Printer Friendly

Prevalence of Vibrio Parahaemolyticus in Various Seafood Consumed in North Cyprus.

INTRODUCTION

Vibrio parahaemolyticus is a human enteropathogenic, sucrose non-fermenting, facultative, and halophilic bacterium that is widely distributed in both marine and estuarine habitats and is present in seafood harvested from aquatic environments worldwide (1, 2). This marine-based enteropathogenic bacterium is responsible for most of the seafood-borne bacterial illnesses leading to gastrointestinal problems such as nausea, vomiting, abdominal cramps and watery or bloody diarrhea sometimes accompanied with fever (3).

The debilitating effects of V. parahaemolyticus are because of the presence of virulence genes ([t.sup.dh] and [t.sup.rh]), type III secretion systems (T3SS1 and T3SS2), clonal serotypes (O3:K6 and its serovariants), and extracellular proteases (4-9). Antibiotic treatment is not usually needed for V. parahaemolytiticus poisoning, however, in cases with prolonged diarrhoea, antibiotic therapy can be given.

A significant number of individuals worldwide depend on seafood as a primary source of valuable nutrients, particularly protein, poly unsaturated fatty acids, vitamins, and minerals (10). Virtually, the nutritional value of seafood has led to its worldwide acceptance and excessive consumption. In recent years, the world consumption per capita of marine and aquaculture fishery products has reached over 20 kg a year (11). Accordingly, European Market Observatory for Fisheries and Aquaculture Products (EUMOFA) reported similar trends in the consumption per capita of 23.9 kg in the EU member states (12).

According to a report released by the Food and Agriculture Organization of the United Nations (FAO), Mediterranean seafood production has increased in the previous decades because of the large production of sea bream and sea bass (13). Concurrently, an epidemiological report has shown that among seafood-borne pathogens, V. parahaemolyticus present a significant threat, and shrimp and finfish are high on the list of the most contaminated seafood (2, 14). Surveying, monitoring, and detecting pathogens in foods are the most important approaches to reduce, control, or prevent foodborne bacterial infections (15). Bacterial infections mostly because of the consumption of fish and shellfish have been attributed to pathogenic Vibrios (16).

In the Turkish Republic of Northern Cyprus (TRNC), like other Mediterranean countries, the most important finfish consumed are sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax). Among shellfish, shrimp is the most widely consumed. To the best of our knowledge, the occurrence of V. parahaemolyticus in the seafood consumed in the TRNC has never been investigated. Hence, the present study aimed to investigate the occurrence of V. parahaemolyticus in retail sea bream (S. aurata L.), European sea bass (D. labrax), and shrimps. The various seafood caught along the coast of the Mediterranean Sea in Famagusta and Kyrenia have also been included in the present study.

MATERIAL and METHODS

Seafood samples in this study include European sea bass (D. labrax), gilt-head sea bream (S. aurata L.), blue whiting (Micromesistius poutassou), marbled spinefoot (Siganus rivulatus), mackerel (Scomber scombrus), and shrimp. These fish varieties were selected because they are widely consumed and are available throughout the year. Seafood samples were taken from major seafood outlets of Famagusta, Kyrenia, Nicosia, and Morphou regions and also directly from the coasts and/or bays of the Famagusta and Kyrenia regions (Table 1). Five randomly selected fishes and 10 randomly selected shrimps (totaling approximately 1 kg) were taken from each shop's daily sold during Summer and early-Fall seasons of 2016. These representative samples were drawn in accordance with standardized procedures for fresh seafood sampling (17, 18). This study was carried out according Helsinki Declaration.

The isolation and identification of V. parahaemolyticus by a conventional culture technique was done in accordance with Food and Drug Administration/Bacteriological Analyses Manual (FDA/BAM) (19). Seemingly, apathogenic and pathogenic bacteria live on the skin, in the gills, and in the intestines of fish. Therefore, the gills and intestines from each fish sample were removed and then separately homogenized in 225 mLof alkaline peptone water (APW) with 3% NaCl for 1 min.

