Sequencing the Parasitic Nematode Contracaecum spp. in Edible Fish(Planiliza Abu)

Abstract

Contracaecum rudolphii Hartwich,1 964 (Nematoda: Ascaridoidea, Anisakidae) is a typical anisakid recorded globally. Consumption of undercooked seafood, raw or infected, contains the nematode larvae that cause human anisakidosis . Fish (Planiliza abu) specimens were obtained from a local market in Karbala, Iraq. This type of fish came from the Razzaza Lake. The prevalence of Contracaecum spp. parasites was done over eight months from June 2022 until January 2023, using ITS1, morphological and molecular analysis of the nematodes Contracaecum spp from fish. Out of 395 fish, 124(31.3%) had visceral infections caused by Contracaecum spp. larval type (L3). The morphological and genetic identification of Contracaecum spp. was validated in the parasitology laboratory of the veterinary medicine college at Kerbala University. The result showed the infection rate in January 2023 was (46.6%) and it dropped in 2022, September with a percentage of (20%). With the use of ITS1 gene, the molecular analysis for Contracaecum spp. was to investigate Contracaecum spp. and to confirm it. However, the nematode count, number of infected fish, and length were all substantially different at the P≤0.05. This study detected the isolate=(a1) at the locus=OP787071 and sequencing the parasites. The isolates were confirmed as Contracaecum rudolphii, isolate a1 internal transcribed spacer 1, partial sequence. In conclusion, molecular genotyping might be a useful technique for identifying the Contracaecum L3 larval species., life-cycle biology, transmission methods, and types of intermediate hosts.

Keywords

• contracaecum rudolphii
• Fish
• ITSI
• zoonosis
• anisakidosis

Supporting Institution

There is no any funding.

Project Number

Date: 23/02/2023, Ref: UOK.VET. PA.2022.05.

Ethical Statement

Ethics required are approved, by the Ethical Committee of the University of Kerbala/ College of Veterinary Medicine.

Thanks

CUKUROVA 10th INTERNATIONAL SCIENTIFIC RESEARCHES CONFERENCE April 2-4, 2023/ Adana, TURKEY.

Sequencing the Parasitic Nematode Contracaecum spp. in Edible Fish (Planiliza Abu)

Abstract

Contracaecum rudolphii Hartwich,1964 (Nematoda: Ascaridoidea, Anisakidae) is a typical anisakid recorded globally. Consumption of undercooked seafood, raw or infected, contains the nematode larvae that cause human anisakidosis. Fish (Planiliza abu) specimens were obtained from a local market in Karbala, Iraq. This type of fish came from the Razzaza Lake. The prevalence of Contracaecum spp. parasites were done over eight months from June 2022 until January 2023, using ITS1, morphological and molecular analysis of the nematodes Contracaecum spp from fish. Of 395 fish, 124 (31.3%) had visceral infections caused by Contracaecum spp. larval type (L3). The morphological and genetic identification of Contracaecum spp. was validated in the parasitology laboratory of the Veterinary Medicine College at Kerbala University. The result showed that the infection rate in January 2023 was (46.6%) and increased while it was (20%) in September 2022. With the use of ITS1 gene, the molecular analysis for Contracaecum spp. was to investigate Contracaecum spp. and to confirm it. However, the nematode count, number of infected fish, and length were all substantially different at the P≤0.05. This study detected the isolate=(a1) at the locus=OP787071 and sequenced the parasites. The isolates were confirmed as Contracaecum rudolphii, isolate a1 internal transcribed spacer 1, partial sequence. In conclusion, molecular genotyping might be a useful technique for identifying the Contracaecum L3 larval species, life-cycle biology, transmission methods, and types of intermediate hosts.

Keywords

• contracaecum rudolphii
• Fish
• ITSI
• zoonosis
• anisakidosis

Project Number

Date: 23/02/2023, Ref: UOK.VET. PA.2022.05.

