Ozonized Water, Microwaves and Freezing Effects on Viability of Encysted Metacercariae in Fish Muscle

International Journal of Veterinary Science
© 2020 by SSRG - IJVS Journal
Volume 6 Issue 1
Year of Publication : 2020
Authors : Hanan, A.A.Elghayaty, Samar, W.A.Tadros
How to Cite?

Hanan, A.A.Elghayaty, Samar, W.A.Tadros, "Ozonized Water, Microwaves and Freezing Effects on Viability of Encysted Metacercariae in Fish Muscle," SSRG International Journal of Veterinary Science, vol. 6,  no. 1, pp. 6-15, 2020. Crossref, https://doi.org/10.14445/24550868/IJVS-V6I1P102


The objective of this study is to evaluate the impact of ozonized water, microwaves cooking and freezing to inactivate the encysted metacercariae (EMC) and evaluate the prevalence of it in both Tilapia and Mugil species fish from Port-Said, Egypt. A total of 275 fishes (150, from Tilapia species and 125, Mugil species) was collected from June 2016 to May 2017, the incidence of EMC was 71.63%,with the highest prevalence in Summer 89.55% followed by Autumn 86.25%, the Spring 60% and the lowest was in Winter 46.03%. The prevalence was 70.66% and 72.8% in Tilapia and Mugil fish respectively. From the biological experimental infestation of chickens 4 species of trematodes were recovered (2 from Tilapia) Metagonimus Yokogawai & Haplorchis Yokogawai. and (2 from Mugil) Heterophyes heterophyes & Heterophyes aequalis. The treatments of infested fish muscle revealed that Microwave cooking at 500 Watt for 2 minutes be sufficient to destroy any EMC in it, also freezing the fish at -4° C to -5° C for 10 days are keeping the fish completely safe to consumer. The treatments of fish muscle by ozonized water, 25 ppm/ 8 minutes and 30 ppm / 10 minutes not completely safe enough to consumer to eat raw fish but the result give that; as the ozone concentration increased and prolonged time the affectivity increased.


Fish, Encystedmetacercare, ozone, Microwave, Freezing.


