Structure of the metazoan parasite communities of haemulid fish (Actinopterygii: Perciformes) in the South Atlantic Ocean: a comparative approach.

Haemulidae represents one of the most diverse, widespread and conspicuous families of Perciformes, in which most species are marine, some brackish and rarely from freshwater. From April 2009 to July 2012, 120 specimens of Conodon nobilis, 60 Orthopristis rubra and 50 Anisotremus virginicus were collected off the State of Rio de Janeiro, Brazil, and analyzed for metazoan parasites. A total of 16 parasite taxa were found on/in A. virginicus, 19 on/in Co. nobilis and 21 on/in O. rubra, in which ectoparasites were most common on A. virginicus and Co. nobilis and endoparasites in O. rubra. All parasites showed aggregate pattern of distribution (discrepancy index values higher than 0.70). Mean abundance, species richness and Brillouin index (diversity) differed among the host species, i.e., O. rubra showed the highest values, followed by A. virginicus and Co. nobilis with intermediate and lowest values, respectively. At infracommunity level was possible to observe high similarity of parasite composition among the three host species. The digeneans Leurodera decora and Monorchis latus, and the acanthocephalan Koronacantha sp. represent new locality records off Brazil. Moreover, five, four and two taxa are reported, for the first time, parasitizing Co. nobilis, A. virginicus and O. rubra, respectively.

Cucullanus pinnai pinnai and C. pinnai pterodorasi (Nematoda Cucullanidae): what does the integrative taxonomy tell us about these species and subspecies classification?

Cucullanus pinnai has been divided in two subspecies (C. pinnai pinnai and C. pinnai pterodorasi) based on the morphology of oesophastome. While C. pinnai pinnai apparently shows low host specificity and broad geographic occurrence, with certain morphological variations, C. pinnai pterodorasi was reported once, parasitizing Pterodoras granulosus. We used an integrative taxonomic approach to evaluate whether or not populations of C. pinnai pinnai from Trychomycterus spegazzinii (Escoipe River, Argentina) and Pimelodus fur (Miranda River, Brazil), and of C. pinnai pterodorasi from Pterodoras granulosus (Miranda River, Brazil) are conspecific. Parasites were observed using light microscopy and genetically characterized based on partial sequences of the 18S and 28S rDNA, ITS1-5.8S-ITS2, and COI mtDNA. Phylogenies were reconstructed and the Generalized Mixed Yule Coalescent (GMYC), Poisson Tree Process (bPTP), and Automatic Barcode Gap Discovery (ABGD) were used for species delimitation purposes. The present samples formed well-supported monophyletic assemblages, corroborating in part the results of morphological analyses; however, they grouped according to geographic origin. Species delimitation suggested conspecificity of C. pinnai pinnai with C. pinnai pterodorasi from Brazil; consequently, the morphology of oesophastome may be an intraspecific variation. Results also indicated that C. pinnai may represent a species complex as samples from Argentina were suggestive of an independent specific entity. However, definitive affirmations are premature, since there is no autapomorphy for separating C. pinnai from Brazil and Argentina and sampling was limited to three host species from two river basins. The phylogenetic reconstructions also confirmed the artificiality of some genera within Cucullanidae.

Review of the parasitic nematodes of marine fishes from off the American continent.

The ichthyofauna of the Atlantic and Pacific coasts off the American continent is very rich. Consequently, a high biodiversity of nematodes parasitizing these vertebrates is also expected. Currently, data on nematode parasites of marine fish off the Americas are fragmented. A review of all adult nematode species reported parasitizing marine fish from off the American continent is herein presented, as well as comments on their patterns of diversity, life cycles and advances in the taxonomic and phylogenetic knowledge. A total of 209 valid species, 19 species inquirendae and 6 dubious records have been recorded, the majority from the fish taxa Eupercaria and Perciformes. The families Sciaenidae, Serranidae and Lutjanidae, as well as the tropical and temperate Atlantic waters, exhibited the highest records of parasitic nematodes. The Cucullanidae, Philometridae and Cystidicolidae were the most speciose families of nematodes, which may be related to technological advances and relatively recent efforts of taxonomists, resulting in description of new taxa and the resolution of taxonomic problems. Numerous taxonomic questions still need resolution and, even though genetic data have been important for this process, the database is very scarce. This is the first review on all currently known nematode species parasitizing marine fish off the Americas and may serve as an important basis of reference for future approaches on these organisms.

