Anisakidosis is a foodborne zoonotic infection induced by members of the family Anisakidae via the consumption of raw or undercooked fish such as sushi and sashimi. Identifying anisakid larval species is critical for the epidemiology and diagnosis of diseases caused by them. This study aimed at identifying Anisakis larvae collected from marine fish in Egyptian waters based on morphological characteristics and molecular analysis. Thirty marine fish coral trout, Plectropomus areolatus, were collected from Hurghada, Red Sea, Egypt, to investigate larval nematodes of the genus Anisakis. The larvae were detected encapsulated in the peritoneal cavity and muscle of the fish host. This examination revealed that anisakid larvae naturally infected 19 fish specimens with a prevalence of 63.33% and a mean intensity of 4.1 ± 0.40. Most of them (68 larvae: 71.57%) were found in the musculature. Morphological and morphometric analyses using light and scanning electron microscopy revealed a head region with a prominent boring tooth, inconspicuous lips, and a characteristic protruded cylindrical mucron. All larvae in this study possessed the same morphology as Anisakis Larval type I. Molecular analysis based on ITS region using maximum likelihood and Bayesian phylogenetic methods confirmed them as Anisakis typica. This is the first study to identify A. typica larvae from the commercial fish coral trout P. areolatus in Egyptian waters using morphological and molecular methods.

The study aimed to perform the molecular identification of Anisakis larvae in commercial fish from the coast of the Canary Islands and to provide data on their infection level for the host and the species of this nematode parasite that we could find in several species of commercial interest in the Canary Archipelago. Fish specimens (n = 172) from the Canary coasts were examined for parasites. In total, 495 larvae were identified; PCR was carried out for the entire ITS rDNA and cox2 mtDNA region, obtaining sixteen sequences for the entire ITS rDNA region and fifteen for the cox2 mtDNA, this being the first contribution of nucleotide sequences of Anisakis species of fish caught from the Canary Islands. An overall prevalence of 25% was obtained in the fish analyzed, and five species of Anisakis were identified, these being Anisakis simplex (s.s.), Anisakis pegreffii, Anisakis physeteris, Anisakis nascettii and Anisakis typica and the hybrid Anisakis simplex x Anisakis pegreffii. The results obtained in this study have relevance for public health, since the pathology will depend on the species of Anisakis, so it is important to know the health status of fish in the waters of the Canary Islands to assure a safer consumption and take adequate measures, in addition to the provision of epidemiological data.

Nematode parasites of the genus Anisakis (Nematoda, Anisakidae) are considered among the most important biological hazards in seafood products worldwide. In temperate and tropical waters, the most common species appears to be Anisakis typica, generally found around the viscera and sporadically in the flesh of various fish host species. This study investigated the infection sites and genetic diversity of A. typica infecting commercial fishes from the South-West Indian Ocean. Largehead hairtail (N = 20) and brushtooth lizardfish (N = 72) fished off Tanzania were inspected for anisakid nematodes by UV-press. A subsample of 168 nematodes were identified by sequence analyses of the cox2 mtDNA gene and ITS region of rDNA. The species A. typica (s.l.) (N = 166), Pseudoterranova ceticola (N = 1) and Anisakis paggiae (N = 1) were molecularly identified. Phylogenetic analysis of A. typica (s.l.) sequences based on both genes, indicated the existence of two distinct phylogenetic lineages forming two well-supported clades. The first clade comprised 12 A. typica specimens including individuals from its type locality (central Atlantic Ocean). The second clade comprising 154 specimens, clustered with reference sequences retrieved from GenBank including one apparently undescribed taxon, i.e., Anisakis sp. 1, and A. typica var. indonesiensis. The two reciprocally monophyletic clades are closely related and correspond to two distinct sister species within A. typica (s.l.), presently indicated as A. typica sp. A and A. typica sp. B. Two and four fixed alternative nucleotide substitutions (SNPs), i.e., diagnostic positions, between the two taxa, respectively, were found at the mtDNA cox2 and the ITS region of rDNA. The genetic data, as well as their occurrence in sympatry, strengthens the hypothesis that the actual specimens represent two distinct gene pools. The occurrence of both A. typica sp. A and A. typica sp. B in the musculature of freshly examined T. lepturus and S. undosquamis, suggests that both species can migrate intra-vitam into the flesh. Although the zoonotic potential of A. typica s.l. is still unclear, the presence of these parasites in the musculature, edible part of the fish, raises health concerns for consumers.

