Do host habitat use and cospeciation matter in the evolution of Oswaldocruzia (Nematoda, Molineidae) from neotropical amphibians?

The genus Oswaldocruzia Travassos, 1917 includes approximately 90 species that are parasitic on amphibians and reptiles around the world, of which 43 occur in the neotropical region. However, molecular data supporting the taxonomic status of most species of the genus are scarce, and their phylogenetic relationships remain unknown. Using specimens of four molineid taxa (Oswaldocruzia belenensis Santos, Giese, Maldonado Jr. and Lanfredi, 2008; Oswaldocruzia chabaudi Ben Slimane & Durette-Desset, 1996, Oswaldocruzia chambrieri Ben Slimane & Durette-Desset, 1996 and Kentropyxia hylae Feitosa, Furtado, Santos and Melo, 2015) from amphibian hosts collected in different regions of Pará, Brazil, we conducted morphological studies, molecular analyses and phylogenies (maximum likelihood and Bayesian inference) using the cytochrome c oxidase subunity I (Cox1) gene. The newly generated sequences were compared with those of ten publicly available Cox1 sequences of Oswaldocruzia from Mexico. Our findings demonstrated significant differences between the sequences of amazonian specimens and sequences from specimens collected in Mexico, and we suggest that host-parasite cospeciation or habitat use might be related to molineid evolution in amphibian hosts. Additionally, this work presents new hosts and new geographical records for species of Oswaldocruzia from the neotropics.

"Revisiting the past": a redescription of Physaloptera retusa (Nemata, Physalopteridae) from material deposited in museums and new material from Amazon lizards.

Physaloptera Rudolphi, 1819 is a genus of nematodes that includes approximately 100 species parasitic in vertebrates around the world. From these, approximately 30 occur in the Neotropical region, with nine reported from neotropical reptiles. Physaloptera spp. are recognized by their distinct morphology of the apical end and characters of the reproductive system. However, despite the fact that the morphological characters for species diagnosis have been firmly established, we frequently find identification problems regarding poorly detailed descriptions and poorly preserved specimens. These may lead to taxonomic incongruencies. Physaloptera retusa (Rudolphi, 1819) is the most common species of the genus and has been reported from several species of neotropical reptiles. Based on our reexaminations of nematode specimens identified as P. retusa from different museum collections, we provide a detailed redescription including the type material, voucher specimens and new specimens recovered currently and showed in this study with new morphological data obtained using light and scanning electron microscopy tools.

A new species of Aplectana Railliet & Henry, 1916 (Nematoda: Cosmocercidae) in the Brazilian Amazon and the taxonomic status of Aplectana longa.

Nematodes of the genus Aplectana Railliet & Henry, 1916 are common parasites of the digestive tract of amphibians and reptiles in the Neotropical region. During a parasite survey on Boana boans (Linnaeus, 1758), we found specimens of nematodes with Aplectana characteristics. We observed a set of characteristics that differs the species of our study from its congeners, and the present study describes a new species of Aplectana parasite of B. boans using light microscopy and scanning electron microscopy. Aplectana pella n. sp. has lateral alae and somatic papillae in males and females; males have equal short spicules, and the gubernaculum is absent. The arrangement of pairs of caudal papillae also differs from other species (2 precloacal,1 adcloacal, and 5 postcloacal+1 unpaired). In females, the vulva is simple, with non-prominent lips, and equatorial. This is the first record of the genus Aplectana parasitizing B. boans and the 58th species described for this genus. Additionally, we added the precloacal papillae pattern of A. delirae, and based on morphological and morphometric characteristics, we propose the reallocation of Aplectana longa to the genus Oxyascaris.

The natural interaction between Myotis nigricans (Schinz, 1821) and its trematodes: A histopathological analysis.