Shrimp samples were thoroughly washed under running water and shucked for sampling according to Cook et al. (20). Samples were comminuted to get a puree representing the whole sample location. A total of 25g puree was taken and then homogenized in 225 mL of APW. The fish and shrimp homogenates were transferred into sterile polythene stomacher bags and incubated in an incubator (Thermo Scientific, Massachusetts, USA) at 37[degrees]C for 24h. After incubation (24 h), 1 mL of each homogenate was taken aseptically using a sterile wooden cotton applicator stick and streaked onto sterile surface thiosulfate-citrate-bile salts-sucrose (TCBS; Liofilchem s.r.l., Teramo, Italy) agar plates. The plates were then incubated at 37[degrees]C for 24h.

Following plate incubation, TCBS plates were checked for suspect colonies that were sucrose non-fermenting with a green or bluish-green color and a dark blue or green center approximately 3-5 mm long indicating the presence of V. parahaemolyticus. Such colonies were carefully selected. The suspect colonies were purified and further characterized by performing catalase and gram-staining tests. Suspect isolates that were positive for catalase and that were gram-negative-stained were selected for biochemical identification and confirmation. Vibrio parahaemolyticus ATCC 17802 was used as control (Figure 1a). Suspect isolates were screened by automated identification and antimicrobial testing (BD Phoenix, Franklin Lakes, USA).

RESULTS

V. parahaemolyticus was not detected in any of the examined fish and shrimp samples, but other gram-negative bacteria were detected in the intestines of sea bass from Kyrenia and sea bream from Morphou regions. Following a series of biochemical tests and by the BD Phoenix Identification Instrument, three bacterial species, including Photobacterium damselae (formerly V. damsela), Providencia rettgeri, and Pseudomonas fluorescens, were confirmed. Two of these bacteria, namely, P. damsalae and P. rettgeri, are pathogenic in both humans and animals. Results for seafood species, locations, and pathogens are presented in Table 2 and suspected bacterial colonies on TCBS agar in Figure 1b.

DISCUSSION

Surveying, monitoring, and detecting pathogens in foods are the most important approaches to reduce, control, or prevent foodborne bacterial infections (15). Bacterial infections mostly because of the consumption of fish and shellfish have been attributed to pathogenic Vibrios (16).

Fortunately, V. parahaemolyticus was not found in any seafood species sampled in our study, although a study from the United States reported elevations in the number of Vibrio infections associated with seafood (16). In Europe, V. parahaemolyticus has been considered to be an emerging foodborne pathogen responsible for most of the recent sporadic and epidemic seafood-borne infections (21, 22). According to Abd-Elghany and Sallam (23), 120 shellfish samples (40 each of shrimp, crab, and cockle) were collected from different fish shops in Mansoura, Egypt and tested for the presence of potentially pathogenic strains of V. parahaemolyticus. The conventional technique as shown by biochemical means showed that 40 (33.3%) out of 120 samples were positive for V. parahaemolyticus. V. parahaemolyticus was found in 4.0% of the winter samples, 13.3% of the spring samples, 18.6% of the summer samples, and 8% of the autumn samples. A significant proportion of shrimps marketed and consumed in Morocco caught in the coastal region of the city of Agadir contained V. parahaemolyticus (24). The absence of V. parahaemolyticus in the shrimps in our study may be because of seasonal variations. However, the results of this study are in agreement with those of a previous study conducted in some European countries where fish samples sourced from France and Great Britain did not contain V. parahaemolyticus (25).

Nonetheless, other aquatic bacterial pathogens, such as P. damselae and P. rettgeri, were found in our fish samples. P. damselae is a pathogen for several species of fish and shellfish. In humans, this bacterium can cause a wide range of infections that may result in necrotizing fasciitis usually with severe clinical consequences. For over two decades, P. damselae has become a threat for several species as well as humans worldwide (26-28). P. rettgeri is one of the major causes of diarrhea in humans. It is also a major source of tetrodotoxin (a potential neurotoxin), predominantly in some Asian countries and recently in Europe, and its abundance in various fish species is widely increasing (29-31).

These bacteria could be isolated because they are also sugar non-fermenting and gram-negative like V. parahaemolyticus and share similar growing conditions in the sea. The incidence of these pathogens could be an alert to maximum exposure to multiple microbial pathogens from seafood (32).

