First report of molecular and phylogenetic analysis of Physaloptera praeputialis in naturally infected stray cats from India.

Nematodes of the genus Physaloptera are globally distributed and infect a multitude of hosts. Their life cycle involves orthopterans and coleopterans as intermediate hosts. The morphological characters alone are inadequate to detect and differentiate Physaloptera spp. from its congeners. Moreover, molecular studies are limited to compare them precisely. The present communication reports the first molecular phylogenetic characterization of feline Physaloptera spp. from India based on mitochondrial cytochrome c oxidase subunit 1 (COX1) and small subunit ribosomal DNA (18S rDNA). The nematodes were first isolated from the stomach of adult stray cats during necropsy examination. Based on the gross and microscopic characters, the worms were identified as P. praeputialis. Morphological identification was further confirmed through PCR targeting the barcode region of the mitochondrial cytochrome c oxidase subunit I (MT-COI) gene, using nematode-specific primers cocktail followed by species specific primers targeting partial COX1 and 18S rRNA genes. Generated sequences were submitted in NCBI GenBank (MW517846, MW410927, MW411349), and phylogenetic trees were constructed using the maximum likelihood method. When compared with other sequences of Physaloptera species across the globe, the present isolates showed 85.6-97.7% and 97.3-99% nucleotide homology based on COX1 and 18S rRNA gene, respectively. BLASTn analysis revealed a strong identity to other Physaloptera spp., and the phylogenetic tree placed all Physaloptera spp. in the same cluster. This study again indicates the usefulness of molecular techniques to substantiate the identity of species that may lack adequate descriptions and impart new insight for the potentially overlooked significance of P. praeputialis infections in felines.

[Human Gongylonema pulchrum infection in esophagus: a case report].

This paper reports a female patient with Gongylonema pulchrum parasitizing in the esophagus, with aims to call for the attention to the role of parasite detection in the diagnosis of human diseases.

Atrophic Proventriculitis Associated with Gravid Females of Tetrameres sp. (Nematoda: Tetrameridae) in the Magellanic Penguin (Spheniscus magellanicus) on the Coast of Brazil.

The present study offers the first description of proventriculitis associated with the presence of gravid female nematodes of the genus Tetrameres (Nematoda: Tetrameridae) in 3 juvenile Magellanic penguins (Spheniscus magellanicus) found dead on the coast of Brazil. This study broadens knowledge on parasites associated with these hosts and the real impact of this association.

The Occurrence of Physaloptera hispida and a Mastophorus Sp. in Pulmonary Vessels of Hispid Cotton Rats (Sigmodon hispidus) from Georgia, U.S.A.

During 2017-2018, a survey for the rat lungworm, Angiostrongylus cantonensis (Nematoda: Metastrongyloidea), in rodents from Piedmont and Lower Coastal Plains physiographic regions of Georgia was conducted. On 4 occasions, a single worm was recovered from the pulmonary vessels of a single cotton rat (Sigmodon hispidis). One of these worms was identified as a Physaloptera sp. and the remaining 3 as a Mastophorus sp. by morphology. No A. cantonensis were found. Physaloptera (Nematoda: Physalopteroidea) and Mastophorus species (Nematoda: Spiruroidea) are stomach parasites of many wild and domestic animals. This is the first report of these species in the pulmonary vessels of a definitive host. To better characterize these parasites, representative specimens were collected from cotton rat stomachs and identified morphologically and molecularly. Based on partial cytochrome c oxidase subunit 1 (COI) gene sequences, Physaloptera hispida from stomachs were identical to the Physaloptera sp. from the pulmonary vessels. The COI sequences from the Mastophorus sp. from the stomach exhibited a higher degree of variability but confirmed that the pulmonary worms were the same Mastophorus species. Furthermore, sequences of Mastophorus from a coastal site clustered separately from a clade of Mastophorus sequences from cotton rats from a Piedmont site. Our data show that adult worms recovered from pulmonary vessels of cotton rats could be either Physaloptera or Mastophorus sp., indicating that these parasitic worms are not always restricted to the stomach and that worms from pulmonary vessels must be carefully examined to obtain a definitive diagnosis of A. cantonensis infection.

First case of human Gongylonema pulchrum infection in Slovenia.