Bats have a wide diversity of digeneans; however, even with the recent increased interest in studies of parasites on these hosts, there are no data on the microscopic alterations of this host-parasite interaction. The present work characterizes and compares the histological aspects of the liver, gallbladder, and intestine of non-parasitized and parasitized Myotis nigricans by digeneans. Ten specimens of Myotis nigricans collected in an urban area of Western Amazonia were analyzed for parasites. The digeneans were removed from the hosts and identified. Tissue samples of the liver, gallbladder, and intestines of parasitized and non-parasitized hosts were collected for histological studies. The gallbladder was observed in repletion and presents mucosa formed by simple epithelium that varies from cubic to cylindrical. The hepatic lobes do not have a classic polyhedral-hexagonal aspect. Variations in basophilia, acidophile, and cytoplasmatic granulations were observed in hepatocytes. The parasitism of the intestinal digeneans was restricted to space delimited by the extensions of villi in high association with the intestinal epithelium, not invading the region of the intestinal glands at the base of the villi. Trematodes maintained attached to the villus by the oral sucker and acetabulum, connected by a "pleat" composed of epithelium and lamina propria layers. We observed no signs of inflammatory processes and cellular defense infiltrates in host tissues. Cytochemistry alterations, size variation, and granular deposits in hepatocytes, enterocytes, and goblet cells were observed. Thus, this report is the first study of the natural parasite-host interaction in the liver, gallbladder, and intestine in M. nigricans in the neotropical region.

Rhabdias paraensis sp. nov.: a parasite of the lungs of Rhinella marina (Amphibia: Bufonidae) from Brazilian Amazonia.

The nematode parasites of Rhinella marina include species of the genus Rhabdias (Rhabdiasidae: Rhabditoidea). The present study describes Rhabdias paraensis sp. nov., which parasitizes the lungs of R. marina in Brazilian Amazonia. Of the more than 70 known species of this genus, 18 are parasites of bufonids, of which, eight are Neotropical. The new species described here is similar to Rhabdias alabialis in the absence of lips is different by the presence of conspicuous cephalic papillae. We describe details of the four rows of pores, which are distributed equally along the whole of the length of the body and connected with hypodermal cells, using histology and scanning electron microscopy. Other histological aspects of the internal structure of this nematode are also described.

Lanfrediella amphicirrus gen. nov. sp. nov. Nematotaeniidae (Cestoda: Cyclophyllidea), a tapeworm parasite of Rhinella marina (Linnaeus, 1758) (Amphibia: Bufonidae).

The family Nematotaeniidae, tapeworms commonly found in the small intestines of amphibians and reptiles, includes 27 recognised species distributed among four genera: Bitegmen Jones, Cylindrotaenia Jewell, Distoichometra Dickey and Nematotaenia Lühe. The taxonomy of these cestodes is poorly defined, due in part to the difficulties of observing many anatomical traits. This study presents and describes a new genus and species of nematotaeniid parasite found in cane toads (Rhinella marina) from eastern Brazilian Amazonia. The cestodes were collected during the necropsy of 20 hosts captured in the urban area of Belém, Pará. The specimens were fixed and processed for light microscopy, scanning electron microscopy (SEM) and three-dimensional (3D) reconstruction. Samples were also collected for molecular analyses. The specimens presented a cylindrical body, two testes and paruterine organs. However, they could not be allocated to any of the four existing nematotaeniid genera due to the presence of two each of dorsal compact medullary testes, cirri, cirrus pouches, genital pores, ovaries and vitelline glands per mature segment. Lanfrediella amphicirrus gen. nov. sp. nov. is the first nematotaeniid studied using Historesin analysis, SEM and 3D reconstruction, and it is the second taxon for which molecular data have been deposited in GenBank.

Morphological diagnosis of Setaria labiatopapillosa in domestic bovids from Marajó Island, Brazil.

The genus Setaria is known worldwide for parasiting wild and domestic mammals, with 43 species registered, of which five occur in the American continent. The objective of this study was to characterize the occurrence, morphology and morphometry of S. labiatopapillosa in B. taurus and B. bubalis in Marajó Island, Brazil. Carcass inspections of 420 cattle were carried out in two slaughterhouses in Pará. The nematodes were found on the white and serous viscera of the small intestine, being cleaned and discussed for analysis by light microscopy and scanning electron microscopy. The morphological and morphometric characteristics are compatible with S. labiatopapillosa parasitizing both species, with a prevalence of 25% (B. bubalis) and 24% (B. taurus). Information obtained by scanning electron microscopy was added to the description of the species, in addition to expanding the distribution of this parasite in the national territory.

