Identification of the avian tracheal trematode Typhlocoelum cucumerinum (Trematoda: Cyclocoelidae) in a host-parasite-environment system: diagnosis, life cycle and molecular phylogeny.

Typhlocoelum cucumerinum is a tracheal parasite of birds widely distributed across the globe. Nevertheless, aspects of the biology of this cyclocoelid are still poorly understood. Herein, we report the finding of T. cucumerinum in definitive and intermediate hosts from an urban waterbody of Brazil. The parasite was initially detected during the necropsy of domestic Muscovy ducks (Cairina moschata) found dead in the locality. Coproparasitological tests in live animals revealed that 12/47 (25.53%) Muscovy ducks and 2/8 (25%) mallards (Anas platyrhynchos platyrhynchos) were infected with T. cucumerinum. Moreover, rediae and metacercariae morphologically similar to T. cucumerinum were found in 3/248 (1.33%) Biomphalaria straminea collected in the same waterbody frequented by the birds. The conspecificity between the adult and the larval stages was confirmed molecularly (100% similarity in Cox-1). Moreover, the phylogenetic position of T. cucumerinum was determined for the first time based on partial fragments of the 28S, Cox-1 and Nad-1 genes. The species grouped with other members of the subfamily Typhlocoelinae with sequences available, but the data obtained do not support the distinctiveness of the genera Typhlocoelum and Tracheophilus. Further studies involving a broader range of species can result in taxonomic rearrangements in Typhlocoelinae.

A new species of Gorgoderina (Digenea: Gorgoderidae) from Rana berlandieri in Los Tuxtlas tropical rainforest, Mexico, with the elucidation of its life cycle.

Species in the genus Gorgoderina Looss, 1902 are parasites of the urinary bladder of amphibians and include around 50 species described globally. Molecular data on species of the genus are scarce, as is the information of their life-cycle patterns. During a survey on the genetic characterization of the frog trematodes in the tropical rain forest of Los Tuxtlas, in the Gulf of Mexico slope of Mexico, specimens of two morphotypes of Gorgoderina were sampled from the Rio Grande leopard frog, Rana berlandieri. One of them represented an undescribed species which is described herein as Gorgoderina rosamondae n. sp., whereas the other one was morphologically very similar to an apparently widely distributed North American species, G. attenuata, which has been previously reported in the same geographical area. Specimens of both morphotypes were sequenced for two nuclear and one mitochondrial genes. Phylogenetic trees corroborated the distinction of the new species, and data on the internal transcribed spacer 2 revealed genetic differences between G. attenuata sequenced from frogs in USA and specimens of Gorgoderina sp. from Los Tuxtlas, indicating the possibility that they also represent an undescribed species. COI sequences showed high genetic divergence values between the new species and Gorgoderina sp. from Los Tuxtlas (8.63-9.99%). Additionally, COI sequences of the larval forms (sporocyst, cercariae and metacercariae) sampled in the same locality from their first and second intermediate hosts (Pisidium sp. and Agriogomphus tumens, respectively) showed conspecificity, and the 3 host life-cycle of the new species was elucidated.

Exploring Neotropical anuran parasites: a morphological, life cycle and phylogenetic study of Catadiscus marinholutzi (Trematoda: Diplodiscidae).

Amphistome species belonging to the genus Catadiscus are poorly studied intestinal trematodes found primarily in Neotropical anurans. Herein, developmental stages of an amphistome species found during herpetological and malacological surveys in a temporary marsh pond from Brazil were subjected to morphological (light and scanning electron microscopy) and molecular analyses. Adult parasites recovered from anurans were identified as Catadiscus marinholutzi. Amphistome cercariae found in the planorbid snails Drepanotrema depressissimum and Drepanotrema lucidum from the same waterbody were used for experimental and molecular studies. Immature parasites, morphologically compatible with members of Catadiscus, were experimentally obtained in laboratory-reared tadpoles. Sequencing of a partial region of 28S rDNA gene of both adult and cercariae revealed 100% similarity between these developmental stages, confirming their conspecificity. Phylogenetic analyses were attempted for the first time to reveal the position of a species of Catadiscus in the superfamily Paramphistomoidea. Catadiscus marinholutzi falls in a virtual polytomy together with other paramphistomoids, which leaves its phylogenetic relationships within the group unclear. Moreover, the high genetic divergence to Diplodiscus spp. (10.06­10.84%) cast doubts on the placement of Catadiscus within Diplodiscidae. Hence the species composition of the Diplodiscidae should be re-evaluated in further studies using a broader spectrum of related taxa.

