Molecular identification and prevalence of plerocercoid larvae (Cestoda: Diphyllobothriidae) in some commercial fish species from Peru.

Diphyllobothriosis, a fish-borne zoonosis in South America, is mainly caused by the Pacific broad tapeworm Adenocephalus pacificus Nybelin, 1931, a parasite of considerable concern in fishery resources due to its impact on public health. A new diphyllobothrid, Diphyllobothrium sprakeri Hernández-Orts et al. Parasites Vectors 14:219, 2021, was recently described from sea lions from the Pacific Coast, but marine fish acting as intermediate hosts are unknown. The objective of this study was to confirm the presence of plerocercoid larvae of Diphyllobothriidae Lühe, 1910 (Cestoda: Diphyllobothriidea) in nine fish species of commercial importance in Peru. Of a total of 6999 fish (5861 Engraulis ringens, 853 Sciaena deliciosa, 6 Sciaena callaensis, 171 Scomber japonicus, 40 Trachurus murphyi, 40 Ariopsis seemanni, 18 Merluccius peruanus, 5 Sarda chiliensis, and 5 Coryphaena hippurus), 183 were infected with plerocercoid larvae, representing a total prevalence of 2.61% and a mean intensity of 3.2. Based on mtDNA cox1 sequences of 43 plerocercoids, a phylogenetic analysis revealed that 41 belong to A. pacificus and two to D. sprakeri. These findings are first molecular data for D. sprakeri larvae, and the infections of E. ringens and T. murphyi by plerocercoid larvae represent the first records of intermediate/paratenic hosts for this species. Hence, the findings of the current study enhance our understanding of the presence of diphyllobothriid species in commercial fish from the Southeastern Pacific Ocean and their potential impact on seafood safety for local human populations.

An Annotated Checklist of Monogeneans (Platyhelminthes, Monogenea) from Aquatic Vertebrates in Peru: A Review of Diversity, Hosts and Geographical Distribution.

Monogeneans are flatworm parasites that infest fish gills primarily but can also infect various other vertebrates, including amphibians, aquatic reptiles, mammals like hippos, and occasionally invertebrates like copepods, isopods, and cephalopods. Despite their remarkable diversity, our knowledge of monogenean parasites in Peru remains significantly limited, resulting in substantial gaps in our comprehension of their taxonomic identities, host associations, and geographic distribution. To address these knowledge deficits, we present an extensively curated checklist of monogeneans associated with aquatic vertebrates in Peru. This comprehensive compilation is derived from meticulous literature surveys, the examination of specimens deposited in both international and national collections, and the inclusion of additional freshly collected specimens. The checklist offers a thorough repository of data encompassing the diversity, host associations, and geographical distribution of these parasites. Taxonomic discrepancies are addressed through a critical review of the existing literature, supplemented by the direct examination of specimens, including type or voucher specimens, deposited within scientific collections. Additionally, we provide data on the DNA sequences of individual taxa. The compiled list comprises records of 358 monogenean species, including 270 valid species and 88 taxa identified at the family or generic level, all reported across 145 host species in Peru. Predominantly, these parasitic species exhibit associations within fish, with 335 infecting teleosts and 20 affecting chondrichthyans. Three monogenean species have been documented as infecting amphibians, namely Mesopolystoma samiriensis, Polistoma sp. and Wetapolystoma almae. Among the monogeneans reported, 141 were found in marine environments and 214 in freshwater environments. The most diverse families were Dactylogyridae and Diplectanidae, comprising 217 and 24 species, respectively. The hosts that harbored the highest number of monogeneans were Pygocentrus nattereri (with 23 species), followed by Stellifer minor (13 spp.) and Triportheus angulatus (11 spp.). We detected many species that do not have any material deposited in a scientific collection due to the loss or deactivation of the collection. These findings represent only a fraction of the potential diversity, considering the wide variety of aquatic vertebrate hosts inhabiting the tropical and subtropical regions of Peru.

Using DNA barcoding to link cystacanths and adults of the acanthocephalan Corynosoma australe of the Southeastern Pacific Ocean (off Peru coast).