In conclusion, V. parahaemolyticus was not detected in any of the examined fish samples taken from different regions of the TRNC. However, seafood consumed in might be a source of other bacterial pathogens, such as P. damselae and P. rettgeri species, because the concentrations of these bacteria were found to be >[10.sup.5] cfu/mL (minimum infective dose) in the intestines of sea bass and sea bream fishes from Kyrenia and Morphou regions, respectively. It is highly recommended to investigate the occurrence and epidemiology of these species in various seafood products because they are pathogenic in both humans and animals.

Ethics Committee Approval: No ethical committee was consulted because no live animal experiment was conducted in this study, consequently no ethical information can be given.

Informed Consent: N/A

Peer-review: Externally peer-reviewed.

Author contributions: Concept--K.S., P.A., S.S.; Design - K.S., P.A., H.I.K.; Supervision - K.S., P.A.; Resource - P.A.; Materials - K.S., S.S., H.I.K.; Data Collection and/or Processing - H.I.K., M.G.; Analysis and/or Interpretation - H.I.K., M.G., G.C.Z.; Literature Search - H.I.K., M.G.; Writing - H.I.K., M.G., G.C.Z.; Critical Reviews - K.S., S.S., P.A.

Acknowledgement: We appreciate the staff of Near East University Hospital Clinical Microbiology Laboratory, for their technical assistance.

Conflict of Interest: The authors have no conflicts of interest to declare.

Financial Disclosure: The authors gratefully acknowledge the project grants no. 2016-04018 from the Near East University Center of Excellence.

REFERENCES

(1.) Iwamoto M, Ayers T, Mahon BE, Swerdlow DL. Epidemiology of seafood-associated infections in the United States. Clinical Microbiol Rev 2010; 23: 399-411. [CrossRef]

(2.) Odeyemi OA. Incidence and prevalence of Vibrio parahaemolyticus in seafood: a systematic review and meta-analysis. Springerplus 2016; 5, 464. [CrossRef]

(3.) Su YC, Liu C. Vibrio parahaemolyticus: a concern of seafood safety. Food Microbiol 2007; 24: 549-58. [CrossRef]

(4.) Okuda J, Ishibashi M, Hayakawa E, Nishino T, Takeda Y, Mukhopadhyay AK, et al. Emergence of a unique O3: K6 clone of Vibrio parahaemolyticus in Calcutta, India, and isolation of strains from the same clonal group from Southeast Asian travelers arriving in Japan. J Clin Microbiol 1997; 35: 3150-5.

(5.) Makino K, Oshima K, Kurokawa K, Yokoyama K, Uda T, Tagomori K, et al. Genome sequence of Vibrio parahaemolyticus: a pathogenic mechanism distinct from that of V cholerae. Lancet 2003; 361: 743-9. [CrossRef]

(6.) Drake SL, Depaola A, Jaykus LA. An overview of Vibrio vulnificus and Vibrio parahaemolyticus. Compr Rev Food Sci Food Saf 2007; 6: 120-44. [CrossRef]

(7.) Mahoney JC, Gerding MJ, Jones SH, Whistler CA. Comparison of the pathogenic potentials of environmental and clinical Vibrio parahaemolyticus strains indicates a role for temperature regulation in virulence. Appl Environ Microbiol 2010; 76: 7459-65. [CrossRef]

(8.) Letchumanan V, Chan KG, Lee LH. Vibrio parahaemolyticus: a review on the pathogenesis, prevalence, and advance molecular identification techniques. Front Microbiol 2014; 5: 705. [CrossRef]

(9.) Caburlotto G, Suffredini E, Toson M, Fasolato L, Antonetti P, Michela Zambon M, et al. Occurrence and molecular characterisation of Vibrio parahaemolyticus in crustaceans commercialised in Venice area, Italy. Intl J Food Microbiol 2016; 2: 39-49. [CrossRef]

(10.) Sudha S, Divya PS, Francis B, Hatha AA. Prevalence and distribution of Vibrio parahaemolyticus in finfish from Cochin (south India). Vet Ital 2012; 48: 269-81.

(11.) Food and Agriculture Organization of the United Nations, FAO. Global per capita fish consumption rises above 20 kilograms a year, 2016, Rome. Available from: http://www.fao.org/docrep/013/i1820e/i1820e00.htm.

(12.) European Market Observatory for Fisheries and Aquaculture Products, EUMOFA. The EU Fish Market, 2016. Available from https://www.eumofa.eu/.

