2012;287:1561021. Statistical analyses were performed using the log-rank test and the resulting P-values were provided. Six, five, three, and one patient had liver dysfunction or liver cirrhosis, diabetes, cancer, and poor nutritional status due to mental disease, respectively. Considering that the patients would call an ambulance on appearance of severe symptoms, which also indicates increased bacilli in the blood, the time of calling for an ambulance was considered as disease onset. At the designated incubation time for each experiment, the mixtures were serially diluted with M9-salt minimal medium and the resultant aliquots were spread on the RCM agar plates. Nat Chem Biol. The overall mortality rate for wound infection approximates 25% but rises to 54% in patients with underlying hepatic disease. Solutions of organic acids (pyruvate, formate, lactate, and acetate [Sigma Aldrich]), cyclic dipeptides (cFP, cPV, and cPT [Bachem Inc.]), and a dipeptide (FP [Sigma Aldrich]) were prepared in the deionized water or DMSO. Debridement at an early stage and, if medical treatment is resisted, enforcing amputation of the lower extremities should be performed immediately to increase chances of patient survival. This suggested that the inhibition of B. vulgatus growth may have been mediated by a direct interaction with V. vulnificus cells and/or an indirect interaction via secreted compounds from V. vulnificus. When the general state is not stabilized by debridement and amputation, it is likely that multiplication of the bacillus has not ceased. This bacterial cell-to-cell interaction was specifically mediated by cFP and ObgE and caused B. vulgatus cells to be highly sensitive to the membrane-disrupting stress(es). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Funding: This work was supported by grants from the Chang Gung Memorial Hospital, Taiwan (Grant Nos. (Fig.9A).9A). The differences in the distribution of LC causes may contribute to the discrepancy in geographical mortality due to VNSSTIs. Analyses of microbiota in the fecal samples of mice that died due to V. vulnificus infection revealed the decreased abundance of bacteria belonged to Bacteroidetes, notably, the species Bacteroides vulgatus. cFP-mediated death of B. vulgatus was accompanied with alterations in membrane integrity, which was discernible in the images taken under the electron microscopes (Fig. Studies evaluating prognostic factors and treatment outcomes of Vibrio vulnificus infections were included. E Effects of other cyclic dipeptides on the survival of B. vulgatus. Yes Po-Yao Chuang and Tien-Yu Yang are co-first authors on this work. Furthermore, depending on intraprocedural observations, amputation of the lower extremities should be performed without delay. The authors declare no conflict of interests. Mortality rates exceeding 40 percent have been reported in case series, . Cellular contents of ObgE were estimated by extrapolating the densitometric readings of the ObgE bands derived from cell lysates to the regression line derived from those of the known concentrations of rObgE (from 0.2 to 10 ng). Potential chimera sequences, which were detected by the bellerophone method [56] were also removed. Department of Orthopaedic Surgery, Chiayi Chang Gung Memorial Hospital, Chiayi, Taiwan, Roles Vibrio Species Causing Vibriosis | Vibrio Illness (Vibriosis) | CDC The viability of all B. vulgatus strains tested in this study decreased in the presence of V. vulnificus, of which reduction appeared to be dependent on the abundance of V. vulnificus in cocultures. Polymerized samples were sectioned (200250 nm) using ultramicrotome, then further thin-sectioned (70 nm) using Leica EM UC-7 (Leica Microsystems) and a diamond knife (Diatome). In the present study, the effect of V. vulnificus on the community structures of gut microbiota in mice was examined. Patients with LC had an almost two fold higher risk (n = 82/132) of mortality compared to those without LC (n = 51/149) and HD (n = 27/101) with RRs of 1.84 (95% CI 1.212.79) and 2.00 (95% CI 1.412.85), respectively (Fig 3A and 3B). Is It Safe To Eat Raw Oysters? What To Know About Vibrio Rise A comparison of thiosulphate-citrate-bile salts-sucrose (TCBS) agar and thiosulphate-chloride-iodide (TCI) agar for the isolation of Vibrio species from estuarine environments. Conceptualization, It is speculated that the ObgE of B. vulgatus may have activity similar to the ObgE*. The effect of cFP on bacterial membrane permeability was determined using a membrane potential-sensitive fluorescent dye, DiSC3(5) (Sigma). Events refers to the number of patients who died. Bethesda, MD 20894, Web Policies Two oligonucleotides, Bv_obgE-over F and Bv_obgE-over R (Additional file: Table S3), were