Nematodes of the genus Gongylonema infect a wide range of mammals worldwide but are only sporadically reported in humans. We describe a case of human infection with Gongylonema pulchrum in a 41-year-old man. The patient extracted the nematode from the submucosa under his tongue and correctly self-diagnosed the infection with the help of the Google search engine. In the laboratory, the collected nematode was confirmed as G. pulchrum microscopically by morphological analysis and genetically by amplifying and sequencing the parasite's rDNA. This is the first report of human G. pulchrum infection in Slovenia.

Incidence of cutaneous habronemosis in Manipuri ponies in India.

Information pertaining to parasitic fauna and parasitic diseases in Manipuri ponies in India is not available. Moreover, no systematic studies have been undertaken on cutaneous habronemosis in Manipuri ponies which is a common skin problem of Manipuri ponies as reported by pony owners. Keeping in the view of the importance of parasitic infections in veterinary health coverage particularly in Manipuri ponies, the present study was planned. A survey of natural cases of cutaneous habronemosis followed by molecular confirmation of species involved and treatments were done. Out of 200 ponies examined, nine cases (4.5%) of cutaneous habronemosis was recorded. Gross examination revealed raised and ulcerated wounds with necrotic tissues covered with yellowish-tan granulation. Histopathological study revealed eosinophilic granuloma and in the center of the granuloma with necrotic debris. Remnants of the Hebronema larvae with infiltrating neutrophils surrounded by proliferating fibrous tissue with numerous eosinophils, macrophages and lymphocytes were also observed. Molecular detection of Habronema sp. was confirmed by semi-nested PCR. Sequence analysis revealed larvae of H. muscae was the common spirurid species responsible for producing cutaneous habronemosis in Manipuri ponies. Subsequently, sequence submitted to NCBI GenBank and accession number obtained (MH038181). Surgical removal of necrotic tissue, ivermectin injection along with antibiotics successfully cured all the lesions in infected ponies.Results confirmed occurrence of cutaneous habronemosis in Manipuri ponies in India.

A New Species of Ascarophis (Nematoda: Cystidicolidae) Parasitizing Clinocottus analis (Pisces: Cottidae) from Baja California, Mexico.

A new species of nematode, Ascarophis morronei n. sp. (Cystidicolidae), is described from the stomach wall of the woolly sculpin Clinocottus analis (Cottidae) collected in the rocky intertidal from northwestern Baja California, Mexico. Collected nematodes were studied using both light and scanning electron microscopy. Sequence fragments for 18S rDNA molecular markers were obtained from the new nematode species, in order to test its position within the family Cystidicolidae under a phylogenetic context. Main characters distinguishing this new species include the reduced labia and the morphology of the eggs, distances of nerve ring and excretory pore from the anterior end, and left spicule of males. The new species described here is the second for the genus Ascarophis reported as adult in the Southern California Bight, and the first one recorded for the fish genus Clinocottus.

A new species of Physaloptera (Nematoda: Spirurida) from Proechimys gardneri (Rodentia: Echimyidae) from the Amazon rainforest and molecular phylogenetic analyses of the genus.

Nematodes of the genus Physaloptera are globally distributed and more than 100 species are known. Their life cycle involves insects, including beetles, cockroaches and crickets, as intermediate hosts. This study describes a new species of Physaloptera and reports molecular phylogenetic analyses to determine its relationships within the family Physalopteridae. Physaloptera amazonica n. sp. is described from the stomach of the caviomorph rodent Proechimys gardneri collected in the Amazon rainforest in the state of Acre, Brazil. The species is characterized by the male having the first and second pair of sessile papillae asymmetrically placed, lacking a median papilla-like protuberance between the third pairs of sessile papillae, differentiated by size and shape of the spicules, while females have four uterine branches. For both nuclear 18S rRNA and MT-CO1 gene-based phylogenies, we recovered Turgida sequences forming a clade nested within Physaloptera, thus making Physaloptera paraphyletic to the exclusion of Turgida, suggesting that the latter may have evolved from the former monodelphic ancestral state to a derived polydelphic state, or that some species of Physaloptera may belong to different genera. Relationships between most taxa within Physaloptera were poorly resolved in our phylogenies, producing multifurcations or a star phylogeny. The star-like pattern may be attributed to evolutionary processes where past simultaneous species diversification events took place. Physaloptera amazonica n. sp. formed an independent lineage, separately from the other species of Physaloptera, thus supporting the status of a new species. However, all molecular data suggested a closer relationship with other Neotropical species. In conclusion, we added a new species to this already largely diverse genus Physaloptera, bringing new insights to its phylogenetic relationships. Further analyses, adding more species and markers, should provide a better understanding of the evolutionary history of physalopterids.

