A multichromatic colorimetric detection method for Vibrio parahaemolyticus based on Fe3O4-Zn-Mn nanoenzyme and dual substrates.

IMPORTANCE: The Fe3O4-Zn-Mn nanomimetic enzyme demonstrates significant importance in dual-substrate colorimetric detection for V. parahaemolyticus, owing to its enhanced sensitivity, selectivity, and rapid detection capabilities. Additionally, it offers cost-effectiveness, portability, and the potential for multiplex detection. This innovative approach holds promise for improving the monitoring and control of V. parahaemolyticus infections, thereby contributing to advancements in public health and food safety.

Case Report: First case of HIV, hepatitis B, hepatitis C, and Vibrio vulnificus coinfection.

Our patient, a 48-year-old man from Guangdong's coastal region, worked selling and processing oysters and other seafood. He started experiencing swelling and pain in his left knee on October 4, 2022, and they got worse over time. The findings of mNGS test showed Vibrio vulnificus infection. The patient had AIDS, hepatitis A and hepatitis B concurrently. He was admitted to the hospital's intensive care unit (ICU) for treatment as his symptoms worsened. We refrained from performing an amputation because the family members expressed a desire to keep the limb. The limb was managed with regular dressing changes, thorough debridement, wound closure, ongoing VSD drainage, and local antibiotic irrigation. The patient's organ function eventually returned to normal, and the systemic infection got better. On November 1, the wound's new granulation tissue had grown well and had gradually crept to cover 80% of the wound. The tissue's blood flow had also improved, indicating a trend of growth and healing.

Enhancing Vibrio vulnificus infection diagnosis for negative culture patients with metagenomic next-generation sequencing.

Objective: To evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) in Vibrio vulnificus (V. vulnificus) infection. Methods: A retrospective analysis of patients with V. vulnificus infection at the Fifth Affiliated Hospital of Sun Yat-Sen University from January 1, 2020 to April 23, 2023 was conducted. 14 enrolled patients were diagnosed by culture or mNGS. The corresponding medical records were reviewed, and the clinical data analyzed included demographics, epidemiology laboratory findings, physical examination, symptoms at presentation, antibiotic and surgical treatment, and outcome. Results: In this study, 78.6% (11/14) patients had a history of marine trauma (including fish stab, shrimp stab, crab splints and fish hook wounds), 7.1% (1/14) had eaten seafood, and the remaining 14.3% (2/14) had no definite cause. Isolation of V. vulnificus from clinical samples including blood, tissue, fester and secreta. 9 cases were positive for culture, 5 cases were detected synchronously by mNGS and got positive for V. vulnificus. 85.7% (12/14) cases accepted surgical treatment, with 1 patient suffering finger amputated. 14 enrolled patients received appropriate antibiotic therapy, and all of them had recovered and discharged. 9 strains V. vulnificus isolated in this study were sensitive to most beta-lactam antibiotics, aminoglycosides, quinolones, etc. Conclusion: Vibrio vulnificus infection is a common water-exposed disease in Zhuhai, which requires identification of a number of pathogens. Of severe infections with unknown pathogen, mNGS can be used simultaneously, and the potential to detect multiple pathogens is of great help in guiding treatment.

Fulminant Overwhelming Necrotizing Vibrio vulnificus Sepsis Secondary to Oyster Consumption.

Vibrio vulnificus is an opportunistic gram-negative rod-shaped bacteria found in warm, low salinity waters. Transmission through open wounds or consumption of contaminated seafood can lead to wound infections, sepsis, and potentially death. A 44-year-old man with a history of poly-substance abuse, cirrhosis, and recent oyster consumption presented to the emergency department in June with acute onset bilateral leg pain associated with rash and fever. Within 6 hours of his arrival, the rash rapidly progressed to large bullae with extensive necrosis ascending to the level of the abdomen, and he developed septic shock. Despite prompt surgical intervention and appropriate antibiotic and resuscitative therapies, the patient had progressive multi-system organ failure and died 7 days after admission. Concurrent necrotizing fasciitis with sepsis secondary to V. vulnificus infection is rare and potentially fatal, as demonstrated in this case.