Parasite communities and their ecological implications: comparative approach on three sympatric clupeiform fish populations (Actinopterygii: Clupeiformes), off Rio de Janeiro, Brazil.

Fish parasite communities can be directly influenced by characteristics of host species. However, little is known about the host-parasite relationships in commercially important fish of the southeastern Atlantic. To address this knowledge gap, a comparative analysis of the parasite communities of three sympatric Clupeiformes was conducted. Cetengraulis edentulus (Engraulidae), Opisthonema oglinum (Clupeidae) and Sardinella brasiliensis (Clupeidae) were collected from an estuarine lagoon near Rio de Janeiro, Brazil. Prevalence, abundance and aggregation were estimated for infrapopulations; richness, diversity, evenness and dominance for infracommunities. The three component communities were compared using both quantitative and qualitative components. Canonical discriminant analysis was used to determine if a host population could be characterised by the component community of its parasites. Multivariate models revealed that host species, a proxy for diet and phylogenetic relationships, was the main factor influencing the composition of parasite infracommunities. Diet was found to be the main factor shaping the communities of endoparasites, in which digeneans were dominant and best indicator of host population. Ectoparasites (copepods, isopods and monogeneans) displayed strong host-specificity with some species restricted to a single host population. The similarity of the component communities of the two clupeid populations demonstrated the influence of host phylogeny. Parasite infracommunities exhibited low diversity and high dominance, with many taxa restricted to a single host species (specialists) and few occurring in more than one (generalists). Host phylogeny and by extension, diet, morphology and coevolution with parasites appear to be important factors in determining the host-parasite relationships of clupeiform fish in the southeastern Atlantic.

Deep in the systematics of Camallanidae (Nematoda): using integrative taxonomy to better understand the phylogeny and consistency of diagnostic traits.

Due to conflicts between classic and molecular systematics of Camallanidae, different data types were used for the first time, to better understand the evolutionary history and taxa consistency within this family. Genetic [18S and 28S rDNA; cytochrome c oxidase subunit I (COI) mtDNA], morphological and life history traits were used to infer phylogenies using Bayesian inference, reconstructed from separated and concatenated datasets. The consistency of tree and morphological traits was evaluated using the consistency index. Characters were mapped on the trees and the phylogenetic informativeness of genetic markers was estimated. Phylogenetic informativeness of 18S provided better resolution for outer nodes, COI for inners and 28S had an intermediate profile. New sequences for two camallanid species were obtained. Phylogenies of genetic and concatenated data largely agreed, showing more divergence in the COI dataset, due to its higher mutation rate vs stable morphology for diagnosing higher taxa. No genus sustained monophyly. The lack of autapomorphy and phylogenetic proximity supported the partition of Batrachocamallanus as synonym of Procamallanus and Spirocamallanus, which should not be considered as subgenera. Although traits of buccal capsule, male tail, habitat, host and biogeographic were highly consistent, intrinsic patterns varied according to different taxa assemblages. Morphological systematics of Camallanidae, based on buccal capsule, is artificial for certain taxa.

Three new species of Anacanthorus Mizelle & Price, 1965 (Monogenea: Dactylogyridae) from Markiana nigripinnis Perugia (Actinopterygii: Characidae) in Pantanal wetlands, Brazil.

