International Journal of One Health

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Vibriosis and Aeromonas Infection in Shrimp:Isolation, Sequencing, and Control


Affiliations
1 Department of Animal Hygiene and Zoonoses, Suez Canal University, Ismailia, Egypt
2 Department of Fish Diseases and Management, Suez Canal University, Ismailia, Egypt
 

Background and Aim: Shrimp is one of the most commonly consumed types of seafood. It is a very nutritious healthy food. Shrimp is low in calories and rich in protein and healthy fats. It also contains a treasure trove of vitamins and minerals. On the negative side, it may be affected by many bacterial diseases which affect its health. Furthermore, it may be incriminated as a vector of foodborne illnesses that range from mild gastrointestinal upset to life-threatening diseases. This study was designed to assess the clinical picture and zoonotic importance of vibriosis and Aeromonas infection in live shrimp and to study the antibacterial effect of citric acid (lemon juice) and acetic acid (vinegar) on these pathogens.

Materials and Methods: A total of 170 live shrimp (Metapenaeus monoceros) samples were collected from Suez City, Egypt. The samples were examined clinically, and then, they were enriched into alkaline peptone water and cultivated on thiosulfate-citrate-bile salts-sucrose agar and ampicillin MacConkey agar for the isolation of Vibrio and Aeromonas species, respectively. The recovered isolates were confirmed biochemically and genotypically using duplex polymerase chain reaction (PCR) and sequencing. The germicidal effects of vinegar and lemon on artificially contaminated shrimp samples with Aeromonas hydrophila and Vibrio parahaemolyticus at different times (0.25, 1, 1.5, and 24 h) and temperatures (5° and 30°C) were studied.

Results: The results revealed that some of the infected shrimp were hypoxic, lethargic with abnormal swimming behavior. In most cases, body appendages, telsons, uropods, and gills took black coloration. In addition, the hepatopancreas appeared soft, swollen, and congested. The prevalence rates of vibriosis in each of the musculature and hepatopancreas were 4.7%, while the prevalence rates of Aeromonas infection in the musculature and hepatopancreas were 11.8% and 11.2%, respectively. Duplex PCR showed that Aeromonas isolates gave double bands: 237 bp specific for gcat and 500 bp specific for 16S rRNA, while Vibrio spp. and Plesiomonas shigelloides isolates gave single band at 500 bp. The effect of organic acid treatment showed that acetic acid (vinegar 5%) had increasing reduction rates that reached its maximum level after 24 h; where it caused (100% inhibition) for A. hydrophila at both temperatures and (33.63% and 60% inhibition) for V. parahaemolyticus at refrigerator and room temperatures, respectively. Moreover, acetic acid was more effective at room temperature than at refrigerator temperature. Concerning the effect of lemon juice (citric acid), it was more effective than acetic acid at short marination (0.25 and 1 h) at both temperatures for the two pathogens. Moreover, lemon was more effective at refrigerator temperature than at room temperature at the same aforementioned time. The difference between the reduction effects of the two acids on both pathogens was statistically significant (p<0.0001).

Conclusion: Overall, the examined shrimp samples were found to be vectors for Vibrio and Aeromonas spp. Application of hygienic measures during handling and cooking of shrimp should be esteemed. The organic acid treatment trial showed that vinegar and lemon juice can be used as a safe and economic method to limit the microbial contamination in seafood.


Keywords

Shrimp, Vibriosis, Aeromonas Infection, Sequencing, Decontamination, Citric Acid, Acetic Acid.
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  • Dayal JS, Ponniah AG, Khan HI, Babu EP, Ambasankar K, Vasagam KP. Shrimps-a nutritional perspective. Curr Sci 2013;104:1487-91.

  • Center for Disease Control and Prevention (CDC a). The Final Update on a Multistate Outbreak of Vibrio parahaemolyticus Infections Linked to Fresh Crab Meat; 2018. Available from: https://www.cdc.gov/media/releases/2018/s0927-vibrio-crab.html. [Last accessed on 2018 Oct 24].

  • Gopinath R. Extracellular Products of Aeromonas hydrophila and its Effects on Indian White Prawn, Penaeus indicus. M.Sc Thesis of Fisheries Science (Mariculture). Central Institute of Fisheries Education, (Deemed University); 1999.

  • Brock JA, Lightner DV. Diseases of Crustacea. In: Kinne O, editor. Diseases of Marine Animals. Vol. 3. Ch. 3. Hamburg: Biologische Anstalt Helgoland; 1990. p. 245-424.

  • Alavandi SV, Vijayank K, Rajendran KV. Shrimp diseases, their prevention and control. Ciba Bull 1995;3:1-17.