There are relatively few studies on parasite fauna of marine fishes in Philippine waters. This study aimed to determine the prevalence of marine ascaridoid infection in Decapterus species in Balayan Bay and Tayabas Bay. A total of 371 fishes belonging to three different species of Decapterus (D. tabl [n = 130], D. macrosoma [n = 121], and D. maruadsi [n = 120]) were collected. Ascaridoid parasite larvae were found in all fish host species, with an overall fish infection rate of 22%. The highest infection rate was observed in D. tabl (27.69%), followed by D. macrosoma (19%), and then D. maruadsi (17.50%). Moreover, a higher prevalence of infection was detected in Tayabas Bay (27.57%) than in Balayan Bay (15.59%). Molecular analyses based on the ITS2 and 18S rRNA gene supported the identification of the larvae into two species: Anisakis typica and Raphidascaris (Ichthyascaris) lophii. This is the first report of the genetic identification of these two helminth parasites in Decapterus fish species in the Philippines. Paucity in the database of Philippine marine fish parasites warrants more research efforts, especially concerning economically important fish species with implications to food safety and food security.

The third stage larvae (L3) of Anisakis typica were detected in 2 species of threadfin bream, Nemipterus hexodon and N. japonicus, from the Gulf of Thailand, and were morphologically and molecularly characterized. Total 100 threadfin breams, 50 Nemipterus hexodon and 50 N. japonicus, were examined with naked eyes after the opening of abdominal cavity with scissors. Almost all infected larvae remained alive and active even the fish were transported for 1-2 days. Anisakid larvae were exclusively distributed in the body cavity and rarely in the liver. The prevalence of A. typica L3 were 68.0% and 60.0% in N. hexodon and N. japonicus and their infection intensities were 3.5 and 4.2 per fish infected each. Morphological and morphometric analysis were performed by viewing specimens under both a light microscope and a scanning electron microscope. Interestingly, the protruded mucron of Anisakis typica under SEM showed a distinct cylindrical shape that differed from the cone shape of A. simplex. The protruded mucron could be used to identify A. typica L3 larvae in the future. A comparison of the ITS1-5.8S-ITS2 rDNA nucleotide sequences of these species revealed high blast scores with A. typica. Conclusively, it was confirmed that A. typica L3 are prevalent in threadfin breams from the Gulf of Thailand, and their morphological and molecular characters are something different from those of other anisakid larvae, including A. simplex and A. pegreffii.

BACKGROUND: In the marine environment, transitional zones between major water masses harbour high biodiversity, mostly due to their productivity and by containing representatives of species characteristic of adjacent communities. With the aim of assessing the value of larval Anisakis as zoogeographical indicators in a transitional zone between subtropical and sub-Antarctic marine currents, larvae obtained from Zenopsis conchifer were genetically identified. Larvae from Pagrus pagrus and Merluccius hubbsi from two adjacent zoogeographical provinces were also sequenced.
RESULTS: Four species were genetically identified in the whole sample, including Anisakis typica, A. pegreffii, A. berlandi and a probably new species related to A. paggiae. Anisakis typica and A. pegreffii were identified as indicators of tropical/subtropical and sub-Antarctic waters, respectively, and their presence evidenced the transitional conditions of the region. Multivariate analyses on prevalence and mean abundance of Anisakis spp. of 18 samples represented by 9 fish species caught south of 35°S determined that host trophic level and locality of capture were the main drivers of the distribution of parasites across zoogeographical units in the South-West Atlantic.
CONCLUSIONS: Most samples followed a clear zoogeographical pattern, but the sample of Z. conchifer, composed mostly of A. typica, was an exception. This finding suggests that population parameters of A. typica and A. pegreffii could differ enough to be considered as a surrogates of the identity of larvae parasitizing a given host population and, therefore, a step forward the validation of the use of larval Anisakis as biological indicators for studies on host zoogeography.

Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica AB517571/DQ116427. The difference of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait.