First Report of Blood Fluke Pathogens with Potential Risk for Emerging Yellowtail Kingfish (Seriola lalandi) Aquaculture on the Chilean Coast, with Descriptions of Two New Species of Paradeontacylix (Aporocotylidae).

Blood flukes are digeneans that infect wild and farmed fish that can cause a severe and potentially lethal disease in farmed fish. These parasites are undetectable in the larval stage based on macroscopic observations in the definitive host with the infection becoming evident when eggs accumulate in the branchial vessels. There are nine known species of the genus Paradeontacylix and seven exclusively parasitize Seriola spp. from several geographical areas. Seriola lalandi aquaculture farms are emerging at various localities in northern Chile. Here, we report, for the first time, two blood fluke species parasitizing S. lalandi in the Southeastern Pacific (Chile). In the laboratory, the gills and heart of fish were removed. The retained blood flukes were separated according to the infection site, fixed in 70% or 95% ethanol for taxonomic and molecular analysis, respectively. Morphometrical differences among the fluke species were evaluated with a principal component analysis (PCA) using proportional body measurements. Phylogenetic trees were constructed based on 28S rDNA, cox1 mDNA using Bayesian inference (BI), and maximum likelihood (ML). Based on morphology, morphometry, and molecular analyses, two new species are proposed: P. humboldti n. sp. from the gills and P. olivai n. sp. from the heart of S. lalandi. Both were clearly distinguished from other species of Paradeontacylix by a combination of morphologic features (posterior tegumental spines, testes arrangement, body size). The genetic distance (based on cox1) among species was >10%. P. humboldti n. sp. and P. olivai n. sp. are sister species (with a common ancestor) independent of P. godfreyi from S. lalandi in Australia. The newly identified parasites may pose a risk to farmed S. lalandi as aporocotylids have been the cause of diseases in farmed fish from other geographical areas. In addition, some cages of S. lalandi are currently maintained in an open circulating system, which could favor the transmission of these parasites (if involved hosts are present in the environment).

Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the deep-sea Eastern Pacific black ghost shark Hydrolagus melanophasma from the Atacama Trench, with comments on host specificity of Calicotyle spp.

We describe Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the cloaca of deep-water Eastern Pacific black ghost sharks, Hydrolagus melanophasma captured as bycatch at a local fishery for Patagonian toothfish Dissostichus eleginoides, (Nototheniidae) in the Atacama Trench using morphological and nucleotide (LSU rRNA and SSU rRNA) data. This new species is differentiated from its congeners by a number of characters, including the absence of a cecal diverticula, the size and shape of the male copulatory organ and the shape of the vagina, as well as by differences in molecular data (SSU rRNA and LSU rRNA). The suitability of some sclerotized structures such as the male copulatory organ (MCO) as a taxonomic character is discussed; specifically, we found that the relationship between MCO and total length exhibit different trends in members of Calicotyle isolated from sharks, skates and chimaeras. Additional efforts to obtain sample of Calicotyle species and further molecular studies based on ribosomal and mitochondrial genes are necessary to clarify the degree of host specificity in this genus. Additionally, this is the first report of a member of Calicotyle to be reported in the Southeastern Pacific Ocean.

Two new species of Acanthocotyle Monticelli, 1888 (Monogenea: Acanthocotylidae), parasites of two deep-sea skates (Elasmobranchii: Rajiformes) in the South-East Pacific.

BACKGROUND: Parasites of deep-sea fishes from the South-East Pacific (SPO) are poorly known. Of c.1030 species of fish found in this area, 100-150 inhabit the deep-sea (deeper than 200 m). Only six articles concerning metazoan parasites of fish from deep-waters of SOP are known, and nine monogenean species have been reported. Currently, ten species are known in Acanthocotyle Monticelli, 1888 (Monogenea) and when stated, all of them are found in shallow waters (10-100 m). Acanthocotyle gurgesiella Ñacari, Sepulveda, Escribano & Oliva, 2018 is the only known species parasitizing deep-sea skates (350-450 m) in the SPO. The aim of this study was the description of two new species of Acanthocotyle from two Rajiformes. METHODS: In September 2017, we examined specimens of two species of deep-sea skates (Rajiformes), Amblyraja frerichsi (Krefft) and Bathyraja peruana McEachran & Myyake, caught at c.1500 m depth off Tocopilla, northern Chile, as a by-catch of the Patagonian tooth fish Dissostichus eleginoides Smitt fishery. Specimens of Acanthocotyle were collected from the skin of the skates. Morphometric (including multivariate analysis of proportional measurements, standardized by total length), morphological and molecular analyses (LSU rRNA and cox1 genes) were performed in order to identify the collected specimens. RESULTS: The three approaches used in this study strongly suggest the presence of two new species in the genus Acanthocotyle: Acanthocotyle imo n. sp. and Acanthocotyle atacamensis n. sp. parasitizing the skin of the thickbody skate Amblyraja frerichsi and the Peruvian skate Bathyraja peruana, respectively. The main morphological differences from the closely related species Acanthocotyle verrilli Goto, 1899 include the number of radial rows of sclerites, the non-discrete vitelline follicles and the number of testes. CONCLUSIONS: The two species of monogeneans described here are the only recorded parasites from their respective host species in the SPO. Assessing host specificity for members of Acanthocotyle requires clarifying the systematics of Rajiformes.

The complete mitochondrial genome and description of a new cryptic species of Benedenia Diesing, 1858 (Monogenea: Capsalidae), a major pathogen infecting the yellowtail kingfish Seriola lalandi Valenciennes in the South-East Pacific.

BACKGROUND: The monogenean Benedenia seriolae parasitizes fishes belonging to the genus Seriola, represents a species complex, and causes substantial impact on fish welfare in aquaculture systems worldwide. This study reports, for the first time, the complete mitochondrial genome of B. humboldti n. sp., a new cryptic species from the South-East Pacific (SEP). METHODS: The mitogenome of B. humboldti n. sp. was assembled from short Illumina 150 bp pair-end reads. The phylogenetic position of B. humboldti n. sp. among other closely related congeneric and confamiliar capsalids was examined using mitochondrial protein-coding genes (PCGs). Morphology of B. humboldti n. sp. was examined based on fixed and stained specimens. RESULTS: The AT-rich mitochondrial genome of B. humboldti is 13,455 bp in length and comprises 12 PCGs (atp8 was absent as in other monogenean genomes), 2 ribosomal RNA genes, and 22 transfer RNA genes. All protein-coding, ribosomal RNA, and transfer RNA genes are encoded on the H-strand. The gene order observed in the mitochondrial genome of B. humboldti n. sp. was identical to that of B. seriolae from Japan but different from that of B. seriolae from Australia. The genetic distance between B. humboldti n. sp. and B. seriolae from Japan was high. Minor but reliable differences in the shape of the penis were observed between Benedenia humboldti n. sp. and congeneric species. CONCLUSIONS: Phylogenetic analyses based on PCGs in association with differences in the shape of the penis permitted us to conclude that the material from the South-East Pacific represents a new species of Benedenia infecting S. lalandi off the coast of Chile. The discovery of this parasite represents the first step to improving our understanding of infestation dynamics and to develop control strategies for this pathogen infecting the farmed yellowtail kingfish, Seriola lalandi, in the South-East Pacific.

Morphological and molecular characterisation of digenean parasites of the Galápagos sheephead Semicossyphus darwini (Jenyns) with the re-description of Labrifer secundus Manter, 1940 (Lepidapedidae) from the Humboldt Current Large Marine Ecosystem.

The Humboldt Current Large Marine Ecosystem (HCLME) is recognised as one of the largest and most productive marine ecosystems. The estimated number of teleost fish species for the Chilean coast influenced by the HCLME reaches c.1,030 but digeneans have been reported for 39 teleost species, i.e. less than 4% of the potential hosts. During a survey of parasites of the Galápagos sheephead wrasse Semicossyphus darwini (Jennyns) (Perciformes: Labridae) from off northern Chile, we obtained representatives of three digenean species, Helicometrina nimia Linton, 1910 (Opecoelidae), Proctoeces humboldti George-Nascimento & Quiroga, 1983 (Fellodistomidae) and Labrifer secundus Manter, 1940 (Lepidapedidae). Labrifer secundus, originally described from Semicossyphus pulcher (Ayres, 1854) off the coast of Baja California, Mexico is re-described. Helicometrina nimia is a common parasite from littoral fishes in the HCLME. Semicossyphus darwini is a new host for these three digenean species.

