Brevilactibacter coleopterorum sp. nov., isolated from the intestine of the dark diving beetle Hydrophilus acuminatus, and Weissella coleopterorum sp. nov., isolated from the intestine of the diving beetle Cybister lewisianus.

A polyphasic taxonomic approach was used to characterize two novel bacterial strains, designated as HDW11T and HDW19T, isolated from intestine samples of the dark diving beetle Hydrophilus acuminatus and the diving beetle Cybister lewisianus, respectively. Both isolates were Gram-stain-positive, facultatively anaerobic and non-motile. Strain HDW11T grew optimally at 30 °C, pH 8 and in the presence of 1% (w/v) NaCl. Strain HDW19T grew optimally at 25 °C, pH 7 and in the presence of 0.3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences and genome sequences revealed that strain HDW11T is a member of the genus Brevilactibacter and is closely related to Brevilactibacter flavus VG341T [with 97.9% 16S rRNA sequence identity and 79.1% average nucleotide identity (ANI)], and that strain HDW19T belongs to the genus Weissella and is closely related to W. koreensis KCTC 3621T (with 98.9% 16S rRNA sequence identity and 79.5% ANI). The major cellular fatty acids of strains HDW11T and HDW19T were C18:1 ω9c and anteiso-C15:0, respectively. The sole respiratory quinone of strain HDW11T was MK-9 (H4). The major polar lipid components of strain HDW11T were diphosphatidylglycerol and phosphatidylglycerol, and the major polar lipid component of strain HDW19T was diphosphatidylglycerol. The genomic DNA G+C content of strains HDW11T and HDW19T were 72.1 and 37.2 mol%, respectively. The results of phylogenetic, phenotypic, chemotaxonomic and genotypic analyses suggest that strain HDW11T represents a novel species within the genus Brevilactibacter, and that strain HDW19T represents a novel species within the genus Weissella. We propose the name Brevilactibacter coleopterorum sp. nov. for strain HDW11T (=KACC 21335T=KCTC 49320T=JCM 33680T) and the name Weissella coleopterorum for strain HDW19T (=KACC 21347T=KCTC 43114T=JCM 33684T).

Weissella sagaensis sp. nov., isolated from traditional Chinese yogurt.

Three Gram-stain-positive bacterial strains, designated X0750T, X0278 and X0401, isolated from traditional yogurt in Tibet Autonomous Region, PR China, were characterized by a polyphasic approach, including sequence analyses of the 16S rRNA gene and three housekeeping genes (pheS, rpoA and recA), determination of average nucleotide identity (ANI) and average amino acid identity (AAI), in silico DNA-DNA hybridization (isDDH), fatty acid methyl ester (FAME) analysis and phenotypic characterization. Strain X0750T was phylogenetically related to the type strains of Weissella hellenica, Weissella bombi, Weissella paramesenteroides, Weissella jogaejeotgali, Weissella thailandensis, Weissella oryzae, Weissella cibaria and Weissella confusa, having 94.4-100 % 16S rRNA gene sequence similarities, 76.7-90.0 % pheS gene sequence similarities, 88.9-99.4 % rpoA gene sequence similarities and 77.6-92.8 % recA gene sequence similarities, respectively. ANI, isDDH and AAI values between strain X0750T and type strains of phylogenetically related species were less than 90.4, 40.9 and 92.8 % respectively, confirming that strain X0750T represents a novel species within the genus Weissella. Based upon the data obtained in the present study, a novel species, Weissella sagaensis sp. nov., is proposed and the type strain is X0750T(=NCIMB 15192T=CCM 8924T=LMG 31184T=CCTCC AB 2018403T).

A polyphasic approach to characterize Weissella cibaria and Weissella confusa strains.

AIM: To study Weissella cibaria and Weissella confusa strains, lactic acid bacteria (LAB) members naturally present in food products, but not yet included in Qualified Presumption of Safety (QPS) list of European Food Safety Authority (EFSA). METHODS AND RESULTS: We carried out a comparative genome analysis of 23 sequenced W. cibaria and 7 W. confusa genomes, in parallel with a physiological and functional characterization of several strains previously isolated from sourdough-like maize bran fermentation. The genome analysis revealed the absence of dedicated pathogenicity factors. Some putative virulence genes found in Weissella genomes were also present in other LAB strains, considered safe by EFSA and commonly used as probiotics. The physiological tests carried out on our strains corroborated the genomic results. Moreover, the following functional traits of interest to application in the food sector were identified: the majority of tested strains displayed high acidification rate, high reducing ability, production of exopolysaccharides (EPS), arabinoxylan degradation ability, growth in the presence of fructo-oligosaccharides (FOS), bile and gastric juice tolerance, and antifungal activity. CONCLUSIONS: These results provide evidence for the possible use of selected strains of W. cibaria and W. confusa in the food sector. SIGNIFICANCE AND IMPACT OF THE STUDY: This polyphasic study adds to the body of knowledge on the functional and applicable characteristics of these controversial species of LAB. This knowledge contributes to design new selected cultures included in the QPS list required for food applications.

