vibrio vulnificus mortality rate

An official website of the United States government. The authors have no conflict of interest to declare. Its frequency and severity increase as hepatic functional reserve (HFR) worsens, and common precipitants include bacterial and viral infections, alcoholism, and surgery [30,31]. Pathogenesis of Vibrio vulnificus | FEMS Microbiology Letters | Oxford Improved Method for Transformation of Vibrio vulnificus by No, Is the Subject Area "Skin tissue" applicable to this article? 10.1074/jbc.M111.301630. ; J.-A.K., B.-R.J., Y.-C.J., K.-S.K., and K.-H.L. Yes Szke I, Pascu C, Nagy E, Ljung A, Wadstrm T. Binding of extracellular matrix proteins to the surface of, Maerz JK, Steimle A, Lange A, Bender A, Fehrenbacher B, Frick JS. Four-week-old female mice were orally injected with cFP as described above, then infected with various doses of V. vulnificus, ranging from 104 to 108 cells (n = 7 mice per each treatment) (open symbol). 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. It suggests that cFP treatment made B. vulgatus membranes more susceptible to membrane permeabilizing stress. (Fig.9A9A and B). A mild case of vibriosis typically includes chills, fever, diarrhea, stomach pain and possibly vomiting. (Fig.1D).1D). Eight patients had disseminated intravascular coagulation (DIC). This should garner the significant attention as premature death costs US$ 238 million, which accounts for 99% of the total V. vulnificus health costs and 75% of the total cost of . Then, we measured the membrane integrity in B. vulgatus cells having different levels of ObgE. The risk ratios estimating the risk of mortality of VNSSTIs in patients with HD compared to those without it of the individual studies were combined using the Mantel-Haenszel method. You JS, Yong JH, Kim GH, et al. Accessibility Multiple intraintestinal signals coordinate the regulation of. Epub 2010 Jan 28. Bethesda, MD 20894, Web Policies Funding acquisition, Risk of mortality of Vibrio vulnificus necrotizing skin and soft tissue infections in patients with hepatic disease compared to those without it: (A) the worlds coastal areas; (B) the western Atlantic coastal areas; and (C) the western Pacific coastal areas. Events refers to the number of patients who died. To obtain pure cultures of bacteria in the mouse guts, the fecal samples collected from seven-week-old female ICR mice were resuspended in PBS. Regarding treatment options, there is no definitive cure, although there is no objection to early intervention with antibiotics and surgery.11, 12 No significant differences in the type of antibiotic, administered dose, or the time of first administration were observed in our study groups. When pathogenic bacteria meet the intestinal microbiota. No language restrictions were placed on the searches or search results. E. coli used for plasmid DNA preparation and conjugal transfer were grown at 37 C in LB medium supplemented with appropriate antibiotics. Densitometric readings of the bands corresponding to each protein in the control and cFP-treated cells were plotted. To examine whether the decrease in B. vulgatus levels in the V. vulnificus-fed mice was caused by an interaction between these two species of bacteria, a strain of B. vulgatus (MGM001; Additional file: Table S2) was isolated as described in Methods. In addition to MGM001, two strains of B. vulgatus were purchased from the American Type Culture Collection (ATCC) and Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ). Peterson KM, Gellings PS. Five patients exhibited stomachache, nausea, and vomiting. After 48 h of incubation in an anaerobic chamber, the viable B. vulgatus cells were counted. From the pooled dataset, nearly half (46.8%) of the patients with VNSSTIs had HD. Antibiotic use for Vibrio infections: important insights from 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. here. Analysis of gastric body microbiota by pyrosequencing: possible role of bacteria other than. A spike last year, which amounted to 74 cases and 17 deaths, was due to the . Rajkowski K.T., Rice E.W. 