Vibrio vulnificus pneumonia with multiorgan failure: a case report and review of the literature.

BACKGROUND: Vibrio vulnificus is a gram-negative bacterium causing three clinical syndromes namely, gastrointestinal symptoms, skin sepsis and primary sepsis. Primary sepsis exhibits mortality rates exceeding 50%, particularly in the immunocompromised. Vibrio vulnificus is transmitted via consumption of contaminated seafood and contaminated seawater skin exposure. We describe a rare case of an immunocompetent male presenting with an atypical Vibrio vulnificus infection, culminating in severe pneumonia requiring intensive care. CASE PRESENTATION: A 46 year old Indian male dockyard worker, a non-smoker and teetotaler, of Indian origin presented to the emergency treatment unit of a tertiary care hospital in Sri Lanka, with fever, productive cough with yellow sputum, pleuritic chest pain and tachypnea for five days. He had no gastrointestinal or skin manifestations. His respiratory rate was 38 breaths/min, pulse rate was 120 bpm, blood pressure was 107/75 mmHg and pulse oximetry was 85% on air. Chest X-ray revealed consolidation of the left lung. Empiric intravenous Piperacillin-tazobactam and Clarithromycin were commenced after obtaining blood and sputum cultures. Over the next 24 h, his oxygen requirement rose and as he required vasopressor support, he was admitted to the intensive care unit. He was intubated and bronchoscopy was performed on day two, which demonstrated thick secretions from left upper bronchial segments. His antibiotics were changed to intravenous ceftriaxone and doxycycline following a positive blood culture report of Vibrio vulnificus. He was ventilated for ten days and his intensive care stay was complicated with a non-oliguric acute kidney injury, with serum creatinine rising up to 8.67 mg/dL (0.81-0.44 mg/dL). He developed mild thrombocytopenia with platelets dropping to 115 × 103 /uL (150-450 × 103/uL) which resolved spontaneously. Vasopressors were weaned off by day eight and the patient was extubated on day ten. He was discharged from intensive care on day twelve and made a full recovery. CONCLUSIONS: Pneumonia itself is an atypical manifestation of Vibrio vulnificus and furthermore, this patient was immunocompetent and did not exhibit the classical gastro-intestinal and skin manifestations. This case highlights the occurrence of atypical Vibrio sp. infections in patients with high exposure risks and the need for early supportive and appropriate antibiotic therapies.

On-site Gram staining that increases a post-test probability of an ominous infection: a case of necrotizing fasciitis caused by Vibrio vulnificus: a case report.

BACKGROUND: Gram staining is a classic but standard and essential procedure for the prompt selection of appropriate antibiotics in an emergency setting. Even in the era of sophisticated medicine with technically developed machinery, it is not uncommon that a classic procedure such as Gram staining is the most efficient for assisting physicians in making therapeutic decisions in a timely fashion. CASE PRESENTATION: A 65-year-old Asian man with alcoholic cirrhosis complicated by esophageal varices was brought to the emergency division of Saga Medical School Hospital in early August, complaining of severe pain, redness, swelling, and purpura of the lower extremities. On physical examination he appeared in a critically ill condition suggestive of deep-seated soft tissue infection, raising a pre-test probability of streptococci, staphylococci, Vibrio sp., or Aeromonas sp. as a causative pathogen. A characteristic of his residency in an estuarine area is that raw seafood ingestion, as documented in this patient prior to the current admission, predisposes those who have a chronic liver disease to a life-threatening Vibrio vulnificus infection. Given the pathognomonic clinical features suggestive of necrotizing fasciitis, our immediate attempt was to narrow down the differential list of candidate pathogens by obtaining clinical specimens for microbiological investigation, thus inquiring about the post-test probability of the causative pathogen. The Gram stain of the small amount of discharge from the test incision of the affected lesion detected Gram-negative rods morphologically compatible with V. vulnificus. After two sets of blood culture, intravenous meropenem and minocycline were immediately administered before the patient underwent emergency surgical debridement. The next day, both blood culture and wound culture retrieved Gram-negative rods, which were subsequently identified as V. vulnificus by mass spectrometry, matrix-assisted laser desorption/ionization. The antibiotics were switched to intravenous ceftriaxone and minocycline. CONCLUSION: The pre-test probability of V. vulnificus infection was further validated by on-site Gram staining in the emergency division. This case report highlights the significance of a classic procedure.

