Detection of the Potential Inactivation of Tetrodotoxin by Lactic Acid Bacterial Exopolysaccharide

Tu, Nguyen Hoang Khue; Dat, Nghe Van; Canh, Le Van; Vinh, Doan Thi Thanh

doi:10.3390/toxins10070288

Cited by 10 publications

(5 citation statements)

References 23 publications

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“…Using the TLC method, a similar result was obtained by Tu et al. (2018), indicating that EPS produced by L. rhamnosus strain was composed with d ‐glucose. In another study, the HPTLC technique revealed the presence of three different types of sugar, namely, d ‐glucuronic acid, fructose, and galactose, in EPS from L. casei B9‐1 and L. plantarum P35 (Shirzad et al., 2018).…”

Section: Resultssupporting

confidence: 80%

Investigation on exopolysaccharide production by Lacticaseibacillus rhamnosus P14 isolated from Moroccan raw cow's milk

Akhtach

Tabia

Bricha

et al. 2021

Journal of Food Science

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Twenty‐four strains were isolated from 50 samples of raw cow's milk originated from different regions of Morocco. After different screening methods, one strain was selected as the highest exopolysaccharide (EPS)‐producing isolate and was identified by 16S rDNA sequencing as Lacticaseibacillus rhamnosus P14. Moreover, the EPS‐producing ability, bacterial growth, and pH of the medium were monitored. The optimization of culture conditions indicated that the high yield of EPS was 685.14 mg/L obtained at 42°C, with lactose as a carbon source. The characterization study showed that the purified EPS consisted of one main fraction that contained 97.67% of carbohydrates. Furthermore, the EPS was identified as a homogeneous polysaccharide, mainly composed of glucose. These results demonstrated the high EPS production ability of the selected L. rhamnosus P14, representing a promising candidate to improve the textural and sensory properties of fermented food.

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Section: Resultssupporting

confidence: 80%

Investigation on exopolysaccharide production by Lacticaseibacillus rhamnosus P14 isolated from Moroccan raw cow's milk

Akhtach

Tabia

Bricha

et al. 2021

Journal of Food Science

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“…Bacterial surface polysaccharides are considered key macromolecules in determining microbe-host interactions through passive forces, electrostatic interactions, and hydrophobic and steric forces [99,100]. Surface polysaccharide production is widely reported in LAB isolates, in particular among members of the Lactobacillus genus [101,102]. Since polysaccharides display a high diversity among LAB [103], they are thought to be involved in determining relevant strain-specific properties for probiotic action, such as the degree of bacterial adhesion to host cells [104–106,60].…”

Section: Discussionmentioning

confidence: 99%

Skin-associated lactic acid bacteria from North American bullfrogs as potential control agents of Batrachochytrium dendrobatidis

et al. 2019

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The fungal pathogen Batrachochytrium dendrobatidis (Bd) is the causative agent of chytridiomycosis and has been a key driver in the catastrophic decline of amphibians globally. While many strategies have been proposed to mitigate Bd outbreaks, few have been successful. In recent years, the use of probiotic formulations that protect an amphibian host by killing or inhibiting Bd have shown promise as an effective chytridiomycosis control strategy. The North American bullfrog (Lithobates catesbeianus) is a common carrier of Bd and harbours a diverse skin microbiota that includes lactic acid bacteria (LAB), a microbial group containing species classified as safe and conferring host benefits. We investigated beneficial/probiotic properties: anti-Bd activity, and adhesion and colonisation characteristics (hydrophobicity, biofilm formation and exopolysaccharide-EPS production) in two confirmed LAB (cLAB-Enterococcus gallinarum CRL 1826, Lactococcus garvieae CRL 1828) and 60 presumptive LAB (pLAB) [together named as LABs] isolated from bullfrog skin.We challenged LABs against eight genetically diverse Bd isolates and found that 32% of the LABs inhibited at least one Bd isolate with varying rates of inhibition. Thus, we established a score of sensitivity from highest (BdGPL AVS7) to lowest (BdGPL C2A) for the studied Bd isolates. We further reveal key factors underlying host adhesion and colonisation of LABs. Specifically, 90.3% of LABs exhibited hydrophilic properties that may promote adhesion to the cutaneous mucus, with the remaining isolates (9.7%) being hydrophobic in nature with a surface polarity compatible with colonisation of acidic, basic or both substrate types. We also found that 59.7% of LABs showed EPS synthesis and 66.1% produced biofilm at different levels: 21% weak, 29% moderate, and 16.1% strong. Together all these properties enhance colonisation of the host surface (mucus or epithelial cells) and may confer protective benefits against Bd through competitive exclusion. Correspondence analysis indicated that biofilm synthesis was LABs specific with high aggregating bacteria correlating with strong biofilm producers, and EPS producers being correlated to negative biofilm producing LABs. We performed Random Amplified Polymorphic DNA (RAPD)-PCR analysis and demonstrated a higher degree of genetic diversity among rod-shaped pLAB than cocci. Based on the LAB genetic analysis and specific probiotic selection criteria that involve beneficial properties, we sequenced 16 pLAB which were identified as Pediococcus pentosaceus, Enterococcus thailandicus, Lactobacillus pentosus/L. plantarum, L. brevis, and L. curvatus. Compatibility assays performed with cLAB and the 16 species described above indicate that all tested LAB can be included in a mixed probiotic formula. Based on our analyses, we suggest that E. gallinarum CRL 1826, L. garvieae CRL 1828, and P. pentosaceus 15 and 18B represent optimal probiotic candidates for Bd control and mitigation.

