Research Department of Plant Biology and Biotechnology, Loyola College, Nungambakkam, Chennai, 600034, Tamil Nadu, India.
Department of Bacteriology, National Institute for Research in Tuberculosis, ICMR, Sathyamoorty road, Chetpet, Chennai-31, Tamil Nadu, India.
Microb Pathog. 2018 Jan;114:239-250. doi: 10.1016/j.micpath.2017.11.054. Epub 2017 Nov 28.
In the last few years, the demand for the tremendous therapeutic applications of indigenous probiotic bacteria from diversified fermented food products has surged. In view of this, the present study was documented to evaluate the anti-tubercular and probiotic properties of coagulase-negative staphylococci (CNS) indigenous to Koozh, a traditional fermented food product of South India. A total of 18 isolates were purified from Koozh, and tested for anti-tubercular activity against Mycobacterium tuberculosis H37Rv using luciferase reporter phage (LRP) assay. Among them, six isolates revealed higher percentage (>90%) of relative light unit (RLU) reduction. These six isolates were further evaluated for their in vitro probiotic attributes using standard protocols. All six staphylococci strains disclosed good probiotic properties. Moreover, Staphylococcus hominis strain MANF2 showed high cell survival percentage (92.2%) at pH 2.0 as well as towards simulated gastric juice (88.51%). Furthermore, strain MANF2 was found to be resistant to bile salt after 24 h of incubation with maximal viability of 5.71 ± 0.02 log cfu/mL, and depicted the deconjugation of bile salt as well. All the isolates exhibited strong auto-aggregation capacity (44.4 ± 1.2-68.1 ± 1.5%), and hydrophobicity against toluene (55.0 ± 1.2-72.0 ± 1.1%). Additionally, strain MANF2 was observed to be highly resistant to phenol (6.27 ± 0.01 log cfu/mL) and lysozyme (81.1 ± 1.6% viability). Most importantly, all six isolates depicted good hypocholesterolemic effect, slight β-galactosidase activity, and moderate proteolytic property. The strains were sensitive to all the tested conventional antibiotics, except Nalidixic acid. In addition to this, all staphylococci strains demonstrated significant DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging, hydrogen peroxide tolerance, and hydroxyl radical scavenging activity in a dose dependent manner, thereby exhibiting the potent antioxidative properties of isolates. The negative results obtained from haemolytic, DNase, and gelatinase tests revealed the non-pathogenicity and safety aspect of these strains. In a nutshell, the present investigation divulges the persuasive anti-tubercular and probiotic properties of staphylococci, particularly strain MANF2, and recommended the further exploitation of Koozh associated CNS in pharmaceutics.
在过去的几年中,对来自各种发酵食品的本土益生菌巨大治疗应用的需求猛增。有鉴于此,本研究记录了评估印度南部传统发酵食品 Koozh 中凝固酶阴性葡萄球菌(CNS)的抗结核和益生菌特性。从 Koozh 中分离纯化了 18 株,并用 luciferase reporter phage (LRP) assay 检测其对 Mycobacterium tuberculosis H37Rv 的抗结核活性。其中,6 株分离株显示出更高比例(>90%)的相对光单位(RLU)减少。进一步使用标准方案评估了这 6 株分离株的体外益生菌特性。所有 6 株葡萄球菌株均显示出良好的益生菌特性。此外,金黄色葡萄球菌 MANF2 株在 pH 2.0 下以及模拟胃液中具有高细胞存活率(92.2%)(88.51%)。此外,在孵育 24 小时后,菌株 MANF2 对胆盐表现出抗性,最大存活率为 5.71±0.02 log cfu/mL,并显示出胆盐的去共轭作用。所有分离株均表现出较强的自聚集能力(44.4±1.2-68.1±1.5%)和对甲苯的疏水性(55.0±1.2-72.0±1.1%)。此外,菌株 MANF2 对苯酚(6.27±0.01 log cfu/mL)和溶菌酶(81.1±1.6%存活率)具有高度抗性。最重要的是,所有 6 株分离株均表现出良好的降胆固醇作用、轻微的β-半乳糖苷酶活性和适度的蛋白水解特性。除了萘啶酸之外,这些菌株对所有测试的常规抗生素均敏感。此外,所有葡萄球菌株均表现出显著的 2,2-Diphenyl-1-picrylhydrazyl (DPPH) 清除、过氧化氢耐受和羟基自由基清除活性,呈剂量依赖性,从而表现出分离株的强大抗氧化特性。溶血、DNase 和明胶酶试验的阴性结果表明这些菌株无致病性和安全性。简而言之,本研究揭示了凝固酶阴性葡萄球菌,特别是 MANF2 株的抗结核和益生菌特性,并建议进一步开发 Koozh 相关 CNS 在制药方面的应用。