Mundtii Strain Research Articles

Isolation and Identification of Multi-Traits PGPR for Sustainable Crop Productivity Under Salinity Stress

High salinity poses a significant threat to arable land globally and contributes to desertification. Growth-promoting rhizobacteria assist plants in mitigating abiotic stresses and enhancing crop productivity through the production of siderophores, exopolysaccharides (EPS), solubilisation of phosphate, indole-3-acetic acid (IAA), and other secondary metabolites. This study aimed to isolate, identify, and characterise bacteria that exhibit robust growth-promoting properties. A total of 64 bacterial isolates from the rhizosphere of Miscanthus sinensis were evaluated for plant growth-promoting (PGP) traits, including IAA, EPS, siderophores, and solubilisation of phosphate. Among them, five isolates were selected as plant growth-promoting rhizobacteria (PGPR) based on their PGP features and identified via 16S rRNA sequencing: Enterococcus mundtii strain INJ1 (OR122486), Lysinibacillus fusiformis strain INJ2 (OR122488), Lysinibacillus sphaericus strain MIIA20 (OR122490), Pseudomonas qingdaonensis strain BD1 (OR122487), and Pseudomonas qingdaonensis strain MIA20 (OR122489), all documented in NCBI GenBank. BD1 demonstrated a higher production of superoxide dismutase (SOD) (17.93 U/mg mL), catalase (CAT) (91.17 U/mg mL), and glutathione (GSH) (0.18 U/mg mL), along with higher concentrations of IAA (31.69 µg/mL) and salicylic acid (SA) (14.08 ng/mL). These isolates also produced significant quantities of amino and organic acids. BD1 exhibited superior PGP traits compared to other isolates. Furthermore, the NaCl tolerance of these bacterial isolates was assessed by measuring their growth at concentrations ranging from 0 to 200 mM at 8-h intervals. Optical density (OD) measurements indicated that BD1 and INJ2 displayed significant tolerance to salt stress. The utilisation of these isolates, which enhances plant growth and PGP traits under salt stress, may improve plant development under saline conditions.

Safety assessment of enterocin-producing Enterococcus strains isolated from sheep and goat colostrum

BackgroundThis study investigates the safety evaluation of enterocin-producing 11 E. mundtii and two E. faecium strains previously isolated from small livestock colostrums. Enterococcus species do not possess Generally Recognized as Safe (GRAS) status. Hence, it is critical to scrutinize enterococci’s antibiotic resistance, virulence characteristics, and biogenic amine production capabilities in order to assess their safety before using them as starter or adjunct cultures.ResultsEnterococcus strains showed susceptibility to medically significant antibiotics. Multiple-drug resistance (MDR) was found in only E. faecium HC121.4, and its multiple antibiotic resistance (MAR) index was detected to be 0.22. The tetL and aph(3')-IIIa were the most commonly found antibiotic resistance genes in the strains. However, E. mundtii strains HC56.3, HC73.1, HC147.1, and E. faecium strain HC121.4 were detected to lack any of the antibiotic resistance genes examined in this study. Only E. mundtii HC166.3 showed hemolytic activity, while none of the strains engage in gelatinase activity. The strains were identified to have virulence factor genes with a low rate. None of the virulence factor genes could be detected in E. mundtii HC26.1, HC56.3, HC73.1, HC165.3, HC166.8, and E. faecium HC121.4. The E. mundtii HC73.2 strain displayed the highest presence of virulence factor genes, namely gelE, efaAfs, cpd, and ccf. Similarly, the E. mundtii HC112.1 strain showed a significant presence of genes efaAfm, ccf, and acm. There was no decarboxylation of histidine, ornithine, or lysine seen in any of the strains. Nevertheless, E. faecium HC121.4 and HC161.1 strains could decarboxylate tyrosine, but E. mundtii HC26.1, HC56.3, HC73.1, HC73.2, HC112.1, HC147.1, HC155.2, HC165.3, HC166.3, HC166.5, and HC166.8 strains only showed a limited capacity for tyrosine decarboxylation. None of the strains possessed the hdc, odc, or ldc genes, but all of them had the tdc gene.ConclusionThe E. mundtii HC56.3 and HC73.1 strains were deemed appropriate for utilization in food production. Using the remaining 11 strains as live cultures in food production activities could pose a possible risk to consumer health.