Three monogenean species, Anacanthorus luquei n. sp., A. scholzi n. sp. and A. cohenae n. sp. are described from the gills of the tetra fish Markiana nigripinnis (Perugia) (Characidae), collected in the Pantanal wetlands, State of Mato Grosso do Sul, Brazil. Among other differences, Anacanthorus luquei n. sp. differs from the most morphologically similar species, based on the structure of the accessory piece as follows: branches with smooth margins (vs with irregular margins in A. cuticulovaginus), without pointed projections at distal end (vs with projections in A. dipelecinus) and with 2 branches (vs 3 in A. quinqueramus). Anacanthorus scholzi n. sp. is most morphologically similar to A. luquei n. sp., differing from it because one of the branches of the accessory piece is bifurcated at the distal portion. Anacanthorus cohenae n. sp. can be differentiated from the congeners based on the combination of the following features: MCO cylindrical and robust with sclerotised flanges on the extremities, accessory piece V-shaped, bearing two branches similar in length and with blunt distal ends, and hooks with a proximal bulb. This is the first parasitological study on M. nigripinnis and, currently, Anacanthorus allocates 88 species infesting characiform fishes in the Neotropical region, including the three new species described here.

An integrative approach assesses the intraspecific variations of Procamallanus (Spirocamallanus) inopinatus, a common parasite in Neotropical freshwater fishes, and the phylogenetic patterns of Camallanidae.

Integrative taxonomy was used to evaluate two component populations of Procamallanus (Spirocamallanus) inopinatus in Brazil and the phylogeny Camallanidae. Parasite populations were collected in the characiform Anostomoides passionis from River Xingu (Amazon basin) and Megaleporinus elongatus from River Miranda (Paraguay basin). Morphology was analysed using light and scanning electron microscopy (SEM). Genetic characterization was based on partial sequences of the 18S and 28S rDNA, and COI mtDNA. Phylogenies were based on 18S and COI due to data availability. Generalized Mixed Yule Coalescent (GMYC), Poisson Tree Process (PTP) and *BEAST were used for species delimitation and validation. SEM revealed for the first time the presence of minute denticles and pore-like structures surrounding the oral opening, phasmids in females and confirmed other important morphological aspects. Statistical comparison between the two-component populations indicated morphometric variations, especially among males. The different component population of P. (S.) inopinatus showed variable morphometry, but uniform morphology and were validated as conspecific by the GMYC, PTP and *BEAST. Some camallanid sequences in GenBank have incorrect taxonomic labelling. Host, environment and geographic aspects seem to be related to some lineages within Camallanidae; however, their real phylogenetic meanings are still unclear.

Dadaytrema oxycephala (Digenea: Cladorchiidae) in definitive host Pimelodus blochii (Pisces: Pimelodidae), with morphological and geographic distribution data in fishes from the South America.

The aim of this study was to describe structural and ultrastructural morphological data of Dadaytrema oxycephala (Diesing, 1836) Travassos, 1931 in Pimelodus blochii Valenciennes, 1840 from western Amazon (Brazil), besides geographic distribution in host fishes from South America. Of 160 P. blochii examined in the rivers Acre and Iaco, 15% were infected by D. oxycephala with mean intensity of 6.5 ± 1.9 and mean abundance of 1.0 ± 5.1. All parasites were found in the intestine of the infected hosts. The structural and ultrastructural characteristics of the developmental stages of D. oxycephala are presented and discussed. Dadaytrema oxycephala has a wide geographic distribution in South America, parasitizing fish species of Characiformes and Siluriformes from Brazil, Venezuela, Argentina, Peru and Paraguay. We elucidated part of life cycle of D. oxicephala in P. blochii, which is a definitive host for this digenean.

Integrative taxonomy of Sprentascaris mahnerti and phylogeny of raphidascaridid nematodes (Nematoda: Ascaridoidea: Raphidascarididae) suggest the monophyly of Sprentascaris.