First report of Gongylonema sp. in a free ranging callitrichid from the Brazilian Atlantic Forest: case report / Primeiro relato de Gongylonema sp. em um calitriquídeo de vida livre proveniente da Mata Atlântica brasileira: relato de caso

This is the first report of parasitism by Gongylonema sp. in a free-ranging callitrichid from the Atlantic forest of Espírito Santo, Brazil. A juvenile male of Geoffroy's marmoset (Callithrix geoffroyi) was euthanized due to poor prognosis, then necropsied. Samples of the tongue were collected for routine histological processing. Microscopically, there were transversal sections of adult nematodes within the epithelial layer of the mucosa of the tongue. Lingual scraping demonstrated a small number of oval embryonated eggs with a thick capsule. The morphology of the adult parasites and the eggs, associated with its location, were compatible with the Spirurida nematode of the genus Gongylonema. Further studies are needed to evaluate the impact of this parasite on free-ranging callitrichid populations.(AU)

Este é o primeiro relato de parasitismo por Gongylonema sp. em um calitriquídeo de vida livre proveniente da Mata Atlântica do Espírito Santo, Brasil. Um sagui-da-cara-branca (Callithrix geoffroyi), macho, jovem, foi eutanasiado, devido a prognóstico desfavorável, e necropsiado. Fragmentos de língua foram coletados para processamento histológico de rotina. Microscopicamente, havia cortes transversais de nematoides adultos na mucosa da língua. Na raspagem da mucosa da língua, foi observada pequena quantidade de ovos larvados ovais com cápsula espessa. A morfologia do parasita adulto e dos ovos, associada à localização do agente, é compatível com nematoide Spirurida do gênero Gongylonema. São necessários estudos adicionais para avaliar o impacto desse parasito nas populações de calitriquídeos de vida livre.(AU)

Diversity of Rhabdochona mexicana (Nematoda: Rhabdochonidae), a parasite of Astyanax spp. (Characidae) in Mexico and Guatemala, using mitochondrial and nuclear genes, with the description of a new species.

Among fish parasitic nematodes Rhabdochona is one of the most speciose genera, with c. 100 species. Twelve congeneric species occur in Mexican freshwater fishes, in a region located between the Nearctic and Neotropical biogeographical regions. Host association and biogeographical history have determined the high species richness of Rhabdochona in Mexico. One of these species, Rhabdochona mexicana, is highly specific to the characid genus Astyanax. Characids are a group of freshwater fish with Neotropical affinity. In this paper, we explore the genetic diversity of R. mexicana through samples obtained from populations of Astyanax spp. across river basins of Mexico and Guatemala. Sequences of one mitochondrial and two ribosomal genes were obtained from 38 individuals and analysed using Maximum Likelihood and Bayesian Inference analysis. Phylogenetic analyses using cox1, and a concatenated alignment of 18S + 28S + cox1 recovered two genetic lineages. One of them corresponded with R. mexicana sensu stricto; this lineage included three reciprocally monophyletic subgroups; the other lineage was highly divergent and represented a putative candidate species. A detailed morphological study was conducted to corroborate the molecular findings. We describe a new species herein and discuss the implications of using molecular tools to increase our knowledge about the diversity of a speciose genus such as Rhabdochona.

Helminth community and host dynamics in northern bobwhites from the Rolling Plains Ecoregion, U.S.A.

One hundred and sixty-one northern bobwhites (Colinus virginianus; hereafter 'bobwhite') were examined from the Rolling Plains ecoregion of Texas and western Oklahoma from 2011 to 2013. Complete necropsies yielded 13 species, of which two are new host (Gongylonema phasianella) and region (Eucoleus contortus) records and three (Dispharynx nasuta, Tetrameres pattersoni and Oxyspirura petrowi) are known to cause morbidity and mortality. Of the species found, Aulonocephalus pennula commonly occurred, Oxyspirura petrowi was intermediate in prevalence, and the remaining species were rare. Species richness was similar compared to studies from the southeastern U.S., but higher than studies from the same region. In addition, 12 of the 13 species were heteroxenous helminths, supporting the theory that heteroxenous helminths in semi-arid regions are more successful than monoxenous helminths. Prevalence and abundance of A. pennula and O. petrowi were higher in adult bobwhites than in juveniles. Abundance of A. pennula and O. petrowi was higher at southern locations compared to northern locations in the study area. Our study is the first to provide a current assessment of the bobwhite helminth community across the Rolling Plains ecoregion of the U.S.

