A rare waterborne outbreak of Bacillus paranthracis in Shandong province, China, 2020: epidemiologic survey, genomic insights, and virulence characteristics.

Bacillus paranthracis, a Gram-positive conditional pathogen of Bacillus cereus group species, is capable of causing foodborne and waterborne illnesses, leading to intestinal diseases in humans characterized by diarrhoea and vomiting. However, documented cases of B. paranthracis infection outbreaks are rare in the world, and the genomic background of outbreak strains is seldom characterized. This study retrospectively analyzed strains obtained from an outbreak in schools, as well as from water systems in peri-urban areas, China, in 2020. In total, 28 B. cereus group isolates were retrieved, comprising 6 from stool samples and 22 from water samples. Epidemiological and phylogenetic investigations indicated that the B. paranthracis isolate from drinking water as the causative agent of the outbreak. The genomic comparison revealed a high degree of consistency among 8 outbreak-related strains in terms of antimicrobial resistance gene profiles, virulence gene profiles, genomic content, and multilocus sequence typing (MLST). The strains related to the outbreak show highly similar genomic ring diagrams and close phylogenetic relationships. Additionally, this study shed light on the pathogenic potential and complexity of B. cereus group through its diversity in virulence genes and mice infection model. The findings highlight the usefulness of B. paranthracis genomes in understanding genetic diversity within specific environments and in tracing the source of pathogens during outbreak situations, thereby enabling targeted infection control interventions.

The role of biofilm in the development and dissemination of ubiquitous pathogens in drinking water distribution systems: an overview of surveillance, outbreaks, and prevention.

A variety of pathogenic microorganisms can survive in the drinking water distribution systems (DWDS) by forming stable biofilms and, thus, continually disseminating their population through the system's dynamic water bodies. The ingestion of the pathogen-contaminated water could trigger a broad spectrum of illnesses and well-being-related obstacles. These waterborne diseases are a significant concern for babies, pregnant women, and significantly low-immune individuals. This review highlights the recent advances in understanding the microbiological aspects of drinking water quality, biofilm formation and its dynamics, health issues caused by the emerging microbes in biofilm, and approaches for biofilm investigation its prevention and suppression in DWDS.

A Whole-Cell Biosensor for Point-of-Care Detection of Waterborne Bacterial Pathogens.

Water contamination by pathogenic bacteria is a major public health concern globally. Monitoring bacterial contamination in water is critically important to protect human health, but this remains a critical challenge. Engineered whole-cell biosensors created through synthetic biology hold great promise for rapid and cost-effective detection of waterborne pathogens. In this study, we created a novel whole-cell biosensor to detect water contamination by Pseudomonas aeruginosa and Burkholderia pseudomallei, which are two critical bacterial pathogens and are recognized as common causative agents for waterborne diseases. The biosensor detects the target bacterial pathogens by responding to the relevant quorum sensing signal molecules. Particularly, this study constructed and characterized the biosensor on the basis of the QscR quorum sensing signal system for the first time. We first designed and constructed a QscR on the basis of the sensing module in the E. coli host cell and integrated the QscR sensing module with a reporting module that expressed an enhanced green fluorescent protein (EGFP). The results demonstrated that the biosensor had high sensitivity in response to the quorum sensing signals of the target bacterial pathogens. We further engineered a biosensor that expressed a red pigment lycopene in the reporting module to produce a visible signal readout for the pathogen detection. Additionally, we investigated the feasibility of a paper-based assay by immobilizing the lycopene-based whole-cell biosensor on paper with the aim to build a prototype for developing portable detection devices. The biosensor would provide a simple and inexpensive alternative for timely and point-of-care detection of water contamination and protect human health.

Identifying septic pollution exposure routes during a waterborne norovirus outbreak - A new application for human-associated microbial source tracking qPCR.