Sprentascaris mahnerti (Nematoda: Raphidascarididae) collected from Loricariichthys labialis (Siluriformes: Loricariidae) in the Pantanal wetlands, State of Mato Grosso do Sul (Brazil), was redescribed using light and scanning electron microscopy (SEM), and genetically characterised along with two other raphidascaridids: Raphidascaroides brasiliensis and Ro. moraveci. Due to the systematic discussion regarding Raphidascaris and Sprentascaris, as well as the poor knowledge about the phylogenetic relationships within Raphidascarididae, phylogenies were reconstructed based on partial sequences of the 18S and 28S nuclear rRNA gene, the nuclear ITS1-5.8S-ITS2 and the cytochrome c oxidase subunit I (cox1) mtDNA. Morphological study of S. mahnerti, confirmed some previously described features, revealed new characteristics and permitted to elucidate some inconsistencies noted in the literature. Morphological and genetic characterisation of S. mahnerti supported its validity. Phylogenetic reconstructions supported the monophyly of Sprentascaris, which has three pairs of interlabial conspicuous cuticular projections as a synapomorphy. The relationships among several lineages of raphidascaridids were unsolved, albeit Goezia and Ichthyascaris formed well-supported monophyletic assemblages, in which the first included species with no relations regarding the habitat of hosts and the geographic origin. The present findings represent one more step towards the understanding of the interrelationships of raphidascaridid nematodes. In this sense, Sprentascaris should be considered valid as an independent lineage from Raphidascaris.

An integrative taxonomic assessment of Procamallanus (Spirocamallanus) huacraensis (Nematoda: Camallanidae), infecting the freshwater catfish Trichomycterus spegazzinii (Siluriformes: Trichomycteridae) in Argentina.

Procamallanus (Spirocamallanus) huacraensis infecting the catfish Trichomycterus spegazzinii from Escoipe River, Salta province (Argentina), is redescribed and genetically characterised for the first time, based on three genetic markers (nuclear 18S and 28S rRNA; cytochrome c oxidase subunit I [cox1] mtDNA). The phylogeny of Camallanidae was also discussed. Morphological evaluation of P. (S.) huacraensis using light and scanning electron microscopy revealed the previously undescribed features: location of deirids, accurate morphology of larvae (L1) and ovijector in females, as well as phasmids in males. Differences were found comparing the newly collected material and the type specimens, probably because the original description lacked detailing. Unfortunately, type specimens of P. (S.) huacraensis were no available for loan. The results of morphological and genetic analyses supported the validity of P. (S.) huacraensis. Inconsistencies regarding the taxonomic identification of species of Camallanidae in GenBank database were noted. Based on the current genetic database of Camallanidae, phylogenetic reconstructions using the 18S rRNA sequences were most consistent, due to the inclusion of higher number of taxa. Procamallanus (S.) huacraensis appeared as sister group of P. (S.) rarus, also isolated from a catfish in a neighbouring region. The order and habitat of hosts were also similar within some well-supported parasite lineages, but without common geographic origin. However, it is still premature to make definitive affirmations regarding the role of such features in the phylogenetic patterns of Camallanidae, given the scarcity of genetic data. The phylogenetic reconstructions also confirmed the artificiality of the morphology-based systematics of the family.

Pathobiology and first report of larval nematodes (Ascaridomorpha sp.) infecting freshwater mussels (Villosa nebulosa, Unionidae), including an inventory of nematode infections in freshwater and marine bivalves.

Little information is available on host-parasite relationships between bivalves and larval nematodes. Herein, we describe nematode larvae (likely stage 2) in the infraorder Ascaridomorpha infecting the foot, intestine, and mantle of a freshwater mussel (Alabama rainbow, Villosa nebulosa [Conrad, 1834]) and detail histopathological changes to infected tissues. A total of 43 live mussels from the South Fork of Terrapin Creek, Alabama, were collected between 2010 and 2014, with 14 sectioned for histopathology and 29 dissected. Of the 14 sectioned mussels, 5 appeared to be uninfected, and 7, 1, and 1 had histozoic infections observed in the foot and intestine, intestine only, and mantle edge and foot, respectively. Twenty-three of 29 (79%) of the mussels dissected were infected by live nematodes, and mean nematode abundance was 8.3 (CL = 5.23-13), with 2 mussels infected with >100 nematodes each. Thus, with a total of 32 of the 43 collected mussels observed with nematodes, overall infection prevalence was 74.4% (CL = 0.594-0.855). The 18S rDNA of this nematode was 99% similar to that of several ascaridids (species of Kathlaniidae Lane, 1914 and Quimperiidae Baylis, 1930) that mature in aquatic/semi-aquatic vertebrates; the recovered 18S phylogenetic tree indicated this nematode from V. nebulosa shares a recent common ancestor with Ichthyobronema hamulatum (Ascaridomorpha: Quimperiidae; GenBank Accession Number KY476351). Pathological changes to tissue associated with these infections comprised focal tissue damage, but a cellular response was not evident. The Alabama rainbow possibly represents an intermediate or paratenic host. Given these results, the nematode is likely not pathogenic under normal stream conditions; however, high intensity infections in the foot could inhibit pedal extension and retraction; which would have demonstrable health consequences to a freshwater mussel. Based on our review of the bivalve mollusc parasite literature, a collective biodiversity of 61 nematodes reportedly exhibit some degree of symbiosis (from commensal to parasitic) with 21 bivalves (28 nematode spp. from 17 marine bivalve spp.; 33 nematode spp. from 4 freshwater bivalve spp.); only four records exist of putatively parasitic nematodes from Unionida. The present study represents the first description of a nematode species that invades the tissues of a Unionidae species.