First report of Melanoides tuberculata (Mollusca: Thiaridae) harboring a xiphidiocercaria in Brazil: A new parasite introduced in the Americas?

Xiphidiocercariae were found in the invasive snail Melanoides tuberculata collected during a malacological survey in Ceará-Mirim, State of Rio Grande do Norte, Northeastern Brazil in November 2018 and submitted to morphological and molecular analyses. The morphology revealed similarities between the larvae here reported for the first time in M. tuberculata from Brazil and other xiphidiocercariae described in thiarid snails from Asia and Africa. Phylogenetic analyses based on 28S and ITS-2 sequences revealed that the larvae correspond to an unidentified species of the family Lecithodendriidae. Aspects related to the morphology and taxonomy of xiphidiocercariae found in M. tuberculata are briefly discussed. It is possible that the parasite here reported is a newly introduced species transmitted by M. tuberculata in the American continent.

The life cycle of Magnivitellinum saltaensis n. sp. (Digenea: Alloglossidiidae) in Salta Province, Argentina.

We describe the alloglossiid trematode Magnivitellinum saltaensis n. sp., a parasite of the characiform fish Psalidodon endy, and its life cycle from Salta, northwest of Argentina. This is the first life cycle described for a species belonging to the genus Magnivitellinum. Cercariae emerged naturally from Biomphalaria tenagophila snails and infected experimentally exposed larvae of Diptera and Ephemeroptera as second intermediate hosts. These larvae in turn were exposed to commercially raised fish, and adults were recovered from characiform albino fish Gymnocorymbus ternetzi. Molecular analysis of natural and experimental adults showed the same genetic sequence for the partial region of 28S rDNA, thus confirming conspecificity. Comparison of these sequences with those published for M. simplex from Mexico showed 1.45% divergence, indicating that the specimens found in Salta belong to a different species, the third described of Magnivitellinum, in agreement with morphological data, geographical location, and host species composition. The new species is distinguished by its small body, vitelline follicles extending from the mid-level of the ventral sucker, Y-shaped excretory vesicle, and presence of papillae around the mouth.

Reproductive compensation in female Palaemonetes argentinus (Decapoda: Natantia) due to Microphallus szidati (Trematoda) infection.

Parasites may affect host demographic characteristics because they can directly or indirectly cause the death of their hosts and/or influence their reproduction. Parasitism is therefore recognized as a factor that influences the composition and structure of populations and communities. One of these behaviours is the compensatory response: the host can compensate for the parasite losses effect, modifying the reproductive effort to enhance fitness. Ovigerus female Palaemonetes argentinus was collected and sorted into two groups according to the degree of development of their embryos: newly spawned embryos and embryos ready to hatch. The number of embryos and their dry weight for each female were determined. All parts of the female body were checked for parasites. The females of P. argentinus were parasitized by Microphalus szidati. We found that parasitized females produce more embryos but had more egg loss during development and the percentage of embryonic loss was higher in the parasitized females than in non-parasitized. Parasitized females produced lighter eggs than those from uninfected females. This supports the compensatory reproduction hypothesis suggested for this species. Parasitism can change life history traits in a way that fecundity can be compensated; this co-evolution between host and parasites will be population or context dependent. Parasites are a functional part of any ecosystem and as our results show, deleting parasites in life history traits and reproduction studies in free living organisms could lead to an incomplete picture of the true processes that happen in nature.

Molecular and Morphological Elucidation of the Life Cycle of the Frog Trematode Langeronia macrocirra (Digenea: Pleurogenidae) in Los Tuxtlas, Veracruz, Mexico.