Rapid, specific and sensitive detection of Vibrio parahaemolyticus in seafood by accelerated strand exchange amplification.

Vibrio parahaemolyticus infectious diseases caused by seafood contamination may be life-threatening to people with weak immunity. The detection of the Vibrio parahaemolyticus pathogen in aquatic foods is critical for reducing the outbreak of human Vibrio parahaemolyticus-associated diseases. In this study, a highly sensitive, specific, and time-saving real-time narrow thermal-cycling amplification detection method was developed based on accelerated strand exchange amplification (ASEA). It can detect cultured Vibrio parahaemolyticus at concentrations as low as 25 CFU mL-1. In addition, for artificially spiked scallop meat, the detection limit was 1.8 × 103 CFU g-1 without pre-culture and 18 CFU g-1 of initial inoculum after 3 h enrichment. The whole assay, starting from DNA extraction, can be completed within 20 min. The ASEA detection method established in this study is an effective tool for the rapid detection of Vibrio parahaemolyticus strains in a large number of seafood samples.

Vibrio vulnificus sepsis after shrimp shelling in a patient with preexisting primary biliary cholangitis: a case report.

BACKGROUND: Vibrio vulnificus is typically present in seawater, fish, and shellfish, and is known to cause severe sepsis, particularly in patients with liver diseases such as cirrhosis. V. vulnificus is one of the most dangerous waterborne pathogens, and infection mainly occurs in western Japan during the summer, with an increased fatality rate. Herein, we report the case of a patient with primary biliary cholangitis and sepsis caused by V. vulnificus infection sustained through shrimp shelling. CASE PRESENTATION: An 82-year-old Japanese Asian woman with no medical history or underlying disease developed redness, swelling, and pain, which extended from the right fingers to the upper arm. A diagnosis of sepsis due to cellulitis was made. Blood culture detected V. vulnificus; thus, minocycline was administered in addition to meropenem. The disease course was uneventful, and the patient was discharged on day 28 of hospitalization. Symptoms in the right upper arm developed 1 day after the patient shelled a large number of shrimp; therefore, the infection route was assumed to be through wounds sustained during shrimp shelling. We suspected liver disease and measured serum anti-mitochondrial M2 antibody levels, leading to the diagnosis of primary biliary cholangitis. CONCLUSIONS: As in this case, small wounds caused by handling fish and shrimp are a potential source of infection. Patients with severe V. vulnificus infection should be thoroughly assessed for the presence of liver diseases such as primary biliary cholangitis.

An Uncommon Case of Necrotizing Fasciitis and Septic Shock Caused by Vibrio vulnificus Infection-Related Freshwater Shrimp Stung.

Vibrio vulnificus is a deadly marine pathogen that can cause necrotizing fasciitis, septic shock, and even death in severe cases. The relatively low incidence and atypical early-stage symptoms may hinder many physicians from carrying out surgical intervention effectively, thus leading to an increase of mortality in infected patients. This article reported a patient who developed necrotizing fasciitis and septic shock after the exposure to freshwater shrimp stabbed on the limb. By reviewing and analyzing previous studies, it was found out that the timing of surgery could have a significant impact on the patients for their necrotizing fasciitis caused by Vibrio vulnificus infection. The mortality among patients undergoing early-stage surgical treatment (≤12 hours from the time of admission) was significantly lower than that of patients undergoing late surgical treatment (>12 hours).

Vibrio vulnificus necrotizing fasciitis with sepsis presenting with pain in the lower legs in winter: a case report.