Monogeneans of the catfish Pimelodus blochii Valenciennes (Siluriformes: Pimelodidae) from the Brazilian Amazon, with a description of a new species of Ameloblastella Kritsky, Mendoza-Franco & Scholz, 2000 (Monogenea: Dactylogyridae).

Ameloblastella amazonica n. sp. (Monogenea: Dactylogyridae) collected on the gills of Pimelodus blochii Valenciennes (Siluriformes: Pimelodidae) from Rivers Acre and Iaco, State of Acre (Brazil) is described. The new species differs from the congeners mainly by the morphology of the dorsal bar, i.e. slightly U-shaped bearing a posteriorly directed flap, arising from the anterior edge of its middle third. The most morphologically similar congeners, A. paranaensis (França, Isaac, Pavanelli & Takemoto, 2003) and A. satoi Monteiro, Kritsky & Brasil-Sato, 2010, resemble A. amazonica n. sp. regarding the general structure of the copulatory complex; however, besides other features, they also differ from the new species because their hooks are similar in size (vs hooks with three different sizes). In addition to the dorsal bar lacking a flap, A. amazonica n. sp. can be differentiated from all congeners based on the combination of the following features: size of hooks (one or two different sizes), shape of the dorsal bar (other than slightly U-shaped), anchor points largely passing the level of superficial root (vs not or slightly passing it), number of MCO rings (other than 3-4) and structure of the articulation between the MCO base and the accessory piece (other than thick, medium-sized, smooth and tubular). Demidospermus leptosynophallus Kritsky & Gutierrez, 1998 and Scleroductus yuncensi Jara & Cone, 1989 were also found infesting P. blochii, both representing new host and locality records.

First description of the male of Philometroides acreanensis and phylogenetic assessment of Philometridae (Nematoda: Dracunculoidea) suggest instability of some taxa.

The male of Philometroides acreanensis, parasitic in the anterior intestine external wall of the freshwater catfish Pimelodus blochii, from the Brazilian Amazon, is described for the first time. Additional data on the morphology of females is given. The new morphological data strengthened the validity of the species as well as its first genetic characterization, using three nuclear genetic markers (18S and 28S of the rDNA and ITS1-58S-ITS2), confirmed the high genetic resemblance of male and female specimens. Philometroides acreanensis shows morphological features of the generic diagnosis of Neophilometroides, Alinema, Philometra and Philometroides. Phylogenetic analyses using sequences of the18S rDNA from representatives of Dracunculoidea confirmed the validity of P. acreanensis and its close relatedness with Alinema rather than with other genera. The validity of Philonemidae was confirmed, as was the monophyly of Philometridae and Clavinema. However, Dentiphilometra, Philometra and Philometroides appear not to be monophyletic. Host taxa, habitat and geographic occurrence seem to have some relationship with the evolutionary traits of certain phylogenetic assemblages of philometrids, which were highly supported in the phylogentic reconstructions. Even though interesting aspects of the phylogeny and taxonomy of Philometridae came to light, further integrative approaches should be used that include additional genetic markers, due to the loose boundaries between some genera as observed here.