Contracaecum larvae: morphological and morphometric retrospective analysis, biogeography and zoonotic risk in the amazon.

In Brazil there are several records of parasitic nematodes of fish with zoonotic potential, especially those belonging to the family Anisakidae. This study considers the morphology, morphometry and prevalence of Contracaecum in Astronotus ocellatus , fish consumed in the Amazon and sold as ornamental and it also performs a retrospective analysis of the diversity of fish with larvae of Contracaecum, in studies carried out in Brazil over a period of 90 years. 40 specimens of A. ocellatus were necropsied, and the nematodes were collected and fixed in 93 parts 70% ethyl alcohol, 5 parts formaldehyde, and 2 parts glacial acetic acid (AFA) for morphological analysis under light microscopy and scanning electron microscopy. Of the 40 fish collected during this work, 27 were parasitized by Contracaecum larvae with a total intensity of 150 larvae. Retrospective analysis of intermediate host diversity for Contracaecum larvae resulted in 16 orders, 49 families, 96 genera, 140 species and a hybrid morphotype. In the retrospective study, half of the fish were from freshwater, with the order Perciformes being the most representative, with 16 families, 30 genera and 37 species. In Brazil, the occurrence of larvae of Contracaecum in fish was reported in 15 of the 26 states, with Rio de Janeiro presenting the most information regarding fish harboring Contracaecum larvae.

Cucullanus marajoara n. sp. (Nematoda: Cucullanidae), a parasite of Colomesus psittacus (Osteichthyes: Tetraodontiformes) in the Marajó, Brazil.

Cucullanus marajoara n. sp. (Cucullanidae) is reported to parasitize Colomesus psittacus (Tetraodontiformes), which is a fish species from the Marajó Archipelago, state of Pará, estuarine region of the Brazilian Amazon. The new species differs from similar species by the presence of a protruding upper lip on the cloacal opening, the distribution of the cloacal papillae: five pre-cloacal papillae pairs and 5 are ventral and located posteriorly to the pre-cloacal sucker and an unpaired papilla is located on the upper cloacal lip and five post-cloacal pairs, and a pair of lateral phasmids located between papillae pairs. Additionally, Cucullanus marajoara n. sp. is compared to other species of this genus described in Brazil, particularly Cucullanus ageneiosus and Cucullanus oswaldocruzi, which both occur in the same zoogeographic region of this study but parasitize fish of a different order (Siluriformes). Cucullanus dodsworthi and Cucullanus brevicaudatus are the only described species parasitizing fish of the order Tetradontiformes in Brazil, and the new species differs from these species by the distribution of the cloacal papillae and the host habitat. The description of Cucullanus marajoara n. sp. adds data to the biodiversity of described parasites that parasitize Tetradontiformes of the estuarine ichthyofauna in the Brazilian Amazon.

Gongylonema Parasites of Rodents: A Key to Species and New Data on Gongylonema neoplasticum.

Specimens of the genus Gongylonema were collected from the gastric mucosa of rodents of Rattus rattus Linnaeus, 1758, and Rattus norvegicus Berkenhout, 1769, collected in urban areas in Belém, Pará, in the eastern Brazilian Amazon. The helminths were processed for analysis using light microscopy and scanning electron microscopy (SEM) techniques and presented taxonomic characteristics of the species Gongylonema neoplasticum. The SEM analyses revealed the presence of 2 developed buccal plates (1 dorsal, 1 ventral), also called interlabia, with a prominent and bifurcated ventral plaque. The occurrence of the bifurcated ventral interlabium had not yet been identified by any other author from G. neoplasticum. As a result of our extensive research on published data on Gongylonema spp., we propose a taxonomic key for species of this genus that parasitize rodents. This is the first record of G. neoplasticum in urban areas of the Brazilian Amazon.

Occurrence of Dipetalonema gracile in a wild population of woolly monkey Lagothrix poeppiigii in the northeastern Peruvian Amazon.