BACKGROUND: Vibrio vulnificus infections develop rapidly and are associated with a high mortality rate. The rates of diagnosis and treatment are directly associated with mortality. CASE PRESENTATION: We describe an unusual case of a 61-year-old male patient with chronic liver disease and diabetes who presented with a chief complaint of pain in both lower legs due to V. vulnificus infection in winter. Within 12 h of arrival, typical skin lesions appeared, and the patient rapidly developed primary sepsis. Despite prompt appropriate antibiotic and surgical treatment, the patient died 16 days after admission. CONCLUSION: Our case findings suggest that V. vulnificus infection should be suspected in patients with an unclear infection status experiencing pain of unknown origin in the lower legs, particularly in patients with liver disease or diabetes, immunocompromised status, and alcoholism.

[Two cases of Vibrio vulnificus primary sepsis].

This article analyzed the medical records of two patients with Vibrio vulnificus primary sepsis who were admitted to the First Affiliated Hospital of Naval Medical University and reviewed the latest literature. On November 6, 2019, a 54-year-old male patient was admitted to the hospital. The patient's lower limbs were red, swollen, and painful with ecchymosis and hemorrhagic bullae after he ate freshwater products. The emergency fasciotomy was performed 3 h after admission, and the multiple organ failure occurred after operation. The patient was given up treatment 24 h after admission. On August 12, 2020, a 73-year-old male patient was admitted to the hospital. He was in shock state on admission and had hemorrhagic bullae on his right lower limb after he ate seafood. At 3 h post admission, he underwent emergency surgical exploration and amputation of right thigh. Six days later, he received negative pressure wound treatment on the stump. On the 13th day post admission, his families forgo the active treatment and he died 15 d after admission. The two cases were both failed to be diagnosed at the first time, and the disease progressed rapidly. Necrotizing fasciitis and multiple organ failure occurred. After the diagnosis was confirmed, timely fasciotomy and high amputation were performed respectively. The microbiological examinations both reported Vibrio vulnificus. Although the 2 cases were not cured successfully, the course of disease and some indexes of patient with early amputation were better than those of patients with fasciotomy. Vibrio vulnificus is widely distributed and frequently detected in fresh water products. The pathogenic pathway is fuzzy and complex, and it is easy to be misdiagnosed. It is necessary to establish the treatment process of Vibrio vulnificus sepsis. Early and aggressive surgical intervention should be carried out as soon as possible, fasciotomy and debridement should be thorough, and the patients with hemorrhagic bullae should be amputated early. Postoperative comprehensive measures are also important for improving the survival rate of patients.

Bacterial coloration immunofluorescence strip for ultrasensitive rapid detection of bacterial antibodies and targeted antibody-secreting hybridomas.

The indirect enzyme-linked immunosorbent assay (ELISA) is the gold standard method for monoclonal antibody (McAb) detection and plays a unique role in the preparation of bacterial antibodies. To solve the laborious issues associated with indirect ELISA, a novel bacterial coloration immunofluorescence strip (BCIFS) for antibody detection using colored bacteria instead of a labeled antibody as the antigen and tracer simultaneously and goat anti-mouse IgG as the test line was developed. The affinity range survey of BCIFS indicated that hybridoma cell cultures of E. coli O157:H7 (D3, E7) and Vibrio parahemolyticus (H7, C9) were detected, which complied with the results of indirect ELISA. Compared with the traditional indirect ELISA, the BCIFS sensitivity for E7 cell cultures, ascites, and purified antibodies was at least 4-fold more sensitive, and the BCIFS cross-reactivity for E7 cell cultures was almost consistent with that of indirect ELISA. In addition, the BCIFS isotypes for E. coli O157:H7 cell cultures and Vibrio parahemolyticus were IgG2a and IgG1, respectively, which were identical to the indirect ELISA. Furthermore, the BCIFS method was confirmed by McAb preparation, effective antibody use, and targeted antibody-secreted hybridoma preparation and screening, which showed excellent performance and substitution of the indirect ELISA method. Combined with methylcellulose semisolid medium, BCIFS offers a novel, easy to operate, rapid preparation method for antigen-specific hybridomas. This is the first report using BCIFS instead of indirect ELISA for bacterial antibody detection and application in different samples, which demonstrates a rapid and powerful tool for antibody engineering.