(13.) Food and Agriculture Organization of the United Nations, FAO. The State of World Fisheries and Aquaculture (SOFIA), 2010, Rome. Available from: http://www.fao.org/docrep/013/i1820e/i1820e00.htm.

(14.) European Commission, EC. Final Report of an Audit Carried out in The United States from 17 March 2015 to 27 March 2015 in Order to Evaluate the Control Systems in Place Governing the Production of Bivalve Molluscs and Fishery Products Derived There from Intended for Export to The European Union, Directorate-General for Health and Food Safety, 12-16, 2015.

(15.) Zhao X, Lin CW, Wang J, & Oh DH. Advances in rapid detection methods for foodborne pathogens. J Microbiol Biotechnol 2014; 24: 297-312. [CrossRef]

(16.) Ronholm J, Lau F, Banerjee SK. Emerging seafood preservation techniques to extend freshness and minimize Vibrio contamination. Front Microbiol 2016; 350: 1-6. [CrossRef]

(17.) International Commission on Microbiological Specifications for Foods, ICMSF. Microorganisms in Foods 2. Sampling for microbiological analysis: Principles and specific applications. Second Edition, London, University of Toronto Press, 192, 1986.

(18.) Canadian Food Inspection Agency (CFIA). Fish Inspection Program Sampling Policy, Standards and Methods Manual Sampling Documents, Canada, 2013. Available from: http://www.inspection.gc.ca/food/fish-and-seafood/manuals/standards-and-methods.

(19.) Kaysner CA, De Paola A, Vibrio cholerae, V. parahaemolyticus, V. vulnificus, and other Vibrio spp, Bacteriological Analytical Manual, 8th ed, Washington, DC, US Food and Drug Administration. Revision A, 1998.

(20.) Cook DW, O'Leary P, Hunsucker JC, Sloan EM, Bowers JC, Blodgett RJ, et al. Vibrio vulnificus and Vibrio parahaemolyticus in US Retail Shell Oysters: A National Survey from June 1998 to July 1999. J Food Prot 2002; 65: 79-87. [CrossRef]

(21.) Baker-Austin C, Stockley L, Rangdale R, Martinez-Urtaza J. Environmental occurrence and clinical impact of Vibrio vulnificus and Vibrio parahaemolyticus: a European perspective. Environ Microbiol Rep 2010; 2, 7-18. [CrossRef]

(22.) Powell A, Baker-Austin C, Wagley S, Bayley A, Hartnell R. Isolation of pandemic Vibrio parahaemolyticus from UK water and shellfish produce. Microb Ecol 2013; 65: 924-7. [CrossRef]

(23.) Abd-Elghany SM, Sallam KI. Occurrence and molecular identification of Vibrio parahaemolyticus in retail shellfish in Mansoura, Egypt. Food Control 2013; 33: 399-405. [CrossRef]

(24.) Kriem MR, Banni B, Bouchtaoui HE, Hamama A, Marrakchi AE, Chaouqy N, et al. Prevalence of Vibrio spp. in raw shrimps (Parapenaeus longirostris) and performance of a chromogenic medium for the isolation of Vibrio strains. Lett Appl Microbiol 2015; 61: 224-30. [CrossRef]

(25.) Davis AR, Capell C, Jehanno D, Nychas GJ, Kirby RM. Incidence of foodborne pathogens on European fish. Food Control, 2001; 12: 67-71. [CrossRef]

(26.) Kim HR, Kim JW, Lee MK, Kim JG. Septicemia progressing to fatal hepatic dysfunction in ancirrhotic patient after oral ingestion of Photobacterium damsel, A Case Report. Infection 2009; 37: 555-6. [CrossRef]

(27.) Serracca L, Ercolini C, Rossini I, Battistini R, Giorgi I, Prearo M. Occurrence of both subspecies of Photobacterium damselae in mullets collected in the river Magra (Italy). Can J Microbiol 2011; 57: 437-40. [CrossRef]

(28.) Rivas AJ, Lemos ML, Osorio CR. Photobacterium damselae subsp. damselae, a bacterium pathogenic for marine animals and humans. Front Microbiol 2013; 4: 283. [CrossRef]

(29.) Kalaitzis JA, Chau R, Kohli GS, Murray SA, Neilan BA. Biosynthesis of toxic naturally-occurring seafood contaminants. Toxicon 2010; 56: 244-58. [CrossRef]

