Host-parasite relationship Octopus tehuelchus - Aggregata valdessensis in Patagonian coast, southwestern Atlantic Ocean.

Histopathology associated with the apicomplexan Aggregata valdessensis in the wild population of Octopus tehuelchus was studied. Moreover, to analyze the impact of the prevalence and infection intensity on the dorsal mantle length, sex, maturity stages, condition index and total hemocyte counts on wild O. tehuelchus, Generalized Linear Models (GLMs) were applied. Kruskal-Wallis (KW) test was used to compare growth parameter (RNA/DNA ratios) in infected and uninfected octopuses. Lesions including hypertrophy of host cells and destruction of the tissue organ architecture as well as the replacement of host tissue by parasites were associated with infection. The infection caused an inflammatory defensive response in form of hemocytic infiltration.. The prevalence of apicomplexan increased with octopus dorsal mantle length (GLM, t = 2.68; p = 0.007). Infection intensity was significantly higher in male octopus (GLM, t = 2.69; p = 0.009) and decreased at higher temperatures in both sexes (GLM, t = -2.42; p = 0.018). The digestive gland condition index did not significantly vary with infection intensity, and no significant relationship between total hemocyte counts and coccidian infection was found (Lineal Model, t = 0.46; p = 0.64). The acid nucleic ratio was significantly lower in octopuses infected by A. valdessensis (Kruskal Wallis test, H = 4.8; p = 0.02), suggesting a detrimental effect on O. tehuelchus growth caused by the coccidian.

Parasitological survey of coastal birds from the Magellanic coast, Southwestern Atlantic Ocean.

Ecto and endoparasites of four species of coastal birds, Haematopus ater, Larus dominicanus, Leucophaeus scoresbii (Charadriiformes), and Lophonetta specularioides (Anseriformes), are reported from Puerto Deseado on the Patagonian coast, Argentina. Only H. ater was infested with lice (Phthiraptera), belonging to 2 species (Ischnocera, Amblycera). A total of 19 helminth species were found parasitizing the coastal birds studied: 4 cestodes (1 Tetrabothriidae, 3 Cyclophyllidea); 11 trematodes (2 Gymnophallidae, 3 Microphallidae, 2 Notocotylidae, 1 Philophthalmidae, 2 Renicolidae, 1 Schistosomatidae); 3 nematodes (1 Anisakidae, 2 Acuariidae); and 1 acanthocephalan (Polymorphidae). Although some isolated records have been previously reported for these birds, the present work provides a parasitofauna study for H. ater, L. scoresbii, and L. specularioides for the first time. Endoparasites reflected the feeding habits of the birds; the parasite assemblage of L. dominicanus was the richest, indicating their wide prey spectrum and the diversity of the habitats frequented. A great species richness of trematodes, whose life cycles are partially known, suggests that L. specularioides feeds upon crustaceans and small bivalves. The blackish oystercatcher H. ater preys upon the limpet Nacella magellanica which hosts two larval trematodes corresponding to the adults found parasitizing it.

Comparative genomics for Agmasoma sp. (Microsporidia) parasitising invasive Carcinus aestuarii and Carcinus maenas in Argentina.

Carcinus spp. are global aquatic invaders and carriers of several parasites, including a taxonomically unrecognised microsporidian recently observed from Argentina. We provide genome drafts for two parasite isolates, one from Carcinus maenas and one from Carcinus aestuarii, and use multi-gene phylogenetics and genome comparison methods to outline their similarities. Their SSU genes are 100 % similar and other genes have an average similarity of 99.31 %. We informally name the parasite Agmasoma carcini, terming the isolates Ac. var. aestuarii and Ac. var. maenas, following the wealth of genomic data available for each. This study follows on from Frizzera et al. (2021), where this parasite was first histologically identified.

Paramonostomum deseado n. sp. (Digenea: Notocotylidae) parasitizing the South American Black Oystercatcher and their atypical life cycle from the Patagonian coast.