Dipetalonema gracile (Rudolphi, 1809) (Filarioidea: Onchocercidae) is one of six species of cavities filarial parasites of Neotropical non-human primates. The present study recorded the occurrence of D. gracile, provides morphological and morphometric data and extends the geographical distribution. Adult filariae were obtained from the thoracic and abdominal cavities of 38 specimens of woolly monkey, which were used for local human consumption, in the northeastern Peruvian Amazon. Male and female filarids were processed and analysed using light and scanning electron microscopy. Details of the cephalic papillae, post-cloacal bands and papillae, vulva, phasmid position and lateral appendages are showed by scanning electron microscopy and is recorded the occurrencce of Lagothrix poeppigii monkey as a new host of this filaria in the Yavari-Mirin river basin, Peruvian Amazon.

Atractis thapari (Nematoda, Atractidae) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí, Brazil.

Atractis thapari Petter, 1966, an atractid nematode, was collected parasitizing the large intestine of tortoises of the species Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) (Testudinidae) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. Taxonomic identification was based on morphological and morphometrical features, and using bright-field and scanning electron microscopy. The present study adds new observations on the morphology of A. thapari, mainly relating to mouth papillae, the excretory pore, deirids, and male and female posterior ends. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) for A. thapari in these two tortoise species were: P = 100%, MI = 154,667, MA = 154,667, RI = 5,500-588,500 (Cc); P = 100%, MI = 93,639, MA = 93,639, RI = 1,000-224,500 (Cd). This report confirms the occurrence of A. thapari in Neotropical region, South America, Brazil, and extends its occurrences to a new host, the tortoise C. carbonarius. Adjustment of host management with the aim of improving hygiene and health conditions is suggested.

Chapiniella variabilis (Nematoda) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí.

Chapiniella variabilis (Chapin, 1924), a strongylid nematode, was collected parasitizing the large intestine of the tortoises Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. The taxonomic identification was based on morphological and morphometric features, using bright-field and scanning electron microscopy. The present study adds new observations on the morphology, mainly relating to the mouth papillae, external and internal leaf-crown elements, excretory pore, deirids and male and female posterior end. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) of C. variabilis in these two tortoise species were: P = 100%, MI = 833.3, MA = 833.3, RI = 500-1,500 (Cc); P = 100%, MI = 472.2, MA = 472.2, RI = 333-500 (Cd). This record expands occurrences of C. variabilis to a new host, C. carbonarius, and to another state in Brazil, in the Neotropical region of South America. Adjustment to host management with the aim of improving hygiene and health conditions is suggested.

Gnathostomatidae nematode parasite of Colomesus psittacus (Osteichthyes, Tetraodontiformes) in the Ilha de Marajó, Brazilian Amazon.

The genus Gnathostoma comprises 17 species, whose adult specimens are found in the stomach serosa of animals that consume raw fish; some species of the genus are zoonotic agents. The present study describes the presence of a nematode (Gnathostomatidae) parasitizing the digestive tract of Colomesus psittacus in the Ilha de Marajó in the eastern Brazilian Amazon. Thirty specimens of C. psittacus were collected in the municipality of Soure, Ilha de Marajó, state of Pará, Brazil, transported to the laboratory, necropsied and the helminths were collected and fixed. Of the 30 fish that were studied, 16.67% were parasitized with nematodes. The nematode larvae found encysted in the intestinal serosa have anterior region with two lips, each with a pair of papillae; a cephalic bulb armed with six rows of discontinuous spines; four cervical sacs; a claviform esophagus; cuticular striations along the body; a simple excretory pore; and a short tail ending in a mucron. These morphological structures are diagnostic characters of the genus Gnathostoma, whose adults parasitize the stomach of carnivorous mammals and, rarely, the stomach of fish. However, fish, amphibians, reptiles, and birds are intermediate hosts of the third-stage larvae (L3), and humans may act as accidental hosts.

Choledocystus elegans (Digenea: Plagiorchiidae) of Leptodactylus paraensis (Amphibia: Leptodactylidae) from the Brazilian Amazon.