The genus LangeroniaCaballero and Bravo-Hollis, 1949, currently contains 6 species of amphibian trematodes distributed in North and Middle America. The type species of the genus, Langeronia macrocirraCaballero and Bravo-Hollis, 1949, occurs in Mexico and is relatively commonly found as a parasite of leopard frogs. However, information regarding its life cycle is lacking. In this paper, we study the life cycle of L. macrocirra in Laguna Escondida, Los Tuxtlas, Veracruz. Definitive hosts (Rana spp.) as well as potential intermediate hosts (gastropods, bivalves, crustaceans, tadpoles, hemipterans, and odonate naiads) were sampled in the locality and studied to search for the presence of adults and larval stages of the trematode. Specimens were morphologically characterized, and some individuals were sequenced for 1 ribosomal gene (28S rRNA) and 1 mitochondrial gene (COI). DNA sequences of the adults obtained from leopard frogs were matched with those of the larval forms in their intermediate hosts (metacercariae, cercariae, and sporocysts) to demonstrate conspecificity. Further, we conducted a detailed study of the tegument of the body surface with scanning electron microscopy to characterize each of the developmental stages of the life cycle of L. macrocirra.

Phylogenetic position of Diplostomum spp. from New World herons based on complete mitogenomes, rDNA operons, and DNA barcodes, including a new species with partially elucidated life cycle.

Diplostomum ardeae Dubois, 1969 has seldom been reported since its description from the great blue heron (Ardea herodias L., 1758) in the USA. Sequences obtained in this study from the barcode region of cytochrome c oxidase 1 (CO1) in diplostomids from black-crowned night heron (Nycticorax nycticorax (L., 1758)) in Puerto Rico matched data from D. ardeae from A. herodias in the type region. We also obtained DNA barcodes from morphologically similar diplostomids from a rufescent tiger heron (Tigrisoma lineatum (Boddaert, 1783)) and from metacercariae from eye lenses of Trachelyopterus galeatus (Linnaeus, 1766) from the Paraná River basin in Argentina and Brazil, respectively. Barcodes matched (97-100% identity) in these South American adult and larval specimens as well as in recently published sequences from metacercariae from 11 other siluriform fishes from the same region. Barcodes from the South American species, which we describe as Diplostomum lunaschiae n. sp., differed from those of D. ardeae by 7.2-9.8%, and the new species differs from D. ardeae in its size, pharynx:oral sucker length ratio, egg:body length ratio, and distribution of vitellaria. As in prior phylogenetic analysis of CO1 sequences, both D. ardeae and D. lunaschiae n. sp. were not associated with Diplostomum. In more character-rich analyses of nuclear rDNA and of mitochondrial genomes, D. ardeae was an early divergent member of clades of species of Diplostomum. Consequently, we continue to consider D. ardeae and D. lunaschiae n. sp. members of Diplostomum, in contrast to recent suggestions that these species may belong to a different genus.

Effects of temperature on the life cycle of Neobenedenia sp. (Monogenea: Capsalidae) from Seriola rivoliana (Almaco jack) in Bahía de La Paz, BCS Mexico.

Seriola rivoliana cultivated in Mexico are infected by Neobenedenia sp. (Monogenea: Capsalidae), resulting in dermal ulceration and subsequent bacterial invasion that can cause fish death. This study assesses the effects of temperature over hatching success, oncomiracidia longevity, and infection success. The experimental design consisted of culturing the parasite at temperatures ranging between 16 and 32 °C. The oncomiracidia infection success, time to sexual maturity, and size at sexual maturity of Neobenedenia sp. were examined only at three temperatures (20 °C, 24 °C, and 30 °C). Experiments were conducted under controlled conditions in the laboratory. The oncomiracidia development was found to be faster at warmer temperatures (4-5 days between 24 and 30 °C) than in colder treatments (7-11 days between 18 and 20 °C). Hatching success and oncomiracidia longevity were higher at 24 °C and 26 °C. At 20 °C, 24 °C, and 30 °C, infection success was greater than 90%. Additionally, the laid eggs were observed at 9, 12, and 15 days at 30 °C, 24 °C, and 30 °C, respectively. The results of this study will allow for improving the temporal schedule of applications of treatments against Neobenedenia sp. by the function of temperatures. In conclusion, it is recommended to treat fish more frequently if the temperature in cultures is higher than 24 °C, because Neobenedenia sp. development is faster. As an alternative, the fish could be moved to deeper and cooler waters.