By way of morphological and molecular analysis we describe a new species of notocotylid, Paramonostomum deseado n. sp., parasitizing Haematopus ater from Argentina and we contribute to elucidate its life cycle. Within this genus, 4 groups can be morphologically distinguished according to body shape: 'Oval', 'Pyriform', 'Elongate', 'Overlong'. The new species belongs to the 'Elongate group', which presents a wide variation in body length (597­4500 µm). The new species, Paramonostomum caeci from Australia, Paramonostomum actitidis from the Caribbean and Paramonostomum alveoelongatum from Russia share the smallest range of body size in this group (<1130 µm). The new species more closely resembles P. actitidis but differs from it by cirrus-sac length, which is shorter in the new species (97­146 vs 280­430 µm in P. actitidis), and egg size which is larger in the new species (25­33 vs 18­20 µm in P. actitidis). Paramonostomum deseado n. sp. uses the limpet Nacella magellanica as both first and second intermediate hosts in which metacercariae encyst inside the redia. This is the first abbreviated cycle described for notocotylid species. Ribosomal RNA sequences provided for adults (ITS1, ITS2 and 28S) and metacercariae inside the rediae (ITS1) support the species identification and the correspondence among stages. Phylogenetic analysis based on 28S placed P. deseado n. sp. close to other Notocotylus spp. and Paramonostomum anatis. Molecular results demonstrate that the hosts involved in the life cycles and the habitat more than morphological differences are determining the phylogenetic relationships in members of Notocotylidae.

A phylogenetically unresolved apicomplexan (APXSc) causing swirl lesions in the Tehuelche scallop, Aequipecten tehuelchus, from the Southwest Atlantic coast.

The study reports a previously unknown apicomplexan (APXSc) parasite infecting wild scallops Aequipecten tehuelchus (d'Orbigny, 1842) from two separate areas (La Tapera and Punta Conos) of the San José gulf, in Patagonia Argentina. Histology, transmission electron microscope, molecular analyses and in situ hybridization were performed to describe the morphology of APXSc, and confirm its phylogenetic status. The prevalence of APXSc infection was 24% and 72% in scallops from La Tapera and Punta Conos, respectively. Seasonal variation was observed for scallops from La Tapera, recording highest prevalence in summer. A positive relationship between the presence of the APXSc and the size of the scallops was observed. A SSU rDNA consensus sequence of 1758 base pairs was generated which has a 94.8% identity to sequences obtained from a pathogenic apicomplexan parasite infecting Ostrea chilensis in New Zealand, but not closely related to other apicomplexans. The asexual reproduction, i.e. merogony, occurs in the Tehuelche scallop whilst the gamogonic and sporogonic stages were absent, suggesting a yet unknown definitive host. Severe host inflammation response involving fibroblast-like hemocytes surrounding the APXSc in the form of granuloma-like "swirls" is characteristic for this apicomplexan infection. Further studies are needed to reveal the life cycle, and presumable pathogenicity of APXSc.

A new species of Auriculostoma (Digenea: Allocreadiidae) in South America: life cycle and phylogenetic relationships.

Auriculostoma is a genus of digenean (Trematoda) whose adults are parasites of Neotropical freshwater fishes. We describe Auriculostoma ocloya n. sp. using morphological and molecular tools, and we elucidated its life cycle, the first known of a species of this genus. The first intermediate host is the bivalve Pisidium ocloya, the second intermediate host is the amphipod Hyalella sp., and the definite hosts are siluriform fishes. The adult presents a single pair of muscular lobes on either side of the oral sucker with a broad base, stretching from ventrolateral to dorsolateral side, a structure also present in the rest of species of the genus. Nevertheless, the new species differs from all congeners by the combination of several traits, and mainly because on the dorsal side "free" ends of the lobes are absent because they are fused. This is the first study to provide sequence data on larval and adult stages of a species of Auriculostoma. Our phylogenetic analysis demonstrated its basal position among species of the genus. Therefore, integrative morphological, molecular, and life cycle data on other South American species of the genus, would contribute to reveal more patterns in the allocreadiid systematics.

Symbionts of invasive and native crabs, in Argentina: The most recently invaded area on the Southwestern Atlantic coastline.

Biological invasions have the capacity to introduce non-native parasites. This study aimed to determine whether the invasive green crab population, Carcinus spp., on the Southwestern Atlantic coast of Argentina harbours any symbionts, and whether these may spillover or spillback between native crabs, Cyrtograpsus altimanus and C. angulatus. Macroscopy, histology, and molecular analyses of some parasites were used to describe and compare their diversity across the three species of crab. We also evaluated the susceptibility of invasive Carcinus spp. to a native digenean, Maritrema madrynense, via experimental infections (exposure and cohabitation). Our results revealed that the green crab pathobiome included similar symbiotic groups to native crabs. This included putative viral, bacterial, and protozoan parasites. Haplosporidium-like observations were recorded in all crab species, and a single green crab was found to be parasitized by an Agmasoma-like microsporidium. Metagenomic analysis of one individual revealed additional symbiotic diversity (46 bacteria, 5 eukaryotic species). The green crabs were infected by more microparasite taxa than the native crabs (5:3). Wild populations of Carcinus spp. were free of metazoan parasites and are shown not to be susceptible to M. madryense under experimental conditions. Our results suggest a reduction/escape of macroparasites (trematode Maritrema madrynense; acanthocephalan Profilicollis chasmagnathi) in invasive Carcinus spp. compared to their native competitors.

Gulls as carriers of antimicrobial resistance genes in different biogeographical areas of South America.

The aim of this communication was to establish if Enterobacterales associated with gulls in Argentina harbored antimicrobial resistance (AMR) genes. We analyzed cloacal swabs in two contrasting areas: Ensenada, Buenos Aires province (26 Larus dominicanus and 22 Chroicocephalus maculipennis) and Puerto Madryn, Chubut province (20 L. dominicanus). In Ensenada, bla CTX-M and mcr-1 genes, were isolated from both gull species, whereas in the Puerto Madryn, only bla CTX-M gene was found. We report for the first time C. maculipennis as carrier of AMR. The finding of AMR in wildlife constitutes a useful tool in evaluating the anthropogenic impact on environmental health.

Cryptic speciation of the zoogonid digenean Diphterostomum flavum n. sp. demonstrated by morphological and molecular data.

Diphterostomum brusinae (Zoogonidae) is a digenean species that has been recorded worldwide parasitizing marine fishes. Several species have been synonymized with D. brusinae because they lack conspicuous morphological differences. However, due to the breadth of its geographic distribution and the variety of hosts involved in the life cycles, it is likely to be an assemblage of cryptic species. Diphterostomum flavum n. sp. is described here as a morphologically cryptic relative of D. brusinae, in the fish Pinguipes brasilianus (Pinguipedidae) off the Patagonian coast, Southwestern Atlantic Ocean, and its life cycle is elucidated through morphology and molecular analysis. This species uses the gastropod Buccinanops deformis (Nassariidae) as first and second intermediate host with metacercariae encysting within sporocysts. They also, however, use the polychaete Kinbergonuphis dorsalis (Onuphidae) as second intermediate host. No morphological differences were found between adults of D. flavum n. sp. and D. brusinae; however, the number of penetration glands of the cercariae, a diagnostic feature, differed (9 vs. 3 pairs), as well as the ITS2 sequences for the two species. This work provides morphological and molecular evidence of cryptic diversification among species described as D. brusinae, in which the only clear differences are in larval morphology and host spectrum. The strict specificity to the snail acting as the first intermediate host and the variety of fishes with different feeding habits acting as definitive hosts support the likely existence of multiple cryptic species around the world.

TITLE: Spéciation cryptique du Digène Zoogonidae Diphterostomum flavum n. sp. démontrée par des données morphologiques et moléculaires. ABSTRACT: Diphterostomum brusinae (Zoogonidae) est une espèce de digène, parasite des poissons marins, qui a été signalée dans le monde entier. Plusieurs espèces ont été synonymisées avec D. brusinae parce qu'elles ne montrent pas de différences morphologiques visibles. Cependant, en raison de l'étendue de sa répartition géographique et de la variété d'hôtes impliqués dans les cycles de vie, il est probable qu'il s'agisse d'un assemblage d'espèces cryptiques. Diphterostomum flavum n. sp. est décrit ici comme une espèce morphologiquement cryptique de D. brusinae, chez le poisson Pinguipes brasilianus (Pinguipedidae) au large de la côte de la Patagonie, dans le sud-ouest de l'océan Atlantique, et son cycle de vie est élucidé par la morphologie et l'analyse moléculaire. Cette espèce utilise le gastéropode Buccinanops deformis (Nassariidae) comme premier et deuxième hôte intermédiaire avec des métacercaires s'enkystant dans les sporocystes. Cependant, elle utilise également le polychète Kinbergonuphis dorsalis (Onuphidae) comme deuxième hôte intermédiaire. Aucune différence morphologique n'a été trouvée entre les adultes de D. flavum n. sp. et D. brusinae, mais le nombre de glandes de pénétration des cercaires, une caractéristique diagnostique, diffère (9 paires contre 3), ainsi que les séquences ITS2 pour les deux espèces. Ce travail fournit des preuves morphologiques et moléculaires de la diversification cryptique parmi les espèces décrites comme D. brusinae, dans lesquelles les seules différences claires sont dans la morphologie des larves et le spectre de l'hôte. La spécificité stricte au mollusque servant de premier hôte intermédiaire et la variété de poissons avec différentes habitudes alimentaires servant d'hôtes définitifs soutiennent l'existence probable de plusieurs espèces cryptiques dans le monde.