Probiotic Potential of Lactobacillus and Enterococcus Strains Isolated From the Faecal Microbiota of Critically Endangered Hangul Deer (Cervus hanglu hanglu): Implications for Conservation Management.

The mammalian gut microbiota plays a crucial role in promoting host health, and lactic acid bacteria (LAB) are commonly employed as probiotics for their beneficial effects. The Hangul deer (Cervus hanglu hanglu), a critically endangered red deer subspecies found in the Indian subcontinent, requires meticulous health management for its conservation. This pioneering study aimed to isolate, identify, and evaluate the in-vitro probiotic functional properties of LAB strains from the faeces of Hangul deer. A total of 27 LAB strains were isolated and identified using 16S rDNA gene sequencing, followed by comprehensive probiotic characterization and safety assessment. Remarkably, four species exhibited robust resistance and survivability against varying pH levels and bile salts, along with high aggregation and co-aggregation capacities. Notably, Lactobacillus acidophilus and Enterococcus mundtii strains displayed antibacterial activities. Safety assessment revealed the absence of hemolytic activity and virulence genes in all four strains. Antibiotic susceptibility testing showed that Lactobacillus acidophilus and Enterococcus casseliflavus were susceptible to all tested antibiotics, while Enterococcus mundtii exhibited resistance to clindamycin, and Enterococcus gallinarum exhibited resistance to erythromycin. These findings suggest that the isolated LAB strains possess advantageous probiotic characteristics and hold potential as dietary supplements for promoting the health and disease management of Hangul deer.

Genomic Insights Into Enterococcus mundtii 203: A Promising Probiotic Candidate Isolated From Camel Feces.

Enterococcus, a common commensal organism in the human gut, exhibits a dual nature with certain strains offering probiotic benefits, while others are associated with nosocomial infections. In this study, we conducted a comprehensive examination of the genome of Enterococcus mundtii strain 203 to assess its probiotic potential and safety profile. The complete genome sequencing, assembly, and annotation were performed, followed by bioinformatics analysis. Our investigation reveals a detailed characterization of the Enterococcus mundtii 203 genome, originally isolated from camel feces, with a size of 3,053,234 bases and a GC content of 38.4%. Importantly, our analysis suggests that this strain poses no risk as a human pathogen due to the absence of antibiotic resistance determinants and virulence factors. The genome harbors a multitude of genes responsible for lactic acid production, bioactive peptide synthesis, adhesion molecule expression, resistance to harsh gut conditions, and enhancement of host metabolism. These findings underline the potential probiotic functionality of Enterococcus mundtii 203, positioning it as a promising candidate. Notably, our study did not identify any sequences related to insertion elements or CRISPR-Cas fragments.

The Enterococcus secretome inhibits the growth of Mycobacterium tuberculosis complex mycobacteria.

Enterococcus mundtii , a commensal intestinal bacterium, was demonstrated to inhibit the growth of some Mycobacterium tuberculosis complex (MTC) species that cause tuberculosis in humans and mammals. To further explore this preliminary observation, we cross-investigated five E. mundtii strains and seven MTC strains representative of four MTC species using a standardized quantitative agar well diffusion assay. All five E. mundtii strains, calibrated at 10 MacFarland, inhibited the growth of all M. tuberculosis strains with various susceptibility profiles, but no inhibition was observed with lower inoculums. Further, eight E. mundtii freeze-dried cell-free culture supernatants (CFCS) inhibited the growth of M. tuberculosis , Mycobacterium africanum, Mycobacterium bovis and Mycobacterium canettii, the most susceptible MTC species (inhibition diameter 25±1 mm), proportionally to CFCS protein concentrations. The data reported here indicate that the E. mundtii secretome inhibited growth of all MTC species of medical interest, which broadens previously reported data. In the gut, the E. mundtii secretome may modulate the expression of tuberculosis, exhibiting an anti-tuberculosis effect, with some protective roles in human and animal health.

Bacteriocin production and technological properties of Enterococcus mundtii and Enterococcus faecium strains isolated from sheep and goat colostrum.

Enterococci are lactic acid bacteria (LAB) that play a role in the aroma formation, maturation, and sensory development of fermented foods such as meat and dairy products. They also contribute to the improvement of the extended shelf life of fermented foods by producing bacteriocin. The aim of this study was to isolate bacteriocin-producing LAB from sheep and goat colostrum, to characterize the bacteriocin-producing strains, and determine the technological properties of the strains. A total of 13 bacteriocin-producing LAB was isolated and identified as 11 Enterococcus mundtii and two Enterococcus faecium. The strains were found to be genetically different from each other by phylogenetic analysis of 16S rRNA gene sequences and random amplified polymorphic-DNA (RAPD-PCR). It has been determined that bacteriocins show activity in a wide pH range and are resistant to heat, lose their activity with proteolytic enzymes and α-amylase, but are resistant to detergents. While the presence of the munKS gene was detected in all of the strains, it was determined that E. faecium HC121.4, HC161.1, E. mundtii HC147.1, HC166.5, and HC166.8 strains contained multiple enterocin genes. Trisin-SDS-PAGE analysis revealed two active protein bands of approximately 5.1 and 5.5kDa in E. faecium HC121.4 and one active protein band with a weight of approximately 4.96kDa in other strains. E. mundtii strains and E. faecium HC161.1 were identified as mundticin KS producers, and E. faecium HC121.4 was defined as an enterocin A and B producer. Except for E. mundtii HC166.8, acid production of strains was found to be slow at 6h and moderate at 24h. None of them showed extracellular proteolytic and lipolytic activities. It was found that the strains had esterase, esterase lipase, leucine arylamidase, acid phosphatase, and naphthol-AS-Bl-phosphohydrolase activities, while protease activities were low and peptidase activities were high. In conclusion, bacteriocin producer 13 Enterococcus strains isolated from sheep and goat colostrum were found to have the potential to be included in starter culture combinations.

Safety Assessment of Fecal, Bacteriocin-Producing Strains Enterococcus mundtii from Horses

Backround: Knowing and optimizing the host microbiota is important regarding the maintenance of horse`s health. For this approach beneficial bacteria have been usually used. However, to be the most effective in use, their safety, tolerability and efficacy needs to be assessed. Therefore, fecal strains Enterococcus mundtii from horses with promising bacteriocinogenic potential have to be evaluated for their safety. Methodology: E. mundtii strains were isolated from feces of 47 horses (n=47; 40 mares and seven stallions), the Norik breed from Muráň in eastern Slovakia. MALDTI-TOF spectrometry and sequencing were used. Phenotypic characteristics were assessed in accordance with those for the reference strain E. mundtii ATCC43186. Biofilm-forming ability was tested using plate assay. For enzyme production commercial tests were applied and virulence factor genes were tested using PCR and primers. Antibiotic profile was tested with diffusion method. In vivo safety was tested using Balb/c breed mice. Results: E. mundtii strains did not produce the enzyme β-glucuronidase; however, most E. mundtii strains produced β-galactosidase. The strains were absent of virulence factors such as gelatinase, aggregation substance, cytolysin A, enterococcal superficial protein, adhesins, hyaluronidase and IS16 element, except efaAfs gene in the strain EMKD 24/1. Six strains were found with lowgrade (0.1 ≤ A570< 1) biofilm - forming ability. E. mundtii were mostly susceptible to antibiotics. Bacteriocinogenic strain EMKD41/3 reached high counts in feces (5.12 ± 0.26 CFU/g log 10) of Balb/c mice during its 30 days application. No mortality of mice was noted during whole period of EMKD41/3 strain application. Conclusion: Bacteriocin-producing strains E. mundtii should not threaten horses because they were mostly susceptible to antibiotics, they were virulence factor genes absent and with low-grade biofilm formation ability. Bacteriocinproducing strain EMKD41/3 even indicates its sufficient implementation in host organism with any side effect.