Xiphidiocercariae from naturally infected Lymnaea columella (Mollusca, Gastropoda) in urban area: morphology and ultrastructure of the larvae and histological changes in the mollusc host / Xifidiocercárias obtidas em Lymnaea columella (Mollusca, Gastropoda) naturalmente infectadas em área urbana: morfologia e ultraestrutura da larva e alterações histológicas do molusco hospedeiro

Abstract Urban agriculture is very common in the cities, as Rio de Janeiro and adjacencies, being important as economic activity. However, this practice can create appropriate conditions for establishment and maintenance of life cycle of many parasites. Lymneid snails can act as intermediate hosts of many trematodes species, including those that infect humans and wild and domestic. In the present study, the snail Lymnaea columella was collected from an urban watercress plantations and the morphology of cercariae released by them was analyzed by light and scanning electron microscopy. The specimens were identified as xiphidiocercariae, belonging to the Cercariae Ornatae group, of the order Plagiorchiiformes. Histological examination of the digestive gland of the infected snails revealed the presence of developing sporocysts, with hemocitary infiltration and metaplasia in the epithelial tissue.

Resumo Agricultura urbana é muito comum nas cidades, como Rio de Janeiro e adjacências, sendo uma importante atividade econômica de subsistência. No entanto, esta prática pode criar condições adequadas para o estabelecimento e manutenção do ciclo de vida de muitos parasitos. Os moluscos limneídeos podem atuar como hospedeiros intermédios de muitas espécies de trematódeos, incluindo aqueles que infectam humanos e animais selvagens e domésticos. No presente estudo, o molusco Lymnaea columella foi coletado de plantações urbanas de agrião e a morfologia das cercárias liberadas por eles foi analisada por microscopia eletrônica de varredura leve e de varredura. As larvas foram identificadas como xifidiocercárias, pertencentes ao grupo das Cercariae Ornatae, Ordem Plagiorchiiformes. Análises histológicas da glândula digestiva dos moluscos infectados evidenciaram esporocistos, com formação de granuloma e metaplasia do tecido epitelial.

Molecular characterization and identification of digenean larval stages in Aylacostoma chloroticum (Prosobranchia: Thiaridae) from a neotropical floodplain.

Digeneans (Trematoda: Digenea) are endoparasites that present a complex life cycle, generally involving an intermediate invertebrate host and a vertebrate host. There is limited information about which species of molluscs may act as intermediate hosts in the upper Paraná River floodplain (UPRF), where Aylacostoma chloroticum can be considered a potential candidate. The study of digeneans in this region is important because some of these parasites are potentially zoonotic, and, therefore, are relevant to public health. However, the correct identification of these organisms during the larval stages is difficult because of the lack of morphologically diagnostic characteristics. The objective of this study was to identify and molecularly characterize the larval stages of digeneans found in A. chloroticum in the UPRF, using the mitochondrial marker of subunit I of cytochrome c oxidase and the 28S nuclear marker. The molluscs were examined in the laboratory and three morphotypes of cercariae were found. DNA was extracted from the specimens obtained and was then amplified and sequenced. The morphotypes exhibited high genetic similarities with Pseudosellacotyla, Paralecithodendrium and Philophthalmus, indicating that these organisms belong to these genera. This is the first record of larval stages of these genera in molluscs collected in the UPRF.

A morphological, molecular and life cycle study of the capybara parasite Hippocrepis hippocrepis (Trematoda: Notocotylidae).

Hippocrepis hippocrepis is a notocotylid that has been widely reported in capybaras; however, the molluscs that act as intermediate hosts of this parasite remain unknown. Furthermore, there are currently no molecular data available for H. hippocrepis regarding its phylogenetic relationship with other members of the family Notocotylidae. In the present study, we collected monostome cercariae and adult parasites from the planorbid Biomphalaria straminea and in the large intestine of capybaras, respectively, from Belo Horizonte, Minas Gerais, Brazil. We subjected them to morphological and molecular (amplification and sequencing of partial regions of 28S and cox-1 genes) studies. Adult parasites collected from the capybaras were identified as H. hippocrepis and the sequences obtained for both molecular markers showed 100% similarity with monostome cercariae found in B. straminea. The sequences obtained for H. hippocrepis were compared with data available in public databases; analysis revealed this species differs from other notocotylids with available sequences (1.5-3.8% with respect to 28S and 11.4%-13.8% with respect to cox-1). On the phylogenetic analyses, H. hippocrepis appeared to be a distinct lineage in relation to other notocotylids. Some ecological aspects related to the infection of capybaras with H. hippocrepis are briefly discussed.