References

1. Abdullah YS, Abdullah SM, Hussein RH, 2021a: Morphology and molecular studies of Contracaecum larvae (Nematoda: Anisakidae) in some fish species from Sulaimani Province, Kurdistan Region, Iraq. Basrah J Agri Sci, 34(1), 93-110.
2. Abdullah YS, Abdullah SMA, Hussein RH, 2021b. Ultramorphology and molecular studies of larvae (Nematoda: Anisakidae) collected in five Cyprinid fish species from Sulaimani Province, Kurdistan Region-Iraq. Helminthologia, 58(1), 41-58.
3. Aguilar-Marcelino L, Bautista-Garfias CR, Zaheer T, Maqsood A, Bamarni SS, Abdullah BH, Faruk AZ, Salman M, Akhtar T, Silva FE, Hussain R, 2022: Potential of Anisakiasis in Foodborne Zoonosis. Pakistan Vet J, 42(4).
4. Aibinu IE, Smooker PM, Lopata AL,2019: Anisakis nematodes in fish and shellfish-from infection to allergies. Int. J Parasit: Parasites and Wildlife, 9, 384-393.
5. Al-Saadi AJ, T Mhaisen F, R Hasan H, 2010: Endoparasites of seven fish species from Al-Hussainia creek, Karbala province, mid Iraq. Basrah J Agricul Sci, 23(2), 139-146.
6. Al-Zubaidy AB, 2009; Prevalence and Densities of Contracaecum sp. Larvae in Liza abu from Different Iraqi Water Bodies. Marine Sci, 20(1).
7. Barson M, Marshall BE, 2004: First record of Contracaecum spp. (Nematoda: Anisakidae) in fish-eating birds from Zimbabwe. J South African Vet Associat, 75(2),74-8.
8. Buchmann K, Mehrdana F, 2016: Effects of anisakid nematodes Anisakis simplex (sl), Pseudoterranova decipiens (sl) and Contracaecum osculatum (sl) on fish and consumer health. Food Waterborne Parasitol, 4, 13-22.