Antimicrobial resistance, biofilm production and invasion of mammary epithelial cells by Enterococcus faecalis and Enterococcus mundtii strains isolated from bovine subclinical mastitis in Brazil

Members of the genus Enterococcus are among the most relevant etiologic agents of bovine clinical and subclinical mastitis, a major problem for the dairy industry. In Brazil, clonal diversity, and multidrug resistance profiles related to bovine infections need further investigation. In this study, 11 bacterial strains recovered from mastitis subclinical cases detected in different farms of São Paulo, Brazil, were identified as Enterococcus faecalis (n = 8) and Enterococcus mundtii (n = 3) by biochemical testing and MALDI-TOF mass spectrometry. Pulsed-field gel electrophoresis categorized the enterococcal isolates into two main clusters (A and B) with similarity ranging from 85 to 100%. The isolates were shown to be resistant tetracycline (73%), erythromycin (73%), quinupristin-dalphopristin (64%), norfloxacin (9%), fosfomycin (9%) and linezolid (9%). Moreover, seven strains (64%) were considered multidrug-resistant. All the isolates were able to produce biofilms when grown in milk for 24 h: 54·54% were classified as moderate producers and 45·45% were weak producers. Interestingly, only two strains (Ef17 and Em42) remained as moderate biofilm producers after 48 h incubation. Moreover, all isolates showed no ability to form biofilm in tryptic soy broth (TSB) after 24 and 48 h incubation. In addition, cytoskeleton components were partially involved in E. faecalis and E. mundtii entry to epithelial cells as demonstrated by induction of actin stress fibre. In conclusion, enterococci isolates recovered from bovine subclinical mastitis were resistant to several classes of antibiotics, showing the ability to form biofilms in milk and invade mammary epithelial cells, suggesting an advantageous feature in mammary gland colonization during mastitis development. In addition, they can spread along the food chain by different routes and eventually constitute a possible threat for public health, including E. mundtii specie.

Identification of cellulolytic lactic acid bacteria from the intestines of laying hens given AKBISprob based on 16S ribosomal ribonucleic acid gene analysis.

Background and Aim:Supplementation of AKBISprob (developed in a previous study) in feed can improve production efficiency and poultry health, especially laying hens. In addition, it can also increase cellulolytic lactic acid bacteria (LAB) in chicken intestines, but these bacteria are still unknown; thus, they need to be identified. This study aimed to identify cellulolytic LAB in the intestines of laying hens administered AKBISprob based on 16S ribosomal ribonucleic acid (16S rRNA) gene analysis.Materials and Methods:The samples used in this study were 13 LAB isolates from the intestines of laying hens that were given AKBISprob 4%. Cellulolytic LAB DNA was isolated and 16S rRNA gene was amplified by polymerase chain reaction, followed by sequencing, bioinformatics analysis, and phylogenetic tree construction.Results:From 10 cellulolytic LAB isolates with a clear zone of >6 mm, four were selected and their DNA was amplified with BaCF and UniB primers ~1500 bp DNA fragments. Of these, the P31H62 isolate was genetically close to Enterococcus hirae strain 1-1X-16 with 92.90% maximum identity, the P33S52 isolate had homology with Enterococcus mundtii strain ZU 26 with 96.76% maximum identity, and the P33S62 isolate was closely related to E. hirae strain SJ3 with 72.96% maximum identity. The phylogenetic tree revealed that the cellulolytic LAB isolates P31H62 and P33S52 were in one cluster closely related to the genus Enterococcus.Conclusion:This study suggests that the isolates P31H62, P33S62, and P33S52 from the intestines of laying hens administered 4% AKBISprob are cellulolytic LAB belonging to the genus Enterococcus.

Utilization of Agro-Industrial Byproducts for Bacteriocin Production Using Enterococcus spp. Strains Isolated from Patagonian Marine Invertebrates.

This work's objective was to determine the antagonist activity of 11 Enterococcus spp. using industrial food wastes as a culture medium. The strains were isolated from invertebrates collected on the Argentinian Patagonia coast and selected by their high antibacterial activity. Phenotypic and genotypic techniques allowed identifying five E. hirae strains, five E. faecium strains, and one E. mundtii strain. The cell-free supernatants displayed inhibitory activity against most of the Gram-positive bacteria tested and Vibrio anguilarum. PCR amplification techniques detected the encoding genes of enterocin P in ten strains, mundtiicin KS in seven strains, enterocin B in six strains, hiracin JM79 in five strains, and enterocin A in three strains. The strains did not show gelatinase or hemolytic activities and were sensitive to gentamicin, kanamycin, streptomycin, tylosine, tetracycline, chloramphenicol and vancomycin. Cheese whey and hot trub derived from beer brewing were used alone or in combination to assay enterocin production. In all cases, the highest inhibitory activities were achieved when mixtures of both byproducts were used as growth medium. The results suggest that the selected strains can produce high levels of enterocins in a low-cost media composed of a mix of cheese whey and hot trub without additional supplementation with carbon or nitrogen sources.

Lilium spp., as unnoticed environmental vector, spreading OptrA-carrying Enterococcus spp.

Flower is an essential element in the human lifestyle but its role in disseminating antimicrobial resistance (AMR) between the environment and humans is unclear. In this study, we screened fresh flowers (Lilium spp.) collected from planting bases, market and florists in Guangzhou China aiming to investigate the prevalence of AMR genes, particularly cfr, optrA and poxtA mediating resistance to linezolid, a first-line drug for the treatment of different Gram-positive bacterial infections. We found 223 Enterococcus isolates consisting of Enterococcus faecalis, Enterococcus faecium and Enterococcus mundtii, and >50% of these isolates exhibited multiple-drug resistance. Additionally, 31 optrA-positive Enterococcus including 22 E. faecalis and 9 E. mundtii strains were recovered, however cfr and poxtA were not detected. The 22 E. faecalis strains were belonged to 7 Multilocus sequence types in which ST202 and ST376 were predominant and 9 E. mundtii strains from the same plantation bases were divided into three PFGE groups. Genetically, the majority of optrA were located on the chromosome and shared similar insertion sites and transpositions mediated by Tn554 family members. Plasmid-bearing optrA were identified in 6 E. faecalis strains where IS1216 family played key roles in horizontal transfer of optrA. These findings emphasize that the prevalence of drug resistant Enterococcus in fresh flowers is a latent danger and increases the risk of AMR dissemination to humans from the environment.

Biological control of Listeria monocytogenes in soil model systems by Enterococcus mundtii strains expressing mundticin KS production

The agricultural practices applied in pre-harvest greatly influence the presence and the levels of microorganisms in fresh produce. Among these, Listeria monocytogenes represents one of the most lethal foodborne pathogens associated with vegetables. The main hypothesis of this work is that bacteriocin producer Enterococcus mundtii strains can be effective against L. monocytogenes in soil. To this purpose, bacteriocin production by E. mundtii WFE3, WFE20 and WFE31, three strains showing a strong bacteriocin activity in terms of inhibitory power and inhibition spectra, was evaluated in sterile extracts from agricultural soil and peat moss, in organic nutrient solution (ONS) and mineral nutrient solution (MNS). ONS supernatants from E. mundtii WFE3 showed the highest inhibition of the strain L. monocytogenes ATCC 19114. Thus, this strain [104 colony forming units (CFU) g−1 dw] was co-inoculated with E. mundtii WFE3 (106 CFU g−1 dw) in the sterile extracts and solutions. A general increase of E. mundtii cell densities and the contemporary decrease of the levels of L. monocytogenes were observed, particularly in ONS. This solution was used to amend autoclaved soil to test in vivo the competition among bacteriocin producer and sensitive strain. L. monocytogenes decreased of almost 1.5 log CFU when E. mundtii was added to soil. The test was also carried out with basil plants showing that the anti-Listeria effect of E. mundtii is exerted during the very first days from inoculation. After 2 days, the levels of NO3−-N in soil increased for all trials, while the concentrations of ammonium and α-amino N decreased. The lowest concentrations of NH4+-N were found in presence of L. monocytogenes. At harvest, the plants were analysed for the presence of E. mundtii and L. monocytogenes, but none of the two bacterial species deliberately added to soil was transferred to plants. Although a biocontrol based on bacteriocin application in soil has to be properly set, this study provides useful insight for the future development of chemical-free agricultural strategies.

Effect of enterocins against methicillin-resistant animal-derived staphylococci.