Notocotylus primulus n. sp. (Trematoda: Notocotylidae) from the crested duck Lophonetta specularioides (Aves, Anatidae) from Patagonian coast, southwestern Atlantic Ocean.

Species in the genus Notocotylus (Trematoda, Notocotylidae) are cosmopolitan parasites of mainly aquatic birds and, to a lesser extent, mammals. In this paper we describe Notocotylus primulus n. sp. parasitizing the Crested Duck, Lophonetta specularioides, from the Patagonian coast, southwestern Atlantic Ocean, based on morphological and molecular data. The new species could be distinguished from its congeners by the combination of several features; among them, the position of the genital pore that is prebifurcal and located close to the posterior margin of the oral sucker, a greater number of both ventral papillae and uterine loops, and a large cirrus sac. Molecular results of the ITS2 sequence support the location of this new species in the genus Notocotylus due to a close relationship with Notocotylus malhamensis (the lower genetic distance of the ITS2 sequences).

A single clonal lineage of transmissible cancer identified in two marine mussel species in South America and Europe.

Transmissible cancers, in which cancer cells themselves act as an infectious agent, have been identified in Tasmanian devils, dogs, and four bivalves. We investigated a disseminated neoplasia affecting geographically distant populations of two species of mussels (Mytilus chilensis in South America and M. edulis in Europe). Sequencing alleles from four loci (two nuclear and two mitochondrial) provided evidence of transmissible cancer in both species. Phylogenetic analysis of cancer-associated alleles and analysis of diagnostic SNPs showed that cancers in both species likely arose in a third species of mussel (M. trossulus), but these cancer cells are independent from the previously identified transmissible cancer in M. trossulus from Canada. Unexpectedly, cancers from M. chilensis and M. edulis are nearly identical, showing that the same cancer lineage affects both. Thus, a single transmissible cancer lineage has crossed into two new host species and has been transferred across the Atlantic and Pacific Oceans and between the Northern and Southern hemispheres.

Histopathological study of Corydoras micracanthus (Siluriformes: Callichthyidae) parasitized by the nematode Procamallanus (Spirocamallanus) pintoi (Camallanidae) from Lesser River, northwestern Argentina, South America

The nematode parasites can cause mechanical injuries such as irritation or atrophy of tissue and occlusions of the digestive tract, blood vessels or other ducts in their hosts. Some species belonging to the family Camallanidae have been the focus of histopathological studies because they lacerate the host´s intestinal wall with their buccal capsule. In this study, we describe the histopathological effects produced by Procamallanus (Spirocamallanus) pintoi in the small fish Corydoras micracanthus from Lesser River, in the northwest of Argentina, South America. The samples were processed with routine histology techniques. At light microscopy, specimens of P. (S.) pintoi were observed occupying the lumen of the gut. The histopathological sections revealed damage to the host intestine with the total destruction of the mucosa and submucosa, observed mainly in the region which is surrounding the buccal capsule of parasites. Moreover, an inflammatory reaction was observed with lymphatic accumulations in the affected tissue. This kind of degenerative and necrotic changes in the mucosa could affect negatively the absorption function of the host intestine, which may have adverse effects of the health status of fish. The present study provides the firs data about the histopathological effects of P. (S.) pintoi in C. micracanthus.

Atypical lesions and infection sites of larval trematodes in marine gastropods from Argentina.

This study documents the lesions caused by 9 digenean larvae parasitizing 6 marine gastropod species from Patagonia, Argentina. Most digeneans produce a lesion where the parthenitae replace the gonadal tissues and also occupy part of the digestive gland. Three atypical lesions and tropism were observed in (1) Trophon geversianus (Muricidae) infected by Renicolidae gen. et sp. 1, where the sporocysts occur inside the gonad acini which retain their general structure due to the persistence of the acinus wall, (2) Nacella magellanica (Nacellidae) infected by Renicolidae gen. et sp. 2, where the gonad is not affected and the sporocysts are located only among the digestive gland tubules, and (3) Siphonaria lessonii (Siphonariidae) infected by Schistosomatidae gen. et sp., where fully developed cercariae occur inside the digestive tubules. At high infection intensities or in double infections, larvae can invade other organs (mantle, foot or gill) and, on occasion, may elicit hemocytic encapsulation.