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). The same cells prepared for the experiments shown in Fig. Medical treatment is initiated according to the following flow chart. Wang RF, Cao WW, Cerniglia CE. 1). The purified recombinant proteins were used to generate polyclonal antibodies by the three immunizations of ICR mice (CrljOri:CD1[ICR]) (100 g of recombinant proteins per immunization) at 3-week intervals. Then, the cell suspensions were added by various bacterial cells, their spent media (SM), or chemical compounds, and were incubated at 37 C. Primary wound presentation is seen in about 25% of all V. vulnificus cases. Study or subgroup on the Y-axis refers to first author and publication year. The percentages of 16S rDNA copies belonged to the phylum Bacteroidetes (C) and the species of B. vulgatus (D) in the fecal samples of V. vulnificus-infected dead mice were compared with those of PBS-treated control mice. Multi-factor regulation of the master modulator LeuO for the cyclic-(Phe-Pro) signaling pathway in, Park NY, Lee KW, Kim KS. Maintenance of normal microflora in GI tracts is important in the host response to pathogens [48]. Of these, we excluded 179 articles that were not clinical studies and 46 that were duplicates or used overlapping datasets. For example, doxycycline has shown intermediate resistant profile in Italy [43] while ceftazidime in the U.S. [44] and ceftriaxone in India [45]. In vivo effects of cFP in mice. RFUs were measured in a fluorometer (EnSpire Multimode Plate Reader, PerkinElmer), for which the temperature was set at 30 C and wavelengths of 622 and 670 nm, for excitation and emission, respectively. Various combinatory ratios of B. vulgatus to V. vulnificus, ranging from 1:1 to 1:10, were prepared in the M9-minimal medium and incubated in a 37 C anaerobic chamber. (Fig.2C).2C). D, E, and F TEM of B. vulgatus cells. Int J Infect Dis. Disruptive changes in their membranes were determined using a fluorescent dye DiSC3(5), which was incorporated into the bacterial membranes. In addition, the incubation of B. vulgatus with a mutant V. vulnificus (llcA), of which production of cFP was estimated to be less than ~ 30% of the wild type [24], resulted in lowered effectiveness in decreasing B. vulgatus survival, compared to that with the same amount of wild-type V. vulnificus (Fig. It is very important for a clinician to recognize the characteristic epidemiology and clinical picture identified in this research and to perform quick diagnosis and treatment according to the flow chart presented in this paper. When a patient in his 60s presents with sharp pain in his/her leg, the following clinical history should be examined: Does he/she have hepatic dysfunction and diabetes as underlying conditions? What does a skin infection look like? Interestingly, a substitution of an amino acid residue in the G domain of E. colis ObgE (ObgE*) resulted in the induction of programmed cell death via an unknown mechanism [32, 46]. Verstraeten N, Knapen WJ, Kint CI, et al. The medians of estimated values of 2[CT(B.vulgatus) CT(Total Bacteria)] were 0.1369 ( 0.0748) and 0.0619 ( 0.0766) in the PBS-treated control (n = 20 mice) and cFP-treated mice (n = 20 mice), respectively (Fig. Fig.7,7, the values of released DiSC3(5) from 0.002% (0.002%) and 0.1% Triton X-100-treated cells (0.1%) were estimated. 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. No, Is the Subject Area "Medical risk factors" applicable to this article? Community structures of microbiome derived from sequencing analysis of 16S rDNAs in the fecal samples of V. vulnificus-infected dead mice and PBS-treated control mice were subjected to PCoA based on Jensen-Shannon distance method. People with vibriosis become infected by consuming raw or undercooked seafood or exposing a wound to seawater. 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. Inclusion in an NLM database does not imply endorsement of, or agreement with, Vibrio vulnificus Infection - StatPearls - NCBI Bookshelf Bumler AJ, Sperandio V. Interactions between the microbiota and pathogenic bacteria in the gut. Yoon SH, Ha SM, Kwon S, et al. 2019;10:1484. Necrotizing soft-tissue infections and sepsis caused by Vibrio vulnificus compared with those caused by Aeromonas species. The effectiveness of debridement and lower extremity amputation decreases over time. The sequences corresponding to the barcode, linker, and primers were removed from the original sequencing reads. FOIA All the Vibrio species tested in this study showed antagonistic effects on B. vulgatus. Vibrio vulnificus infection is a highly fatal disease resulting from the consumption of raw or undercooked seafood and exposure to seawater containing the organism. Unauthorized use of these marks is strictly prohibited. Experiments were performed using 4- or 7-week-old female ICR mice (CrljOri:CD1[ICR], OrientBio, Korea). Are certain abnormalities observed on his/her skin? Supervision, Combining clinical history with a medical examination is helpful in predicting V. vulnificus infection. After unincorporated dyes were discarded, the cells were resuspended in the same volume of 5 mM HEPES. No, Is the Subject Area "Soft tissue infections" applicable to this article? The site is secure. Patients with HD contracted VNSSTIs were found to be two or more times (RR = 2.61, 95% CI = 2.143.19) as likely to die compared with those without HD. Vibrio vulnificus (V. vulnificus), a naturally occurring Gram-negative bacterium found in estuarine and marine environments throughout the world, is extremely virulent and can cause three types of infections: (1) acute gastroenteritis, (2) primary septicemia, and (3) necrotizing wound infections [13]. Yes Hepatic disease and the risk of mortality of Vibrio vulnificus necrotizing skin and soft tissue . The intestinal microbiota: antibiotics, colonization resistance, and enteric pathogens. 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. The Brazilian Journal of Infectious Diseases. The resuspension was briefly centrifuged to eliminate fecal debris and the supernatants were retrieved. Clipboard, Search History, and several other advanced features are temporarily unavailable. cFP was able to induce the cell death of B. vulgatus, a major constituent of the gut microbiota (Figs. The purpose of this prospective study was to clarify the clinical characteristics between death and survival NF . Vibrio vulnificus infection: a persistent threat to public health Ma LS, Narberhaus F, Lai EM. Statistical analysis was performed using the Students t-test and the resulting P-value was provided. In this study, we sought to define the characteristic epidemiology of V. vulnificus infection and clarify its diagnosis at the global level. Therefore, this pathogen should interact with them to overcome the potential defense mechanisms directly or indirectly derived from the gut microbiota, while interacting with the hosts via timely produced virulence factors. Before [11] performed an analysis on another series of 269 cases and found that more than 50% of these patients had HD. If this does not happen, further amputation of the lower extremity (reamputation) should proceed promptly within 8h. If detection and treatment are not performed early in the course of infection, the mortality rate is very high. Reduced levels of B. vulgatus were observed in all of the V. vulnificus-fed mice in each set of experiments (Mann-Whitney U test, P = 0.050): 52.5% 9.5% in the control vs. 33.2% 6.5% in the infected mice in experiment 1; and 36.4% 1.5% in the control vs. 27.8% 1.8% in the infected mice in experiment 2 (Additional file: Table S1). Vibrio vulnificus DNA Load and Mortality - PMC - National Center for 9). B Release of DiSC3(5) from cFP-treated B. vulgatus. (Fig.8A).8A). Hepatic disease and the risk of mortality of Vibrio vulnificus - PLOS It does not alter the appearance, taste, or odor of oysters. Flow chart from diagnosis to treatment of Vibrio vulnificus. Studies evaluating prognostic factors and treatment outcomes of Vibrio vulnificus infections were included. Ultimately, we identified 12 and 6 studies that fulfilled the eligibility criteria for the systematic review for questions 1 and 2, respectively. National Library of Medicine Dewachter L, Verstraeten N, Fauvart M, Michiels J. 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). 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. Next, a medical examination should be performed. It has been previously reported that a trial of obgE gene deletion was not successful in many bacterial species [33]. Although abnormalities such as low blood pressure and bleeding tendencies indicate a poor prognosis, debridement should be performed within 4h to maximize patient survival. The National Shellfish Sanitation Program, recognized by the Food and Drug Administration, ensures that shellfish-producing states are following safety guidelines, including certification . Formal analysis, Background: Vibrio vulnificus is a rare but life-threatening infection that effects the population near warm coastal areas. Investigation, 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]). Conceptualization, The total RNAs were extracted from B. vulgatus cells treated with 4 mM cFP for 0, 2, and 4 h. Then, cDNAs were produced through the reverse transcriptase reaction and subjected to quantitative PCR using primer sets specific to obgE, recA, or cidA (Additional file: Table S3). Unable to load your collection due to an error, Unable to load your delegates due to an error. In addition, the mortality rate in HD patients with VNSSTIs was 53.9%, which was considerably higher than 16.1% of non-HD patients. Early diagnosis and treatment are critical to saving patients' lives. The surviving mice were enumerated for 24 h, and their numbers were plotted against incubation time. https://doi.org/10.1371/journal.pone.0223513.g003. Raw oysters can be a risk for infections. These results evidencing the increased susceptibility of cFP-treated cells to Triton X-100 suggested that cFP caused the membranes of B. vulgatus to be permeable, thus easily disruptive. Statistical analysis of the taxonomically classified sequencing data was processed using the EzBioClaud. Chen C, Yang X, Shen X. Does he/she have stomachache, nausea, or vomiting? The mortality rate in HD patients with VNSSTIs was 53.9% (n = 292/542), which was considerably higher than the mortality rate of 16.1% (n = 99/615) in non-HD patients. If V. vulnificus is detected in a TCBS agar medium culture, diagnosis is quick and accurate.16 If a Gram-negative banana-shaped organism is detected by Gram staining, the possibility of V. vulnificus infection is very high. Fifteen micrograms of protein lysates of B. vulgatus, which had been exposed to 0 or 4 mM cFP for 4 h, were fractionated in SDS-PAGE and subjected to western blotting analysis using anti-ObgE and anti-RecA polyclonal antibodies. Four patients died without surgery because of unstable general health. In fact, no patient was treated only with antibiotics in our study. The estimated cellular contents of ObgE were increased from 1.3 to 2.5 ng ObgE per g of cell lysate by carrying pBAD-obgEBv, and those of B. vulgatus cells harboring pBAD-obgEBv were further increased to 3.1, 4.9, and 9.3 ng ObgE per g of cell lysate if cells were incubated with 0.01%, 0.1%, and 1.0% arabinose, respectively. Lee K, Jeong JE, Kim IH, Kim KS, Ju BG. A principal coordinate analysis (PCoA) showed that the microbial compositions in most fecal samples of the V. vulnificus-fed mice were distinct from those of the control mice (permutational multivariate analysis of variance [PERMANOVA], P = 0.024) (Fig. Lee KJ, Jeong CS, An YJ, et al. In contrast, the titration of rObgE with 400 M of another cyclic dipeptide, cPT did not show any signature for the inter-molecular interaction. Cyclo(phenylalanine-proline) induces DNA damage in mammalian cells via reactive oxygen species. PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples. https://doi.org/10.1371/journal.pone.0223513.s003. When infected with the highest dose of V. vulnificus (i.e., 108 cells), mice under both conditions died equally and quickly within 24 h. Similarly, the mice infected with the lowest dose of V. vulnificus (i.e., 104 cells) showed similar patterns of survival, regardless of the exogenous addition of cFP. The patient's general condition usually stabilizes after removal of the affected tissue. Although fever, sharp limb pain, and limb abnormalities were observed in all patients, necrosis of the extremities, a defining characteristic of V. vulnificus infection, was identified only in one patient. Compared to the control, 0% of SM (the fresh LBS medium only), the addition of SM resulted in a decrease in B. vulgatus viability in a dose-dependent manner. A mutant (icmF) deficient in one of the T6SS components of V. vulnificus, i.e., IcmF (the intracellular multiplication factor F [25];), was constructed and mixed with B. vulgatus cells. This finding is very interesting since a signal used by a pathogen to regulate its virulence factors is also utilized to change the gut environments to favor its pathogenicity by specifically transducing the signal to a group of gut commensals and removing these bacteria from the guts. Differences were considered significant if P-values were < 0.01. 