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Original Article | Open Access | Eur. J. Med. Health Sci., 4(5), 153-162 | doi: 10.34104/ejmhs.022.01530162

Occurrence of Vibrio Species in Marine Sources Surroundings Bonaire, Dutch Caribbean

Harish C Gugnani* Mail Img ,
Quincy Chopra ,
Alicia Currie ,
Trevor Ostrer ,
Willem Wiegersma ,
Palabh Ray

Abstract

There is a lack of information on the presence of vibrios in the marine environment in the Caribbean. The aim of this study was to determine the occurrence of Vibrios in the coastal waters of Bonaire. Fifty samples of marine water collected at different depths from various sources around Bonaire were examined for the presence of vibrios. Species identification was confirmed by KB007 HiVibrioTM, Identification Kit, and TOFEL-MALDI. Forty of the samples contained Vibrio alginolyticus, 33 yielded V. parahaemolyticus and 29 showed the presence of V. vulnificus / V. cholerae. Regarding total colony counts in the sample, 47.4% of the colonies were V. alginolyticus, 35.2% were V. parahaemolyticus, and 17.4% represented V. vulnificus / V. cholerae. Further, of the 25 surface samples from various sites, 14 had a colony count percentage of 50% or greater number of V. alginolyticus.  Another 10 sites had a colony count percentage of 50% or greater for V. parahaemolyticus; three of them had a colony count percentage of 50% or greater for V. vulnificus / V. cholerae. The present study constitutes the first study of its kind providing evidence of the prevalence of pathogenic Vibrio species, viz.  V. alginolyticus, V. parahaemolyticus, and V. vulnificus / V. cholerae in marine water from the Dutch Caribbean. 

INTRODUCTION

Vibrios are Gram negative, curved rods that are oxidase positive and facultative anaerobes. They are abundant in aquatic environment (Thompson et al., 2004). Among the more than seventy different Vibrio species inhabiting marine, estuarine, and freshwater ecosystems, eight are recognized as clinically import-ant, namely, V. cholerae, V. parahaemolyticus, V. vulnificus, V. mimicus, V. alginolyticus, V. harveyi, V. metschnikovii, and V. cincinnatiensis (Farmer, 2006). These organisms are halophilic (salt-loving) and thus most species reat species require salt in the growth media to indicate their presence (Murray et al., 2013). Toxigenic strains of V. cholerae 01 have been detected by PCR from the Latin American cho-lera epidemic (Fields et al., 1992). According to the CDC estimates, vibriosis causes 80,000 illnesses each year in the United States, about 52,000 of these illnesses resulting from eating contaminated food in the United States annually (CDC 24/7). Toxigenic Vibrio cholerae are found after O1 in water and seafood, Haiti (Hill et al., 2011). The species clinically most significant for humans are V. cholerae, V. vulnificus, and V. parahaemolyticus, and within these three species, there are different sero-groups (Thompson et al., 2004). There are currently 200 serogroups of V. cholerae identified and it is important to note that not all sero groups have been found to be pathogenic (Daniel and Shefaei, 2004). Of great importance, however, are V. cholerae O1 and O139 since these serogroups produce the cholera toxin that are responsible for epidemic Cholera (Dutta et al., 2013; Murray et al., 2013) require salt in the growth media to indicate their presence (Murray et al., 2013). According to the CDC estimates, vibriosis causes 80,000 illnesses each year in the United States, about 52,000 of these illnesses resulting from eating contaminated food in the United States annually (CDC 24/7). Toxigenic Vibrio cholerae O1 in water and seafood, Haiti (Hill et al., 2011). The species clinically most significant for humans are V. cholerae, V. vulnificus, and V. para-haemolyticus, and within these three species, there are different serogroups (Thompson et al., 2004). 