The occurence and spread of animal-derived methicillin resistant staphylococci (MRS) worldwide is acurrent problem, especially due to their increasing incidence in food animals and their products, with possible contamination of food consumers and handlers. Staphylococci isolated from animals (n = 123) were identified with MALDI-TOF mass spectrometry and screened for methicillin/oxacillin/cefoxitin resistance (MR) using the disk diffusion method. Twenty-three phenotypically MRS strains were analysed using PBP2' Latex Agglutination Test Kit to confirm the phenotypic MR and PCR was performed for mecAgene detection; mecA gene positive strains were furtherly confirmed by means of sequencing. The susceptibility of MRS to 11 partially-purified enterocins (Ent) produced by E. faecium, E. durans and E. mundtii strains of animal, feed/food and environmental origin was checked using agar spot tests. Out of 23 MRS, PBP testing confirmed MR in 17 strains. Three Staphylococcus epidermidis and oneS. vitulinus weremecA positive. The majority of MRS, including two mecA gene-positive strains S. epidermidis R44/1 and P3/Tr2a, were susceptible to the tested enterocins, mainly to Ent7420, EntA(P)/EK13, Ent412, Ent55 and Ent9296 (in the range 100 - 12,800 AU/mL). The most susceptible strains appeared to be the mecA gene-positive S. epidermidis SE R44/1 and SE P3/Tr2a strains, inhibited by eight enterocins out of 11 tested (100-200 AU/mL). Only four strains (including mecA gene positive S. epidermidis SE P3/Tr1 and S. vitulinus SV K12PL/1) were resistant to the tested antimicrobial substances. These results indicate that the enterocins used offer a promising option for prevention and treatment of bacterial infection caused by MRS in animals.

Draft Genome Sequence of Enterococcus mundtii SCPM-O-B-8398 (E28), Isolated from Fermented Milk in the Moscow Region, Russian Federation.

ABSTRACTWe report the draft genome sequence of the bacteriocin-producing Enterococcus mundtii strain SCPM-O-B-8398 (E28), which was isolated from fermented milk in the Moscow region, Russian Federation.

Aislamiento de una cepa de Enterococcus mundtii bacteriocinogénico proveniente de Hemiodema spectabilis (pepino de mar)

En este estudio se evaluó la actividad antibacteriana de una cepa de Enterococcus mundtii tw278 productora de bacteriocina aislada del contenido intestinal de Hemiodema spectabilis (pepino de mar), recolectado en la costa patagónica de la Argentina. La cepa se identificó mediante pruebas bioquímicas y análisis filogenético del gen ARNr 16S. Además, se detectó el gen estructural que codifica para la mundticina KS mediante técnicas de PCR. La investigación de los factores de virulencia reveló que la cepa de E. mundtii tw278 no presentó actividad gelatinasa ni hemolítica y fue susceptible a todos los antibióticos analizados, excepto la cefalotina. La máxima actividad inhibitoria se logró al final de la fase logarítmica cuando se utilizó el caldo MRS como medio de cultivo a 35 °C. Luego de 12 h de incubación, el sobrenadante libre de células (SLC) alcanzó un título de 163 840 unidades arbitrarias por mililitro contra la cepa indicadora de Listeria innocua ATCC 33090. El SLC exhibió actividad contra todas las cepas de Listeria ensayadas, Enterococcus faecalis ATCC 29212, enterococos resistentes a vancomicina (Van A, Van B y Van C), Lactobacillus plantarum TwLb 5 y Vibrio anguilarum V10. Este sería el primer estudio que informa el aislamiento de una cepa bacteriocinogénica de E. mundtii aislada del contenido intestinal de Hemiodema spectabilis.

In vivo bioluminescence imaging of the spatial and temporal colonization of lactobacillus plantarum 423 and enterococcus mundtii ST4SA in the intestinal tract of mice

BackgroundLactic acid bacteria (LAB) are major inhabitants and part of the normal microflora of the gastrointestinal tract (GIT) of humans and animals. Despite substantial evidence supporting the beneficial properties of LAB, only a few studies have addressed the migration and colonization of probiotic bacteria in the GIT. The reason for this is mostly due to the limitations, or lack of, efficient reporter systems. Here we describe the development and application of a non-invasive in vivo bioluminescence reporter system to study, in real-time, the spatial and temporal persistence of Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA in the intestinal tract of mice.ResultsThis study reports on the application of the firefly luciferase gene (ffluc) from Photinus pyralis to develop luciferase-expressing L. plantarum 423 and E. mundtii ST4SA, using a Lactococcus lactis NICE system on a high copy number plasmid (pNZ8048) and strong constitutive lactate dehydrogenase gene promoters (Pldh and STldh). The reporter system was used for in vivo and ex vivo monitoring of both probiotic LAB strains in the GIT of mice after single and multiple oral administrations. Enterococcus mundtii ST4SA reached the large intestine 45 min after gavage, while L. plantarum 423 reached the cecum/colon after 90 min. Both strains predominantly colonized the cecum and colon after five consecutive daily administrations. Enterococcus mundtii ST4SA persisted in faeces at higher numbers and for more days compared to L. plantarum 423.ConclusionsOur findings demonstrate the efficiency of a high-copy number vector, constitutive promoters and bioluminescence imaging to study the colonization and persistence of L. plantarum 423 and E. mundtii ST4SA in the murine GIT. The system allowed us to differentiate between intestinal transit times of the two strains in the digestive tract. This is the first report of bioluminescence imaging of a luciferase-expressing E. mundtii strain to study colonization dynamics in the murine model. The bioluminescence system developed in this study may be used to study the in vivo colonization dynamics of other probiotic LAB.