Isolation and characterization of high exopolysaccharide-producing Weissella confusa VP30 from young children's feces.

BACKGROUND: Lactic acid bacteria (LAB) are known to have a significant ability to colonize the human intestinal tract and adhere to the surface of intestinal epithelial cells. Among the various lactic acid bacteria, exopolysaccharide (EPS) producing strains are known to provide a variety of health benefits for their hosts (e.g. anti-inflammatory, antioxidant, antitumor and stress tolerant effects). Recently, EPSs and EPS-producing lactic acid cultures have gained interest within the food industry and are playing important roles as biothickeners and texturizing agents due to their hydrocolloidal nature. In this study, 156 lactic acid bacterial strains isolated from fecal samples of healthy young children were screened and evaluated for active EPS-production capability. RESULTS: Among the various human origin lactic acid flora isolated, Weissella confusa VP30 showed the highest EPS productivity and its EPS producing properties were characterized under various cultural conditions in this research. To document the safety of W. confusa VP30, antibiotic resistance, hemolytic, and ammonia production properties were evaluated in addition. No significant negative results were observed. The maximum EPS production by W. confusa VP30 was 59.99 ± 0.91 g/l after 48 h of cultivation in media containing 10% sucrose, far exceeding EPS production by other bacterial strains reported elsewhere. Based on gel permeation chromatography (GPC), the molecular weight of EPS produced by W. confusa VP30 was 3.8 × 106 Da. Structural analysis of the released EPS fraction by 13C and 1H nuclear magnetic resonance (NMR) spectroscopy revealed that W. confusa VP30 can produce dextran with glucose units linked with 96.5% α (1 → 6) glycosidic bonds and 3.5% α (1 → 3) branches. CONCLUSION: The high EPS production capability and safety of W. confusa VP30 justify food industry consideration of this cell strain for further evaluation and potential industrial use.

Weissella cryptocerci sp. nov., isolated from gut of the insect Cryptocercus kyebangensis.

A taxonomic study of a Gram-stain-positive, rod-shaped, non-motile, non-spore-forming, catalase-negative bacterium, isolated from the gut of an insect, Cryptocercus kyebangensis collected from the mountainous area of Seoraksan, Yangyang-gun, Republic of Korea, was conducted. Its 16S rRNA gene sequence showed high similarity values to Weissella ghanensis LMG 24286T (95.9 %), Weissella beninensis 2L24P13T (95.9 %), Weissella fabalis M75T (95.7 %) and Weissella fabaria 257T (95.7 %). The phylogenetic tree indicated that the novel organism formed a cluster with W. ghanensis LMG 24286T, W. beninensis 2L24P13T, W. fabalis M75T and W. fabaria 257T. The G+C content was 41.1 mol% on the basis of the whole-genome sequence. Polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, two unidentified aminophospholipids, one unidentified phospholipid and four unidentified lipids. The major cellular fatty acids were C18 : 1 ω9c, C16 : 0, C14 : 0, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and summed feature 8 (C16 : 1 ω7c and/or C16 : 1 ω6c). The cell-wall peptidoglycan was of A4α type with the interpeptide bridge of Gly-d-Glu. Based on these results, strain 26KH-42T could be classified as a novel species of the genus Weissella, for which the name Weissellacryptocerci sp. nov. is proposed. The type strain is 26KH-42T (=KACC 18423T=NBRC 113066T).

Novel lactic acid bacteria isolated from the bumble bee gut: Convivina intestini gen. nov., sp. nov., Lactobacillus bombicola sp. nov., and Weissella bombi sp. nov.

Twelve isolates of lactic acid bacteria (LAB) were obtained in the course of a bumble bee gut microbiota study and grouped into four matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry clusters. Comparative 16S rRNA gene sequence analysis revealed that cluster 1 isolates, represented by strain LMG 28288(T), are most closely related to Lactobacillus apis (97.0% sequence similarity to that of L. apis LMG 26964(T)). Cluster 2 isolates represented by strain LMG 28290(T) are most closely related to Weissella hellenica (99.6% sequence similarity to that of W. hellenica LMG 15125(T)). The single cluster 3 and 4 isolates had identical 16S rRNA gene sequences which were 94.8% similar to that of Leuconostoc mesenteroides subsp. mesenteroides LMG 6893(T), their nearest phylogenetic neighbour. A polyphasic taxonomic study additionally including comparative pheS sequence analysis, DNA-DNA hybridization experiments, DNA G+C content analysis, (GTG)5-PCR fingerprinting and a biochemical characterization, demonstrated that cluster 1 isolates represent a novel Lactobacillus species for which we propose the name Lactobacillus bombicola sp. nov. with LMG 28288(T) (= DSM 28793(T)) as the type strain; and that cluster 2 isolates represent a novel Weissella species for which we propose the name Weissella bombi sp. nov. with LMG 28290(T) (= DSM 28794(T)) as the type strain. Cluster 3 and 4 isolates, in contrast, represented a very distinct, novel taxon that could be distinguished from members of the genera Leuconostoc and Fructobacillus, its nearest phylogenetic neighbours, by its cellular morphology, non-fructophilic metabolism and DNA G+C content. We therefore classify both isolates into a novel species representing a novel LAB genus for which the name Convivina intestini gen. nov., sp. nov. is proposed with LMG 28291(T) (= DSM 28795(T)) as the type strain.

