Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Gomti Nagar Extension, Near Malhaur Railway Station, Lucknow, 226028, India.
Department of Microbiology, Dr. Ram Manohar Lohia Institute of Medical Sciences, Vibhuti Khand, 226010, Lucknow, India.
Probiotics Antimicrob Proteins. 2021 Jun;13(3):739-750. doi: 10.1007/s12602-020-09724-w. Epub 2020 Nov 14.
The past decade has brought a significant rise in antimicrobial resistance, and the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter species) have considerably aggravated a threat to public health, causing nosocomial infections worldwide. The objective of the current study was to isolate novel probiotic strain with antimicrobial activity against multidrug-resistant ESKAPE pathogens. For this purpose, eighteen breastfed infant faeces were collected and lactic acid bacteria (LAB) with antagonistic activity were isolated. Out of 102 anaerobic LAB isolated, only nine exhibited inhibitory activity against all ESKAPE pathogens. These selected nine isolates were further characterized for their probiotic attributes such as lysozyme tolerance, simulated gastrointestinal tolerance, cellular auto-aggregation and cell surface hydrophobicity. Bile salt deconjugation and cholesterol-lowering capacity was also determined. Among all nine, isolate LBM220 was found to possess superior probiotic potential. Confirmatory identification of isolate LBM220 was done by both 16S rRNA sequence analysis and mass spectrometric analysis using MALDI-TOF. Based on BLAST result, isolate LBM220 was identified as Lactobacillus gasseri. Phylogenetic analysis of Lactobacillus gasseri LBM220 [accession number MN097539] was performed. Also, detailed safety evaluation study of Lact. gasseri LBM220 showed the presence of intrinsic antibiotic resistance and the absence of hemolytic, DNase, gelatinase and toxic mucinolytic activity. Time kill assay was also performed to confirm the strong kill effect of Lact. gasseri LBM220 on all six multidrug resistant ESKAPE pathogens. Thus, Lact. gasseri LBM220 can be utilized and explored as potential probiotic with therapeutic intervention.
过去十年，抗菌药物耐药性显著上升，ESKAPE 病原体（屎肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌属）对全球的医院感染造成了严重威胁。本研究的目的是分离具有抗多重耐药 ESKAPE 病原体活性的新型益生菌株。为此，收集了 18 份母乳喂养婴儿的粪便，并分离出具有拮抗活性的乳酸菌（LAB）。在分离出的 102 株厌氧 LAB 中，只有 9 株对所有 ESKAPE 病原体均具有抑制活性。对这 9 株具有抑制活性的分离株进行了进一步的鉴定，以确定其益生菌属性，如溶菌酶耐受性、模拟胃肠道耐受性、细胞自动聚集和细胞表面疏水性。还测定了胆汁盐去共轭和降胆固醇能力。在这 9 株中，LBM220 分离株被发现具有优越的益生菌潜力。通过 16S rRNA 序列分析和 MALDI-TOF 质谱分析对 LBM220 分离株进行了确认鉴定。根据 BLAST 结果，LBM220 分离株被鉴定为格氏乳杆菌。对 Lactobacillus gasseri LBM220 [登录号 MN097539]进行了系统发育分析。此外，对 Lact. gasseri LBM220 的详细安全性评估研究表明，该菌存在固有抗生素耐药性，且无溶血、DNase、明胶酶和有毒黏液溶解活性。时间杀伤试验也证实了 Lact. gasseri LBM220 对所有 6 株多重耐药 ESKAPE 病原体的强烈杀伤作用。因此，Lact. gasseri LBM220 可作为具有治疗干预作用的潜在益生菌进行利用和探索。