Shrimp AHPND Causing Vibrio anguillarum Infection: Quantitative Diagnosis and Identifying Antagonistic Bacteria.

Acute hepatopancreatic necrosis disease (AHPND) is one of the most common and serious diseases in shrimp aquaculture. Relevant works have focused on the gut microbiota-disease relationship when serious AHPND occurs. In contrast, little is known about how the gut microbiota responds to pathogen infection over AHPND progression, whereas this knowledge is fundamental to uncover the etiology of AHPND. Here, we explored the temporal succession of shrimp gut microbiota during Vibrio anguillarum (a causal pathogen of AHPND) challenge. The successful infection of V. anguillarum was confirmed by linearly increased abundance of the pathogen in the shrimp gut over AHPND progression. V. anguillarum infection caused an irreversible disruption in the shrimp gut microbiota, of which infection and hours post infection (hpi) respectively constrained 6.2% and 10.2% of variation in the data. Furthermore, the predicted functional pathways involved in immunity and metabolism significantly decreased, while those facilitating infectious diseases significantly enriched in the infected shrimp. Intriguingly, after ruling out the effect of background changes in gut microbiota, we identified 20 infection-discriminatory taxa that could be served as independent variables for accurately (89.4%) diagnosing V. anguillarum infection, even at the early infection stage, i.e., 24 hpi. Using a consensus network, we identified several Vibrio and Pseudoalteromonas taxa that directly antagonized V. anguillarum, following the Darwin's niche theory. This is one of the few attempts to identify gut bioindicators for diagnosing pathogen infection. In addition, the antagonistic commensals of V. anguillarum might be the candidate probiotics for preventing AHPND.

Heatwave-associated Vibrio infections in Germany, 2018 and 2019.

BackgroundVibrio spp. are aquatic bacteria that prefer warm seawater with moderate salinity. In humans, they can cause gastroenteritis, wound infections, and ear infections. During the summers of 2018 and 2019, unprecedented high sea surface temperatures were recorded in the German Baltic Sea.AimWe aimed to describe the clinical course and microbiological characteristics of Vibrio infections in Germany in 2018 and 2019.MethodsWe performed an observational retrospective multi-centre cohort study of patients diagnosed with domestically-acquired Vibrio infections in Germany in 2018 and 2019. Demographic, clinical, and microbiological data were assessed, and isolates were subjected to whole genome sequencing and antimicrobial susceptibility testing.ResultsOf the 63 patients with Vibrio infections, most contracted the virus between June and September, primarily in the Baltic Sea: 44 (70%) were male and the median age was 65 years (range: 2-93 years). Thirty-eight patients presented with wound infections, 16 with ear infections, six with gastroenteritis, two with pneumonia (after seawater aspiration) and one with primary septicaemia. The majority of infections were attributed to V. cholerae (non-O1/non-O139) (n = 30; 48%) or V. vulnificus (n = 22; 38%). Phylogenetic analyses of 12 available isolates showed clusters of three identical strains of V. vulnificus, which caused wound infections, suggesting that some clonal lines can spread across the Baltic Sea.ConclusionsDuring the summers of 2018 and 2019, severe heatwaves facilitated increased numbers of Vibrio infections in Germany. Since climate change is likely to favour the proliferation of these bacteria, a further increase in Vibrio-associated diseases is expected.

Programmable receptors enable bacterial biosensors to detect pathological biomarkers in clinical samples.

Bacterial biosensors, or bactosensors, are promising agents for medical and environmental diagnostics. However, the lack of scalable frameworks to systematically program ligand detection limits their applications. Here we show how novel, clinically relevant sensing modalities can be introduced into bactosensors in a modular fashion. To do so, we have leveraged a synthetic receptor platform, termed EMeRALD (Engineered Modularized Receptors Activated via Ligand-induced Dimerization) which supports the modular assembly of sensing modules onto a high-performance, generic signaling scaffold controlling gene expression in E. coli. We apply EMeRALD to detect bile salts, a biomarker of liver dysfunction, by repurposing sensing modules from enteropathogenic Vibrio species. We improve the sensitivity and lower the limit-of-detection of the sensing module by directed evolution. We then engineer a colorimetric bactosensor detecting pathological bile salt levels in serum from patients having undergone liver transplant, providing an output detectable by the naked-eye. The EMeRALD technology enables functional exploration of natural sensing modules and rapid engineering of synthetic receptors for diagnostics, environmental monitoring, and control of therapeutic microbes.