(30.) Tu N, Tu Q, Tung H, Hieu D, Romero-Jovel S. Detection of tetrodotoxin-producing Providencia rettgeri T892 in Lagocephaluspufferfish. World J Microbiol Biotechnol 2014; 30: 1829-35. [CrossRef]

(31.) Yotsu-Yamashita M, Gilhen J, Russell RW, Krysko KL, Melaun C, Kurz A, et al. Variability of tetrodotoxin and of its analogues in the red-spotted newt, Notophthalmus viridescens (Amphibia: Urodela: Salamandridae). Toxicon 2012; 59: 257-64. [CrossRef]

(32.) Tortorello ML. Indicator organisms for safety and quality-uses and methods for detection: mini Review. J AOAC Int 2003; 86, 1208-17.

Hafizu Ibrahim Kademi (1), Grace Charles Zebere (2), Meryem Guvenir (3), Perihan Adun (1), Serdar Susever (3), Kaya Suer (4)

(1) Department of Food Engineering, Near East University School of Engineering, Nicosia, Cyprus

(2) Department of Food Technology, Kaduna Polytechnics, Kaduna, Nigeria

(3) Health Sciences Vocational School, Near East University, Nicosia, Cyprus

(4) Department of Clinical Microbiology and Infectious Diseases, Near East University School of Medicine, Nicosia, Cyprus

ORCID IDs of the authors: H.I.K.: 0000-0002-9141-1803; G.C.Z.: 0000-0001-5865-0880; M.G.: 0000-0002-9702-9947; P.A.: 0001-1000-00010001; S.S.: 0000-0002-4988-5634; K.S.: 0000-0002-2565-3425

Corresponding Author: Kaya Suer

E-mail: kaya.suer@neu.edu.tr

Received: 14.01.2018

Accepted: 27.06.2018

DOI: 10.5152/cjms.2018.338
TABLE 1. Sampling regions in TRNC and number of primary samples taken

Region     Fish species                         Number of primary
                                                samples

Famagusta  Sea bass                              5
           Sea bream                             5
           Shrimp                               10
           Catch of the day: Mackerel            5
           Catch of the day: Marbled spinefoot   5
Kyrenia    Sea bass                              5
           Sea bream                             5
           Shrimp                               20
           Catch of the day: Blue whiting        5
Nicosia    Sea bass                              5
           Sea bream                             5
           Shrimp                               30
Morphou    Sea bass                              5
           Sea bream                             5
           Shrimp                               30
           Catch of the day: Mackerel            5

TABLE 2. Incidence of bacterial pathogens in seafood consumed in the
TRNC

Region     Seafood            Number of samples positive/
                              number of samples examined

Famagusta  Sea Bass           0/5
           Sea Bream          0/5
           Shrimp             0/10
           Mackerel           0/5
           Marbled spinefoot  0/5
Kyrenia    Sea Bass           1/5
           Sea Bream          0/5
           Shrimp             0/20
           Blue whiting       0/5
Nicosia    Sea Bass           0/5
           Sea Bream          0/5
           Shrimp             0/30
Morphou    Sea Bass           0/5
           Shrimp             0/30
           Sea Bream          1/5
           Mackerel           0/5

Region     Identified Pathogen      Concentration of
                                    pathogen (cfu/mL)

Famagusta

Kyrenia    Providencia rettgeri     >[10.sup.5]

Nicosia

Morphou

           Photobacterium damselae  >[10.sup.5]
COPYRIGHT 2018 AVES
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2018 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:Original Article
Author:Kademi, Hafizu Ibrahim; Zebere, Grace Charles; Guvenir, Meryem; Adun, Perihan; Susever, Serdar; Suer
Publication:Cyprus Journal of Medical Sciences
Date:Aug 1, 2018
Words:2914
Previous Article:Patient Satisfaction with Enhanced Recovery after Colorectal Surgery: A Cross-Sectional Analytical Study.
Next Article:Comparison of the Effects of the Sixth and Seventh TNM Staging on Survival in Operable Non-Small Cell Lung Cancer.
Topics:

Terms of use | Privacy policy | Copyright © 2020 Farlex, Inc. | Feedback | For webmasters