9. D’amelio S, Cavallero S, Dronen NO, Barros NB, Paggi L, 2012: Two new species of Contracaecum Railliet and Henry, 1912 (Nematoda: Anisakidae), C. fagerholmi n. sp. and C. rudolphii F from the brown pelican Pelecanus occidentalis in the northern Gulf of Mexico. System Parasitol, 81, 1-16.
10. D'amelio S, Barros NB, Ingrosso S, Fauquier DA, Russo R, Paggi L, 2007: Genetic characterization of members of the genus Contracaecum (Nematoda: Anisakidae) from fish-eating birds from west-central Florida, USA, with evidence of new species. Parasitol, 134(7), 1041-1051.
11. Decruyenaere P, Van de Maele B, Hulstaert E, Van Vlierberghe H, Decruyenaere J, Lapeere H, 2022: IgE-Mediated Gastroallergic Anisakiasis with Eosinophilic Oesophagitis: A Case Report. Acta Clin. Belgica Int. J. Clin. Lab. Med, 77, 396–399.
12. Dinas S, Diakou A, Vasiliadis K, Chaintoutis SC, Massa E, Konstantinou GN, Totsi A, Xakis A, Papavasiliou C, 2024: First Case of Human Anisakiosis in Greece: Acute Invasive Infection Mimicking Peritoneal Malignancy. Pathogens, 13(2),149.
13. Golden O, Caldeira AJR, Rangel LF, Santos MJ, 2022: Seafood safety and food‐borne zoonoses from fish: Examining the risk of Anisakis in the Portuguese Population and Consumer Risk Perceptions of Fish Consumption. Efsa J, 20, e200409.
14. Hamouda AH, Younis AE, 2022: Molecular characterization of zoonotic anisakid Contracaecum spp. larvae in some fish species from Lake Nasser, Egypt. Aquacul Res, 53(6), 2548-2561.
15. Hirosawa T, Sakamoto T, Shimizu T, 2020: Gastric Anisakiasis. Am J Med Sci, 360, 318–319.
16. İnnal D, Stavrescu-Bedivan M, Öztürk MO, Özmen Ö, 2020: First record of Contracaecum rudolphii Hartwich, 1964 in Carassius gibelio (Bloch, 1782) from Turkey. Aqua Sci Engin, 35(1), 1-5.
17. Jawad M, Ridha ZM, Alali F, Saeed EL, Alhesnawi ASM, 2022: Distribution and Identification of the Parasitic Nematode Contracaecum spp. in Planiliza Abu (Heckel, 1843: Mugiliformes, Mugilidae) From Razzaza Lake, Karbala Province, Iraq. Bozok Vet Sci, 3(2), 40-46.
18. Kibet CJ, Zhao WT, Sarwa H, 2021: Occurrence and molecular identification of larval Contracaecum spp. in Labeobarbus intermedius and Clarias gariepinus from Lake Tana, Ethiopia. Sci Parasitol, 22(1), 1-12.
19. Martínez KAM, García CM, Delgado AF, Güitrón LC, Saravia DO, Montañez MS, Maya JJM, Rubio JM, Villalobos N, 2022: Parasite identification in mullet fish (Mugil curema) from Chautengo Lagoon, Guerrero, Mexico, based on morphology and molecular analysis. Experim Parasitol, 240, 108335.
20. Mattiucci S, Paoletti M, Olivero-Verbel J, Baldiris R, Arroyo-Salgado B, Garbin L, Navone G, Nascetti G, 2008: Contracaecum bioccai n. sp. from the brown pelican Pelecanus occidentalis (L.) in Colombia (Nematoda: Anisakidae): morphology, molecular evidence and its genetic relationship with congeners from fish-eating birds. System Parasitol, 69, 101-121.
21. Mattiucci S, Sbaraglia GL, Palomba M, Filippi S, Paoletti M, Cipriani P, Nascetti G, 2020: Genetic identification and insights into the ecology of Contracaecum rudolphii A and C. rudolphii B (Nematoda: Anisakidae) from cormorants and fish of aquatic ecosystems of Central Italy. Parasitol Res, 119, 1243-1257.
22. Mohammad ZAA, Hbaiel MK, 2019: Morphological and Molecular Study of Contracaecum Larvae with a New Record of Contracaecum Septentrionale in Al-Sanaf Marsh Southern Thi-Qar Province, Iraq. Indian J Pub Heal Res and Develop, 10 (10).
23. Mohammadi F, Jolodar A, Mesbah M, Razi Jalali MH, 2021: Molecular identification of the zoonotic parasites Contracaecum spp. and Anisakis pegreffii (Nematoda: Anisakidae) isolated from Mesopotamichthys sharpeyi and Barbus grypus in the Shadegan Marsh of Khuzestan Province. Iranian J Fish Sci, 20(6), 1674-83.
24. Peck R, Olsen C, Devore JL, 2015: Introduction to statistics and data analysis. Fifth ed. Cengage Learning. Canada: Nelson Education, Ltd.
25. Pekmezci GZ, Onuk EE, Bolukbas CS, Yardimci B, Gurler AT, Acici M, Umur S, 2014: Molecular identification of Anisakis species (Nematoda: Anisakidae) from marine fishes collected in Turkish waters. Vet Parasitol, 201(1-2), 82-94.
26. Pons-Bordas C, Hazenberg A, Hernandez-Gonzalez A, Pool RV, Covelo P, Sánchez-Hermosin P, López A, Saavedra C, Fraija-Fernández N, Fernández M, Aznar FJ, 2020: Recent increase of ulcerative lesions caused by Anisakis spp. in cetaceans from the north-east Atlantic. J Helminthol, 94,e127.
27. Shamsi S, 2019: Parasite loss or parasite gain? Story of Contracaecum nematodes in antipodean waters. Parasite Epidemiol Cont, 4, e00087.
28. Shamsi S, Barton DP, 2023: A critical review of anisakidosis cases occurring globally. Parasitol Res, 26, 1-3.
29. Shamsi S, Butcher AR, 2011: First Report of Human Anisakidosis in Australia. Med J Aust, 194, 199–200.
30. Shamsi S, Gasser RB, Beveridge I, 2011: Mutation scanning-coupled sequencing of nuclear ribosomal DNA spacers as a tool for the specific identification of different Contracaecum (Nematoda: Anisakidae) larval types. Molecul Cell Probes, 25(1), 13-18.
31. Shamsi S, Norman R, Gasser R, Beveridge I, 2009: Genetic and morphological evidences for the existence of sibling species within Contracaecum rudolphii (Hartwich, 1964) (Nematoda: Anisakidae) in Australia. Parasitol Res, 105, 529-538.
32. Shamsi S, Suthar J, 2016: A revised method of examining fish for infection with zoonotic nematode larvae. Int J Food Microbiol, 227, 13-16.
33. Shamsi S, Turner A, Wassens S, 2018: Description and genetic characterization of a new Contracaecum larval type (Nematoda: Anisakidae) from Australia. J Helminth, 92(2), 216-222.
34. Shibata K, Yoshida Y, Miyaoka Y, Emoto S, Kawai T, Kobayashi S, Ogasawara K, Taketomi A,2020: Intestinal Anisakiasis with Severe Intestinal Ischemia Caused by Extraluminal Live Larvae: A Case Report. Surg Case Rep, 6, 4–8.
35. Szostakowska B, Fagerholm HP, 2007: Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland. J Parasitol, 93(4), 961-964.
36. Younis AE, Saad AI, Rabei JM, 2017: The occurrence of Contracaecum sp. larvae (Nematoda: Anisakidae) in four teleostean species from Lake Nasser, Egypt: morphological and molecular studies. J Basic Appli Zool, 78, 1-3.
37. Yusni E, Batubara A, Frantika C, 2022: Detection of endoparasites in mackerel tuna (Euthynnus affinis) in north Sumatra province, Indonesia. Iraqi J Vet Sci, 36(2), 519-524.
38. Zhu X, Gasser RB, Podolska M,Chilton NB, 1998: Characterisation of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. Int J Parasitol, 28(12), 1911-1921.