Draft Genome Sequences of Enterococcus mundtii Strains Isolated from Beef Slaughterhouses in Kenya.

ABSTRACTWe present here draft genome sequences of Enterococcus mundtii strains K7-EM, P2-EM, C11-EM, and H18-EM, which were isolated from slaughterhouse equipment, carcasses, and personnel of small- and medium-sized beef slaughterhouses in Kenya.

Genome Sequence of Enterococcus mundtii EM01, Isolated from Bombyx mori Midgut and Responsible for Flacherie Disease in Silkworms Reared on an Artificial Diet.

ABSTRACTThe whole genome sequence of Enterococcus mundtii strain EM01 is reported here. The isolate proved to be the cause of flacherie in Bombyx mori. To date, the genomes of 11 other E. mundtii strains have been sequenced. EM01 is the only strain that displayed active pathological effects on its associated animal species.

Probiotic Enterococcus mundtii Isolate Protects the Model Insect Tribolium castaneum against Bacillus thuringiensis.

Enterococcus mundtii strains isolated from the larval feces of the Mediterranean flour moth Ephestia kuehniella show antimicrobial activity against a broad spectrum of Gram-positive and Gram-negative bacteria. The in vitro probiotic characterization of one isolate revealed a high auto-aggregation score, a hydrophilic cell surface, tolerance for low pH, no hemolytic activity, and susceptibility to all tested antibiotics. We used the red flour beetle Tribolium castaneum, an established model organism, for the in vivo characterization of one probiotic E. mundtii isolate from E. kuehniella larvae. Tribolium castaneum larvae were fed orally with the probiotic isolate or the corresponding supernatant and then infected with either the entomopathogen Bacillus thuringiensis or Pseudomonas entomophila. Larvae exposed to the isolate or the supernatant showed increased survival following infection with B. thuringiensis but not P. entomophila. Heat treatment or treatment with proteinase K reduced the probiotic effect of the supernatant. However, the increased resistance attracts a fitness penalty manifested as a shorter lifespan and reduced fertility. T. castaneum has, pending on further research, the potential as an alternative model for the pre-screening of probiotics.

Tyrosine decarboxylase activity of Enterococcus mundtii: new insights into phenotypic and genetic aspects.

SummaryFew information is available about the tyraminogenic potential of the species Enterococcus mundtii. In this study, two plant‐derived strains of E. mundtii were selected and investigated to better understand the phenotypic behaviour and the genetic mechanisms involved in tyramine accumulation. Both the strains accumulated tyramine from the beginning of exponential phase of growth, independently on the addition of tyrosine to the medium. The strains accumulated also 2‐phenylethylamine, although with lower efficiency and in greater extent when tyrosine was not added. Accordingly, the tyrosine decarboxylase (tyrDC) gene expression level increased during the exponential phase with tyrosine added, while it remained constant and high without precursor. The genetic organization as well as sequence identity levels of tyrDC and tyrosine permease (tyrP) genes indicated a correlation with those of phylogenetically closer enterococcal species, such as E. faecium, E. hirae and E. durans; however, the gene Na+/H+ antiporter (nhaC) that usually follow tyrP is missing. In addition, BLAST analysis revealed the presence of additional genes encoding for decarboxylase and permease in the genome of several E. mundtii strains. It is speculated the occurrence of a duplication event and the acquisition of different specificity for these enzymes that deserves further investigations.