Safety assessment of Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA determined in trials with Wistar rats

Colonization of Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA in the gastro-intestinal tract was determined by using Wistar rats as model. The strains were administered through intragastric gavage over 14 days. FISH with strain-specific oligonucleotide probes indicated that Lact. plantarum 423 adhered to the surfaces of the ileum and the cecum. Enterococcus mundtii ST4SA, on the other hand, adhered to the surfaces of the cecum and colon. Results obtained by DGGE have shown that strains 423 and ST4SA excluded Enterobacteriaceae, but not lactic acid bacteria, from the cecum and colon. No signs of perforation of epithelial cells by strains 423 and ST4SA were detected. The spleen and liver appeared healthy and blood counts were normal, suggesting that the strains are not pathogenic. Both strains produce antimicrobial peptides active against a number of pathogens and may be considered as probiotics

Molecular insights into probiotic mechanisms of action employed against intestinal pathogenic bacteria

CITATION: van Zyl, W. F.; Deane, S. M. & Dicks, L. M. T. 2020. Molecular insights into probiotic mechanisms of action employed against intestinal pathogenic bacteria. Gut Microbes, 12(1). doi:10.1080/19490976.2020.1831339The original publication is available at https://www.tandfonline.com/loi/kgmi20Gastrointestinal (GI) diseases, and in particular those caused by bacterial infections, are a major cause of morbidity and mortality worldwide. Treatment is becoming increasingly difficult due to the increase in number of species that have developed resistance to antibiotics. Probiotic lactic acid bacteria (LAB) have considerable potential as alternatives to antibiotics, both in prophylactic and therapeutic applications. Several studies have documented a reduction, or prevention, of GI diseases by probiotic bacteria. Since the activities of probiotic bacteria are closely linked with conditions in the host’s GI-tract (GIT) and changes in the population of enteric microorganisms, a deeper understanding of gut-microbial interactions is required in the selection of the most suitable probiotic. This necessitates a deeper understanding of the molecular capabilities of probiotic bacteria. In this review, we explore how probiotic microorganisms interact with enteric pathogens in the GIT. The significance of probiotic colonization and persistence in the GIT is also addressed.https://www.tandfonline.com/doi/full/10.1080/19490976.2020.1831339Publishers versio

Isolation and characterization of lytic proteus virus 309

Proteus mirabilis is frequently associated with complicated urinary tract infections (UTIs) and is the main cause of catheter-associated urinary tract infections (CAUTIs). Treatment of such infections is complicated and challenging due to the biofilm forming abilities of P. mirabilis. If neglected or mistreated, infections may lead to life-threating conditions such as cystitis, pyelonephritis, kidney failure, and bacteremia that may progress to urosepsis. Treatment with antibiotics, especially in cases of recurring and persistent infections, leads to the development of resistant strains. Recent insights into phage therapy and using phages to coat catheters have been evaluated with many studies showing promising results

Corrigendum: A Review: The Fate of Bacteriocins in the Human Gastro-Intestinal Tract: Do They Cross the Gut–Blood Barrier?

The intestinal barrier, consisting of the vascular endothelium, epithelial cell lining, and mucus layer, covers a surface of about 400 m2. The integrity of the gut wall is sustained by transcellular proteins forming tight junctions between the epithelial cells. Protected by three layers of mucin, the gut wall forms a non-permeable barrier, keeping digestive enzymes and microorganisms within the luminal space, separate from the blood stream. Microorganisms colonizing the gut may produce bacteriocins in an attempt to outcompete pathogens. Production of bacteriocins in a harsh and complex environment such as the gastro-intestinal tract (GIT) may be below minimal inhibitory concentration (MIC) levels. At such low levels, the stability of bacteriocins may be compromised. Despite this, most bacteria in the gut have the ability to produce bacteriocins, distributed throughout the GIT. With most antimicrobial studies being performed in vitro, we know little about the migration of bacteriocins across epithelial barriers. The behavior of bacteriocins in the GIT is studied ex vivo, using models, flow cells, or membranes resembling the gut wall. Furthermore, little is known about the effect bacteriocins have on the immune system. It is generally believed that the peptides will be destroyed by macrophages once they cross the gut wall. Studies done on the survival of neurotherapeutic peptides and their crossing of the brain–blood barrier, along with other studies on small peptides intravenously injected, may provide some answers. In this review, the stability of bacteriocins in the GIT, their effect on gut epithelial cells, and their ability to cross epithelial cells are discussed. These are important questions to address in the light of recent papers advocating the use of bacteriocins as possible alternatives to, or used in combination with, antibiotics

A Review: The Fate of Bacteriocins in the Human Gastro-Intestinal Tract: Do They Cross the Gut–Blood Barrier?