The objective of this study is to use DNA barcoding to link cystacanths and adults belonging to the acanthocephalans Corynosoma australe found in the Southeastern Pacific Ocean off the coast central from Peru. We sampled three species of commercial fish (Paralichthys adspersus (Steindachner), Paralabrax humeralis (Valenciennes), and Cheilodactylus variegatus (Valenciennes)) and two South American sea lions, Otaria byronia, stranded on the beaches of the city of Huacho and Barranca, Lima province. A total of 509 acanthocephalan larvae were found in the body cavity of 95 fish (prevalence 54.28%, total mean intensity 8.64). A total of 127 adult worms were found in the large intestine from two South American sea lions (P= 100%, MI= 63.5). A total of 203 larvae from P. humeralis were isolates (P=65.71%; MI= 8.83; MA=5.8), 235 (P=54.29%; MI= 12.37; MA= 6.71) from C. variegatus, and 71 (P=42.86%; MI= 4.73; MA= 2.03) from P. adspersus. All adult and larval specimens were morphologically identified as C. australe. They were generated cytochrome c oxidase subunit 1 (cox1) gene sequences of specimens and were compared with available data from GenBank. Molecular phylogenetic analysis supported our morphological identification, where the Peruvian isolates formed a clade with other isolates of C. australe from other countries of the American continent. Of the sequences obtained, two haplotypes were detected and were not identical with previous reports. Based on both DNA barcoding and morphological analyses, our finding represents the first molecular data of C. australe from Peru and the report of Cheilodactylus variegatus as a new paratenic host on the central coast, extending the knowledge and distribution range of this acanthocephalan in Southeastern Pacific Ocean.

Molecular identification and epidemiological data of Anisakis spp. (Nematoda: Anisakidae) larvae from Southeastern Pacific Ocean off Peru.

The objective of this study is to determine the infection status of nematode larvae and record epidemiological molecular data in commercial fish from the southeast Pacific off the central coast of Peru. Anisakiasis is a fish-borne zoonosis caused by Anisakis larvae, parasites of relevance in the fishery resources that have negative impact on public health. Between January 2012 to December 2014, 345 specimens of four fish species (Trachurus symmetricus murphyi, Scomber japonicus peruanus, Merluccius gayi peruanus and Seriolella violacea) were examined for Anisakis sp. larvae. A total of 997 Anisakis sp. larvae were found in the body cavity of 196 fish (total prevalence 53.7%, total mean intensity 5.08). After morphological analysis, 958 (96.08%) larvae were identified as Type I and 39 (3.92%) as Type II. Specimens were identified by molecular analysis of the mitochondrial cytochrome c oxidase subunit II (cox2) gene, confirming that A. pegreffii is the predominant species and the most important agent of human anisakiasis off the Peru Central Coast. In addition, we revealed the occurrence of A. physeteris (s.l.) in S. japonicus peruanus (P = 18.0%; MI = 2.17). Therefore, the results obtained in the present study improve the knowledge of the occurrence of Anisakis species in the commercial fish from the Southeastern Pacific Ocean, highlighting the importance of considering a potential hazard for humans and the necessity of further research in other fishes of greater preference by the Peruvian population.

Molecular and morphological identification of Adenocephalus pacificus (Cestoda) isolated from South American sea lion Otaria byronia stranded on the northern Peruvian coasts.

The most frequent etiologic agent of diphyllobothriosis in South America and the only one confirmed by molecular data in human cases in Peru is Adenocephalus pacificus (syn. Diphyllobothrium pacificum). This cestode is transmitted by ingestion of the plerocercoids found in marine fish, causing a parasitic zoonosis. The objective of the present study was to identify two cestodes isolated from two specimens of the South American sea lion (Otaria byronia) stranded on the beaches of Huacho and Barranca cities, located on the northern Peruvian coasts, in the department of Lima. Tapeworms were confirmed by morphological characteristics due to the presence of transverse papilla-like tegumental protuberances in proglottids and small sized eggs, as well as by sequencing of the partial cytochrome c oxidase subunit 1 (mtDNA-COI) gene that are congruent with additional available A. pacificus sequences. Even though sea lions in Peru are distributed along the coast and in areas of difficult access, generally located in protected natural areas, the fortuitous finding represented an opportunity to confirm the presence of A. pacificus in South American sea lions. This report of tapeworm A. pacificus could allow future monitoring of the occurrence and geographical distribution of this causative agent in epidemiological studies, since it is one of the main species of zoonotic importance in Peru.

