Abstract

In the authors’ previous study, four unique Lactobacillus strains (Lactobacillus plantarum Dad-13, Lactobacillus plantarum Mut-7, Lactobacillus plantarum T-3 and – Received Lactobacillus paracasei SNP-2) from Indonesian fermented foods and healthy feces have been studied as probiotic agents. In the current study, antibiotic resistance phenotypes of the highlighted Lactobacillus plantarum and Lactobacillus paracasei against eight antibiotics (amoxicillin, tetracycline, erythromycin, clindamycin, chloramphenicol, streptomycin, kanamycin, ciprofloxacin) and antibiotic resistance genes of these strains were investigated. The bacterial antibiotic susceptibility to eight antibiotics was assessed using disk diffusion method. Genome sequencing was carried out using NovaSeq 6000 sequencing platform. Genome was annotated using Rapid Annotation using Subsystem Technology v.2.0. Each group of the predicted products of resistance genes was further aligned using multiple sequence comparison by log- expectation and their functions were verified using comprehensive antibiotic resistance database 2020. All strains showed resistance to aminoglycoside and ciprofloxacin but sensitive to amoxicillin, clindamycin and erythromycin. Resistance to chloramphenicol and tetracycline varied within the strains. Two strains were sensitive and others were intermediate resistance to chloramphenicol. One strain was resistant to tetracycline, while the other three strains demonstrated intermediate resistance to the antibiotic. Genome sequence of the four strains verified the presence of the tetracycline, β-lactamase and ciprofloxacin resistance genes as well as multidrug resistance efflux systems. Occurrence of the resistance genes was correlated to the phenotype results, except for amoxicillin and aminoglycosides. Rapid Annotation using Subsystem Technology annotation showed that all Lactobacillus strains did not include transposable elements, gene transfer agents and plasmid linked functions; thus, horizontal transfer of the antibiotic resistance genes unlikely occurred. read more

Abstract

Adhesion capacity is considered one of the selection criteria for probiotic strains. The purpose of this study was to determine the adhesion properties of two candidate probiotics, Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7. The evaluation included the hydrophobicity of the cell surface using microbial adhesion to hydrocarbons (MATH), autoaggregation, and the adhesion of L. plantarum Dad-13 and L. plantarum Mut-7 to the intestinal mucosa of Sprague Dawley rat, followed by genomic analysis of the two L. plantarum strains. L. plantarum Dad-13 and L. plantarum Mut-7 showed a high surface hydrophobicity (78.9% and 83.5%) and medium autoaggregation ability (40.9% and 57.5%, respectively). The exposure of both isolates to the surface of the rat intestine increased the total number of lactic acid bacteria on the colon compartment, from 2.9 log CFU/cm2 to 4.4 log CFU/cm2 in L. plantarum Dad-13 treatment and to 3.86 log CFU/cm2 in L. plantarum Mut-7 treatment. The results indicate the ability of two L. plantarum to attach to the surface of the rat intestine. The number of indigenous E. coli in the colon also decreased when the compartment was exposed to L. plantarum Dad-13 and Mut-7, from 2.9 log CFU/cm2 to 1 log CFU/cm2. Genomic analysis revealed that both strains have genes related to adhesion properties that could play an important role in increasing the adherence of probiotics to the intestinal mucosa such as gene encoding fibronectin-binding protein, chaperonin heat shock protein 33 (Hsp33), and genes related to the capsule and cell wall biosynthesis. Based on these findings, we believe that L. plantarum Dad-13 and L. plantarum Mut-7 have adhesion properties to the intestinal mucosa in the rat intestine model system. The present research will be essential to elucidate the molecular mechanism associated with adhesion in our two probiotic strains. read more

Abstract

This study aimed to isolate lactic acid bacteria from Halloumi cheese produced by Mazaarat Artisan cheese Indonesia, to evaluate its potential as a probiotics, and to identify it based on molecular characteristics. The lactic acid bacteria were isolated from fresh milk, curd, and the Halloumi cheese and their morphology and physiology were identified. Screening of the lactic acid bacteria isolates for probiotics candidates was mainly based on the resistance of simulated gastric juice, bile salts, antibacterial activities against pathogens, and the adhesion properties by hydrophobicity test. The selected lactic acid bacteria strains as probiotics candidates were then identified molecularly using 16 S ribosomal RNA. From this study, 13 lactic acid bacteria isolates were obtained. Among the 13 isolates, K3 isolate was the most potent as a probiotics candidate. This isolate had high tolerance for simulated gastric juice and bile salts, good ability to protect the gut against pathogenic bacteria since it has antibacterial activities and had ability to adhere to human epithelial cells. Based on molecular identification, this probiotics candidate was identified as Lactobacillus plantarum. read more