New Species of Gyrodactylus von Nordmann, 1832 from Native Fish from Chiapas, Mexico, Studied by Morphology and Molecular Analyses.

BACKGROUND: Three new species of Gyrodactylus von Nordmann, 1832 are described from Poeciliopsis hnilickai and Tlaloc labialis (syn. Profundulus candalarius and Profundulus labialis) from the state of Chiapas, Mexico. Gyrodactylus chiapaneco n. sp. is described from T. labialis. Gyrodactylus guatopotei n. sp. was found to infect P. hnilickai. Gyrodactylus tlaloci n. sp. is described from T. labialis. The new species were characterised morphologically (haptoral hooks) and molecularly (sequences of the Internal Transcribed Spacer region (ITS1-5.8S-ITS2) of rDNA). PURPOSE: During a survey in March 2017 in the state of Chiapas, individuals of Tlaloc labialis were collected by electrofishing from Arroyo Ojo de Agua and from Río Hondo, Ixtapa, Río Las Lajas and Poeciliopsis hnilickai from Río Hondo, Ixtapa and Río Cacao. Three new species of Gyrodactylus were found and are described herein based on morphological and genetic analysis. METHODS: Attachment hooks were excised and partially digested with a proteolytic solution to remove tissue enclosing the haptoral armature. Measurements and pictures of the haptoral hard structures were taken and were compared with other Gyrodactylus species found on poeciliids and profundulids. The bodies from which haptoral hook were removed were stored individually in 95% ethanol at - 20 °C for molecular analyses. Sequences of the Internal Transcribed Spacer region of rDNA were obtained for each species and were compared with 26 Gyrodactylus spp. The genetic divergence among species of Gyrodactylus was estimated using uncorrected "p" distances with the program MEGA. RESULTS: The haptoral hook of Gyrodactylus chiapaneco n. sp. are similar to G. poeciliae from Venezuela; those from Gyrodactylus guatopotei n. sp. resemble the sclerites of G. turnbulli from Singapore and G. pictae from Trinidad, and the haptoral hooks of Gyrodactylus tlaloci n. sp. are comparable to those of G. pseudobullatarudis and G. zapoteco from Mexico; most of these parasites were described from poeciliid fishes, except G. zapoteco which was recently described from Profundulus oxacae from Oxaca, Mexico. The sequences of the three new species were aligned with sequences of 26 other Gyrodactylus spp. Maximum likelihood and Bayesian inference analyses indicated that the three new taxa are members with well-supported lineages-the new taxa are the first species of Gyrodactylus to be described from Poeciliopsis hnilickai and Tlaloc labialis from Chiapas. CONCLUSION: Gyrodactylus chiapaneco n. sp., Gyrodactylus guatopotei n. sp. and Gyrodactylus tlaloci n. sp. are described based on morphological and molecular analyses. Two of the three new species of Gyrodactylus described here, and both roughly fall morphologically and molecularly into the group of parasites harbouring slender marginal hooks: G. chiapaneco n. sp. and G. tlaloci n. sp. both possess marginal sclerites with slim shafts and group phylogenetically with other gyrodactylids of similar marginal hook morphology. Gyrodactylus guatopotei n. sp. groups with gyrodactylids possessing marginal hooks with quite elongated shafts, most of which have been found infecting poeciliids in the Antilles (e.g. G. pictae and G. turnbulli). Poeciliid, profundulid and goodeid fishes are morphologically and ecologically similar and nowadays live in sympatry or close geographical distance in Mesoamerica.

Morphological and molecular analyses of Pseudomazocraes sulamericana n. sp., Pseudomazocraes selene Hargis, 1957, Cemocotyle carangis (MacCallum, 1913) and Zeuxapta seriolae (Meserve, 1938) (Monogenea: Mazocraeidea) from carangid fishes in the south-western Atlantic Ocean.