Comparative immunogenicity of outer membrane protein K and whole-cell antigens of Vibrio parahaemolyticus for diagnosis.

The immunogenicity of soluble outer membrane protein K (OmpK)- small ubiquitin-like modifier, OmpK inclusion bodies, formalin, and heat-killed Vibrio parahaemolyticus cells were prepared and studied in a mouse model. The results of whole-cell ELISA and Western blot (WB) revealed that the serum against soluble OmpK and OmpK inclusion bodies reacted only with homologous V. parahaemolyticus. Furthermore, recombinant OmpK proteins were not recognized by the serum against whole-cell V. parahaemolyticus antigens. Unexpectedly, the serum against formalin and heat-killed V. parahaemolyticus reacted broadly with homologous (an immunization strain) and heterologous (non-immunization strains) V. parahaemolyticus and Vibrio species. The WB results revealed that the serum against the two V. parahaemolyticus whole-cell antigens primarily reacted with proteins that were approximately 100, 70, 36, 28, and 22 kDa in the cell lysates from different Vibrio strains, rather than the recombinant OmpK. The 70 and 28 kDa proteins exhibited specificity to Vibrio species, while the 22 kDa protein was more specific to V. parahaemolyticus. This study showed the limitation of recombinant OmpK to prepare diagnostic antibodies and revealed several specific Omps of Vibrio sp. and V. parahaemolyticus that were promising in diagnosis and vaccine development.

[Non-cholera Vibrio species - currently still rare but growing danger of infection in the North Sea and the Baltic Sea]. / Nicht-Cholera-Vibrionen ­ derzeit noch seltene, aber wachsende Infektionsgefahr in Nord- und Ostsee.

BACKGROUND: The abundance of non-cholera Vibrio spp. in the aquatic environment shows a positive correlation with water temperatures. Therefore, climate change has an important impact on the epidemiology of human infections with these pathogens. In recent years large outbreaks have been repeatedly observed during the summer months in temperate climate zones. OBJECTIVE: To inform medical professionals about the potentially life-threatening diseases caused by non-cholera Vibrio spp. MATERIAL AND METHODS: Review of the current literature on infections with non-cholera Vibrio spp. in general and on the epidemiological situation in Germany in particular. RESULTS: Non-cholera Vibrio spp. predominantly cause wound and ear infections after contact with contaminated seawater and gastroenteritis after consumption of undercooked seafood. As there have not been mandatory notification systems for these pathogens in Germany up to March 2020, a high number of unreported cases must be assumed. Immunosuppressed and chronically ill patients have a much higher risk for severe courses of diseases. If an infection with non-cholera Vibrio spp. is suspected anti-infective treatment should be promptly initiated and surgical cleansing is often necessary for wound and soft tissue infections. CONCLUSION: Due to the ongoing global warming an increased incidence of human infections with non-cholera Vibrio spp. must be expected in the future. Medical professionals should be aware of these bacterial pathogens and the potentially life-threatening infections in order to enable timely diagnostics and treatment.

The bane of coastal marine environment: A fatal case of Vibrio vulnificus associated cellulitis and septicaemia.

Vibrio vulnificus is a Gram negative motile bacterium known to cause fatal septicaemia and wound infection. It is commonly associated with the consumption of under-cooked seafood or exposure to marine environment. We report a case of a 55 year old male patient, who was presented with right lower limb cellulitis and septicaemia due to V. vulnificus. V. vulnificus infection in India are rare. However, increasing reports of V. vulnificus from India recommends considering the pathogen while dealing necrotising fasciitis especially in the proximity of marine environment.