used to amplify a 1185 bp DNA fragment containing obgE ORF from the genomic DNA of B. vulgatus MGM001. The number of dead mice was counted for 24 h. Results were expressed as the mean standard deviation (SD) or median SD in the three independent experiments. official website and that any information you provide is encrypted 2016;371:20150504. Three strains of B. vulgatus were anaerobically incubated in the M9-minimal medium with V. vulnificus. Although there is a wide range of HD types, the stages and damage to the liver are usually consistent. These results suggested that the death of B. vulgatus might be mediated by cFP secreted to SM. Bacterial colonies were processed to amplify their 16S rDNA fragments using the 27F and 1492R primers and the purified PCR products were subjected to DNA sequencing analysis using the 785F and 907R primers (Macrogen, Korea). As a library, NLM provides access to scientific literature. Vibrio vulnificus can cause a vast spectrum of diseases, such as gastroenteritis, cellulitis, necrotizing fasciitis, and sepsis. Mice were orally injected with cFP and they were infected at 12 h post-injection of cFP with various doses of V. vulnificus, ranging from 104 to 108 cells (n = 7 mice per each treatment) (open symbol). Vibrio vulnificus induces the death of a major bacterial species in the Although patients in the death group were stable at the first medical examination, treatment was delayed because their family doctors initially diagnosed them with cellulitis/phlegmon and designed a corresponding treatment plan. Yes Most infections occur from May through October when water temperatures are warmer. Inclusion in an NLM database does not imply endorsement of, or agreement with, History of raw seafood consumption, which was reported by the family, was ambiguous, and the time from consuming uncooked seafood to the development of symptoms differed among patients; some patients may have had an incubation period of several days. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. From an effort to obtain pure cultures of microorganisms from mouse guts, another bacterial species, P. goldsteinii belonging to Bacteroidetes and several Lactobacillus species belonging to Firmicutes have been isolated (mouse gut microorganism (MGM) isolates; Additional file: Table S2). The presence of underlying diseases such as liver dysfunction and diabetes and the presence of symptoms such as warmth, sharp limb pain, and skin abnormalities are characteristic features and aid in diagnosis. At 6 h of incubation, CFUs of the MGM strains were enumerated and presented with the percentage of each control (the same cultures but without treatment of V. vulnificus (A) or cFP (B)). (Fig.8C).8C). A single-amino-acid substitution in Obg activates a new programmed cell death pathway in. Vibrio vulnificus causes both foodborne and wound infections throughout the world and in the United States it carries the highest death rate of any foodborne disease agent .According to estimates from both the U.S. Centers for Disease Control and the Food and Drug Administration, there are 50 foodborne cases per year in the U.S. serious enough . This report clarifies the diagnostic criteria for V. vulnificus infection, thereby aiding in early diagnosis worldwide. The seawater temperature along the coast of the Kyushu district is approximately 10C from January to March, rising gradually by 10C in April and May to exceed 20C, where it remains until September before declining gradually. After 24 h of anaerobic incubation at 37 C, the growth pattern of each colony was observed. Alternatively, there are marked differences in the antibiotic resistance profile of V. vulnificus worldwide [42]. The arrow indicates the effective concentration (EC50) of Triton X-100 to release 50% of 0.1%. Therefore, the levels of B. vulgatus decreased approximately 2.2-fold in mice injected with cFP (P = 0.006, two-sided Students t-test). Four patients died without surgery because of unstable general health. analyzed the data; J.-A.K. At 0.5 h in a fluorometer, Triton X-100 (0.002% or 0.1%) was added to each well and the RFUs were continuously measured for another 0.5 h. The released fluorescence from the Triton X-100-treated B. vulgatus cells was obtained by subtracting the values of basal RFU (averaged RFUs for 10 min before treatment of Triton X-100) from the maximal RFU (averaged RFUs for 10 min after treatment of Triton X-100). Doctors may need to amputate a patient's legs or arms to remove dead or infected tissue. Dewachter L, Deckers B, Martin E, et al. Department of Diagnostic Radiology, Chiayi Chang Gung Memorial Hospital, Chiayi, Taiwan. Bacterial cells were fixed with 2% glutaraldehyde-paraformaldehyde in 0.1 M phosphate buffer (PB, pH 7.4) for 24 h and then with 1% OsO4 dissolved in 0.1 M PB for 1.5 h. Fixed cells were dehydrated through an eight-stepped ascending gradual series of ethanol (from 50% to 100%), infiltrated with isoamyl acetate, and placed in the Critical Point Dryer (Leica EM CPD300). Recognition of the characteristic epidemiology and clinical features of this disease is important, and positive debridement should be performed on suspicion. However, infected patients have typical characteristics. Limb abnormalities were observed on visual examination. FOIA Membranes were blocked with 5% skim milk in TBST for 30 min and incubated overnight at 4C with each polyclonal antibody (1:5000, [vol/vol]), followed by alkaline phosphatase-conjugated rabbit anti-mouse IgG (1:5000, [vol/vol]). The mortality rate in this study was 58.3% (7/12), in line with previous findings.10 Although the system of emergency centers and intensive care units in our country has substantially improved of late, this infection remains troubling because patients can die within two days, as in our study. A decrease in B. vulgatus CFUs in incubations with cFP was evidenced to be due to the actual death of bacterial cells, upon fluorescence microscopy observation of the B. vulgatus cells stained with the live/dead-indicating dyes (Fig. Materials and Methods: In accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic search of PubMed, Embase, and Cochrane Library databases by two independent reviewers was reported. Further studies to understand and clarify the risk factors or mechanisms of disease, controlling and/or reversing them, and finding a clinical pathway that can lower the mortality are crucial. This study was approved by the ethics committee (Institutional Review Board) of the Chang Gung Memorial Hospital in Taiwan (reference number: 103-7038C). For quantitative analysis, each cell lysate was subjected to western blotting along with the known concentrations of rObgE (Fig. The patient's general condition should be stabilized after treatment. These results lead us to speculate the possible involvement of ObgE in the observed membrane disruption in B. vulgatus exposed to cFP. Chang Gung University College of Medicine, Taoyuan, Taiwan, Yes Vibrio vulnificus infections in high-risk individuals have a 50 percent fatality rate. ObgE is a conserved GTPase among a wide-range of bacterial species, and it is reported to be essential for their viability by mediating the stringent response, ribosomal assembly, DNA replication and morphological differentiations including a sporulation process in spore-forming bacteria [31, 44, 45]. Sassone-Corsi M, Raffatellu M. No vacancy: how beneficial microbes cooperate with immunity to provide colonization resistance to pathogens. Transmission through open wounds or consumption of contaminated seafood can lead to wound infections, sepsis, and potentially death. Validation, As described in Fig. No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, Corrections, Expressions of Concern, and Retractions, https://doi.org/10.1371/journal.pone.0223513. Department of Orthopaedic Surgery, Chiayi Chang Gung Memorial Hospital, Chiayi, Taiwan, To determine the effective concentration of Triton X-100 causing the lysis of 50% B. vulgatus cells (EC50), various concentrations of Triton X-100 were exposed to DiSC3(5)-associated B. vulgatus. Since the B. vulgatus ObgE specifically interacted with cFP and its cellular contents increased in cFP-treated cells, the role of ObgE in membrane disruption was examined. PMCID: PMC8901877 DOI: 10.3346/jkms.2022.37.e79 Abstract Background: Vibrio vulnificus infection is a highly fatal disease resulting from the consumption of raw or undercooked seafood and exposure to seawater containing the organism. The taxonomic classification of each read derived from the experiments 1 and 2 was assigned against the EzTaxon-e database deposited in EzBioCloud [57], which contains 16S rRNA gene sequence of type strains that have valid published names and representative species-level phylotypes of either cultured or uncultured entries in the GenBank database with complete hierarchical taxonomic classification from the phylum to the species. A foodborne pathogen, Vibrio vulnificus, encounters normal microflora inhabiting the gut environments prior to causing fatal septicemia or gastroenteritis and should overcome the barriers derived from the gut commensals for successful infection. Vibrio infection; Vibrio necrotizing; Vibrio sepsis; Vibrio septicemia; meta-analysis; waterborne pathogens. Freshly grown cells of B. vulgatus were washed twice and resuspended in the M9-salt minimal medium [59] to contain cells at an OD595 nm of 1.0.
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