Parasites in two coexisting bivalves of the Patagonia coast, southwestern Atlantic Ocean: The Puelche oyster (Ostrea puelchana) and false oyster (Pododesmus rudis).

The purpose of this study was to compare the parasites of two coexisting bivalves, the edible Puelche oyster (Ostrea puelchana) and the false oyster (Pododesmus rudis) that lives attached to O. puelchana shells, and to investigate their host specificity. Samples from wild populations, 465 O. puelchana and 131 P. rudis, were collected seasonally during two years in the San José Gulf (northern Patagonia, Argentina) and were processed using standard histological techniques. To increase the natural low prevalences of Bonamia spp. and Perkinsus spp. that are present in wild populations, an in situ experiment was performed by maintaining captive sentinel bivalves at high densities inside a plastic mesh bag to enhance parasite transmission. Polymerase chain reaction (PCR) assays were used to test for apparent Bonamia sp. infections among captive sentinel O. puelchana specimens (n = 80), and Ray's fluid thioglycollate medium (RFTM) assays and histological immunoassays tested for apparent Perkinsus sp. infections among captive sentinel P. rudis specimens (n = 100). Despite histological observations that revealed the presence of microcells resembling Bonamia sp. infecting hemocytes of some Puelche oysters, PCR assays did not confirm that parasite identification. Among captive sentinel P. rudis that showed histological evidence of Perkinsus sp. infections, neither RFTM nor immunoassays confirmed such parasites. Ostrea puelchana from wild populations were occasional hosts for both Rickettsia-like organism (RLOs) and Urastoma-like turbellarians. In contrast, six parasite taxa infected P. rudis from coexisting populations, including RLOs, Urastoma-like turbellarians, an intracellular gregarine species, Nematopsis-like oocysts, an unidentified coccidian and a Perkinsus qugwadi-like protozoan. These results demonstrated specific infection patterns of the identified parasites in relation to their hosts.

Life Cycle of the Parasite Profilicollis chasmagnathi (Acanthocephala) On the Patagonian Coast of Argentina Based On Morphological and Molecular Data.

This study verifies the identity of adult specimens of the parasite Profilicollis chasmagnathi (Acanthocephala, Polymorphidae) recovered from kelp gulls Larus dominicanus (Aves, Laridae), and cystacanths found in crabs Cyrtograpsus altimanus (Crustacea, Decapoda) from the southwestern Atlantic coast. The life cycle of this parasite is elucidated in the intertidal zone of Patagonia, Argentina, based on morphological and molecular data. Preferences by size and sex of the intermediate host and seasonal variation of this parasite are provided, contributing to the knowledge of this host-parasite association.

Redescription and life cycle of the monorchiid Postmonorcheides maclovini Szidat, 1950 (Digenea) from the Southwestern Atlantic Ocean: Morphological and molecular data.

The adult monorchiid, Postmonorcheides maclovini Szidat, 1950, digenean parasite of the Patagonian blennie Eleginops maclovinus (Cuvier) (Eleginopidae) from Puerto Deseado (47° 45' S, 65° 55' W), Argentina, was characterized and its life cycle elucidated. P. maclovinus is the only species of the genus Postmonorcheides, proposed by Szidat (1950) from Tierra del Fuego province (~54° S), Argentina. This digenean uses the Patagonian blennie as definitive host, and the intertidal bivalve Lasaea adansoni (Gmelin) (Lasaeidae) as both first and second intermediate hosts (metacercariae encyst inside sporocysts), being the first record of this clam as intermediate host of trematode parasites. The cercaria may, in addition to encysting in the sporocyst, emerge and presumably infect other intermediate hosts. This is the second report of a monorchiid species with metacercariae encysting inside the sporocyst. Adults were found parasitizing the fish stomach, pyloric caeca and intestine with a prevalence of 100%; sporocysts with cercariae and/or metacercariae were found parasitizing the gonad of the bivalve with a prevalence of 2.78%. The cercariae possess a well-developed tail and eye-spots are absent. The ITS1 sequence from the adult digeneans found in the Patagonian blennie, identified as P. maclovini, was found to be identical to the ITS1 sequences obtained both from sporocysts containing cercariae and encysted metacercariae found in L. adansoni.

Revision of the family Gymnophallidae Odhner, 1905 (Digenea) based on morphological and molecular data.

