Spatial and Temporal Persistence of Fluorescent Lactiplantibacillus plantarum RS-09 in Intestinal Tract

Abstract

The beneficial effects of the probiotic strain Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) are based on its adherence and colonization ability in the gut. However, little is known about the migration and long-term gut colonization of the strain. This study evaluated the gut colonization modes of Lactiplantibacillus plantarum RS-09 to identify the strain with long-term gut colonization potential. We established CFDA/SE-labeled RS-09 to study the temporal and spatial distribution of RS-09 in the intestine as well as to analyze its persistence in different parts of the intestine by flow cytometry. This study has shown that the RS-09 strain maintains strong adhesion abilities under acid (pH 2.5) and base (pH 8.5) conditions. In addition, CFDA/SE can be used as an indicator for the labeling of L. plantarum RS-09 in the intestinal tract in vivo. We established a growth kinetics model of RS-09 to elucidate its persistence in the intestine. In vivo persistence experiments showed that the persistence rate of RS-09 was the highest in the cecum (69.5%) and the lowest in the duodenum (12.8%) at 96 h. After 20 days, RS-09 was predominantly localized in the cecum and colon steadily. These studies provide new insights into the long-term persistence of L. plantarum in the gastrointestinal tract. The CFDA/SE label system may be used to study the in vivo colonization dynamics of other probiotic strains.

Carbon dots labeled Lactiplantibacillus plantarum: a fluorescent multifunctional biocarrier for anticancer drug delivery

“…CDs are currently well established as cell imaging agents owing to their tunable fluorescence emission, brightness, and low toxicity ( Hallaji et al, 2022 ). Findings of the present study support convenience of CDs as a simple, effective and economic alternative to fluorescent dyes and metal-based quantum dots ( Hu et al, 2016 ; Xing et al, 2018 ; Zhao et al, 2022 ) as well as fluorescent proteins expressed by genetically engineered bacteria ( Han et al, 2015 ; Stojanov et al, 2021 ) for bacterial identification and tracking.…”

“…In this context, we present herein an innovative fluorescent L. plantarum (LP) as trackable hybrid drug biocarrier for potential colorectal cancer (CRC) therapy using prodigiosin (PG) as anticancer agent. LP, formerly Lactobacillus plantarum , was selected as biocarrier because of its recognized safety, tolerance toward GIT environment as well as adhesion and cytotoxicity to CRC cells ( Ahmad et al, 2018 ; Zhao et al, 2022 ). HILP was used for further improved probiotic safety and keeping properties, given that heat inactivation may not significantly affect the surface characteristics of Lactobacillus spp.…”

“…and their bioactivity ( Ciandrini et al, 2017 ). Although organic dyes ( Zhao et al, 2022 ), metal-based quantum dots ( Butrimienė et al, 2022 ) and fluorescent proteins produced by genetically engineered probiotic cells ( Han et al, 2015 ) were commonly documented as probiotic labeling agents, fluorescent carbon dots (CDs) were used in the present study because of their sustainability, cytocompatibility, non-toxicity, and high fluorescence stability ( Nandi et al, 2015 ; Unnikrishnan et al, 2020 ). CDs are a relatively new type of fluorescent nanomaterials with excellent photoluminescence ( Jing et al, 2023 ).…”

Assessment of the safety and probiotic properties of Roseburia intestinalis: A potential “Next Generation Probiotic”

“…CFDA-SE is a fluorescent dye with many advantages such as strong fluorescence, low toxicity, and good stability. Several researchers ( Lee et al, 2004 ; Wang et al, 2019 , 2021 ; Zhao et al, 2022 ) have used this method to tag and track bacteria. R. intestinalis was detected in all intestinal sites 2 h after administration, whereas after 48 h, R. intestinalis was predominantly located in the cecum, and not present in the colon.…”

“…To investigate the characteristics of R. intestinalis colonization of the GIT, we used a previously described method ( Zhao et al, 2022 ). R. intestinalis was collected during the logarithmic growth phase, stained with CFDA-SE, and resuspended in PBS at a concentration of 1 × 10 10 CFU/ml.…”