2021 Feb;107(2):E12-E17. Most studies of V. vulnificus pathogenicity have focused on the bacterial effects on model animals at the levels of cells, tissues, or organs. As a negative control, fecal samples from healthy mice treated with V. vulnificus-free PBS were also collected at each experimental set (two animals in each set). Department of Life Science, Sogang University, 35 Baekbeom-Ro, Mapo-Gu, Seoul, South Korea. The overall pooled proportion of individuals who died in the HD group was 53.9% (n = 292/542) while in the non-HD group 16.1% (n = 99/615) of the individuals died after contracting VNSSTIs. C and D Comparison of relative abundance of Bacteroidetes and Bacteroides vulgatus in the mouse fecal samples. Two patients converted to amputation during debridement itself. Upon the addition of 0.002% Triton X-100, B. vulgatus cells treated with these dipeptides produced 4mM dipeptide values (562, 572, and 561 RFUs in treatment with FP, cPT, and cPV, respectively) with similar ranges to that of the control cells (608 RFUs). rObgE in the eluted fractions (1 ml) was concentrated using Amicon Ultra-15 10K (Millipore) and its buffer was exchanged with that used for an ITC assay. Various concentrations of freshly grown V. vulnificus were resuspended in PBS and injected intragastrically into seven mice per set. The values of 0 mM cFP and 4 mM cFP were estimated at 605 ( 64) and 727 ( 80) RFUs, which was equivalent to 29% ( 2.3%) and 43% ( 3.7%) of the total fluorescence initially incorporated into the cells, respectively. How you get necrotizing fasciitis The bacteria most commonly enter the body through a break in the skin. Any read containing multiple ambiguous nucleotides, low-quality score (average score < 25), or shorter than 300 bp were discarded. Ferreira RB, Gill N, Willing BP, et al. To circumvent the difficulty in finding the active compound(s), B. vulgatus was challenged by various bacterial species and its survival was examined (Fig. B Effect of cFP on mouse lethality caused by V. vulnificus. Livak KJ, Schmittgen TD. Chassaing B, Koren O, Carvalho FA, Ley RE, Gewirtz AT. Quality control (QC) of the amplified DNA was assessed on a Bioanalyzer 2100 (Agilent) using a DNA 7500 Chip, and pyrosequencing was carried out with a GS Junior Sequencing system (Roche). Nine bacterial isolates showed more than 97% identity to the corresponding regions of 16S rDNAs of B. vulgatus, P. goldsteinii, L. johnsonii, L. reuteri, L. murinus, or L intestinalis, and were named by MGM001 to 009 (Additional file: Table S2). Fig.2C.2C. Furthermore, the regulatory role of cFP at another level of V. vulnificus-host interactions was identified in this study. An official website of the United States government. At 6 h post-exposure to cFP, B. vulgatus CFUs were enumerated as described in Fig. D Effects of Phe (F), Pro (P), and a dipeptide of Phe-Pro (F-P) on the survival of B. vulgatus. Nonetheless, LC with or without decompensation is still a critical stage associated with a variety of major complications [29]. These results suggested that the death of B. vulgatus might be mediated by cFP secreted to SM. B. vulgatus cells treated with FP, cPT, or cPV showed a slightly lowered basal RFU (averaged RFUs for 10 min before Triton X-100 treatment) compared to the control and much lower maximal RFU (averaged RFUs for 10 min after Triton X-100 treatment) than the cFP-treated cells. The numbers of viable and dead cells, which were shown as green and red cells, respectively, were counted and their average numbers per microscope field were obtained through at least eight observations. I'll be live on a #NTWC special today at 10am CDT discussing #hurricane safety and providing some survival tips. Clotting abnormalities were detected in all patients, and 70% exhibited the diagnostic criteria of DIC. The concept of LC has changed from being a static and irreversible entity to a dynamic disease with reversible stages [27,28]. To examine the mortality of the mice infected with V. vulnificus in the presence of cFP, four-week-old female ICR mice (CrljOri:CD1[ICR]), which have been starved for 12 h, were intragastrically treated with cFP (110 g cFP per gram of mouse) for 12 h prior to V. vulnificus infection. High salinity relay as a postharvest processing strategy to reduce, Matsumoto K., Ohshige K., Fujita N., et al. 