In June 2017, the Pennsylvania Department of Health (PADOH) was notified of multiple norovirus outbreaks associated with 179 ill individuals who attended separate events held at an outdoor venue and campground over a month period. Epidemiologic investigations were unable to identify a single exposure route and therefore unable to determine whether there was a persistent contamination source to target for exposure mitigation. Norovirus was detected in a fresh recreational water designated swimming area and a drinking water well. A hydrogeological site evaluation suggested a nearby septic leach field as a potential contamination source via ground water infiltration. Geological characterization revealed a steep dip of the bedrock beneath the septic leach field toward the well, providing a viral transport pathway in a geologic medium not previously documented as high risk for viral ground water contamination. The human-associated microbial source tracking (MST) genetic marker, HF183, was used as a microbial tracer to demonstrate the hydrogeological connection between the malfunctioning septic system, drinking water well, and recreational water area. Based on environmental investigation findings, venue management and local public health officials implemented a series of outbreak prevention strategies including discontinuing the use of the contaminated well, issuing a permit for a new drinking water well, increasing portable toilet and handwashing station availability, and promoting proper hand hygiene. Despite the outbreaks at the venue and evidence of ground water contamination impacting nearby recreational water and the drinking water well, no new norovirus cases were reported during a large event one week after implementing prevention practices. This investigation highlights a new application for human-associated MST methods to trace hydrological connections between multiple fecal pollutant exposure routes in an outbreak scenario. In turn, pollutant source information can be used to develop effective intervention practices to mitigate exposure and prevent future outbreaks associated with human fecal contaminated waters.

Survival time of Leptospira kirschneri serovar Grippotyphosa under different environmental conditions.

Leptospirosis is a re-emerging zoonotic disease of high medical importance that affects humans worldwide. Humans or animals acquire an infection with pathogenic leptospires either by direct contact with infected animals or by indirect contact to contaminated environment. Survival of Leptospira spp. in the environment after having been shed via animal urine is thus a key factor to estimate the risk of infection, but not much is known about the tenacity of pathogenic leptospires. Here, the survival time of both a laboratory strain and a field strain of L. kirschneri serovar Grippotyphosa in animal urine and their tenacity while drying was investigated and compared at different temperatures (15°C-37°C). Leptospira spp. are also often found in rivers and ponds. As the infection risk for humans and animals also depends on the spreading and survival of Leptospira spp. in these environments, the survival of L. kirschneri serovar Grippotyphosa was investigated using a 50-meter-long hose system simulating a water stream. Both strains did not survive in undiluted cattle or dog urine. Comparing different temperatures and dilution media, the laboratory strain survived the longest in diluted cattle urine with a slightly alkaline pH value (3 days), whilst the field strain survived in diluted dog urine with a slightly acid pH value up to a maximum of 24 h. Both strains did not survive drying on a solid surface. In a water stream, leptospires were able to move faster or slower than the average velocity of the water due to their intrinsic mobility but were not able to survive the mechanical damage caused by running water in the hose system. From our results we conclude, that once excreted via animal urine, the leptospires immediately need moisture or a water body to survive and stay infectious.

[Waterborne outbreaks in Iceland - analysis of scale and causes]. / Vatnsbornar hópsýkingar á Íslandi - greining á umfangi og ástæðum

PURPOSE: Clean drinking water is essential for public health. The cause of waterborne outbreaks is most often faecal contamination of water from animals or humans. The objective of this resarch was to collect available information on waterborne outbreaks in Iceland for the twenty year period, 1998-2017. Incident of faecal and pathogenic pollution in samples where also collected even though rarely followed by registered outbreak. METHODS: Data are obtained from laboratory databases, the Directorate of Health, reports and interviews with the relevant surveillance authorities and epidemiologists. RESULTS: The results show that for the period investigated fifteen waterborne outbreaks were registered, all in small water supplies, many of which served transitent population, tourists and summerhouse dwellers. About 500 illnesses were confirmed and 8000 people affected. Other research have shown that around 10% of illnesses in waterborne outbreaks are registered so it can be estimated that on average 250 people have been taken ill every year because of contaminated drinking water. Analysis of monitoring water quality data show that on average 50 water supplies, or about 5% of the Icelandic registered water supplies have contained faecal matter every year. The most frequent cause of waterborne outbreak were poor design and inadequate maintainance of water intakes. CONCLUSIONS: It is likely that waterborne outbreaks are more numerous than are registered in official reports, especially concerning small water supplies. It also seems that the local heath authorities are often not informed of incidents of non-compliance. It is important to improve registration, information exchange between parties, epidemiological surveys and follow up of outbreaks due to drinking water to gather lessons learned. Water quality at small water supplies needs to be improved with risk-based approach and risk management.

A nosocomial cluster of Roseomonas mucosa bacteremia possibly linked to contaminated hospital environment.