Weissella confusa bacteremia in a liver transplant patient with hepatic artery thrombosis.

A 54-year-old man with a history of nonalcoholic steatohepatitis and hepatocellular carcinoma presented 2 months after an orthotopic liver transplant with fever and abdominal pain. Two weeks earlier, he had an hepatic artery thrombosis and a biliary stricture, for which an hepatic artery stent and a biliary stent were placed. Laboratory workup was significant for leukocyte count of 7800/mcL with 92% segmented neutrophils, hemoglobin 9.4 g/dL, alanine aminotransferase 98 U/L, aspartate aminotransferase 72 U/L, alkaline phosphatase 358 U/L, albumin 2.8 mg/dL, and total bilirubin 1.6 mg/dL. A computed tomography scan of the abdomen and pelvis revealed multiple small fluid collections in the liver consistent with bilomas, and an hepatic angiogram showed complete occlusion of the common hepatic artery. Two sets of blood cultures were positive for an organism initially identified by MicroScan(®) analysis as an α-hemolytic Streptococcus species that was resistant to vancomycin. Further testing confirmed the organism as Weissella confusa 2 days later. W. confusa is a gram-positive coccobacillus that may be misidentified as a Lactobacillus when cultured. It is commonly found in sewage, carrots, sugar cane, fermented foods, and intestinal flora. Although only 4 cases of clinical infection with W. confusa have been described previously, W. confusa has been isolated from the stool of liver transplant patients, and may be an underreported cause of infection owing to improper identification. As it can cause clinical infection in these immunosuppressed hosts, identification of this organism is paramount because it is vancomycin resistant, and incorrect identification could lead to improper antimicrobial selection and ultimately worsened patient morbidity or mortality.

Weissella confusa: an unexpected cause of vancomycin-resistant gram-positive bacteremia in immunocompromised hosts.

We report the first case of Weissella confusa bacteremia in an allogeneic hematopoietic stem cell transplant patient. After engraftment and discharge, the patient returned with fever and graft failure and was started on an empiric regimen of aztreonam and vancomycin. A blood culture grew an alpha-hemolytic, gram-positive coccus forming pairs and chains, originally thought to be a viridans Streptococcus and a skin contaminant. The isolation of the organism from multiple blood cultures, and the presence of vancomycin resistance prompted identification and additional susceptibility testing. The RapID(™) Str panel, which has W. confusa in its database, provided multiple incorrect identifications. The MicroScan WalkAway 96 SI, using PC-20 or -29 panels, also did not identify this bacterium, because it is not in their database. The organism was identified as W. confusa by 16S rDNA sequencing. Antibiotic susceptibility determination by Etest revealed vancomycin resistance and daptomycin susceptibility. Therapy was changed to daptomycin, and the infection resolved. Additionally, W. confusa sepsis, with multiple positive blood cultures, developed in a patient in the burn unit at our medical center. The patient's blood cultures remained positive until vancomycin was discontinued and daptomycin therapy initiated. Infections with vancomycin-resistant, gram-positive cocci are emerging among immuno compromised hosts. Under appropriate circumstances, clinicians need to request that the laboratory perform susceptibility testing and accurate identification, by nucleic acid sequencing if necessary. Sequencing of 16S rDNA is an important tool in the accurate identification of unusual pathogens.

Bacteraemia caused by Weissella confusa at a university hospital in Taiwan, 1997-2007.

Human infections caused by Weissella confusa are rarely reported. Ten patients with bacteraemia caused by W. confusa who were treated at a tertiary-care hospital in Taiwan during 1997-2007 were studied. All isolates were initially misidentified as various Lactobacillus and Leuconostoc species by two commercial automated identification methods, and were confirmed to be W. confusa by 16S rRNA sequencing analysis. MICs of these isolates for ten antimicrobial agents were determined by the agar dilution method. The characteristics of these patients included underlying malignancy (n = 4), presence of a central catheter (n = 6), surgery within the previous 3 months (n = 4) and concomitant polymicrobial bacteraemia (n = 5, 50%). Mortality was directly attributed to bacteraemia in two patients. All isolates exhibited high trimethoprim-sulphamethoxazole and ceftazidime MICs (≥ 128 mg/L) and were inhibited by linezolid, daptomycin, ceftobiprole and tigecycline at 4, 0.12, 2 and 0.12 mg/L, respectively. In conclusion, W. confusa should be included in the list of organisms causing bacteraemia in immunocompromised hosts. Novel antibiotics, including daptomycin, moxifloxacin, doripenem and tigecycline, exert good activity against W. confusa.