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). Because V. vulnificus infection is very rare and follows an aggressive course, it is thought that death may occur in patients even before diagnosis. Subsequently, the time between disease onset and death was investigated. Vibrio vulnificus is a gram-negative bacterium that can cause serious wound infections, septicemia, and diarrhea. Accessibility The authors thank Prof. S-J Park and Dr. J Kim (Yonsei University College of Medicine) for SEM and TEM analysis. Any antibiotic that is effective against Gram-negative bacilli can be used. (Promega) and passing a gel filtration AKTA-FPLC system (Superdex 200, GE Healthcare) at a flow rate of 0.3 ml/min in a buffer containing 20 mM HEPES (pH 7.5), . This suggested the possible role of V. vulnificus-originating and V. vulnificus-secreted substance(s) in the death of B. vulgatus. Microbiome composition data derived from two sets of experiments with the mice infected with V. vulnificus (Additional file: Table S1) showed a decrease in the abundance of P. goldsteinii in the V. vulnificus-fed mice, compared to the control PBS-fed mice (Mann-Whitney U test, P = 0.050): from 1.81% ( 0.96%) to 0.67% ( 0.18%) in experiment 1; and 0.91% ( 0.41%) to 0.35% ( 0.003%) in experiment 2. No patient was treated only with antibiotics. Fig.6B)6B) to the resuspensions of the isolated gut commensals, only P. goldsteinii experienced decreased survival, while the survival of Lactobacillus species (i.e., L. johnsonii, L. reuteri, L. murinus, and L. intestinalis) was not decreased. Kim S, Covington A, Pamer EG. Analysis of 1000 type-strain genomes improves taxonomic classification of Bacteroidetes. The transcript contents of each gene were normalized with the gap transcript levels. Isothermal titration calorimetry (ITC) experiments were performed by titrating the recombinant ObgE (40 M) with 400 M cFP at 25C. e0223513. C TEM of P. goldsteinii cells. In summary, VNSSTIs is an important public health problem and is becoming more critical because of global warming [46]. Vibrio vulnificus & Wounds | Vibrio Illness (Vibriosis) | CDC Liver dysfunction seems to affect the susceptibility and prognosis of VNSSTIs in multiple ways. 2016;4. HHS Vulnerability Disclosure, Help Interactions between pathogens and gut commensals have been reported in diverse pathogens in model systems. Because V. vulnificus multiplies rapidly and increases to one million in 2h in a blood vessel, a patient's condition can deteriorate rapidly.7 Although skin necrosis accompanied by a blood blister is observed after several hours, numerous bacilli will be present in this blister and treatment will no longer be effective.8, 9. Two distinct clinical syndromes have been identified, depending on the portal of entry (wound infections and fatal septicemias). Three strains of B. vulgatus were anaerobically incubated in the M9-minimal medium with V. vulnificus. Relationship between mortality rate and time to debridement and lower extremity amputation. As a result, several regulations have been placed to reduce risk through sanitation. For comparisons of the viable cells of each species in the monocultures, these were also prepared to contain either B. vulgatus only (designated as 1:0) or V. vulnificus only (designated as 0:1, 0:2, and 0:5). Next, the cell-free supernatant derived from the LBS broth grown by V. vulnificus to the stationary phase (spent medium, SM) was prepared and various volumes of the filter-sterilized SM were added to B. vulgatus (Fig. Although V. vulnificus multiplies in approximately 10min at 20C, its multiplication is controlled below 15C.3 Moreover, it is believed that approximately one million bacilli are required to cause infection via oral ingestion.4 For V. vulnificus infection to materialize, uncooked and V. vulnificus bacilli that grew at a seawater temperature of at least 20C must have been consumed by the patients via raw fresh fish or shellfish that ingested this bacilli, which in turn condensed in the digestive organs of these fish. Interestingly, different antibiotic regimens did not have any impact on outcomes, however, delayed surgical intervention after the first 12 or 24 hours was associated with higher mortality rates (OR, 2.64; 95% CI, 1.39-5.0; p = 0.003 and OR, 2.99; 95% CI, 1.54-5.78; p = 0.001, respectively).

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