An integrative taxonomy approach was followed to analyse morphological and molecular characters of the monogenean species Pseudomazocraes selene, Cemocotyle carangis and Zeuxapta seriolae; specimens were collected from the gills of the carangid fishes Selene vomer, Caranx latus and Seriola lalandi caught off the coasts of the states of Rio de Janeiro and Santa Catarina, Brazil. The research revealed the presence of Pseudomazocraes sulamericana n. sp., which can be differentiated from other congeners by the shape of clamps mid-sclerite possessing a ventral piece bifurcated at the end, with large and right-angled edges of almost the same length, and by the shape of larval hooks and rounded terminal lappet. New genetic sequences include partial 28S and 18S rDNA genes for all species, ITS1 and 5.8S rDNA for Zeuxapta seriolae and Cemocotyle carangis, and ITS2 and mtDNA cox1 for C. carangis. The phylogenetic concatenated analysis based on partial 28S rDNA and 18S rDNA sequences confirmed the position of C. carangis and Z. seriolae within the Heteraxinidae. The previous phylogenetic position of Chauhaneidae was discussed based on morphological studies and it is now confirmed by molecular data that Chauhaneidae is the sister group of Allodiscocotylidae and Protomicrocotylidae.

Xiphidiocercariae from naturally infected Lymnaea columella (Mollusca, Gastropoda) in urban area: morphology and ultrastructure of the larvae and histological changes in the mollusc host.

Urban agriculture is very common in the cities, as Rio de Janeiro and adjacencies, being important as economic activity. However, this practice can create appropriate conditions for establishment and maintenance of life cycle of many parasites. Lymneid snails can act as intermediate hosts of many trematodes species, including those that infect humans and wild and domestic. In the present study, the snail Lymnaea columella was collected from an urban watercress plantations and the morphology of cercariae released by them was analyzed by light and scanning electron microscopy. The specimens were identified as xiphidiocercariae, belonging to the Cercariae Ornatae group, of the order Plagiorchiiformes. Histological examination of the digestive gland of the infected snails revealed the presence of developing sporocysts, with hemocitary infiltration and metaplasia in the epithelial tissue.

Exploring the genetic diversity of Tylodelphys (Diesing, 1850) metacercariae in the cranial and body cavities of Mexican freshwater fishes using nuclear and mitochondrial DNA sequences, with the description of a new species.

Members of the genus Tylodelphys Diesing, 1850 are endoparasites of fish-eating birds, particularly ciconiids, anhingids, and podicipedids across the globe. Metacercariae of Tylodelphys spp. were collected from the cranial and body cavities of freshwater fishes in central and northern Mexico; adults were recovered from the intestine of two species of freshwater diving birds of the family Podicipedidae, commonly known as grebes, in two locations of central Mexico. Specimens were sequenced for two molecular markers, the internal transcribed spacers (ITS1 and ITS2) plus 5.8S gene of the nuclear ribosomal DNA and of the cytochrome c oxidase subunit 1 from mitochondrial DNA. The genetic divergence among the 25 samples (16 metacercariae and 9 adults) and between the newly sequenced specimens and those deposited in the GenBank were estimated. Maximum likelihood and Bayesian inference analyses inferred with each data set revealed the existence of five genetic lineages. Eight metacercariae analyzed in this study were nested in two divergent lineages previously recognized as Tylodelphys sp. 5 and Tylodelphys sp. 6 (sensu Locke et al., Int J Parasitol, 45:841-855, 2015). Five adult specimens recovered from the intestine of the least grebe (Tachybaptus dominicus Linnaeus, 1766) in Tecocomulco Lake, Hidalgo State, nested in a single clade with other sequences identified previously as Tylodelphys aztecae, expanding its distribution range in other areas of central Mexico. The isolates of the metacercariae found in the cranial cavity of the shortfin silverside, Chirostoma humboldtianum Valenciennes, 1835 from Zacapu Lake in central Mexico formed a monophyletic lineage and were recognized as an undescribed species of Tylodelphys. The lack of adult specimens of this lineage in our samples prevented a formal description. However, the metacercariae collected in the cranial cavity of the silverside, Chirostoma jordani Woolman, 1894 and the adult specimens recovered from the intestine of the western grebe, Aechmophorus occidentalis (Lawrence, 1858) from Cuitzeo Lake formed a monophyletic clade, allowing us to link both stages of the life cycle and to describe this as a new species, Tylodelphys kuerepus n. sp. The new species represents the eighth species of the genus described in the Americas and the fourth in the Nearctic region. We briefly discuss the ecological associations between the metacercariae and their second intermediate hosts in relation to the genetic diversity patterns uncovered in our study.