[1] Fish and fishery products hazards and controls guidance (4th Ed.). Gainesville: University of FloridaFDA (Food and Drug Administration). 2011.
[2] Panel on Biological Hazards (BIOHAZ) Scientific opinion on risk assessment of parasites in fishery products. (European Food Safety Authority). EFSA J, 8(4):1543 Section 4, 2010.
[3] Publically Available International Foodborne Outbreak Database Guelph, ON: Public Health Agency of Canada, Laboratory for Foodborne Zoonoses;PAIFOD March 2016.
[4] Phan, VT, Ersbøll, AK, Bui, TQ, Nguyen, HT, Murrell, D and Dalsgaard A: Fish-borne zoonotic trematodes in cultured and wild-caught freshwater fish from the Red River Delta, Vietnam. Vector Borne Zoonotic Dis. 10, 861–866.2010.
[5] World health organization: Report of Joint WHO/FAO workshop on food-borne trematode infections in Asia. HaNoi, Vietnam, 26-28 November, WHO, WPRO, 1-58, 2004.
[6] Ibanez, J. G.; Rodrigo M.-Mondragon; M. T. and Moran-M.:"Laboratory Experiments on the Electrochemical Remediation of the Environment. Part 7: Micro scale Production of Ozone" Journal of Chemical Education. 82 (10): 1546.2005.
[7] Corona-Vasquez B.; Samuelson, A.; Rennecker, J.L. and Mariñas, B.J.): Inactivation of Cryptosporidium parvum oocysts with ozone and free chlorine. Water Res., 36, 4053-4063, 2002.
[8] Maier,I. and Chu,T.: Use of ozone for inactivation of bacteria and virus in cryostats. J of Cytology and Histology, 7. (3), 428-433,2016.
[9] Marino, M.; Maifreni, M.; Baggio,A. and Innocente,N."inactivation of food borne bacteria biofilms by aqueous and gaseous ozone" ,Front.Microbiol.9:2024.2018
[10] Karaca, Hakan and Velioglu, Y. Seadat,"Ozone application in fruit vegetable processing",J.food reviews International. 23(1), 91- 106,2007.
[11] O’Donnel, C. ; Tiwari,B.K.; Cullen,P.J and Rice,R.G. "Status and trends of ozone in food processing in C.O’Donell; B.K.,Tiwari; P.J. Cullen and K.G. Rice (Eds)".Ozone in food processing Oxford U K:Wiley-Blackwell.P,312. 2012.
[12] Agniezka, J.B.; Nowak, A. and Smigielski,K):"Ozone in food industry: principles of ozone treatment, mechanisms of action and applications: an overview .J. Critical Reviews in Food Science and Nutrition,58(13): pp.2176-2201, 2018.
[13] Dehkordi, B. M. and Zokaie, N “Extension of Fish Shelf Life by Ozone Treatment.” International Journal of Environmental, Chemical, Ecological, Geological and Geophysical Engineering 4, 2.pp161,2010.
[14] Kumar, S., and Saurabh,S."Concepts and Applications of Microwave Engineering". PHI Learning Pvt. Ltd. p. 3.ISBN 8120349350.2014.
[15] Heddleson, R.A. & Doores, S. "Factors affecting microwave heating of foods and destruction of food born pathogen"-A review J. Food Protection., Vol., 57(11), pp.1025-1037. 1994.
[16] Zitzewitz, W. Paul. The Handy Physics Answer Book. Visible Ink, 2nd ed., Brownstowen,MI, USA. 2011.
[17] "Fish & Fisheries Products Hazards & Controls Guide. 2nd ed.Washington,D.C. FDA,Office of Seafood. 276 p. 1998.
[18] "WHO Expert Committee on Parasitic Zoonoses. Geneva: WHO. WHO Techn Rep Ser No 637.1979.
[19] Hamed, M.G.E. and Elias, A.N. "Effect of food processing methods upon Survival of the trematode Heterophyes spp. in flesh of mullet caught from brackish Egyptian waters". J. Food Sci. 35:386. 1970.
[20] Syme, J.D. Fish and fish inspection 2nd Ed. PP. 95-115, lewis& Co.ltd. London, U.K, 1966.
[21] Garcia,L.S.Diagnostic Medical Parasitology 4th Ed. ASM Press,Washington, D.C., USA.2001.
[22] El-Nafar,M.K. and El-Shahawi,G.A.:"Studies on the metacercariae of the Nile fishes at El-Minia Province ,A.R.E".Assuit Vet.Med.J.15 (30)46-58, 1986.
[23] Eissa, I.A.M.; Mona, M.A.; Hala, M.E. and Aly, S.M."Yellow grub disease among Oreochromis niloticus in relation to food quality and pathological alterations". SC.V.M.J, IV (2):179-194.2001.
[24] Yamaguti, S.: Systema helminthum V. I, the digenetic trematodes. Part I& II Inter science publishers Inc. New York and London.1958.
[25] Raef, A. M.: Role of marine fish in transmission of some parasites to animals and birds. Ph.D. Thesis (Parasitology), Fac. Vet. Med. Zag. Univ.1994.
[26] El-Amine, M.M.M.T.:Studies on the diseases resulting from encysted metacercariosis in some fresh water fishes. M.V.Sc. Thesis (Fish diseases and management), Fac. Vet. Med. Suez Canal Univ. 2001.