Roseomonas, a genus of pink-pigmented glucose non-fermentative bacteria, has been associated with various primary and hospital-acquired human infections; however, to our knowledge, its nosocomial transmission has never been reported. Clinical and epidemiological investigations were carried out after two cases of R. mucosa bacteremia occurred in our hospital in 2018. Environmental samples were taken of environmental surfaces prone to water contamination in the wards and cultured. The two clinical isolates and all environmental isolates that showed growth of pink colonies were identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry and 16S rRNA gene sequencing. Pulse-field gel electrophoresis (PFGE) was performed and fingerprinting software was used to analyze the DNA restriction patterns and determine their similarity. Two patients who developed R. mucosa bacteremia had received care from the same treatment team. Of 126 environmental samples, five showed growth of R. mucosa. Using 80% similarity as the cut-off, PFGE analysis revealed that the isolates from the two patients' blood cultures and three environmental isolates belonged to the same clone. The hospital water environment was contaminated with the same clone of R. mucosa that caused bacteremia in the two patients, suggesting nosocomial transmission linked to contaminated environment. Increased vigilance is needed to monitor the emergence of Roseomonas in healthcare settings.

Estimates of healthcare utilisation and deaths from waterborne pathogen exposure in Ontario, Canada.

Burden of disease analyses can quantify the relative impact of different exposures on population health outcomes. Gastroenteritis where the causative pathogen was not determined and respiratory illness resulting from exposure to opportunistic pathogens transmitted by water aerosols have not always been considered in waterborne burden of disease estimates. We estimated the disease burden attributable to nine enteric pathogens, unspecified pathogens leading to gastroenteritis, and three opportunistic pathogens leading primarily to respiratory illness, in Ontario, Canada (population ~14 million). Employing a burden of disease framework, we attributed a fraction of annual (year 2016) emergency department (ED) visits, hospitalisations and deaths to waterborne transmission. Attributable fractions were developed from the literature and clinical input, and unattributed disease counts were obtained using administrative data. Our Monte Carlo simulation reflected uncertainty in the inputs. The estimated mean annual attributable rates for waterborne diseases were (per 100 000 population): 69 ED visits, 12 hospitalisations and 0.52 deaths. The corresponding 5th-95th percentile estimates were (per 100 000 population): 13-158 ED visits, 5-22 hospitalisations and 0.29-0.83 deaths. The burden of disease due to unspecified pathogens dominated these rates: 99% for ED visits, 63% for hospitalisations and 40% for deaths. However, when a causative pathogen was specified, the majority of hospitalisations (83%) and deaths (97%) resulted from exposure to the opportunistic pathogens Legionella spp., non-tuberculous mycobacteria and Pseudomonas spp. The waterborne disease burden in Ontario indicates the importance of gastroenteritis not traced back to a particular pathogen and of opportunistic pathogens transmitted primarily through contact with water aerosols.

Leptospirosis: a neglected tropical zoonotic infection of public health importance-an updated review.

Leptospirosis is a zoonotic and waterborne disease worldwide. It is a neglected, reemerging disease of global public health importance with respect to morbidity and mortality both in humans and animals. Due to negligence, rapid, unplanned urbanization, and poor sanitation, leptospirosis emerges as a leading cause of acute febrile illness in many of the developing countries. Every individual has a risk of getting infected as domestic and wild animals carry leptospires; the at-risk population varies from the healthcare professionals, animal caretakers, farmers and agricultural workers, fishermen, rodent catchers, water sports people, National Disaster Response Force (NDRF) personnel, people who volunteer rescue operations in flood-affected areas, sanitary workers, sewage workers, etc. The clinical manifestations of leptospirosis range from flu-like illness to acute kidney failure (AKF), pneumonia, jaundice, pulmonary hemorrhages, etc. But many rare and uncommon clinical manifestations are being reported worldwide. This review will cover all possible updates in leptospirosis from occurrence, transmission, rare clinical manifestations, diagnosis, treatment, and prophylactic measures that are currently available, their advantages and the future perspectives, elaborately. There are less or very few reviews on leptospirosis in recent years. Thus, this work will serve as background knowledge for the current understanding of leptospirosis for researchers. This will provide a detailed analysis of leptospirosis and also help in finding research gaps and areas to focus on regarding future research perspectives.

Epidemiological Aspects of Escherichia albertii Outbreaks in Japan and Genetic Characteristics of the Causative Pathogen.