The trematodes are parasites of the several vertebrates including amphibians, however the knowledge about of the taxonomy these parasites is still confuse. The trematode Choledocystus elegans was found in the small intestine of the Leptodactylus paraensis in eastern Amazon and presents the following characteristics: several pointed tegumentary spines, papillae on the outer and inner edges of the oral and ventral suckers, a round, well-developed cirrus sac, a well-developed cirrus, oblique testicles, a ovary right side, uterine loops extending between the testicles, follicular vitellaria distributed throughout the body, starting at the genital pore region and caeca close the end of the body. For the first time, this study identified C. elegans parasitizing L. paraensis and describes morphological aspects never characterized using light and scanning electron microscopy.

Rhinella marina (Amphibia: Bufonidae) Versus Rhabdias paraensis (Nematoda: Rhabdiasidae): Expanding the View on a Natural Infection.

Amphibian and reptile lungs are frequently infected with Rhabdias parasites, and this condition ultimately leads to reduced survival, performance, and growth because of granulomatous inflammation, nodule formation, and nematodal pneumonia onset. Here we investigate the histopathological features of naturally infected Rhinella marina by the lung nematode Rhabdias paraensis. A total of 10 host animals were captured in peridomiciliar areas in the eastern Brazilian Amazon, and anatomic-histological analyses were performed on both the infected and non-infected lungs of these amphibians. Helminths were usually found within the secondary and primary septa of infected lungs whereas parasites were not detected within vessels or adhering to tissues. In addition, we observed discrete erythrocytes, diapedesis foci, few granulocytes and erythrocytes in the interseptal spaces, discrete cell infiltration, and a small number of melanomacrophages, and no granulomas or cysts were observed. New aspects related to changes in tissue and helminth-host interactions are discussed for the relationship of R. paraensis × Rhi. marina from the Amazon region.

Morphological characterization of Eustrongylides sp. larvae (Nematoda, Dioctophymatoidea) parasite of Rhinella marina (Amphibia: Bufonidae) from Eastern Amazonia.

Absctract Eustrongylides spp. nematodes have birds as final hosts and uses other vertebrates as intermediate/paratenic host (fish, amphibians and reptiles) and have zoonotic potential. In amphibians, the larvae may be located in the subcutaneous tissues, liver and mesentery, between the muscle fibres, especially in the lower limbs. Rhinella marina, which is widely observed in Brazil, has exhibited complex diversity in its helminth fauna, reflecting the unique habitat of the Amazon biome. For the first time, this study describes the morphological aspects of third-stage larvae of Eustrongylides sp. in Rhinella marina from Santa Cruz do Ararí, Marajó Archipelago, Eastern Amazonia, using light and scanning electron microscopy.

Ortleppascaris sp. and your host Rhinella marina: A proteomic view into a nematode-amphibian relationship.

The success of the helminth-host relationship depends on a biochemical molecular arsenal. Perhaps the proteome is the largest and most important set of this weaponry, in which the proteins have a crucial role in vital processes to the parasite/host relationship, from basic metabolism and energy production to complex immune responses. Nowadays, the bioproducts expressed by the parasites are under the "spotlight" of immunoassays and biochemical analysis in helminthology, especially in proteomic analysis, which has provided valuable information about the physiology of the infecting agent. Looking into this point of view, why not turn to the infected agent as well? This study characterised the proteomic profile of fluid-filled fibrous cysts of encapsulated Ortleppascaris sp. larvae in the hepatic parenchyma of their intermediate host, the amphibian Rhinella marina. The proteins were separated by two-dimensional electrophoresis and identified by MS with the aid of Peptide Mass Fingerprint. A total of 54 molecules were analysed in this system, revealing a complex protein profile with molecules related to basic metabolic processes of the parasite, energy production, oxi-reduction and oxidative stress processes as well as molecules related to the host response. This study contributes to proteomic studies of protein markers of the development, infectivity, virulence and co-existence of helminths and their hosts.

