tolerance levels in simulated bile salts and acids were determined. The cytotoxic effects of the LAB strain in RAW264.7 cells were also evaluated. For in vivo evaluation, 24 BALB/c mice were orally administered control and trial diets for 14 days. Genomic analyses of this strain's bacteriocin configuration, stress response system and multidrug resistance genes were annotated to validate in vitro and in vivo results. In vitro antimicrobial results show that the cells and CFS of S. thermophilus KLDS 3.1003 could inhibit both pathogens with the former being more effective (P < 0.05). In addition, its cell-free supernatant (CFS) could inhibit the growth of both pathogens, with catalase treatment having the highest effect against it. More so, after 3 h of incubation, survivability levels of S. thermophilus KLDS 3.1003 were significantly high (P < 0.05). LPS-induced RAW264.7 cell activities were also significantly reduced by 108-109 CFU mL-1 of S. thermophilus KLDS. In vivo, significant weight losses were inhibited in the TSTEC group compared to the TSTSA group (P < 0.05). Moreover, pathogen-disrupted blood biochemical parameters like HDL, LDL, TP, TG, AST, ALT and some minerals were restored in the respective prevention groups (TSTEC and TSTSA). Genomic analyses showed that S. thermophilus KLDS 3.1003 has bacteriocin-coding peptides, which accounts for its antimicrobial abilities in vitro and in vivo. S. thermophilus KLDS 3.1003 is also endowed with intact genes for acid tolerance, salt-resistance, cold and heat shock responses and antioxidant activities, which are required to promote activities against the selected foodborne pathogens. This study showed that S. thermophilus KLDS 3.1003 has the genomic capacity to inhibit foodborne pathogens' growth in vitro and in vivo, thus qualifying it as a potential probiotic, antimicrobial and bio-therapeutic candidate.

摘要

食源性疾病(FBDs)仍然是一个全球性的挑战,最近的研究表明,乳酸菌(LAB)菌株可以有助于预防和治疗这些疾病。本研究调查了嗜热链球菌 KLDS 3.1003 的遗传特性,作为一种潜在的益生菌和促进健康的 LAB 菌株,以及其在体外和体内对两种食源性病原体的活性。在体外,测定了其抗菌活性和在模拟胆盐和酸中的耐受水平。还评估了 LAB 菌株对 RAW264.7 细胞的细胞毒性作用。对于体内评估,将 24 只 BALB/c 小鼠口服对照和试验饮食 14 天。对该菌株的细菌素结构、应激反应系统和多药耐药基因进行了基因组分析,以验证体外和体内结果。体外抗菌结果表明,嗜热链球菌 KLDS 3.1003 的细胞和 CFS 可以抑制两种病原体,前者更有效(P <0.05)。此外,其无细胞上清液(CFS)可以抑制两种病原体的生长,其中过氧化氢酶处理的效果最高。更重要的是,经过 3 小时的孵育,嗜热链球菌 KLDS 3.1003 的存活率水平显著升高(P <0.05)。LPS 诱导的 RAW264.7 细胞活性也被 108-109 CFU mL-1 的嗜热链球菌 KLDS 显著降低。在体内,与 TSTEC 组相比,TSTSA 组的体重显著减轻(P <0.05)。此外,在相应的预防组(TSTEC 和 TSTSA)中,破坏的血液生化参数如 HDL、LDL、TP、TG、AST、ALT 和一些矿物质得到了恢复。基因组分析表明,嗜热链球菌 KLDS 3.1003 具有细菌素编码肽,这解释了其在体外和体内的抗菌能力。嗜热链球菌 KLDS 3.1003 还具有完整的耐酸、耐盐、耐冷和耐热冲击以及抗氧化活性基因,这些基因是促进对所选食源性病原体活性所必需的。本研究表明,嗜热链球菌 KLDS 3.1003 具有在体外和体内抑制食源性病原体生长的基因组能力,因此有资格成为一种潜在的益生菌、抗菌和生物治疗候选物。

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