Multiplex PCR for the identification of Anisakis simplex sensu stricto, Anisakis pegreffii and the other anisakid nematodes

Ascaridoid nematodes are widespread in marine fishes. Despite their major socioeconomic importance, mechanisms associated to the fish-borne zoonotic disease anisakiasis are still obscure. RNA-Seq and de-novo assembly were herein applied to RNA extracted from larvae and dissected pharynx of Hysterothylacium aduncum (HA), a non-pathogenic nematode. Assembled transcripts in HA were annotated and compared to the transcriptomes of the zoonotic species Anisakis simplex sensu stricto (AS) and Anisakis pegreffii (AP). Approximately 60,000,000 single-end reads were generated for HA, AS and AP. Transcripts in HA encoded for 30,254 putative peptides while AS and AP encoded for 20,574 and 20,840 putative peptides, respectively. Differential gene expression analyses yielded 471, 612 and 526 transcripts up regulated in the pharynx of HA, AS and AP. The transcriptomes of larvae and pharynx of HA were enriched in transcripts encoding collagen, peptidases, ribosomal proteins and in heat-shock motifs. Transcripts encoding proteolytic enzymes, anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides were up-regulated in AS and AP pharynx. This study represents the first transcriptomic characterization of a marine parasitic nematode commonly recovered in fish and probably of negligible concern for public health.

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Anisakis physeteris and Pseudoterranova decipiens in the Mugil curema fish caught in Tumaco, Colombia

Revista Mvz Córdoba ◽

10.21897/rmvz.1781 ◽

2020 ◽

pp. e1781

Author(s):

Jenniffer Alejandra Castellanos Garzon ◽

Rubén Mercado P ◽

Sebastián Peña F ◽

María Carolina Pustovrh R ◽

Liliana Salazar M

Keyword(s):

Sensu Stricto ◽

Anisakis Simplex ◽

Anisakis Pegreffii ◽

Pseudoterranova Decipiens ◽

Pcr Multiplex ◽

Contracaecum Osculatum ◽

Hysterothylacium Aduncum ◽

Anisakis Physeteris

Objetivo. Identificar nematodos de la familia Anisakidae en el pez de consumo Mugil curema. Materiales y métodos. Para este estudio, se recolectaron 16 peces Lisa (M. curema) del puerto de Tumaco, una ciudad en la costa colombiana del Pacífico. La identificación morfológica de las larvas se realizó mediante taxonomía clásica y se calculó el porcentaje de infestación de larvas. Para la identificación molecular, se realizó una PCR múltiple con cebadores para las especies Anisakis physeteris, Pseudoterranova decipiens, Anisakis simplex sensu stricto, Contracaecum osculatum, Hysterothylacium aduncum y Anisakis pegreffii. Resultados. La revisión taxonómica permitió la identificación de larvas de tipo II del género Anisakis y larvas del género Pseudoterranova. Las larvas se aislaron principalmente del intestino, donde se encontró que el 94% de los peces estaban parasitados por nematodos anisakidos. La PCR multiplex permitió la identificación de la especie A. physeteris (Larva tipo II) y P. decipiens. Conclusiones. Este estudio en el primer reporte del nematodo Anisakidae en Tumaco, Colombia. Estos resultados proporcionan una justificación convincente para un estudio adicional sobre la familia Anisakidae en Colombia, como un problema de salud pública.

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Molecular characterization and comparison of four Anisakis allergens between Anisakis simplex sensu stricto and Anisakis pegreffii from Japan

Molecular and Biochemical Parasitology ◽

10.1016/j.molbiopara.2013.05.006 ◽

2013 ◽

Vol 190 (1) ◽

pp. 23-26 ◽

Cited By ~ 12

Author(s):

Karl Marx A. Quiazon ◽

Kosuke Zenke ◽

Tomoyoshi Yoshinaga

Keyword(s):

Molecular Characterization ◽

Sensu Stricto ◽

Anisakis Simplex ◽

Anisakis Pegreffii

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Molecular identification of Anisakis simplex sensu stricto and Anisakis pegreffii (Nematoda: Anisakidae) from fish and cetacean in Japanese waters

Parasitology International ◽

10.1016/j.parint.2006.07.001 ◽

2006 ◽

Vol 55 (4) ◽

pp. 267-271 ◽

Cited By ~ 67

Author(s):

Azusa Umehara ◽

Yasushi Kawakami ◽

Toshihiro Matsui ◽

Jun Araki ◽

Akihiko Uchida

Keyword(s):

Molecular Identification ◽

Sensu Stricto ◽

Anisakis Simplex ◽

Anisakis Pegreffii ◽

Japanese Waters

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A comparison of the hydrolase activities of excretory-secretory products and somatic extracts from fish parasitic nematodes, Anisakis simplex sensu stricto and Anisakis pegreffii larvae

Journal of fish pathology ◽

10.7847/jfp.2014.27.1.025 ◽

2014 ◽

Vol 27 (1) ◽

pp. 25-33 ◽

Cited By ~ 1

Author(s):

Chan-Hyeok Jeon ◽

Seong Wi ◽

Jeong-Ho Kim

Keyword(s):

Sensu Stricto ◽

Anisakis Simplex ◽

Parasitic Nematodes ◽

Anisakis Pegreffii ◽

Secretory Products

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Molecular identification of Anisakis species from Pleuronectiformes off the Portuguese coast

Journal of Helminthology ◽

10.1079/joh2005325 ◽

2006 ◽

Vol 80 (1) ◽

pp. 47-51 ◽

Cited By ~ 42

Author(s):

J.F. Marques ◽

H.N. Cabral ◽

M. Busi ◽

S. D'Amelio

Keyword(s):

Restriction Fragment Length Polymorphism ◽

Sibling Species ◽

Length Polymorphism ◽

Fragment Length Polymorphism ◽

Sensu Stricto ◽

Anisakis Simplex ◽

Solea Senegalensis ◽

Portuguese Coast ◽

Anisakid Nematodes ◽

Restriction Fragment Length

AbstractAnisakid nematodes belonging to the Anisakis simplex complex are highly prevalent in several fish species off the coast of Portugal and are an important zoonotic problem in the Iberian Peninsula. Two reproductively isolated sibling species of the Anisakis simplex complex were identified from Pleuronectiformes inhabiting the Portuguese coast using restriction fragment length polymorphism (RFLP). Recombinant genotypes corresponding to presumptive Anisakis simplex sensu stricto and Anisakis pegreffii hybrids were also detected by this technique, as well as the species Anisakis typica. Although 25 species of Pleuronectiformes were investigated, Anisakis spp. larvae were only found in seven: Arnoglossus imperialis, Arnoglossus laterna, Lepidorhombus boscii, Citharus linguatula, Platichthys flesus, Dicologlossa cuneata and Solea senegalensis. The occurrence of hybrids in relatively sedentary fishes such as the Pleuronectiformes suggests that the Portuguese coast may constitute an area of hybridization and, therefore, is of particular interest for the study of the process of hybridization and speciation for these anisakids.

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