Community structure of metazoan parasites from Pimelodus blochii in two rivers of the Western Brazilian Amazon: same seasonal traits, but different anthropogenic impacts.

The present investigation evaluated the influence of seasonality and locality on the structure of the parasite community of the catfish Pimelodus blochii. A total of 160 fish were collected from two rivers in the State of Acre, western Brazilian Amazon: 80 fish in River Acre and 80 in River Iaco, with 40 in each season (rainy and drought). The overall prevalence was 78.7% and 1461 parasite specimens (adults and larvae) were allocated in 22 taxa: 5 of Monogenea, 10 of Nematoda, 3 of Digenea, 1 of Cestoda and 3 of Crustacea. In drought season, Procamallanus (Spirocamallanus) pimelodus and Demidospermus peruvianus were more prevalent in River Acre and Iaco, respectively. The parasite diversity (Brillouin index) as well as the prevalence and abundance of the monogeneans D. peruvianus, D. striatus, Demidospermus sp. and Ameloblastella sp. were higher in River Acre. The parasite community structure were dissimilar among rivers, and seasonally in River Iaco. These results suggest that environmental traits may overshadow seasonal influences on the parasite community structure, which may be related to the higher anthropization in River Acre. Furthermore, seasonality exerted less influence on the parasite community than expected, probably because the two rivers have different hydrological traits than those of other rivers in the Brazilian Amazon. New host and locality records expanded our knowledge of parasite biodiversity of P. blochii.

Redescription of the nematode parasites of lizards: Strongyluris oscari Travassos, 1923 (Heterakidae) from Brazil and Pharyngodon mamillatus (Linstow, 1897) (Pharyngodonidae) from Egypt.

Strongyluris oscari Travassos, 1923, parasite of Topidurus torquatus (Wied-Neuwied, 1820) (Tropiduridae) from Brazil and Pharyngodon mamillatus (Linstow, 1897) of Chalcides ocellatus (Forskal, 1775) (Scincidae) from Egypt, were redescribed based on light and scanning electron microscopy, and re-evaluation of specimens from parasitological collections. The following features were accurately observed in S. oscari for the first time: structure of lips and pharyngeal valve, presence and distribution of somatic papillae, number and arrangement of caudal papillae, presence of inner sclerotised structures supporting the ventral sucker and the ventral region in tail of males, and structure of the posterior anal lip in females. The unpaired papilla on the inferior edge of the ventral sucker in males of S. oscari is a cuticular groove instead of a true papilla. Regarding the morphology of P. mamillatus the following features were given in details for the first time: structure of the cephalic end in both male and female, location of amphids, position of phasmids in females and the accurate morphology of caudal bursa in males, including the distribution of caudal papillae and the genital cone morphology. Strongyluris oscari and P. mamillatus have no intraspecific morphological variations, but wide range on biometric features. These variations may be accounted by host/environment attributes, since they exhibit wide host and geographical spectra. The present results increased the morphological knowledge of P. mamillatus and S. oscari, strengthening their taxonomic validity.

Redescription and first genetic characterisation of Procamallanus (Spirocamallanus) macaensis Vicente & Santos, 1972 (Nematoda: Camallanidae), including re-evaluation of the species of Procamallanus (Spirocamallanus) from marine fishes off Brazil.

Newly collected specimens of Procamallanus (Spirocamallanus) macaensis Vicente & Santos, 1972 from the intestine of Paralonchurus brasiliensis (Steindachner), off Rio de Janeiro, Brazil, are redescribed and genetically characterised. Additionally, all congeners deposited in the Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC) parasitic in marine fishes of the South Atlantic, including types of P. (S.) macaensis, were re-evaluated. The following features are described for the first time in P. (S.) macaensis: morphology and arrangement of cephalic structures, shape of deirids and location of phasmids. The position of the excretory pore, the number and arrangement of caudal papillae in males, the structure of the spicules and of tail end in both males and females are rectified. Most specimens deposited in the CHIOC identified as P. (S.) pereirai Annereaux, 1946 were transferred to P. (S.) macaensis and others were designated as Procamallanus (S.) sp. Procamallanus (S.) cruzi Guimarães, Cristófaro & Rodrigues, 1976 is considered a species inquirenda due to its poor description and the lack of match of its original description with the type-material re-examined. Moreover, several taxonomic problems were noted after observations of the specimens (mostly poorly preserved), including inadequate morphological reports as well as misidentifications. Phylogenies inferred using sequences of the SSU rDNA from camallanids (Nematoda: Camallanidae) mostly generated weakly supported clades; however, Camallanus Railliet & Henry, 1915 and Procamallanus Baylis, 1923 do not seem to be monophyletic. Based on the present results and the lack of molecular data, it would be pertinent to adopt the widely-used classification for the subgenera of Procamallanus.