Zoonotic pathogen Escherichia albertii has been identified as the cause of several human disease outbreaks; however, factors such as the general symptoms and incubation period of E. albertii infection have yet to be defined. Therefore, we aimed to determine the unique aspects of E. albertii outbreaks in Japan and to examine the genetic characteristics of the causative pathogen. We studied all known E. albertii outbreaks that occurred in Japan up until 2015, which consisted of five confirmed outbreaks and one putative outbreak (Outbreaks 1-6). Outbreaks were re-examined based on personal communications between researchers in prefectural and municipal public health institutes, and through examination of any published study conducted at the time. Draft genome sequences of outbreak-associated E. albertii isolates were also generated. The most common symptom displayed by patients across the six episodes was watery diarrhea (>80%), followed by abdominal pain (50-84%) and fever (37.0-39.5°C) (26-44%). The estimated average incubation period of E. albertii infection was 12-24 h. We assumed that most of the outbreaks were foodborne or waterborne, with restaurant foods, restaurant water, and boxed lunches being the suspected transmission vehicles. Three of the six outbreak-associated E. albertii isolates possessed intact ETT2 regions, while the remaining isolates contained disrupted ETT2-encoding genes. Virulence gene screening revealed that more than half (44/70) of the tested genes were present in all 5 strains examined, and that each of the strains contained more than 1 gene from 14 out of the 21 groups of virulence genes examined in this study. The five E. albertii strains were classified into four of the five known phylogroups. Therefore, we determined that multiple E. albertii genotypes in Japan have the potential to cause outbreaks of diarrhea, abdominal pain, and/or fever following infection of a human host.

Point-of-Care Strategies for Detection of Waterborne Pathogens.

Waterborne diseases that originated due to pathogen microorganisms are emerging as a serious global health concern. Therefore, rapid, accurate, and specific detection of these microorganisms (i.e., bacteria, viruses, protozoa, and parasitic pathogens) in water resources has become a requirement of water quality assessment. Significant research has been conducted to develop rapid, efficient, scalable, and affordable sensing techniques to detect biological contaminants. State-of-the-art technology-assisted smart sensors have improved features (high sensitivity and very low detection limit) and can perform in a real-time manner. However, there is still a need to promote this area of research, keeping global aspects and demand in mind. Keeping this view, this article was designed carefully and critically to explore sensing technologies developed for the detection of biological contaminants. Advancements using paper-based assays, microfluidic platforms, and lateral flow devices are discussed in this report. The emerging recent trends, mainly point-of-care (POC) technologies, of water safety analysis are also discussed here, along with challenges and future prospective applications of these smart sensing technologies for water health diagnostics.

Pseudo-outbreak of Mycobacterium chimaera through aerators of hand-washing machines at a hematopoietic stem cell transplantation center.

We identified a waterborne pseudo-outbreak of Mycobacterium chimaera in our stem cell transplantation center, which likely resulted from biofilm on the aerators of the handwashing machines in each patient's room. Regular replacement of faucet parts can prevent biofilm formation and pseudo-outbreaks of M. chimaera through aerators.

Tularemia as a waterborne disease: a review.

Francisella tularensis is a Gram-negative, intracellular bacterium causing the zoonosis tularemia. This highly infectious microorganism is considered a potential biological threat agent. Humans are usually infected through direct contact with the animal reservoir and tick bites. However, tularemia cases also occur after contact with a contaminated hydro-telluric environment. Water-borne tularemia outbreaks and sporadic cases have occurred worldwide in the last decades, with specific clinical and epidemiological traits. These infections represent a major public health and military challenge. Human contaminations have occurred through consumption or use of F. tularensis-contaminated water, and various aquatic activities such as swimming, canyoning and fishing. In addition, in Sweden and Finland, mosquitoes are primary vectors of tularemia due to infection of mosquito larvae in contaminated aquatic environments. The mechanisms of F. tularensis survival in water may include the formation of biofilms, interactions with free-living amoebae, and the transition to a 'viable but nonculturable' state, but the relative contribution of these possible mechanisms remains unknown. Many new aquatic species of Francisella have been characterized in recent years. F. tularensis likely shares with these species an ability of long-term survival in the aquatic environment, which has to be considered in terms of tularemia surveillance and control.

Detection of antimicrobial-resistance diarrheagenic Escherichia coli strains in surface water used to irrigate food products in the northwest of Mexico.

Water contamination by pathogenic bacteria is a global public health problem. Contamination of surface water utilized to irrigate food products, or for human consumption, causes outbreaks of foodborne and waterborne disease. Of these, those caused by diarrheagenic Escherichia coli (DEC) strains present substantial morbidity and mortality. The aim of this study was, therefore, to investigate the microbiological quality of surface water and the presence of DEC strains in different water bodies. A total of 472 water samples were collected from irrigation canal, dam, river, and dike water bodies from January through December 2015 in Sinaloa, a State located in Northwestern Mexico. Our studies demonstrated that 47.0% (222/472) of samples contained thermotolerant coliforms above permissive levels whereas E. coli strains were isolated from 43.6% (206/472). Among these E. coli isolates, DEC strains were identified in 14% (29/206) of samples including in irrigation canal (26/29) and river water (3/29) collected from the northern (83%) and central area (17%). Isolated DEC strains were classified as enteroaggregative E. coli (EAEC) 34.4% (10/29), enteropathogenic E. coli (EPEC) 31.0% (9/29), diffuse adherent E. coli (DAEC) 27.5% (8/29), and enterotoxigenic E. coli (ETEC) 6.8% (2/29). Moreover, 90% of isolated DEC strains exhibited resistance to at least one commonly prescribed antibiotic in Mexico whereas 17% were multi-drug resistant. In conclusion, the presence of DEC strains in surface water represents a potential source for human infection, and thus routine monitoring of DEC in surface water and other indirect affected areas should be considered at northwestern Mexico.

Investigation of healthcare infection risks from water-related organisms: Summary of CDC consultations, 2014-2017.

OBJECTIVE: Water exposures in healthcare settings and during healthcare delivery can place patients at risk for infection with water-related organisms and can potentially lead to outbreaks. We aimed to describe Centers for Disease Control and Prevention (CDC) consultations involving water-related organisms leading to healthcare-associated infections (HAIs). DESIGN: Retrospective observational study. METHODS: We reviewed internal CDC records from January 1, 2014, through December 31, 2017, using water-related terms and organisms, excluding Legionella, to identify consultations that involved potential or confirmed transmission of water-related organisms in healthcare. We determined plausible exposure pathways and routes of transmission when possible. RESULTS: Of 620 consultations during the study period, we identified 134 consultations (21.6%), with 1,380 patients, that involved the investigation of potential water-related HAIs or infection control lapses with the potential for water-related HAIs. Nontuberculous mycobacteria were involved in the greatest number of investigations (n = 40, 29.9%). Most frequently, investigations involved medical products (n = 48, 35.8%), and most of these products were medical devices (n = 40, 83.3%). We identified a variety of plausible water-exposure pathways, including medication preparation near water splash zones and water contamination at the manufacturing sites of medications and medical devices. CONCLUSIONS: Water-related investigations represent a substantial proportion of CDC HAI consultations and likely represent only a fraction of all water-related HAI investigations and outbreaks occurring in US healthcare facilities. Water-related HAI investigations should consider all potential pathways of water exposure. Finally, healthcare facilities should develop and implement water management programs to limit the growth and spread of water-related organisms.

Integrons and antibiotic resistance genes in water-borne pathogens: threat detection and risk assessment.

Antibiotic-resistant genes (ARGs) are regarded as emerging environmental pollutants and pose a serious health risk to the human population. Integrons are genetic elements that are involved in the spread of ARGs amongst bacterial species. They also act as reservoirs of these resistance traits, further contributing to the development of multi-drug resistance in several water-borne pathogens. Due to inter- and intra-species transfer, integrons are now commonly reported in important water-borne pathogens such as Vibrio, Campylobacter, Salmonella, Shigella, Escherichia coli and other opportunistic pathogens. These pathogens exhibit immense diversity in their resistance gene cassettes. The evolution of multiple novel and complex gene cassettes in integrons further suggests the selection and horizontal transfer of ARGs in multi-drug resistant bacteria. Thus, the detection and characterization of these integrons in water-borne pathogens, especially in epidemic and pandemic strains, is of the utmost importance. It will provide a framework in which health authorities can conduct improved surveillance of antibiotic resistance in our natural water bodies. Such a study will also be helpful in developing better strategies for the containment and cure of infections caused by these bacteria.

Seasonal influence, enteropathogenic microbial load and diarrhoeal enigma in the Gangetic Delta, India: Present scenario and health implications.

BACKGROUND: Environmental diarrheagenic enteropathogens, effect of surrounding attributes and disease dynamicity remains far from being conclusively explored. Population flux, poor sanitation and hygienic practice poses potential health threat in diarrhoea endemic tropical countries like India. We aim to identify environmental attributes, seasonality of water-borne enteropathogens and health risk assessment off the river Ganges. METHODS: A yearlong sampling data generated from three sites on either sides of the River was analysed and implications have been reported. Immediately after sample collection, physico chemical and bacterial indices were measured at the sampling site and laboratory respectively, followed by further statistical analysis of the findings. RESULTS: Annual variation of physico-chemical indices viz., temperature 18°C-36°C, pH 7.49-8.67, conductivity 215-468µS/cm and turbidity 25.6-593 NTU was recorded in the riverine water samples. High temperature and turbidity were recorded in the summer and monsoon at all sites. High bacterial dispersion has been positively correlated with turbidity and temperature variation (P<0.01; P<0.1) as we report TBC 103-105CFU/ml, TCC 103-104CFU/ml and CVC 4-212CFU/ml, with higher distribution in the monsoon and reverse in the winter. This suggests that the bacterial pool proliferates at higher temperature whereas turbidity enhances their survival providing the substratum for the bacterial pool. CVC could be positively correlated with conductivity which implies that ionic content of water augments the Vibrio load. Adaptive capability of Vibrios to sustain in very low saline riverine setting seems to be assisted by turbid water coupled with nutrient rich organic matter. CONCLUSION: Our present work establishes the interplay of seasonal variants on the dynamicity of enteropathogenic bacteria in flowing aquatic ecosystem. It also categorises the existing microbial threats in the Ganga River to help monitor the conventional as well as emerging diarrhoeal pathogens to reduce diarrheal recurrences.

Bio-fabrication of silver nanoparticles by Pseudomonas aeruginosa: optimisation and antibacterial activity against selected waterborne human pathogens.

Multiple drug resistance and treatment of contaminated water has become a serious issue in past years. Silver nanoparticles (AgNPs), being bactericidal, non-toxic, cheap and environment friendly behaviour, have drawn attention to overcome these problems. This study has been designed to synthesise AgNPs from Pseudomonas aeruginosa. AgNPs formation was confirmed by colour change and UV-vis spectroscopy. Furthermore, Fourier transform infrared spectroscopy peaks demonstrated the presence of capped proteins as reducing and stabilising agent. Transmission electron microscopy micrograph revealed spherical shape AgNPs with the size ranging between 10 and 20 nm. Antibacterial activity of AgNPs was evaluated against the most prevalent waterborne pathogens enterohaemorrhagic Escherichia coli and Salmonellae typhimurium. Moreover, the antibacterial activity of AgNPs was tested for the treatment of contaminated water which showed attenuation in bacterial load within 8 h as demonstrated by growth kinetics data. Furthermore, AgNPs did not exhibit haemolytic effects on human red blood cells (RBCs) even at 100 mg L-1 concentration of AgNPs. The results herein suggest that AgNPs synthesised by P. aeruginosa under optimised conditions exhibit microbicidal property against waterborne pathogens and having no toxic effect on human RBCs. These AgNPs could be employed for treatment of contaminated water after process optimisation.

Vibrio vulnificus in aquariums is a novel threat to marine mammals and public health.

Vibrio vulnificus is a Gram-negative, curved, obligate halophilic marine bacterium that exclusively exists in coastal seawaters. Previous studies revealed that V. vulnificus is one of the most dangerous foodborne zoonotic pathogens for human beings. However, it remains unknown whether marine mammals can be infected by V. vulnificus. In May 2016, a captive spotted seal (Phoca largha) died due to septicemia induced by V. vulnificus. Upon post-mortem examination, V. vulnificus was isolated, identified, and named as BJ-PH01. Further analysis showed that BJ-PH01 belongs to biotype 1 and the Clinical genotype. Furthermore, we performed an epidemiological investigation of V. vulnificus in six aquariums in northern China. As a result, V. vulnificus was successfully isolated from all investigated aquariums. The positive rates ranged from 20% to 100% in each investigated aquarium. During the investigation, 12 strains of V. vulnificus were isolated, and all 12 isolates were classified into biotype 1. Eleven of the 12 isolates belonged to the Clinical genotype, and one isolate belonged to the Environmental genotype. All 12 isolated V. vulnificus strains showed limited antibiotic resistance. Overall, our work demonstrated that V. vulnificus is frequently distributed in aquariums, thus constituting a threat to captive marine mammals and to public health.