There are currently 200 serogroups of V. cholerae identified and it is important to note that not all serogroups have been found to be pathogenic (Daniel and Shefaei 2004). Of great importance, however, are V. cholerae O1 and O139 since these serogroups pro-duce the cholera toxin that are responsible for epi-demic Cholera (Dutta et al., 2013; Murray et al., 2013) require salt in the growth media to indicate their presence (Murray et al., 2013). Toxigenic strains of V. cholerae 01 have been detected by PCR from the Latin American cholera epidemic (Fields et al., 1992). According to the CDC estimates, vibriosis causes 80,000 illnesses each year in the United States, about 52,000 of these illnesses resulting from eating contaminated food in the United States annu-ally (CDC 24/7). Toxigenic Vibrio cholerae O1 in water and seafood, Haiti (Hill et al., 2011). The species clinically most significant for humans are V. cholerae, V. vulnificus, and V. parahaemolyticus, and within these three species, there are different sero-groups (Thompson et al., 2004). There are currently 200 serogroups of V. cholerae identified and it is important to note that not all serogroups have been found to be pathogenic (Daniel and Shefaei, 2004). Of great importance, however, are V. cholerae O1 and O139 since these serogroups produce the cholera toxin that are responsible for epidemic Cholera (Dutta et al., 2013; Murray et al., 2013) media to indicate their presence (Murray et al., 2013). Toxi-genic strains of V. cholerae 01 have been detected by PCR from the Latin American cholera epidemic (Fields et al., 1992). According to the CDC esti-mates, vibriosis causes 80,000 illnesses each year in the United States, about 52,000 of these illnesses resulting from eating contaminated food in the Uni-ted States annually (CDC 24/7). Toxigenic Vibrio cholerae O1 in water and seafood, Haiti (Hill et al., 2011). The species clinically most significant for humans are V. cholerae, V. vulnificus, and V. para-haemolyticus, and within these three species, there are different serogroups (Thompson et al., 2004). There are currently 200 serogroups of V. cholerae identified and it is important to note that not all serogroups have been found to be pathogenic (Daniel and Shefaei, 2004). Of great importance, however, are V. cholerae O1 and O139 since these serogroups produce the cholera toxin that are responsible for epidemic Cholera (Dutta et al., 2013; Murray et al., 2013) media to indicate their presence (Murray et al., 2013).

Toxigenic strains of V. cholerae 01 have been detected by PCR from the Latin American cholera epidemic (Fields et al., 1992). According to the CDC estimates, vibriosis causes 80,000 illnesses each year in the United States, about 52,000 of these illnesses resulting from eating contaminated food in the United States annually (CDC, 24/7). Toxigenic Vibrio cholerae O1 in water and seafood, Haiti (Hill et al., 2011). The species clinically most significant for humans are V. cholerae, V. vulnificus, and V. para-haemolyticus, and within these three species, there are different serogroups (Thompson et al., 2004). There are currently 200 serogroups of V. cholerae identified and it is important to note that not all serogroups have been found to be pathogenic (Daniel and Shefaei, 2004). Of great importance, however, are V. cholerae O1 and O139 since these serogroups produce the cholera toxin that are responsible for epidemic Cholera (Dutta et al., 2013; Murray et al., 2013). Recently, Vibrio bacteria received media attention when 33 cases of Vibrio vulnificus infection were reported in Florida, Texas, Mississippi, and Maryland (Cheshire, 2014). The fishing industry was also affected as the contamination of oysters with Vibrio parahaemolyticus resulted in the closure of Samish Bays Oyster Harvest in the State of Washing- ton from early August until September 30th, 2014  (http:/www.foodsafetynews.com/2014)[ Vibrio infect-ions are often contracted through ingestion of conta-minated food, usually raw or undercooked sea-food (Rosenbergh et al., 2007; DePaula et al., 2010). The pH range for the growth of Vibrio is significant because these bacteria do not survive in normal sto-mach (Murray et al., 2013). Under the reduced stomach acid secretion (higher pH), there is risk of acquiring a Vibrio infection with symptoms of severe gastroenteritis (Murray et al., 2013). Vibrio infect-ions contracted through an open wound initially result in severe pain at the injury site leading to num-bness and if medical attention is delayed, septicaemia may develop (Hoffman et al., 2012). It is in this context that investigation on occurrence of patho-genic Vibrios in marine sources is of medical and public health importance. Oliver et al. (1982) in a study of sampling water from North Carolina to Georgia found V. vulnificus, V. cholerae, V. mimicus, and V. fluvial in the water, on the sediment and on the marine life. A study from Saudi Arabia (Elhadi 2013) found V. alginolyticus, V. parahaemolyticus, V. vulnificus, V. mimicus and V. cholerae non-O1/ non-O139 in the coastal waters. Isolation of Vibrio parahemolyticus, V. vulnificus. and V. fluvialis from frozen imported raw fish been reported from Egypt (Khalil et al., 2014). Another study performed in Chesapeake Bay found V. vulnificus in water samples obtained during the summer months but not in the winter months (Wright et al., 1996). These studies indicate the Vibrio bacteria are autochthonous in various locations specifically in the coastal waters of the US, Gulf of Mexico, Americas, and the Cari-bbean. So far there are no reported studies document-ing the presence of Vibrio species in coastal waters of the Dutch Caribbean. 

