Safety Assessment of Indigenous Probiotic Strain Lactobacillus plantarum Mut-7 Using Sprague Dawley Rats as a Model

Abstract

Lactobacillus plantarum Mut-7 is a probiotic candidate isolated from gatot, traditional fermented cassava from Java, Indonesia. This study aimed to evaluate safety aspects of high dose consumption of L. plantarum Mut-7 (10 11 CFU/ml/day) on Sprague Dawley rats for 21 days. Twenty four female rats were randomly divided into 4 groups; initial condition group (P.0), control group (P.1), skim milk group (P.2) and probiotics group (P.3). All groups followed adaptation phase of 7 days, followed by treatment phase of 21 days for P.1, P.2 and P.3. The results showed that supplementation of high dose of L. plantarum Mut-7 did not have detrimental effects on general health, organ weight, hematology and histology parameters of treated rats. Feed intake and body weight showed no significant difference between groups. L. plantarum Mut-7 can survive in gastrointestinal tract of rats, resulting in an increased population of L. plantarum in the fecal matter and the digesta of treated rats. Bacterial translocation of L. plantarum Mut-7 was not detected in the blood and organ of treated rats as confirmed by rep-PCR with BOXAIR primer and further 16S RNA gene sequencing analysis. Twenty-six isolates from blood and organs of treated rats had low-level similarity (<75%) to that of L. plantarum Mut-7, with 10 isolates were further analyzed and found that none of them belong to L. plantarum. Although this study was limited to the use of animal study, the findings are useful to support the safety assessment of the use of L. plantarum Mut-7 as a probiotic according to the abovementioned parameters.

Safety assessment of the indigenous probiotic strain <i>Lactiplantibacillus plantarum</i> subsp. <i>plantarum</i> Kita-3 using Sprague–Dawley rats as a model

“…Additionally, the Alignment program in BioEdit software was chosen to complete the matching procedure. The Basic Local Alignment Search Tool approach was used to detect similar results for the DNA sequences that were comparable to or identical to the DNA sequence data for genes from the GenBank of the National Center for Biotechnology Information (NCBI) database [14] . The DNA sequence was applied as reference data to design the phylogenetic tree.…”

“…Bacterial DNA amplification using repetitive-polymerase chain reaction (rep-PCR) analysis has been acknowledged as a straightforward inexpensive PCR approach suitable for characterizing LAB, such as Lactobacillus , to the strain level when compared to other PCR techniques, such as pulsed-field gel electrophoresis and amplified fragment length polymorphism [13] . A previous safety assessment study for the probiotic strain Lactobacillus plantarum Mut-7 identified 26 bacterial isolates from the blood and organs of rats, which were identified using rep-PCR [14] .…”

Adhesion Properties of Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 on Sprague Dawley Rat Intestine

“…Lactobacillus plantarum Dad-13 does not translocate in the organs and blood. Furthermore, Ikhsani et al [49] reported that high-dose supplementation with L. plantarum Mut-7 does not have detrimental effects on general health, organ weight, hematology, and histological parameters in rats. Further, the bacterial translocation of L. plantarum Mut-7 was not detected in the blood or organs.…”

Genomic insight of two indigenous probiotics Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 from different origins of Indonesian fermented foods

“…L. plantarum Dad-13 could increase the population of Lactobacilli and inhibit the growth of Enterobacteriaceae (Rahayu et al 2016); L. plantarum Mut-7 could increase the population of Lactobacillus in fecal and could survive in intestine (Fitrianingthias et al 2018). The safety assessment of two L. plantarum also has been studied (Rahayu et al 2019;Ikhsani et al 2020). The whole study indicated that those strains are promising local probiotics for Indonesian population.…”