Patterns of genetic variation and life history traits of Zeuxapta seriolae infesting Seriola lalandi across the coastal and oceanic areas in the southeastern Pacific Ocean: potential implications for aquaculture.

BACKGROUND: The monogenean, Zeuxapta seriolae, is a host-specific parasite that has an extensive geographical distribution on its host, Seriola lalandi, and is considered highly pathogenic in farmed fish. In recent years, developing cultures of S. lalandi in different coastal localities in Southeastern Pacific Ocean (SEP) have been affected by moderate and heavy infections of this parasite, attributed to contagion from wild to farmed fish. Here, we evaluated the pattern of genetic variations and biological traits of Z. seriolae in a spatial and temporal scale across its geographical distribution in SEP to determine its genetic status and biological traits, which could affect its transmission dynamics from wild to farmed fish. METHODS: Wild fish and their parasites were sampled from fisheries in the northern Chilean coast (NCC: 24°S-30°S) and Eastern islands (JFA: ca 33°S; 80°W) between 2012 and 2014. Fragments of 816 bp of the cytochrome c oxidase subunit I (COI) gene was sequenced for 112 individuals from NCC and 63 from JFA and compared using AMOVA. Prevalence and intensity of Z. seriolae were calculated for each area. The parasite body size, fecundity and size at sexual maturity were estimated for 177 parasites from NCC and 128 from JFA, and significant differences were evaluated using GLM. RESULTS: Geographical genetic structuring was detected for Z. seriolae across SEP, with a population in NCC and the other in JFA, both with the same high haplotype diversity. Neutrality tests and mismatch analyses indicated that both Z. seriolae populations are stable. Parasite biological traits such as fecundity, body size, and size at sexual maturity, and population parameters varied significantly between geographical areas. CONCLUSION: Two genetic groups of Z. seriolae were detected in wild fish across SEP. Because of the seasonal migration of wild host and temporal contact with farming, quantifying the genetic diversity and level of gene flow or isolation between parasite populations is useful for fish health management in farming. The smallest size of sexual maturity in parasites from NCC is predictive of shorter life cycles, and their high genetic diversity suggests high evolutionary potential and high transmission of this parasite to farmed hosts.

Parapedocotyle prolatili gen. n. et sp. n., a representative of a new subfamily of the Diclidophoridae (Monogenea), a gill parasite of Prolatilus jugularis (Teleostei: Pinguipedidae) from Chile.

Parapedocotylinae, a new subfamily, is proposed to accommodate the gen. n. et sp. n. Parapedocotyle prolatili (Monogenea: Diclidophoridae), a gill parasite of the Pacific sandperch, Prolatilus jugularis (Valenciennes) (Pinguipedidae) from northern Chile (30 degrees 56'S; 71 degrees 20'W). Among the Diclidophoridae Cerfontaine, 1895, the species of the Pedocotylinae Yamaguti, 1963 are unique by bearing the first pair of clamps (most posteriorly) in a haptoral projection. Pedocotyle MacCallum, 1913, the only genus in the Pedocotylinae, is characterised by the first pair of clamps non-pedunculate, modified and non-functional, without accessory suckers, and clamps of pairs 2-4 being pedunculate and functional. In contrast, the first pair of clamps in Parapedocotyle is well developed and functional at the terminal end of a long haptoral appendix and having clamp pairs 2-4 pedunculated, modified and apparently not functional. Seminal receptacle is preovarian in Parapedocotyle in opposition to its postovarian position in Pedocotyle. These differences justified the erection of the new subfamily Parapedocotylinae. The new subfamily is also supported by genetic analyses (18S rDNA, 28S rDNA and cox1 sequences) demonstrating that the Pedocotylinae and Parapedocotylinae belong to different clades in the Diclidophoridae.