The intestinal barrier, consisting of the vascular endothelium, epithelial cell lining, and mucus layer, covers a surface of about 400 m2. The integrity of the gut wall is sustained by transcellular proteins forming tight junctions between the epithelial cells. Protected by three layers of mucin, the gut wall forms a non-permeable barrier, keeping digestive enzymes and microorganisms within the luminal space, separate from the blood stream. Microorganisms colonizing the gut may produce bacteriocins in an attempt to outcompete pathogens. Production of bacteriocins in a harsh and complex environment such as the gastro-intestinal tract (GIT) may be below minimal inhibitory concentration (MIC) levels. At such low levels, the stability of bacteriocins may be compromised. Despite this, most bacteria in the gut have the ability to produce bacteriocins, distributed throughout the GIT. With most antimicrobial studies being performed in vitro, we know little about the migration of bacteriocins across epithelial barriers. The behavior of bacteriocins in the GIT is studied ex vivo, using models, flow cells, or membranes resembling the gut wall. Furthermore, little is known about the effect bacteriocins have on the immune system. It is generally believed that the peptides will be destroyed by macrophages once they cross the gut wall. Studies done on the survival of neurotherapeutic peptides and their crossing of the brain–blood barrier, along with other studies on small peptides intravenously injected, may provide some answers. In this review, the stability of bacteriocins in the GIT, their effect on gut epithelial cells, and their ability to cross epithelial cells are discussed. These are important questions to address in the light of recent papers advocating the use of bacteriocins as possible alternatives to, or used in combination with, antibiotics

Soymilk bio-enrichment by indigenously isolated riboflavin-producing strains of Lactobacillus plantarum

CITATION: Bhushan, B. et al. 2020. Soymilk bio-enrichment by indigenously isolated riboflavin-producing strains of Lactobacillus plantarum. LWT, 119. doi:10.1016/j.lwt.2019.108871The original publication is available at https://www.sciencedirect.com/journal/lwtLactobacilli (n = 68) isolated from human feces and fermented milk products were screened for the production of riboflavin (vitamin B2) by culturing into riboflavin assay medium (RAM). Cell-free culture supernatants from positive isolates (BBC32A, BBC32B, BBC33 and BIF43) were transferred onto RAM agar pre-seeded with Enterococcus faecalis MTCC2729 (a riboflavin auxotroph). The enhanced growth of B2-auxotroph confirmed the bioavailability of produced vitamin. Isolate BBC32B produced the highest riboflavin (319 ± 36 μg/l), followed by BBC33 (304 ± 91 μg/l), BBC32A (276 ± 8 μg/l) and BIF43 (257 ± 91 μg/l). All four isolates contained riboflavin genes ribG, ribB, ribA and ribH. Sequencing of DNA fragments amplified from the 16S–23S rRNA intergenic spacer region and areas flanking the 23S rRNA gene grouped these isolates within the species Lactobacillus plantarum. Identifications were confirmed by sequencing 1300-bp of amplified 16S rDNA fragments. Fermentation of soymilk by single cultures of L. plantarum BBC32B, BBC33 and BIF43 yielded 49.05%, 38.97% and 35.94% riboflavin enrichment respectively, which is more than 18.75% recorded in literature for Lactobacillus fermentum MTCC8711. Maximum levels of riboflavin were obtained within 12 h of fermentation in soymilk. Lactobacillus plantarum BBC32B may be used as starter culture for developing of riboflavin-enriched soymilk.https://www.sciencedirect.com/science/article/pii/S0023643819312137?via%3DihubPublishers versio

Procjena probiotičkih svojstava bakterije Enterococcus mundtii, njezino preživljavanje u bozi i proizvodnja bakteriocina in situ