  • Teng T, Liang L, Chen K, Xi B, Xie J, Xu P, et al. Isolation, identification and phenotypic and molecular characterization of pathogenic Vibrio vulnificus isolated from Litopenaeus vannamei. PLoS One 2017;12:e0186135.

  • Center for Disease Control and Prevention (CDC b). Vibrio Species Causing Vibriosis; 2018. Available from: https://www.cdc.gov/vibrio/faq.htm. [Last accessed on 2018 Oct 24].

  • Public Health of England (PHE). UK Standards for Microbiology, Investigations ID 19: Identification of Vibrio and Aeromonas species; 2015. Available from: https://www.gov.uk/government/publications/smi-id-19-identification-of-vibrio-species. [Last accessed on 2018 Jan 24].

  • Lightner DV. A Handbook of Shrimp Pathology and Diagnostic Procedures for Diseases of Cultured Penaeid Shrimp. Baton Rouge, La, USA: World Aquaculture Society; 1996.

  • Harris LJ, Owens L. Production of exotoxins by two luminous Vibrio harveyi strains known to be primary pathogens of Penaeus monodon larvae. Dis Aquat Org 1999;8:11-22.

  • Zaki VH, Abdelkhalek NK, Shakweer MS. Environmental conditions/bacterial infections relationship and their impact on immune parameters of cultured Fenneropenaeus indicus with special reference to in-vitro antibiotic susceptibility. Int J Fish Aquat Stud 2016;4:51-8.

  • Li P, Kinch LN, Ray A, Dalia AB, Cong Q, Nunan LM, et al. Acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus strains maintain an antibacterial Type VI secretion system with versatile effector repertoires. Appl Environ Microbiol 2017;83:1-17.

  • Selvin J, Ninawe AS, Meenatchi R, Kiran GS. Control of Pathogenic Vibrios in Shrimp Aquaculture using Antiinfectives from Marine Natural Products. México, Nuevo León: Conference: Nutrición Acuícola: Investigación y Desarrollo, Universidad Autónoma de Nuevo León, San Nicolás de los Garza; 2015. p.102-41.

  • Batra P, Mathur P, Misra MC. Aeromonas spp.: An emerging nosocomial pathogen. J Lab Physicians 2016;8:1-4.

  • FDA. Electronic Code of Federal Regulations (ECFR). Part 184-Direct Food Substances Affirmed as Generally Recognized as Safe; 2018. Available from: https://www.fda.gov/food/ingredientspackaginglabeling/GRAS/. [Last accessed on 2018 Nov 24].

  • Ricke SC. Perspectives on the use of organic acids and short chain fatty acids as antimicrobials. Poult Sci 2003;82:632-9.

  • Vinus NS, Tewatia BS. Organic acids as alternatives to antibiotic growth promoters in poultry. Pharm Innov 2017;6:164-9.

  • Animal Research Regulation; 2010. Available from: https://www.legislation.nsw.gov.au/#/view/regulation/2010/425. [Last accessed on 2019 Apr 12].

  • Noga EJ. Fish Disease Diagnosis and Treatment. 2nded. USA: Mosby-Yearbook, Inc. Watsworth Publishing Co.; 2010.

  • Yagoub SO. Isolation of Enterobacteriaceae and Pseudomonas spp. From raw fish sold in fish market in Khartoum state. J Bacteriol Res 2009;1:85-8.

  • Martin-Carnahan A, Joseph SW. Genus I. Aeromonas stanier 1943, 213al. In: Brenner DJ, Krieg NR, Staley JT, Garrity GM, editors. Bergey’s Manual of Systematic Bacteriology. 2nd ed., Vol. 2. Part B. New York: Springer; 2005. p. 557-78.

  • Janda JM, Abbott SL. The genus Aeromonas: Taxonomy, pathogenicity, and infection. Clin Microbiol Rev 2010;23:35-73.

  • Farmer JJ 3rd, Janda JM, Brenner FW, Cameron DN, Birkhead KM. Genus I. Vibrio Pacini 1854, 411al. In: Brenner DJ, Krieg NR, Staley JT, Garrity GM, editors. Bergey’s Manual of Systematic Bacteriology. 2nd ed., Vol. 2. Part B. New York: Springer; 2005. p. 494-546.

  • Soumet C, Ermel G, Fach P, Colin P. Evaluation of different DNA extraction procedures for the detection of Salmonella from chicken products by polymerase chain reaction. Lett Appl Microbiol 1994;19:294-8.

  • Mendes-Marques CL, Hofer E, Leal NC. Development of duplex-PCR for identification of Aeromonas species. Rev Soc Bras Med Trop 2013;46:355-7.

  • Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 1991;173:697-703.

  • Shirazinejad A, Ismail N, Bhat R. Lactic acid as a potential decontaminant of selected foodborne pathogenic bacteria in shrimp (Penaeus merguiensis de man). Foodborne Pathog Dis 2010;7:1531-5.

  • Mathur P. The Effect of Lime Juice on Vibrio parahaemolyticus and Salmonella Enterica Inactivation during the Preparation of Raw Fish Dish Ceviche. MSc. Thesis, Graduate School-New Brunswick Rutgers, The State University of New Jersey; 2012.

  • George Brown College Tutoring and Learning Centre. Percentage Change; 2014. Available from: http://www.georgebrown.ca/uploadedFiles/TLC/_documents/Percentage%20Change.pdf. [Last accessed on 2017 Aug 20].

  • Biju VN, Gunalan B. Prevalence of Vibrio infection in Penaeus (litopenaeus) vannamei farms. Int J Sci Invent Today 2016;5:485-93.

  • Cheng AC, Shiu YL, Chenn BJ, Huynh TJ, Liu CH. Isolation and identification of pathogenic bacterium Aeromonas veronii from ornamental shrimp Caridina cf. babaulti. J Fish Soc Taiwan 2016;43:273-83.

  • Taylor HH, Taylor EW. Gills and lungs: The Exchange of Gases and Ions. In: Harrison FW, Humes AG, editors. Microscopic Anatomy of Invertebrates 10. New York: Wiley-Liss; 1992. p. 203-93.

  • Ali MN, Abo-Esa JF. Study on some causative agents’ infection in Red Sea shrimp (Penaeus semiculcatus) in summer season. Egypt J Aquat Biol Fish 2007;11:645-56.

  • El-Bouhy ZM, Abdelrazek FA, Hassanin ME. A contribution on vibriosis in shrimp culture in Egypt. Egypt J Aquat Res 2006;32:443-56.

  • Abraham TJ, Sasmal D, Dash G, Nagesh TS, Das SK, Mukhopadhayay SK, et al. Epizootology and pathology of bacterial infections in cultured shrimp Penaeus monodon Fabricius 1798 in West Bengal, India. Indian J Fish 2013;60:167-71.

  • Aly SM, El-Attar AA. Pathological studies on some bacterial pathogens of cultured shrimp. Assiut Vet Med J 2001;45:298-331.

  • El-Hadi N, Radu S, Chen CH, Nishibuchi M. Prevalence of potentially pathogenic Vibrio species in the seafood marketed in Malaysia. J Food Prot 2004;67:1469-75.

  • Gopal S, Otta SK, Kumar S, Karunasagar I, Nishibuchi M, Karunasagar I, et al. The occurrence of Vibrio species in tropical shrimp culture environments; implications for food safety. Int J Food Microbiol 2005;102:151-9.

  • Koralage MS, Alter T, Pichpol D, Strauch E, Zessin KH, Huehn S. Prevalence and molecular characteristics of Vibrio spp. isolated from preharvest shrimp of the North Western Province of Sri Lanka. J Food Prot 2012;75:1846-50.

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

  • Kriem MR, Banni B, El Bouchtaoui H, Hamama A, El Marrakchi A, 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.

  • Odeyemi OA. Incidence and prevalence of Vibrio parahaemolyticus in seafood: A systematic review and meta-analysis. Springerplus 2016;5:464.

  • Popovic NT, Skukan AB, Dzidara P, Coz-Rakovac R, Strunjak-Perovic I, Kozacinski L, et al. Microbiological quality of marketed fresh and frozen seafood caught off the Adriatic coast of Croatia. Vet Med 2010;55:233-41.

  • Vivekanandhana G, Hatha AA, Lakshmanaperumalsamy P. Prevalence of Aeromonas hydrophila in fish and prawns from the seafood market of Coimbatore, South India. Food Microbiol 2005;22:133-7.

  • Khamesipour F, Moradi M, Noshadi E, Shahraki MM. Detection of the prevalence of Aeromonas hydrophila in shrimp samples by polymerase chain reaction (PCR) and cultural method in Iran. J Biol Environ Sci 2014;4:47-52.

  • Janda JM, Abbott SL, McIver CJ. Plesiomonas shigelloides revisited. Clin Microbiol Rev 2016;29:349-74.

  • Klatte JM, Daswtjerdi MH, Clark K, Harrison CJ, Grigorian F, Stahl ED. Hyper acute infectious keratitis with Plesiomonas shigelloides following traumatic lamellar corneal laceration. Pediatr Infect Dis J 2012;31:1200-1.

