Enteropathogenic vibrios: A Growing Worldwide Public Health Risk

November 24, 2021

WHAT ARE THE ENTEROPATHOGENIC VIBRIOS?

Members of the genus Vibrio are Gram-negative mobile rods, usually producing oxidase and catalase. Over 100 Vibrio species have been described and around a dozen of these have been demonstrated to cause infections in humans (Codex, 2010; Hartnell et al., 2019). The species most associated with foodborne infections include V. parahaemolyticus, V. vulnificus and V. cholerae (FDA-2012).

Pathogenic V. parahaemolyticus strains carry virulence genes coding for the thermostable direct hemolysin (tdh) or the tdh-related hemolysin (trh), or both. Virulent strains represent less than 3 % of all V. parahaemolyticus strains isolated from the environment or foods (EFSA, 2012).

The pathogenic mechanisms of V. vulnificus have not been clearly elucidated, and its virulence appears to be multifaceted; Therefore, all strains are considered virulent (Codex, 2010).

The only serogroups of pathogenic V. cholerae currently recognized as causing significant cholera outbreaks are O1 (mostly the Inaba and Ogawa serotypes) and O139 (FDA,2012; Codex, 2010). The key determinants of V. cholerae virulence, the cholera toxin and the toxin co-regulated pilus are parts of horizontally acquired Mobile Genetic Elements (Kumar et al.,2020). Non-O1/O139 V. choleraestrains are less pathogenic (EFSA, 2012).

WHAT ARE THE RISKS FOR THE CONSUMERS?

Enteropathogenic vibrios’ virulence are responsible for the gastrointestinal symptoms, that is, abdominal cramps, nausea, vomiting, fever, mild to severe watery diarrhea observed during the infection (FDA, 2012).They mainly induce huge losses of ions and water from the epithelium cells of the human intestines.

The gastroenteritis is generally mild or moderate and self-limiting in healthy people. However, in susceptible people (immunocompromised states, diabetes, AIDS, etc.), the infections can be more severe and require hospitalization as possibly leading to septicemia and death.

V. vulnificus and V. parahaemolyticus also can cause wound infections either through wounds resulting from handling fish, crustaceans, or shellfish, or when a pre-existing wound is exposed to contaminated marine or estuarine waters. Beyond inflammation at the wound site, the infection can become systemic, with affected people developing fever, altered mental status, and hypotension. Resulting lesions frequently require surgical debridement or amputation (FDA, 2012).

Ingestion of large numbers (10 ⁶ - 10 ⁹) of Vibrio cells is usually needed to develop an infection. However, even the initial contamination level could be low on a product contaminated through sea water, the risk of infection increases rapidly due to the high growth rate of Vibrio species, especially at inappropriate storage conditions (EFSA, 2012). A correlation exists between probability of infection and warmer months when water temperatures are greater than 15°C (FDA, 2012).

Vibrio is recognized as a primary cause of bacterial gastroenteritis associated with seafood consumption in many areas of the world, including Asia and the U.S. The number of infections related to these pathogens has increased since 1990 (Codex, 2010; EFSA, 2012). There is a growing body of evidence to suggest that climate change, coupled to epidemiological and demographic factors are increasing the geographical spread as well as incidence of these pathogens from foodborne sources (Hartnell et al., 2019).

KEY FIGURES: 

The US CDC estimated that around 50,000 foodborne Vibrio infections occur each year in the USA and that hospitalization and death rates range from 22,5% / 0,9 % for Vibrio parahaemolyticus (86% foodborne) up to 91,3 % / 34,8 % for V. vulnificus (47% foodborne) (Scallan et al., 2011). Compared to other major foodborne pathogens, the number of Vibrio infections also appears to be steadily increasing; Compared with 2016–2018, the US Vibrio incidence in 2019 (0,9 cases / 100,000) increased significantly (79%) (CDC, 2020).

HOW ARE ENTEROPATHOGENIC VIBRIOS TRANSMITTED?

V. parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus are common residents of tropical and temperate marine environments; Their distribution is strongly associated with water salinity. V. cholerae occupies freshwater and brackish habitats, mostly introduced through human feces, whereas V. vulnificus is present in environments with intermediate levels of salinity and V. parahaemolyticus shows preference by more saline environments and may be present in offshore waters (EFSA, 2012). Epidemic V. cholerae is confined to developing countries with warm climates (Codex, 2010).

Vibrios are concentrated in the gut of filter-feeding molluscan shellfish, such as oysters, clams and mussels, where they can multiply (EFSA, 2012).

Typically, Vibrio infections are initiated from exposure to seawater, consumption of raw or undercooked seafood produce (Hartnell et al., 2019) or drinking waters contaminated with V. cholerae (Codex, 2010; FDA, 2012).

WHAT ARE ALL FOOD INDUSTRIES AFFECTED BY ENTEROPATHOGENIC VIBRIOS?