Therefore, this study aims to provide data on the pre-sence of Vibrio species in the marine environment surrounding Bonaire in the southern region of the Caribbean. This study is particularly important be-cause the local economy of Bonaire relies heavily on sightseeing activities that include diving and snor-kelling as well as on the local fishing industry. It is imperative to be aware of the presence of Vibrio bacteria and their potential effects on the marine ecology, Finally, the study is significant for the health of tourists and locals, who may acquire Vibrio wound infection after sustaining injures while con-tacting coral and submerged rock formations (Poutier et al., 2012).

METHODOLOGY

Sampling

A total of 50 samples of marine water were collected in 15 mL sterile culture tubes from various sources around Bonaire at different depths, with water temp-erature and pH values recorded for each sample. Twenty-five of these samples originated from surface waters (referred as surface samples), around Bonaire. Scuba diving was performed to obtain 13 samples collected from 20 feet to 50 feet deep (referred to as shallow samples) and 12 samples from a depth of 60 to 130 feet (referred to as deep samples) from many of the sites from where surface sample originated.

Measurement of pH, temperature, and depth of samples

Temperature and depth measurements were accura-tely measured for each water sample using a dive computer. The pH of the sample was measured uti-lizing a standard pH paper.

Preparation of medium for isolation of Vibrios

A selective and differential medium called CHRO MagarTM Vibrio purchased from Dalynn Biologicals, Calgary, Alberta was used. The four common species of Vibrio, V. alginolyticus, V. parahaemolyticus, V. cholerae and V. vulnificus are differentiated on the CHROMagar plate by colony colour; V. alginolyticus appears colour less or opaque white, V. parahaemo-lyticus appears mauve, and V. cholerae/ V. vulnificus both appear blue on this medium. The medium was prepared as per CHROMagarTM Vibrio protocol. A quantity of 37.3 Grams of the medium was mixed into 0.5L of sterile boiling water in a 500 mL Erle-nmeyer flask. This mixture was allowed to cool before it was poured into sterile polystyrene petri dishes (100 mm X 10 mm). After the media had solidified, the dishes were stored inverted at 4OC.

Incubation temperature

At the commencement of this project, it was nece-ssary to determine the desired incubation temper-ature.  One ml quantities of three water samples were inoculated on plates of CHROMagarTM  with the help of a sterile glass spreader and plates incubated at 370C in the Laboratory of Department of Health, Bonaire (Bonlab) and observed for growth of vib-rions after 24-30 hours of incubation. These were compared to samples from the same sites that were similarly inoculated and incubated at room temp-erature (25-300C) in the laboratory of SJSM. Growth was comparable on the plates; therefore, incubation at room temperature in SJSM Lab was used for all other samples.

Processing of samples

The petri dishes were allowed to come to room tem-perature and 1.0 mL of each sample was placed on the plate using sterile plastic 3 mL pipettes. A glass spreader, sterilized using isopropyl alcohol and flame, was used to evenly spread the samples over the plates. The plates were then incubated inverted at 30OC for 24 hours. To determine if incubating at 30OC was comparable to the standard 37OC four identical water samples were incubated at BonLab at 370C for 24 hours and compared. Post incubation the number of colony growths on the plates were record-ed. One additional plate with no sample applied was used as a control for this study and incubated at BonLab at 370C to assess possible contamination of the plates during preparation. Finally, two water samples were tested: one from tap water supplying each household and another sample of inland water consisting of overflow effluent from the islands salt drying fields. To determine the total bacterial count, one ml quantities of each of four representative samples including one each collected from the north (Oil Slick), south (Chez Hines), east (Sorobon) and west (Yellow submarine) areas of the island were plated onto a standard nutrient agar (non-specific media) plate and incubated at 30OC for 24 hrs. Each individual growing colony was considered to have originated from one bacterium and the bacterial count calculated. Gram staining was performed for three samples.  One was of blue colony of the sample from Karels site on CHROMagarTM (Vibrio chol-erae/ Vibrio vulnificus). Three were of the isolated colonies on Nutrient agar of the samples from Oil slick, Sorobon and Yellow submarine sites. Data ana-lysis included constructing graphs and pie charts to summarize the raw colony counts from the various collection sites.  First, a graph depicting the number of sampling sites that containing relative amounts of each V. alginolyticus, V. parahaemolyticus and V. vulnificus / V. cholerae was constructed Nutrient agar. Lastly, a graph depicting numbers of colonies of the various Vibrio bacteria that divides the sites based on direction (north, south, east, and west) was constructed.