Contracaecum larvae: morphological and morphometric retrospective analysis, biogeography and zoonotic risk in the amazon / Larva de Contracaecum: análise retrospectiva morfológica e morfométrica, biogeografia e risco zoonótico na amazônia

Abstract In Brazil there are several records of parasitic nematodes of fish with zoonotic potential, especially those belonging to the family Anisakidae. This study considers the morphology, morphometry and prevalence of Contracaecum in Astronotus ocellatus , fish consumed in the Amazon and sold as ornamental and it also performs a retrospective analysis of the diversity of fish with larvae of Contracaecum, in studies carried out in Brazil over a period of 90 years. 40 specimens of A. ocellatus were necropsied, and the nematodes were collected and fixed in 93 parts 70% ethyl alcohol, 5 parts formaldehyde, and 2 parts glacial acetic acid (AFA) for morphological analysis under light microscopy and scanning electron microscopy. Of the 40 fish collected during this work, 27 were parasitized by Contracaecum larvae with a total intensity of 150 larvae. Retrospective analysis of intermediate host diversity for Contracaecum larvae resulted in 16 orders, 49 families, 96 genera, 140 species and a hybrid morphotype. In the retrospective study, half of the fish were from freshwater, with the order Perciformes being the most representative, with 16 families, 30 genera and 37 species. In Brazil, the occurrence of larvae of Contracaecum in fish was reported in 15 of the 26 states, with Rio de Janeiro presenting the most information regarding fish harboring Contracaecum larvae.

Resumo No Brasil existem vários registros de nematóides parasitos de peixes com potencial zoonótico, especialmente aqueles pertencentes à família Anisakidae. Este estudo considera a morfologia, morfometria e prevalência de Contracaecum em Astronotus ocellatus, peixe consumido na Amazônia e vendido como ornamental, e também realiza uma análise retrospectiva da ictiofauna parasitada com larvas de Contracaecum, em estudos realizados no Brasil, durante um período de 90 anos. 40 espécimes de A. ocellatus foram necropsiados, e os nematóides foram coletados e fixados em 93 partes de álcool etílico 70%, 5 partes de formaldeído e 2 partes de ácido acético glacial (AFA) para análise morfológica em microscopia de luz e microscopia eletrônica de varredura. Dos 40 peixes coletados durante este trabalho, 27 estavam parasitados por Contracaecum, com intensidade total de 150 larvas. Análises retrospectivas da diversidade de hospedeiros intermediários para larvas de Contracaecum resultaram em 16 ordens, 49 famílias, 96 gêneros,140 espécies e um morfotipo híbrido. No estudo retrospectivo, metade dos peixes eram de água doce, sendo a ordem Perciformes a mais representativa, com 16 famílias, 30 gêneros e 37 espécies. No Brasil, a ocorrência de larvas de Contracaecum em peixes foi relatada em 15 dos 26 estados, com o Rio de Janeiro apresentando mais informações sobre peixes portadores de larvas de Contracaecum.

Histopathologic aspects in Plagioscion squamosissimus (HECKEL, 1940) induced by Neoechinorhynchus veropesoi, metacestodes and anisakidae juveniles.

Plagioscion squamosissimus (Heckel 1840), a fish endemic to the Amazon Basin and commonly known as the "silver croaker", plays an important role in the ecology and economy of Pará State, Brazil. Knowledge of host-parasite relationships is important to understanding the role of parasites in the control of natural host populations. This work describes histopathological aspects caused by several common intestinal parasites found during a helminthological survey of fish in northern Brazil. We observed a high prevalence of helminth infection, especially by J3 nematode juveniles of the family Anisakidae and metacestodes of the family Protocephalidae (both with 100% prevalence). An external capsule surrounded each juvenile with numerous juveniles inside sac-like structures formed of connective tissue. Inflammation was observed to be caused by infection of metacestodes, reaching the intestinal muscularis mucosa. Neoechinorhynchus veropesoi (38% prevalence) was found in the small intestine of P. squamosissimus, invading the mucosa, submucosa, and internal muscularis of the intestine causing intense inflammation. Histopathology of host-parasite relationships in fish has been rare, and the pathology of parasites in P. squamosissimus is described herein.