Foods associated with Vibrio infections include crayfish, lobster, shrimp, fried mackerel, mussel, tuna, seafood salad, raw oysters, clams, steamed/boiled crabmeat, scallops, squid, sea urchin, and sardines. 

These products include either raw and partially treated seafood (heat treatment, high pressure) or even thoroughly treated seafood products later exposed to cross-contamination through contaminated food production environments.

Improper refrigeration of seafood products contaminated with Vibrio may allow the rapid proliferation of bacterial cells, due to their high growth rates, with a corresponding increased risk of infection. Vibrio can survive for extended periods in chilled and frozen foods (EFSA, 2012, 2021; FDA, 2012).

HOW CAN ENTEROPATHOGENIC VIBRIOS BE PREVENTED AND CONTROLLED IN THE FOOD INDUSTRY?

The food safety concerns associated with enteropathogenic vibrios and seafood have led to the development by scientific or regulatory bodies of specific risk assessments and potential risk management strategies for their control (FDA, 2005, 2019, 2021; Codex, 2010; EFSA, 2012; FAO, 2016).

All this supports either specific regulations concerning seafoods products (21CFR part 123) or the broader requirements of EU and US regulations i) for the prevention of adulteration (EU 178/2002; Federal Food, Drug & Cosmetic Act,1938; FDA-FSMA, 2011) and ii) for the mandatory implementation of HACCP- based control systems and Good Hygiene/Manufacturing practices (FDA 21CFR 1 et al.; EU 852/2004).

HOW CAN THE PRESENCE OF ENTEROPATHOGENIC VIBRIOS BE DETECTED IN THE FOOD INDUSTRY?

EU and US regulations further require that food business operators shall perform microbiological testing as appropriate when they are validating or verifying the effectiveness of their HACCP-based control procedures and good hygiene practices (EU 852/2004, 2073/2005; FDA-FSMA, 2011).

Then, even without any Vibrio formal regulatory microbiological criteria (EFSA, 2012; Hartnell et al, 2019; FDA, 2021b), microbiological monitoring for enteropathogenic Vibrio is often put in place by regulatory agencies (EU 625/2017) and by food business operators (FDA, 2019; FCD, 2020) primarily for customer protection but additionally for the prevention of both mandatory product recall (FDA-FSMA, 2011; EU 178/2002) and legal prosecution.

Different microbiological methods either traditional or molecular for the detection, the enumeration, the species identification and the screening for Vibrio virulence determinants have been elaborated to support the management of the enteropathogenic vibrios at different stages, from the monitoring of Vibrio in harvest-areas to the investigation of Vibrio outbreaks, including the post-harvest process verification and the end-product testing. 

They include both:

• national or international standard methods (ISO 21872:2017 and 2020, Hartnell et al., 2019, FDA, 2004, 2017; US-EPA, 2010, FAO, 2016),
• and validated (AOAC) alternative methods developed for the same purposes. These methods additionally bring ease of use and/or reduced times to results compared to the standard methods, offering flexibility to the management of the business flows.

bioMérieux provides the food safety managers with actual microbiological solutions for the effective management of enteropathogenic Vibrio risks along the Food Chain.

BIOMÉRIEUX SOLUTIONS AND PRODUCTS

Sample and culture media preparation:

• DILUMAT^® gravimetric diluter
• SMASHER^®  lab blender
• MASTERCLAVE^® automated media preparator

Traditional Culture media: Large range of traditional culture media

• REF 04189 Alkaline Peptone Water (APW) For the selective enrichment and cultivation of Vibrio species - 10 x 10 ml tubes
• REF AEB153182 TCBS For the detection of Vibrio in food - 500g

Chromogenic culture media compliant with ISO 21872-1:2017:

• CHROMID® Vibrio Agar (VID) can be used as second medium of the laboratory own choice, in parallel to TCBS, as it is recommended by the ISO 21872-1:2017 standard and the International Journal of Food Microbiology 157 (2012) 189–194 (Rosec J.Ph et al.) 

BIOBALL® - Standardized Strains for food applications:

- BIOBALL® A small freeze dried water-soluble Certified Reference Material containing a precise number of viable micro-organisms for your Microbiological Quality Controls

- BIOBALL® LUMINATE 2.0 Green Fluorescent Protein Strains (GFP), Genetically Modified Microorganisms (GMM) to distinguish from natural contaminants

Identification:

• API^® biochemical galleries: API 20E, API Rapid 20E, ID32E, Rapid ID32E  
• VITEK^® 2 GN automated biochemical system
• VITEK^® MS MALDI-TOF technology

REFERENCES

CDC. Preliminary Incidence and Trends of Infections with Pathogens Transmitted Commonly Through Food - Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 2016–2019. MMWR, 2020, 69 (17): 509-514.