  • Levin RE. Plesiomonas shigelloides-An aquatic foodborne pathogen: A review of its characteristics, pathogenicity, ecology, and molecular detection. Food Biotechnol 2008;22:189-202.

  • Akaylı T, Zeybek Z. Bazı akvaryum baliklarinda Plesiomonas shigelloides enfeksiyonu üzerinde bir araştirma. E.U. J Fish Aquat Sci 2005;22:31-4.

  • Oxley AP, Shipton W, Owens L, McKay D. Bacterial flora from the gut of the wild and cultured banana prawn, Penaeus merguiensis. J Appl Microbiol 2002;93:214-23.

  • Yoshino Y, Kitazawa T, Kamimura M, Tatsuno K, Ota Y, Yotsuyanagi H. Pseudomonas putida bacteremia in adult patients: Five case reports and a review of the literature. J Infect Chemother 2011;17:278-82.

  • Akram F, Pietroni MA, Bardhan PK, Bibi S, Chisti MJ. Prevalence, clinical features, and outcome of Pseudomonas bacteremia in under-five diarrheal children in Bangladesh. ISRN Microbiol 2014;2014:469758.

  • Hashish NM, Abbass AA, Amine AE, Kanan S. Pseudomonas aeruginosa in swimming pools. Cogent Environ Sci 2017;3:1328841.

  • Adebayo-Tayo BC, Odu NN, Igiwiloh NJ, Okonko IO. Microbiological and physicochemical level of fresh catfish (Arius hendelotic) from different markets in Akwa Ibom state, Nigeria. N Y Sci J 2012;5:46-52.

  • Ardura A, Linde AR, Garcia-Vazquez E. Genetic detection of Pseudomonas spp. in commercial Amazonian fish. Int J Environ Res Public Health 2013;10:3954-66.

  • Craun GF, Brunkard JM, Yoder JS, Roberts VA, Carpenter J, Wade T, et al. Causes of outbreaks associated with drinking water in the United States from 1971 to 2006. Clin Microbiol Rev 2010;23:507-28.

  • Lingham T, Besong S, Ozbay G, Lee JL. Antimicrobial activity of vinegar on bacterial species isolated from retail and local channel catfish (Ictalurus punctatus). J Food Process Technol 2012;S11:1.

  • Mata L, Vives M, Vicente G. Extinction of Vibrio cholerae in acidic substrata: Contaminated fish marinated with lime juice (ceviche). Rev Biol Trop 1994;42:479-85.

  • Rodrigues A, Sandström A, Cá T, Steinsland H, Jensen H, Aaby P, et al. Protection from cholera by adding lime juice to food-results from community and laboratory studies in Guinea-Bissau, West Africa. Trop Med Int Health 2000;5:418-22.

  • Tomotake H, Koga T, Yamato M, Kassu A, Ota F. Antibacterial activity of citrus fruit juices against Vibrio species. J Nutr Sci Vitaminol (Tokyo) 2006;52:157-60.

  • Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12:564-82.

  • Suradi K, Wulandari E, Permatasari I. The effect of concentrations of lime juice (Citrus aurantifolia) toward acceptability and storage life of culled layer hens meat. Sci Pap Ser D Anim Sci 2015;58:339-41.

  • Priyadarshini S, John S, Iyer P. Antimicrobial activity and characterization of microflora of vinegar preparations developed from peels and fruit of sweet lime. Eur J Biotechnol Biosci 2014;2:42-5.

  • Bang W, Drake MA. Acid adaptation of Vibrio vulnificus and subsequent impact on stress tolerance. Food Microbiol 2005;22:301-9.

  • Mine S, Boopathy R. Effect of organic acids on shrimp pathogen, Vibrio harveyi. Curr Microbiol 2011;63:1-7.

  • Koo J, DePaola A, Marshall DL. Impact of acid on survival of Vibrio vulnificus and Vibrio vulnificus phage. J Food Prot 2000;63:1049-52.

  • Merrell DS, Bailey C, Kaper JB, Camilli A. The toxR-mediated organic acid tolerance response of Vibrio cholerae requires ompU. J Bacteriol 2001;183:2746-54.

  • Wong HC, Peng PY, Han JM, Chang CY, Lan SL. Effect of mild acid treatment on the survival, enteropathogenicity, and protein production in Vibrio parahaemolyticus. Infect Immun 1998;66:3066-71.

  • Yeung SM, Boor KJ. Effects of acid stress on Vibrio parahaemolyticus survival and cytotoxicity. J Food Prot 2004;67:1328-34.