Parapharyngodon hugoi n. sp., a new nematode (Oxyuroidea: Pharyngodonidae) of the tree frog Trachycephalus typhonius (Linnaeus) from the Brazilian Pantanal, including a key to the congeners from amphibians of the American continent.

A new nematode Parapharyngodon hugoi n. sp. (Oxyuroidea: Pharyngodonidae) is described parasitising the large intestine of the tree frog Trachycephalus typhonius (Linnaeus) (Anura: Hylidae) from the wetlands of Pantanal, State of Mato Grosso do Sul, Brazil. The new species exhibits a unique structure of the posterior cloacal lip in males, which is supported by a rigid V-shaped structure. Parapharyngodon hylidae parasitic in hylid frogs, including T. typhonius, from Mexico, is the most similar congener to P. hugoi n. sp. but is distinguished from the new species by the presence of a gubernaculum (vs absence), by the lateral alae in males ending far anterior to cloacal opening (vs near to it) and because in gravid females the ovaries encircle the oesophageal corpus. Additionally, the new species differs from its congeners as well as from species of Thelandros Wedl, 1862, a very closely related genus, by the combination of features such as spicule length, number of caudal papillae, morphology of the anterior cloacal lip, which is echinate, and position of ovaries. The geographical distribution of hosts seems to play an important role in the speciation process of Parapharyngodon spp.; however, due the lack of molecular data this issue along with the validity of both Thelandros and Parapharyngodon are still questions to be solved in the future, after improvement of the genetic database. A key to the species of Parapharyngodon parasitic in amphibians from the American continent is provided.

An integrated phylogenetic analysis on ascaridoid nematodes (Anisakidae, Raphidascarididae), including further description and intraspecific variations of Raphidascaris (Sprentascaris) lanfrediae in freshwater fishes from Brazil.

Genetic and morphological variations in two component populations of Raphidascaris (Sprentascaris) lanfrediae collected in the intestine of Geophagus argyrosticus and G. proximus (Cichlidae) from States of Pará and Amapá, Brazil, respectively, were explored for the first time. A phylogenetic study including two genes (18S and 28S of the rDNA) plus morphological and life history traits of "anisakid-related" nematodes (Anisakidae, Raphidascarididae) was also performed in order to clarify taxonomic and systematic issues related to these taxa. Gene alignments were subjected to maximum likelihood (ML) and Bayesian Inference (BI), and combined data of the genetic and morphological datasets was subjected to maximum parsimony (MP) analysis. Despite of the subtle differences in the morphology (mainly in male caudal papillae) and morphometry between specimens of R. (S.) lanfrediae from the two different hosts and from the type material of the species, no genetic variation was found among representatives of the newly collected material. This find may represent an example of gene-environment interactions, similar to that recently observed for Raphidascaroides brasiliensis. Phylogenetic reconstructions indicated the paraphyly of Anisakidae represented by two subfamilies, i.e., Anisakinae and Contracaecinae and the monophyly of Raphidascarididae. Analysis of the combined datasets revealed that some morphological traits may represent apomorphic characters of Raphidascarididae and Anisakidae, whereas others are highly homoplastic and some may be interpreted with careful to avoid errors. The results support the premise that taxonomists should consider Anisakidae and Raphidascarididae as separate families, and only two subfamilies of Anisakidae, i.e., Anisakinae and Contracaecinae.