Boza is a low-pH and low-alcohol cereal-based beverage produced in the Balkan Peninsula. Barley was cooked and prepared according to a traditional recipe and inoculated with Enterococcus mundtii ST4V (a potential probiotic and bacteriocin-producing strain), commercially produced boza, Saccharomyces cerevisiae, and a combination of strain E. mundtii ST4V and Saccharomyces cerevisiae. Fermentation was carried out at 37 °C for 3 h. The organoleptic properties of fermented products were evaluated by a qualified taste panel. No significant differences in rheological properties were observed, suggesting that E. mundtii ST4V had no effect on the quality of the final product. Microbial cell numbers remained relatively unchanged during one week of storage. The preservative properties of bacteriocin ST4V were evaluated by contaminating boza with Lactobacillus sakei DSM 20017. Changes in microbial populations were monitored by using classical microbiological methods, PCR with species-specific primers and denaturing gradient gel electrophoresis (DGGE). Adsorption of bacteriocin ST4V to target cells is pH-dependent, with the highest adsorption (88 %) recorded at pH=8.0 and pH=10.0. Maximum adsorption of bacteriocin ST4V (75 %) to Enterococcus faecalis and Listeria innocua was recorded at 25 to 37 °C. Growth of E. mundtii ST4V was inhibited only by a few antibiotics and anti-inflammatory medicaments, suggesting that the strain may be used as a probiotic by individuals receiving medical treatment.Boza je napitak od žitarica, male pH-vrijednosti i neznatnog udjela alkohola, koji se priprema na Balkanu. U istraživanju je upotrijebljena boza pripremljena kuhanjem ječma prema tradicionalnom receptu, inokulirana bakterijskim sojem Enterococcus mundtii ST4V (koji proizvodi bakteriocin i ima probiotička svojstva), bozom proizvedenom na komercijalan način, kvascem Saccharomyces cerevisiae i kombinacijom bakterijskog soja i kvasca. Tim stručnjaka ispitao je organoleptička svojstva proizvoda dobivenih nakon 3 sata fermentacije pri 37 ºC. Nije utvrđena značajna razlika u reološkim svojstvima dobivenih proizvoda, što upućuje na zaključak da inokulacija sojem E. mundtii ST4V nije utjecala na kakvoću proizvoda. Čak ni nakon tjedan dana skladištenja nije se promijenio broj stanica mikroorganizama u proizvodima. Da bi se procijenio zaštitni učinak bakteriocina ST4V, ispitana je boza inokulirana sojem Lactobacillus sakei DSM 20017, pa je mikrobna populacija praćena klasičnim mikrobiološkim metodama, reakcijom polimeraze (PCR) sa specifičnim početnicama i elektroforezom u gradijentu denaturirajućega gela (DGGE). Adsorpcija bakteriocina ST4V na ciljane stanice ovisi o pH-vrijednosti, s najvećom adsorpcijom (88 %) zabilježenom pri pH=8 i pH=10. Maksimalna adsorpcija bakteriocina (75 %) na stanice Enterococcus faecalis i Listeria innocua zabilježena je u rasponu temperature od 25 do 37 ºC. Rast bakterije E. mundtii ST4V inhibirali su samo neki antibiotici i protuupalni lijekovi, što upućuje na to da taj soj ima probiotički učinak i na pacijente koji primaju određenu terapiju

Variation in the provision and practice of implant-based breast reconstruction in the UK: Results from the iBRA national practice questionnaire

Introduction The introduction of biological and synthetic meshes has revolutionised the practice of implant-based breast reconstruction (IBBR) but evidence for effectiveness is lacking. The iBRA (implant Breast Reconstruction evAluation) study is a national trainee-led project that aims to explore the practice and outcomes of IBBR to inform the design of a future trial. We report the results of the iBRA National Practice Questionnaire (NPQ) which aimed to comprehensively describe the provision and practice of IBBR across the UK. Methods A questionnaire investigating local practice and service provision of IBBR developed by the iBRA Steering Group was completed by trainee and consultant leads at breast and plastic surgical units across the UK. Summary data for each survey item were calculated and variation between centres and overall provision of care examined. Results 81 units within 79 NHS-hospitals completed the questionnaire. Units offered a range of reconstructive techniques, with IBBR accounting for 70% (IQR:50–80%) of participating units' immediate procedures. Units on average were staffed by 2.5 breast surgeons (IQR:2.0–3.0) and 2.0 plastic surgeons (IQR:1.0–3.0) performing 35 IBBR cases per year (IQR:20-50). Variation was demonstrated in the provision of novel different techniques for IBBR especially the use of biological (n = 62) and synthetic (n = 25) meshes and in patient selection for these procedures. Conclusions The iBRA-NPQ has demonstrated marked variation in the provision and practice of IBBR in the UK. The prospective audit phase of the iBRA study will determine the safety and effectiveness of different approaches to IBBR and allow evidence-based best practice to be explored

Common variants at theCHEK2gene locus and risk of epithelial ovarian cancer

Genome-wide association studies have identified 20 genomic regions associated with risk of epithelial ovarian cancer (EOC), but many additional risk variants may exist. Here, we evaluated associations between common genetic variants [single nucleotide polymorphisms (SNPs) and indels] in DNA repair genes and EOC risk. We genotyped 2896 common variants at 143 gene loci in DNA samples from 15 397 patients with invasive EOC and controls. We found evidence of associations with EOC risk for variants at FANCA, EXO1, E2F4, E2F2, CREB5 and CHEK2 genes (P ≤ 0.001). The strongest risk association was for CHEK2 SNP rs17507066 with serous EOC (P = 4.74 x 10(-7)). Additional genotyping and imputation of genotypes from the 1000 genomes project identified a slightly more significant association for CHEK2 SNP rs6005807 (r (2) with rs17507066 = 0.84, odds ratio (OR) 1.17, 95% CI 1.11-1.24, P = 1.1×10(-7)). We identified 293 variants in the region with likelihood ratios of less than 1:100 for representing the causal variant. Functional annotation identified 25 candidate SNPs that alter transcription factor binding sites within regulatory elements active in EOC precursor tissues. In The Cancer Genome Atlas dataset, CHEK2 gene expression was significantly higher in primary EOCs compared to normal fallopian tube tissues (P = 3.72×10(-8)). We also identified an association between genotypes of the candidate causal SNP rs12166475 (r (2) = 0.99 with rs6005807) and CHEK2 expression (P = 2.70×10(-8)). These data suggest that common variants at 22q12.1 are associated with risk of serous EOC and CHEK2 as a plausible target susceptibility gene.Other Research Uni