  • Oliver JD, Pruzzo C, Vezzulli L, Kaper JB. Vibrio species. In: Doyle MP, Buchanan RL, editors. Food Microbiology: Fundamentals and Frontiers. 4th ed. Washington, DC: ASM Press; 2013. p. 401-39.

  • Koo J, Marshall DL, DePaola A. Antiacid increases survival of Vibrio vulnificus and Vibrio vulnificus phage in a gastrointestinal model. Appl Environ Microbiol 2001;67: 2895-902.

  • Khan SH, Iqbal J. Recent advances in the role of organic acids in poultry nutrition. J Appl Anim Res 2016;44:359-69.

  • Dittoe DK, Ricke SC, Kiess AS. Organic acids and potential for modifying the avian gastrointestinal tract and reducing pathogens and disease. Front Vet Sci 2018;5:216.

  • Rimoldi S, Gliozheni E, Ascione C, Gini E, Terova G. Effect of a specific composition of short- and medium-chain fatty acid 1-Monoglycerides on growth performances and gut microbiota of gilthead sea bream (Sparus aurata). Peer J 2018;6:e5355.


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  • Vibriosis and Aeromonas Infection in Shrimp:Isolation, Sequencing, and Control

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Authors

Hanaa Mohamed Fadel
Department of Animal Hygiene and Zoonoses, Suez Canal University, Ismailia, Egypt
M. M. Maather El-Lamie
Department of Fish Diseases and Management, Suez Canal University, Ismailia, Egypt

Abstract


Background and Aim: Shrimp is one of the most commonly consumed types of seafood. It is a very nutritious healthy food. Shrimp is low in calories and rich in protein and healthy fats. It also contains a treasure trove of vitamins and minerals. On the negative side, it may be affected by many bacterial diseases which affect its health. Furthermore, it may be incriminated as a vector of foodborne illnesses that range from mild gastrointestinal upset to life-threatening diseases. This study was designed to assess the clinical picture and zoonotic importance of vibriosis and Aeromonas infection in live shrimp and to study the antibacterial effect of citric acid (lemon juice) and acetic acid (vinegar) on these pathogens.

Materials and Methods: A total of 170 live shrimp (Metapenaeus monoceros) samples were collected from Suez City, Egypt. The samples were examined clinically, and then, they were enriched into alkaline peptone water and cultivated on thiosulfate-citrate-bile salts-sucrose agar and ampicillin MacConkey agar for the isolation of Vibrio and Aeromonas species, respectively. The recovered isolates were confirmed biochemically and genotypically using duplex polymerase chain reaction (PCR) and sequencing. The germicidal effects of vinegar and lemon on artificially contaminated shrimp samples with Aeromonas hydrophila and Vibrio parahaemolyticus at different times (0.25, 1, 1.5, and 24 h) and temperatures (5° and 30°C) were studied.

Results: The results revealed that some of the infected shrimp were hypoxic, lethargic with abnormal swimming behavior. In most cases, body appendages, telsons, uropods, and gills took black coloration. In addition, the hepatopancreas appeared soft, swollen, and congested. The prevalence rates of vibriosis in each of the musculature and hepatopancreas were 4.7%, while the prevalence rates of Aeromonas infection in the musculature and hepatopancreas were 11.8% and 11.2%, respectively. Duplex PCR showed that Aeromonas isolates gave double bands: 237 bp specific for gcat and 500 bp specific for 16S rRNA, while Vibrio spp. and Plesiomonas shigelloides isolates gave single band at 500 bp. The effect of organic acid treatment showed that acetic acid (vinegar 5%) had increasing reduction rates that reached its maximum level after 24 h; where it caused (100% inhibition) for A. hydrophila at both temperatures and (33.63% and 60% inhibition) for V. parahaemolyticus at refrigerator and room temperatures, respectively. Moreover, acetic acid was more effective at room temperature than at refrigerator temperature. Concerning the effect of lemon juice (citric acid), it was more effective than acetic acid at short marination (0.25 and 1 h) at both temperatures for the two pathogens. Moreover, lemon was more effective at refrigerator temperature than at room temperature at the same aforementioned time. The difference between the reduction effects of the two acids on both pathogens was statistically significant (p<0.0001).

Conclusion: Overall, the examined shrimp samples were found to be vectors for Vibrio and Aeromonas spp. Application of hygienic measures during handling and cooking of shrimp should be esteemed. The organic acid treatment trial showed that vinegar and lemon juice can be used as a safe and economic method to limit the microbial contamination in seafood.


Keywords


Shrimp, Vibriosis, Aeromonas Infection, Sequencing, Decontamination, Citric Acid, Acetic Acid.

References