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“…EPS was collected from bacterial cultures by using ethanol precipitation method with modification. 26,33,34 Basically, the cell free supernatant of bacterium was mixed with absolute ethanol with the ratio 1:2 at 4°C for 24 hours. After that, the mixture was centrifuged at 12,000 rpm for 30 minutes at 4°C.…”

Section: Eps Extractionmentioning

confidence: 99%

“…The study also characterized EPS because EPS involving the TTX toxicity involvement. 26 By detecting carbohydrate using Phenol-sulfuric acid assay, the concentration of EPS was determined (Figure 6). The standard line had the equation y=0.0108x+0.0466 with R2=0.984 >0.97, which meant this standard curve was reliable and can be used for calculating the concentration of EPS.…”

Section: Eps Detectionmentioning

confidence: 99%

“…Some bacteria, such as Pseudomonas and Vibrio species, produce TTX, which also produces EPS. EPS involved TTX toxicity 26 that might provide a protective barrier around the bacteria and help them colonize specific environments. It is important to note that further research is needed to fully understand the role of exopolysaccharides in the production and distribution of tetrodotoxin, and to determine the potential implications for human health and the environment.…”

Section: Introductionmentioning

confidence: 99%

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Isolation, Identification, and Characterization of Tetrodotoxin Producing Bacillus amyloliquefaciens B1 Originated from Carcinoscorpius rotundicauda

Le¹,

Doan²,

Le³

et al. 2023

J. Pure Appl. Microbiol.

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This study isolated and identified Bacillus amyloliquefaciens B1 from Carcinoscorpius rotundicauda by carrying out the 16S rRNA sequence analysis, reconstructing the phylogenetic tree based on the Environment for Tree Exploration (ETE3) v3.1.1 belonging to the GenomeNet. By an indirect competitive enzyme-labeled immunoassay, B1 could produce tetrodotoxin (TTX) in MRS was more highly than LB media. After purification, TTX producing ability in B1 could be detected in ELISA assay, high performance liquid chromatography (HPLC). The gel permeation chromatography and gas chromatography were applied to determine the molecular weight of EPS and the concentration of glucose in EPS. The results indicated the highest molecular weight of exopolysaccharides (EPS) estimated 1.33 × 106 Da consisted of glucose (150.09 µg/g). TTX yield was proportional to EPS production in the bacterium. The antimicrobial activities of EPS were determined by agar well diffusion method. Diameter of inhibition zone (mm) of Bacillus amyloliquefaciens EPS on the test microorganisms. The EPS could inhibit against Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Vibrio parahaemolyticus ATCC 17802 and Micrococcus luteus ATCC 10240. In silico prediction, TTX might interact with Bacillus amyloliquefaciens via the extracellular domain of noncanonic ABC-type transporter from gram positive bacteria. TTX might also interact with peptidase S54, mistic, metal binding protein of Bacillus subtilis and tryptophan-rich sensory protein of Bacillus cereus. This study provides the understanding of TTX producing Bacillus amyloliquefaciens B1 isolated from Carcinoscorpius rotundicauda.

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Detection of the Potential Inactivation of Tetrodotoxin by Lactic Acid Bacterial Exopolysaccharide

Cited by 10 publications

References 23 publications

Investigation on exopolysaccharide production by Lacticaseibacillus rhamnosus P14 isolated from Moroccan raw cow's milk

Investigation on exopolysaccharide production by Lacticaseibacillus rhamnosus P14 isolated from Moroccan raw cow's milk

Skin-associated lactic acid bacteria from North American bullfrogs as potential control agents of Batrachochytrium dendrobatidis

Isolation, Identification, and Characterization of Tetrodotoxin Producing Bacillus amyloliquefaciens B1 Originated from Carcinoscorpius rotundicauda

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