A New Species of Mymarothecium tantaliani n. sp (Monogenea: Dactylogiridae) in the Gills of Gamitana Colossoma macropomum (Cuvier) from Madre de Dios, Peru.

INTRODUCTION: Members of the genus Mymarothecium Kritsky, Boeger & Jégu, 1996 infect characiform fish in the Amazon River Basin. Although the Peruvian Amazon hosts rich fauna of fish, the taxonomic diversity of parasitic organisms such as Mymarothecium has been few explored. A new species of Mymarothecium in Colossoma macropomum Cuvier, from the southeast of the Peruvian Amazon, is described. METHODS: Monogeneans were isolated from gills of C. macropomum, fixed in formaldehyde in 4% solution. For morphological examinations were stained with Gomori's trichrome and others were clarified with Hoyer's medium. RESULTS: Mymarothecium is the third species of the genus recorded in C. macropomum. These new species have been characterized by a slightly sigmoid male copulatory organ (MCO) and the accessory piece with sinuous distal rod with hook-shaped process and a bifurcation in the middle part. Furthermore, the anteromedial projection of the ventral rod has a small conspicuous filament in both anchors. CONCLUSIONS: With the description of this new species, eight species of Mymarothecium have been reported in fish of the Serrasalmidae family originating in the Amazon.

Molecular Identification of Adenocephalus pacificus (Cestoda) from Three Human Cases in Lima Province, Peru.

The Pacific tapeworm Adenocephalus pacifcus (syn. Diphyllobothrium pacificum) is a causative agent of diphyllobothriosis occurred in Pacific coast of South America, mainly in Peru. Source of infections are traditional meal from raw or undercooked marine fish such as "cebiche". We confirmed 3 new cases, one including scolex and the other two headless. A strobila 46 cm long without scolex was discharged from an 8-year-old boy before treatment. Specimens were confirmed morphologically by presence of tegumental protuberances on proglottids and small sized eggs. Partial sequence of cytochrome c oxidase subunit 1 gene was congruent with A. pacificus sequences.

Caracterización del patrón proteico y perfil antigénico de larvas L 3 de anisákidos de importancia médica aisladas de peces comerciales de la costa peruana / Characterization of the protein pattern and antigen profile of L 3 larvae of medical importance in anisakids isolated from commercial fish of the peruvian coast

RESUMEN Introducción. La anisakidosis es una zoonosis causada por la ingestión accidental de larvas (L3) de anisákidos. Objetivo. Caracterizar el patrón proteico y perfil antigénico de la L3 de Anisakis simplex s.l. (tipo I), A. physeteris s.l. (tipo II) y Contracaecum osculatum s.l. aisladas de peces comerciales. Métodos. Se realizó una corrida electroforética en SDS-PAGE de los antígenos somáticos. Se inmunizó conejos experimentalmente y se evaluó por EITB. Resultados. El patrón proteico de Anisakis tipo I mostró 12 bandas, 18 Anisakis tipo II y C. osculatum 13, con las bandas 10 y 35 kDa específicas para Anisakis tipo II, 28 y 77 para C. osculatum no presentes en Anisakis tipo I. Conclusión. Se determinó bandas inmunogénicas específicas para Anisakis tipo I con las proteínas de peso molecular 11, 14, 25 y 40 kDa, para el tipo II de 9, 10, 12, 35 y 41 kDa, y C. osculatum 13, 15, 17, 30 y 47 kDa.

ABSTRACT Introduction. Anisakidosis is a zoonosis caused by accidental ingestion of anisakid larvae (L3). Objective. To characterize the protein pattern and antigenic profile of the L3 of Anisakis simplex s.l. (type I), A. physeteris s.l. (type II) and Contracaecum osculatum s.l. commercial3 fish isolated. Methods. An SDS-PAGE electrophoretic run of the somatic antigens was performed. Rabbits were immunized experimentally and evaluated by EITB. Results. The protein pattern of Anisakis type I showed 12 bands, 18 Anisakis type II and C. osculatum 13, with bands 10 and 35 kDa specific for Anisakis type II, 28 and 77 for C. osculatum, not present in Anisakis type I. Conclusion. Specific immunogenic bands were determined for Anisakis type I with the molecular weight proteins 11, 14, 25 and 40 kDa, for type II of 9, 10, 12, 35 and 41 kDa and C. osculatum 13, 15, 17, 30 and 47 kDa.