Oxidase testing of Vibrio isolates

The CHROMagarTM Vibrio protocol suggested using oxidase testing to provide further proof of Vibrio identification.  Oxidase testing was performed on one white colony, one pink colony and one blue colony. These were inoculated onto oxidase discs and a blue colour change indicated a positive oxidase reaction. The fourth oxidase test was conducted as a control to determine that the oxidase test materials themselves were accurate and was performed on a sample from the nutrient agar. This sample showed Gram positive, catalase positive cocci and coagulase negative. It was presumed to be either S. epidermidis or S. sapro-phyticus, both being oxidase negative.

KB007 HiVibrioTM Identification Kit testing and confirmation by TOFEL-MALDI

In a final attempt to further determine greater speci-ficity of the various Vibrio bacteria isolated from the CHROM TM Vibrio, five fresh samples were utilized in KB007 HiVibrioTM Identification Kit.  Four of the samples were completed together, the samples were first plated on CHROMagarTM Vibrio to isolate Vibrio bacteria, and these were then inoculated on to nutria-ent agar and allowed to grow.  After incubation, the samples were then inoculated into the wells of the testing kit following KB007 HiVibrioTM Identi-fication Kit protocols.  The fifth sample was plated onto CHROMagarTM Vibrio and then directly ino-culated into the wells of the testing kit. Further Confirmation by TOFEL-MALDI was accomplished in Division of Microbiology, Department of Micro-biology, PGI, Chandigarh, India. The number of colonies of pathogenic Vibrio species in samples from different locations in Bonaire (with their pH, depth, and temperature of the samples) is provided in Table 1. Number of colonies of pathogenic Vibrio species in samples from different locations in Bona-ire (with their pH, depth, and temperature of the samples) are reported in Table 2. The average temp-erature for the surface, shallow and deep samples was 82.20F±0.9, 79.80F±0.6 and 80.30F±0.8 respect-ively.  The average pH for surface and shallow samples was 7.1±0.3 whereas that for all deep samples was 7 (Table 1) Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio vulnificus/Vibrio chol-erae were recovered on CHROMagarTM plates ino-culated with marine water samples from almost all the sites tested.  There were only two samples that did not yield any vibrio growth. The control tested for plate contamination during their production was negative thus confirming that the colonies that grew were in fact bacteria from the test samples. To confirm that the islands drinking water supply was safe from Vibrios, a plate was inoculated with a simple tap water sample; it did not show growth of any Vibrio species. The number of samples positive for pathogenic vibrio species in depicted in Fig. 1.  It was found that 40 of the samples contained V. alg-inolyticus, 33 contained V. parahaemolyticus and 29 yielded blue colonies representing either V. vulnificus or V. cholerae. It should be noted here that differ-entiation between V. vulnificus and V. cholerae could not be performed with the Vibrio identification kit. The plates were further examined for the number of various colonies present in the water samples accor-ding to sites of collection of the samples. The sample obtained from oil slick (west) had approximately 80 distinct colonies present 

(http://www.cdc.gov/features/ds.foodnet%202012). (http:www.food safetynews.com/2014/). The sample obtained from Sorobon (east) had approximately 120 distinct colonies. Samples from Chez Hines (south) and Yellow Submarine (west) yielded too numerous colonies to count. Fig. 2 shows the percentages of the Vibrio species present at the surface of the water at different sites. It was determined that 47.4% of the colonies were V. alginolyticus, 35.2% were V. parahaemo-lyticus and 17.4% represented V. vulnificus/ V. cho-lerae. Next, to assess a breakdown of the number of various colonies per sample plate, the percentages for each sample were examined in consideration to the depth of the sample collection. 25 surface samples from various sites and among these 14 of them had a colony count percentage of 50% or greater number of V. alginolyticus.  Further, there were 10 sites that had a colony count percentage of 50% or greater for V. parahaemolyticus, 6 sites that had a colony count percentage of 50% or more for V. alginolyticus, 4 sites had a colony count percentage of 50% or more for V. parahaemolyticus and 2 sites (Alice in Won-derland and Margate Bay) had a col-ony count of 50% or more for V. vulnificus/ V. cholerae, and finally there were 3 sites, viz, Chez Hines, Karels, and Sweet Dreams that had a colony count per-centage of 50% or greater for V. vulnificus/ V. cho-lerae.  There was one site (The Lake) that did not grow any Vibrio species. Lastly regarding the sam-ples from deep sites, it was observed that 5 sites contained a colony count percentage of 50% or more for V. alginolyticus, 4 sites had a colony count per-centage of 50% or more for V. parahaemolyticus. 

There were 2 sites, the lake and Berri reef that had a colony count percentage of 50% or more for V. cholerae. Once again there was one site (Acquarius) that did not grow any Vibrio. Finally, regarding the total colony counts from each site delineating the prevalence of the various Vibrio bacteria according to geographic direction, the East and North, the grossly demonstrated V. alginolyticus and V. paraha-emolyticus as the predominant species. Although differentiation between V. vulnificus and V. cholerae was not possible due to limited testing facility, we confirmed that these colonies as well as colonies of other two species, viz. V. alginolyticus and V. para-haemolyticus were also Gram negative and oxidase positive, the characteristics of Vibrio species (Murray et al., 2003). The colony counts in the West were too small to determine any predominant species. The present investigation constitutes the first study of its kind from the Dutch Caribbean, evidencing the prevalence of Vibrio species, viz.  V. alginolyticus, V. parahaemolyticus, and V. vulnificus /V. cholerae in marine water in and around Bonaire. Coexistence of these species in water samples from some of the sites is a noteworthy observation. The presence of these organisms in the marine water was not unexpected due to their ubiquitous nature (Depaula et al., 2010; Elhad, 2013; Oliver et al., 1982). Vibrio fulnicus, an opportunistic human pathogen is commons in estua-rine environment existing as symbiont of oysters (Wright et al., 1996). Ingestion of oysters can result in septecaemia in 95% of persons with underlying chronic diseases, resulting in more than 59% morta-lity (Wright et al., 1996). Several Vibrio species are of great interest to the health departments of most countries since the presence of a few pathogenic strains can cause significant morbidity and mortality to those who are exposed and susceptible (Ho Ho 2021; http://www.cdc.gov/features/dsfoodnet 2012/;  http:/www.foodsafetynews.com/2014/08/; Oliver, 2005). Monitoring of Vibrio bacterial infection among sea foods is particularly important to reduce this risk (De Paula et al., 2010; Cheshire, 2014; Romesh et al., 2014). Thus, due to the ubiquitous nature of Vibrio species, proper food safety techniques should be used when consuming seafood because Vibrios can be detected even in frozen seafood (Khalil et al., 2014). Two studies from the Caribbean also found Vibrio alginolyticus and Vibrio vulnificus present on corals (Cervino et al., 2008; Morrow et al., 2012). 

An increase in yellow blotch/band disease in corals caused by the rise in water temperature was also observed in some parts of the Caribbean including Bonaire (Cervino et al., 2008). It is note-worthy that the sample from Sorobon where the water temp-erature is 4OC higher than that in other sites showed significantly elevated bacterial counts for V. para-haemolyticus and V. alginolyticus. Interestingly, the shallow waters (depth of 1 to 2 feet) of the effluent overflow from the salt evaporation fields yielded higher counts of V. alginolyticus than that of V. vulnificus was also observed in North Carolina during drought periods when salinity values increase (Drake, 2008). Vibrio vulnificus and V. parahaemo-lyticus are known to be very pathogenic for humans (Poirier et al., 2012). It is interesting to note that pan-demic Vibrio parahemolyticus from UK water and shellfish produce (Powel et al., 2013). Bonaire poss-esses water-processing facilities that desalinate sea-water to provide drinking water since there are no natural water reservoirs on the island. We performed a test on this water to assess for Vibrio content and but did not detect any, possibly because this water would not have encountered wastewaters. When the inhabitants of the island use inland waters as a water source, as in other islands, weather disturbances can cause contaminations and cholera outbreaks as seen in Haitis past (CDC 24/7. Saving lives and protec-ting people). There is also a greater risk of infections when lacerations and open wounds become exposed to Vibrio bacteria. The increased number of scuba diving and snorkeling by tourists as well as local fishermen being exposed to the coastal waters with open skin injuries pose a significant health risk for infection and septicaemia (http://www.cdc.gov/fea tures/dsfoodnet2012/). It is important for the local healthcare authorities to be aware of the presence of potentially pathogenic Vibrio species present in the local waters.

CONCLUSION

This study supports that Vibrios exist as autoch-thonous species throughout the coastal waters rang-ing from the colder water of the north-eastern US to the Gulf. Of the species under investigation, the most prevalent was Vibrio alginolyticus in both number of sites and number of colonies; Vibrio alginolyticus was also the most prevalent organism (38%) in a study performed in Saudi Arabia (Elhadi, 2013). This is significant observation because alginolyticus ca-uses Yellow Blotch disease among coral species (Elhadi, 2013). Bonaire corals may already be at risk for developing this disease based on the amount of Vibrio alginolyticus present. The spread of Vibrio infections in response to climate change and extreme heat waves has been observed in several countries in the North Atlantic and North Sea regions, including the Caribbean countries. Increased water temperat-ures caused by atmospheric warming and increased salinity gradients caused by sea-level rise raise con-cerns about the effect of climate change on the geo-graphic range of Vibrio vulnificus, a species asso-ciated with oyster and shellfish and occurring in the US southeast, and the potential for increased exp-osure risk of human infections and (Froelich and Oliver, 2013; Deeb et al., 2018). A noteworthy pub-lication is the report of the 0ccurrence and virulence properties of Vibrio and Salinivibrio isolates from tropical lagoons of the southern Caribbean Sea (Fernández-Delgado et al., 2017). Of particular con-cern to human health are Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus, though most environmental strains are not pathogenic. Our study has established the presence of Vibrio species in the coastal waters of Bonaire. The rising preval-ence of pathogenic Vibrio species in marine waters has led to a surge of Vibrio infections in humans. Another particularly noteworthy observation revealed in the literature search is the first-time reporting of Vibrio jasicida and V. rotiferianus at UK shellfish sites (Harrison et al., 2022).

Table 1: Number of colonies of pathogenic Vibrios species in marine water samples (with data on the pH, depth, and temperature of the samples) in different locations in Bonaire.

The mean and standard deviation (SD for depth and temperature of different types of water were as follows: Surface water - pH: mean = 7.1, SD + 0.3 Depth: mean = 0, SD = 0. Temperature: mean = 82.2, SD = 0.9, Shallow water - Depth: Mean =40.4, SD =9.7, Temperature: Mean =79.8. SD = 0.6. (There was no variation in pH), Deep water – Depth: Mean = 95 there -8, SD = 15.9, Temperature: Mean = 80.3, SD = 0-8 (There in the parenthesis is no variation in pH).

Table 2: Number of colonies of pathogenic Vibrio species in samples from different locations in Bonaire (with their pH, depth, and temperature of the samples.

Fig. 1: This graph depicts the number of samples positive for pathogenic Vibrio species.

Fig. 2: Bar graph showing the percentages of the Vibrio species present at the surface of the water at different sites.

In this figure, the X-axis contains the name of the site where the sample was collected and the Y-axis is the percentage of each species based on the number of colonies counted in that sample. V. alginolyticus is represented by green, V. parahaemolyticus is repre-sented by pink, and V. vulnificus/ V. cholera is repre-sented by blue

ACKNOWLEDGEMENT

We are grateful to Prof. Kallol Guha, President, and CEO of Saint James Schools of Medicine (SJSM) for providing the facilities, and to Dr. Branca Filipoivc, Director of Research (SJSM) for approving the res-earch project related to this project. Prof. Kallol kindly funded the trip of Harish C Gugnani from Delhi to Chandigarh to conduct a part of the study at PGI, Chandigarh. We also wish to thank the staff of Bon-Lab, Kralendijk for allowing us to use their equipment during this project.

CONFLICTS OF INTEREST

The authors have no conflict of interest with any individual or organization.

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Article Info:

Academic Editor
Dr. Abduleziz Jemal Hamido, Deputy Managing Editor (Health Sciences), Universe Publishing Group (UniversePG), Haramaya, Ethiopia.

Received

August 14, 2022

Accepted

September 23, 2022

Published

October 2, 2022

Article DOI: 10.34104/ejmhs.022.01530162

Corresponding author

Harish C Gugnani*

Emeritus Professor, Saint James School of Medicine (SJSM), Anguilla (BWI); and Professor of Microbiology, V.P. Chest Institute, University of Delhi, Delhi-110007, India.

Cite this article

Gugnani HC, Chopra Q, Currie A, Ostrander T, Wiegersma W, and Ray P. (2022). Occurrence of Vibrio species in marine sources surroundings Bonaire, Dutch Caribbean. Eur. J. Med. Health Sci., 4(5), 153-162. https://doi.org/10.34104/ejmhs.022.01530162 

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