Morphology and molecular characterization hold hands: clarifying the taxonomy of Hysterothylacium (Nematoda: Anisakidae) larval forms.

During the last decade, many morphotypes of larval forms of Hysterothylacium have been proposed; however, their boundaries are taxonomically unclear. We compare newly collected material with data available for Hysterothylacium larvae, based on genetics and morphology, in order to evaluate their relationships and simplify their classification. The previously described Hysterothylacium larval type IV, type V, and type X were found in several marine fishes from Brazil. A new Hysterothylacium larval type XV, parasitic in Elops saurus from State of Sergipe, north-eastern Brazil, was proposed based on its genetic singularity and the presence of numerous spines throughout the body. Based upon phylogenetic reconstruction using the ribosomal DNA (rDNA) ITS2 sequences along with morphological traits, the following third stage larvae were considered valid (synonyms are inside parenthesis) type III (= types VII and VIII), type V (= types VI, XII, and XIII), and type X (= type XIV). Despite the lack of molecular data, Hysterothylacium larval type II is considered as synonym of type I because of their identical morphology. The morphometry of Hysterothylacium third stage larvae proved to be weak as a diagnostic tool due to its high rates of variation. Phylogenetic reconstruction using the concatenated sequences of rDNA ITS1-5.8S-ITS2 and the mitochondrion DNA (mtDNA) cox2 revealed that the sampled Hysterothylacium larval types V may represent three different species and H. deardorffovestreetorum may represent two different taxa. Furthermore, H. deardorffovestreetorum is considered species inquirenda due to its problematic description and diagnosis which are based only on larvae. A key to Hysterothylacium larvae from marine fishes is provided.

A survey of nematodes of the genus Cucullanus Müller, 1777 (Nematoda, Seuratoidea) parasitic in marine fishes off Brazil, including description of three new species.

A taxonomic survey of six nematode species (including three new taxa) from the genus Cucullanus Müller, 1777, parasites of marine fishes off the Brazilian coast, is provided. Nematodes were studied using light and scanning electron microscopy (SEM). Cucullanus gastrophysi n. sp. parasitic in Lophius gastrophysus Miranda Ribeiro differs from its congeners by the combination of the following features: shape and number of sclerotized structures in the oesophastome (a pair of lateral elongate structures and a single small reniform one), position of deirids and excretory pore (both anterior to oesophagus base), spicule length and spicule/body length ratio (0.97-1.29 mm and 6.5-10.5%, respectively), morphology and length of gubernaculum (V-shaped, 107-135 µm long). Cucullanus protrudens n. sp. from Pagrus pagrus (Linnaeus) has the cloacal lips broadly protruded, which differentiates it from several species of Cucullanus; other features, e.g., the length of spicules and gubernaculum (400-415 µm and 91-103 µm, respectively), arrangement of caudal papillae and position of excretory pore (slightly posterior to oesophagus-intestine junction) also characterize this species. Cucullanus pseudopercis n. sp. from Pseudopercis semifasciata (Cuvier) has deirids and excretory pore posterior to the oesophagus-intestine junction, which distinguishes the species from most of the congeners; furthermore, the arrangement of caudal papillae in combination with the length of spicules and gubernaculum (1.0-1.5 mm and 178-196 µm, respectively) separate this species from other taxa. Newly collected specimens of C. cirratus Müller, 1777 (type species of the genus) from Urophycis brasiliensis (Kaup), C. pedroi from Conger orbignianus Valenciennes (type host of the species) and C. genypteri Sardella, Navone & Timi, 1997 from Genypterus brasiliensis Regan, were studied as well. Comparisons between newly collected samples and the taxonomic data available for each respective species revealed features that were not previously mentioned (e.g. presence of unpaired cloacal papilla, detailed morphology of cloacal lips), as well as negligible differences in morphometry and caudal papillae arrangement. Observations on the type material of C. carioca suggested affinities with the genus Dichelyne Jägerskiöld, 1902; however, the poor preservation of these specimens does not allow further conclusions. Cucullanus rougetae is considered to be a species inquirenda.