Helminth Infracommunities of the Montezuma's Frog Lithobates montezumae (Anura: Ranidae) in the Nearctic-Neotropical Transition Zone of Mexico.

Thirty specimens of the Montezuma's frog Lithobates montezumae were collected in San Pedro Tlaltizapán, in the Nearctic-Neotropical transition zone of Mexico, in April 2013, in order to establish the helminth infracommunity structure harbored by this amphibian host. A total of 5,493 individual helminths were obtained, representing 6 species (3 trematodes and 3 nematodes). The depauperate structure of the helminth infracommunities established for the studied frogs herein (richness = 2.4; mean abundance = 183.1; Brillouin's diversity = 0.42) fits with those described for the 6 Ranidae species studied in the Nearctic. Three of the 6 species of helminths recovered are generalists, 2 of them with the highest values for prevalence ( Falcaustra mexicana) and mean abundance ( Renifer sp.). The parasite recruitment process (ingestion) determining mean richness in the helminth infracommunities studied here (ingestion) is shared with those reported for the 9 Mexican frog species for which helminthological records exist; however, in this case, mean abundance was determined by directly penetrating species. The use of aquatic habits by this anuran species likely explains why the composition of their helminth infracommunities was mainly constituted of helminths acquired in this environment (5 of 6 species). Finally, the heterogeneity of their taxonomic composition and abundance of helminth species indicate the unpredictable nature of these host-parasite associations.

Link Between the Adult and the Metacercaria of Clinostomum heluans Braun, 1899 (Trematoda: Clinostomidae) Through DNA Sequences, and its Phylogenetic Position Within the Genus Clinostomum Leidy, 1856.

The phylogenetic position of Clinostomum heluans Braun, 1899 within the genus Clinostomum Leidy, 1856 is reported in this study based on sequences of the barcoding region of the mitochondrial cytochrome c oxidase subunit 1 gene ( COX1). Additionally, molecular data are used to link the adult and the metacercariae of the species. The metacercariae of C. heluans were found encysted infecting the cichlid fish Australoheros sp. in Minas Gerais, Brazil, whereas the adults were obtained from the mouth cavity of the Great White Egret, Ardea alba, in Campeche, Mexico. The COX1 sequences obtained for the Mexican clinostomes and the Brazilian metacercaria were almost identical (0.2% molecular divergence), indicating conspecificity. Similar molecular divergence (0.2-0.4%) was found between sequences of C. heluans reported here and Clinostomum sp. 6 previously obtained from a metacercaria recovered from the cichlid Cichlasoma boliviense in Santa Cruz, Bolivia. Both maximum likelihood and Bayesian inference analyses unequivocally showed the conspecificity between C. heluans and Clinostomum sp. 6, which form a monophyletic clade with high nodal support and very low genetic divergence. Moreover, tree topology reveals that C. heluans occupies a basal position with respect to New World species of Clinostomum, although a denser taxon sampling of species within the genus is further required. The metacercaria of C. heluans seems to be specific to cichlid fish because both samples from South America were recovered from species of this fish family, although not closely related.

Morphological and morphometric study of pre-ovigerous and post-ovigerous adults of Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae)

The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of postovigerous adults (rs: 0.774; p< 0.05), the body length and the length of uterus (rs: 0.839; p< 0,01) and between the ovary width and egg length (rs: 0.777; p p<0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.

Morphological and morphometric study of pre-ovigerous and post-ovigerous adults of Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae)

The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of postovigerous adults (rs: 0.774; p< 0.05), the body length and the length of uterus (rs: 0.839; p< 0,01) and between the ovary width and egg length (rs: 0.777; p p<0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.(AU)