[27] Hegazi,M.A;,Hassan,A.T. Al-Nashar, T.M; Abo-Elkhier,O.Iand El- Lessi,F.M.:" Encysted metacercariae of family Heterophyidae in infected fish in Dakahila Governorate, an endemic focus in Egypt" .J.Egypt .Soc.Parasitol.44 (3):547-58. 2014.
[28] Saleh, R.S.; Abou-Eisha, A.M.; Hanaa, M.F. and Yosra,A.H.: Occurrence of encysted metacercariae of some zoonotic trematodes in freshwater fishes and their public health significance in Port-Said Province. Cairo International Conventation Center, 24-26 October, pp., 341-351.2009.
[29] Abou-Eisha, A.M.; Saleh,R.E.; Fadel,H.M.andYoussef, E.M. "Role of freshwater fishes in epidemiology of some zoonotic trematodes in Ismailia Province", S.C.V.M.J., XIII (2) 2008.
[30] El-Assal, F.M. and Mohamed,N.M."Impact of fish with metacercariae on the public health, at Cairo district, Egypt". International research Journal of Public and Environmental Health.5 (6).pp.72-82.2018.
[31] Arafa, M.I; Shaheen, M.S. and Monib, M.M. "Studies on some Clinostomatid metacercariae from Tilapia Nilotic in Assiut Governorate". Assiut Vet. Med. J. 51. (107)218-227, 2005.
[32] Abd-El-Rahman, A. M. M. "Studies on prevailing parasitic diseases among some fresh water fishes caused by digenetic trematodes" SC.V.M.J. VIII (1), PP 13- 24, 2005.
[33] Abd El-Samie, E K.: Epidemiological studies on some fish-borne parasites. M.V.SC. Thesis (Parasitology). Fac.Vet. Med. (Kafr EL-Sheikh), Tanta Univ. 2003.
[34] Hany, M.E and Attef, M. E.: "Preliminary observations on infection ofbrackish and fresh water fish by heterophyid encysted metacercariae in Egypt". Parasitol. Res. 103:971-977.2008.
[35] Gamal, A. T. "Some studies on metacercaria infection in oreochromisniloticus in Assiut Governorate and their role in transmission of some trematodes toDogs" .Ass.Univ.Bull.Envirom.Res.l.12.1, 63-79. 2009.
[36] Noga, E.N.: Fish diseases (diagnosis and treatment) Blackwell Science Ltd., PapernaU.S.A.PP 378. 2000.
[37] Weiping, ShengJinFeng,LiuXiaoling"Effect of ozone on the viability of Clonorchis sinensis metacercariae in vitro".J. Southern Agriculture.44 (1), pp.150-154, 2013.
[38] Ali, H.B. and Al-Mahmoud,S.S "Pathological effect of Cryptosporidium mansonio ocysts isolated from fish and treated with infra-red, microwave and ozone in common carp" .IraqJ.Vet.Science, 23.1.187-192. 2009.
[39] Panupan S., Thidarut B.1, Jiraporn S., RatchadawanA., Pranee. , Panaratana R., ParichartB., Apiporn S., Surasit A., Narong K.,WatcharinL., Nisana N, , Puangrat Y., Aung PhyoW,, SukhonthipK., Wu Z., BenjamabhornP., Atchara A., SirintipB."Simplified Techniques for Killing the Carcinogenic, Opisthorchis Viverrini Metacercariae in Cyprinid Fish" Asian Pac J Cancer Prev, 18 (6), 1507-1511,2017.
[40] Vidańćek, S.1.; De Las Heras, C.;Solas, M.T.;García, M.L.; Mendizábal,A.;Tejada, M"Viability and antigenicity of anisakis simplex after conventional and microwave heating at fixed temperatures" .J.Food Protection74 (12):2119-26. 2011.
[41] Pel'gunov AN, Riabov IN, FilippovaAIu: Use of microwave ovens in the prevention of opisthorchiasis. Med. Parazitol (Mosk).Jul-Sep ;( 3):42-5. 2005
[42] Mahmoud, N.A.:Parasitic infestation of some native Species of fishes in Cairo markets with special reference to parasites transmissible to man and animals. M. V. Sc (Zoonosis) Fac. Vet. Med. Cairo University. 1983.
[43] Abdallah, K.F.; Hamadto, H.H.; El-Hayawan, I.A.; El-Motayam, M.H.; Ahmed Wel-A."Effect of different temperatures on viability of seven encysted metacercariae recovered from fresh water fishes in Qualyobia, Egypt", J. Egypt Soc. Parasitol. Aug; 39(2):413-20, 2009.
[44] El-Zawawy L.A. ": Viability and infectivity of encysted metacercariae (EMC) of Pygidiopsis genata in fish". J. Egypt Soc.Parasitol. Dec; 38(3):945-56.2008.
[45] Fan,P.C. "Viability of metacercariae of Clonorchis snensis in frozen or salted fresh water fish". International J. Parasitology, 28,603-605. 1998.
[46] Fried, B. and Peoples,R, C. "Effects of various physical and chemical factors on excystation of the encysted metacercariae of Echinostoma Caproni." J. Parasitology, 93(6):1388-1391. 2007.
[47] Youssef,T.H; Haefnawy,Y.A; Khalifa,R. and Mahmoud,A.E."Effect of freezing and chilling on the viability and infectivity of the metacercriae of Haplorchis pumilio and Prohemistomum vivax".AssiutVet.M.J.Vol, 62(148), 164-167. 2016.
[48] Gill, C. O.: Microbiology of frozen foods. In Sun (Ed.), D. W. Handbook of Frozen Food Processing and Packaging (pp. 85-100).Taylor & Francis. England, 2006.