This paper reviews the family Gymnophallidae, recognizing as valid seven genera; four within the subfamily Gymnophallinae: Gymnophallus Odhner, 1900 (syn. Meiogymnophallus Ching, 1965), Paragymnophallus Ching, 1973, Pseudogymnophallus Hoberg, 1981, and Bartolius Cremonte, 2001, and three in the Parvatrematinae: Parvatrema Cable, 1953, Lacunovermis Ching, 1965, and Gymnophalloides Fujita, 1925. Specimens representing one species of each available genus were chosen from those well-described and non controversial species, for which strong morphological information was available, and used for molecular studies (ITS1-5.8S-ITS2-28S rDNA strands were sequenced). The presence or absence of a pars prostatica differentiates between the 2 subfamilies, Gymnophallinae and Parvatrematinae. The characters used to differentiate genera are: location of the ovary (pre-, post- or inter-testicular), size and location of the genital pore (inconspicuous and located at the anterior margin of the ventral sucker, or conspicuous and located at some distance from the anterior margin of ventral sucker), presence of caecal pockets, and presence or absence of ventral pit (a muscular structure which can be either well-developed, similar in size and musculature to the ventral sucker, or be poorly developed). The characters previously used to distinguish among genera that actually should be considered to separate species include: shape of tegument spines (broad, sharp or serrated), presence of lateral projections on the oral sucker (also called papillae or lips), shape of the seminal vesicle (unipartite or bipartite), shape of the prostatic duct (elongate or oval), presence of papillae on the genital pore, shape of the genital atrium (tubular, wide, oval), shape of the vitellaria (follicular in a variable degree, paired or single), shape of the excretory vesicle (V or Y), and extension of uterus (restricted to forebody, at hindbody or extending in both). Additionally, some of these characters may vary with the age of worm. The morphological and molecular information obtained in this study provided strong support for recognizing seven valid genera in the family Gymnophallidae.

A new species of Haplosporidium Caullery & Mesnil, 1899 in the marine false limpet Siphonaria lessonii (Gastropoda: Siphonariidae) from Patagonia.

A new species of Haplosporidium Caullery & Mesnil, 1899 parasitising the pulmonate gastropod Siphonaria lessonii Blainville in Patagonia, Argentina, is described based on morphological (scanning and transmission electron microscopy) and sequence (small subunit ribosomal RNA gene) data. Different stages of sporulation were observed as infections disseminated in the digestive gland. Haplosporidium patagon n. sp. is characterised by oval or slightly subquadrate spores with an operculum that is ornamented with numerous short digitiform projections of regular height, perpendicular to and covering its outer surface. The operculum diameter is slightly larger than the apical diameter of the spore. Neither the immature nor mature spores showed any kind of projections of the exosporoplasm or of the spore wall. Regarding phylogenetic affinities, the new species was recovered as sister to an undescribed species of Haplosporidium Caullery & Mesnil, 1899 from the polychaete family Syllidae Grube from Japanese waters. The morphological characters (ornamentation of the operculum, spore wall structure, shape and size of spores, and the lack of spore wall projections) corroborate it as an as yet undescribed species of Haplosporidium and the first for the phylum in marine gastropods of South America. Siphonaria lessonii is the only known host to date.

A new species of gymnophallid (Digenea) and an amended diagnosis of the genus Gymnophalloides fujita, 1925.

Morphological and molecular evidence suggest that specimens formerly described as Lacunovermis sp. from Nacella (Patinigera) spp. (Patellogastropoda: Patellidae) belong to a new species of Gymnophalloides Fujita, 1925 . Based on the new information, they are identified as Gymnophalloides nacellae n. sp. The new species differs from Gymnophalloides tokiensis, Gymnophalloides seoi , and Gymnophalloides heardi mainly through the presence of a group of papillae located on the ventral surface between oral and ventral suckers. A detailed morphological study revealed the lack of pars prostatica, a character previously reported in G. seoi , which is why it was formerly placed in the Gymnophallinae. Molecular information proved that G. nacellae is close to G. seoi , being nestled together with Parvatrema representatives. This molecular information, along with the absence of pars prostatica, allows these 2 genera to be placed in Parvatrematinae. An amended diagnosis of Gymnophalloides is provided. Histological sections of mantle epithelium of the limpet show metacercariae attached by their oral and ventral suckers in a similar manner to G. seoi in its host, the oyster Crassostrea gigas . Tissue reaction includes cells of outer mantle epithelium being stretched by sucker attachment, hemocyte infiltration of connective tissue between mantle epitheliums, and abnormal calcareous deposition on the inner surface of the shell.