CODEX Alimentarius. Guidelines on the application of general principles of food hygiene to the control of pathogenic Vibrio in seafood. CAC/GL 73-2010. 2010. 14p.

EFSA. Scientific Opinion on the minimum hygiene criteria to be applied to clean seawater and on the public health risks and hygiene criteria for bottled seawater intended for domestic use. EFSA Journal 2012;10(3): 85 pp.

EFSA (European Food Safety Authority) and ECDC (European Centre for Disease Prevention and Control). The European Union One Health 2019 Zoonoses Report. EFSA Journal 2021;19(2):6406, 286 pp.

EU Regulation 178/2002 laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety.

EU Regulation 852/2004 on the hygiene of foodstuffs.23 p.

EU REGULATION 625/2017 on official controls and other official activities performed to ensure the application of food and feed law.

FAO (Food and Agriculture Organization of the United Nations) - WHO (World Health Organization). Selection and application of methods for the detection and enumeration of human-pathogenic halophilic Vibrio spp. in seafood. 2016. 91p.

FCD (Fédération du Commerce et de la Distribution). Critères microbiologiques applicables à partir de 2021 aux marques de distributeurs, marques premiers prix et matières premières dans leur conditionnement initial industriel. 2020.62p.

FDA Food Safety Modernization Act (FSMA) – Public Law to amend the Federal Food, Drug, and Cosmetic Act with respect to the safety of the food supply. 2011. 89 p.

FDA. Bad Bug Book. Foodborne Pathogenic Microorganisms and Natural Toxins. 2012.
. Vibrio parahaemolyticus. 4 p.
. Vibrio vulnificus. 4 p.
. Vibrio cholerae serogroups O1 and O139. 4 p.

FDA. 21 CFR 123. Fish and fishery products.2020. 9 p.

FDA. Seafood HACCP and the FDA Food Safety Modernization Act: Guidance for industry, 2017. 12 p.

FDA. Quantitative Risk Assessment on the Public Health Impact of Pathogenic Vibrio parahaemolyticus In Raw Oysters. 2005. 29p.

FDA. National Shellfish Sanitation Program (NSSP) - Guide for the Control of Molluscan Shellfish, 2019 Revision. 502 p.

FDA a. Guidance for the Industry: Fish and Fishery Products Hazards and Controls Guidance, Fourth Edition. 2021. 542 p.

FDA b. Guidance for the Industry: Fish and Fishery Products Hazards and Controls Guidance, Fourth Edition. Appendix 5: FDA and EPA safety levels in regulations and guidance. 2021: 14p. 

FDA. 21CFR Parts 1, 11, 16, 106, 110, 114, 117, 120, 123, 129, 179, and 211. Food Safety Modernization Act. Current Good Manufacturing Practice, Hazard Analysis, and Risk-Based Preventive Controls for Human Food. 2015.80 (180): 55908-56168.

FDA BAM (Bacteriological Analytical Manual) - Chapter 9 - Vibrio. 2004. 41 p.

FDA BAM - Chap 28 - Detection of enterotoxigenic Vibrio cholerae. 2017. 8 p.

Federal Food, Drug & Cosmetic Act. To prohibit the movement in interstate commerce of adulterated and misbranded food, drugs, devices, and cosmetics, and for other purposes. 1938.

Hartnell RE et al. A pan-European ring trial to validate an International Standard for detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus in seafoods. J Food Microbiol. 2019, 2;288:58-65. 

ISO 21872-1:2017 Microbiology of the food chain - Horizontal method for the determination of Vibrio spp - Part 1: detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus.

(expected in 2021) Project ISO 21872-1:2017/AMD1 Microbiology of the food chain—Horizontal method for the determination of Vibrio spp -Part1:Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholera and Vibrio vulnificus – Amendment1:Inclusion of performance testing for ASPW, TCBS and SNA
 
ISO/TS 21872-2:2020 Microbiology of the food chain - Horizontal method for the determination of Vibrio spp. - Part 2: Enumeration of total and potentially enteropathogenic Vibrio parahaemolyticus in seafood using nucleic acid hybridization.

Kumar A. et al. Vibrio Pathogenicity Island-1: The Master Determinant of Cholera Pathogenesis. Front. Cell. Infect. Microbiol., 2020, 10 : Article N°561296.

Rosec J.Ph et al. The international standard ISO/TS 21872–1 to study the occurence of total and pathogenic Vibrio parahaemolyticus and Vibrio cholerae in seafood: ITS improvement by use of a chromogenic medium and PCR. International Journal of Food Microbiology 157 (2012) 189–194

Scallan E., et al. Foodborne illness acquired in the United State, major pathogens. Emerging Infectious Diseases, 2011; 17:7–15.

U.S. Environmental Protection Agency. Standard analytical protocol for Vibrio cholerae O1 and O139 in Drinking Water and Surface Water., Washington, DC, EPA/600/R-10/139, 2010. 56 p.