Gongylonema infection of wild mammals in Japan and Sardinia (Italy).

The gullet worms, classical Gongylonema pulchrum and newly differentiated Gongylonema nepalensis, are prevalent in various mammals in Japan and Sardinia, Italy, respectively. The former species is cosmopolitan in distribution, dwelling in the mucosa of the upper digestive tract of a variety of domestic and wild mammals, and also humans. At present, the geographical distribution of G. nepalensis is known in Nepal and Sardinia, with the nematode having been recorded from the oesophagus of water buffaloes (Nepal), cattle, sheep, goats and wild mouflon (Sardinia). To clarify their natural transmission cycles among domestic and wild mammals, the present study analysed the ribosomal RNA gene (rDNA) and mitochondrial cytochrome c oxidase subunit 1 gene (cox1) of worms of various origins: G. pulchrum worms from sika deer, wild boars, Japanese macaques, and feral alien Reeves's muntjacs in Japan, and G. nepalensis worms from a red fox and a wild boar in Sardinia. Although the internal transcribed spacer (ITS) regions of rDNA and partial cox1 nucleotide sequences of G. pulchrum from native wild mammals in Japan were distinct from those of the worms in cattle, the worms from feral alien Reeves's muntjacs showed the cattle-type ITS genotype and cox1 cattle-I and II haplotypes. The rDNA and cox1 nucleotide sequences of G. nepalensis from a red fox in Sardinia were almost identical to those of the worms from domestic and wild ruminants on the island. The ecological interaction between domestic and wild mammals and their susceptibility to different Gongylonema spp. must be considered when trying to elucidate this spirurid's transmission dynamics in nature.

Molecular identification of Physaloptera sp. from wild northern bobwhite (Colinus virginianus) in the Rolling Plains ecoregion of Texas.

Physaloptera spp. are common nematodes found in the stomach and muscles of mammals, reptiles, amphibians, and birds. Physaloptera spp. have a complicated life cycle with multiple definitive hosts, arthropod intermediate hosts, aberrant infections, and possible second intermediate hosts or paratenic hosts. For example, Physaloptera sp. larvae have been found within the tissues of wild northern bobwhite quail (Colinus virginianus), and it is suspected that quail may serve as paratenic or secondary hosts of these parasites. However, because it is not known what role quail play in the life cycle of Physaloptera spp. and descriptions of Physaloptera spp. larvae are limited, molecular tools may be beneficial when identifying these helminths. In this study, we generated primers using universal nematode primers and obtained a partial mitochondrial cytochrome oxidase 1 (COX 1) sequence. Morphological identification of Physaloptera sp. in bobwhite was confirmed via polymerase chain reaction (PCR), and a phylogenetic tree was constructed using the maximum likelihood method. BLAST analysis revealed a strong identity to other Physaloptera spp. and the phylogenetic tree placed all Physaloptera spp. in the same cluster. We also documented a marked increase in Physaloptera infections in bobwhite from 2017 to 2018, and the similarity of these parasites to Onchocerca volvulus and Wuchereria bancrofti may give insight into the increased prevalence we observed. This study demonstrates the usefulness of molecular techniques to confirm the identity of species that may lack adequate descriptions and provides new insight for the diagnosis and potentially overlooked significance of Physaloptera sp. infections of bobwhite in the Rolling Plains ecoregion of Texas.

Diversity and prevalence of gastrointestinal parasites in two wild Galago species in Gabon.

In this study, we characterize the diversity and estimated infection levels of gastrointestinal parasites circulating in two galago species, Galago demidoff and G. thomasi in two sites situated in the Southeastern forests of Gabon. Our study reveals that eleven parasites including nine helminthes (Ascaris spp., Ankylostoma spp., Dicrocoelium spp., Gongylonema spp., Oesophagostomum spp., Lemuricola spp., Strongyloides spp. Trichostrongylus spp. and Trichuris spp.) and two protozoans (Balantidium spp. and Entamoeba spp.) may infect Galago spp. with high infection rates. The results show that: a very similar parasite spectrum is found in both host species; all the taxa identified were previously observed in other Primate species and/or Man. They also show that age, gender and forest type may influence infection rates and/or parasite diversity found in a particular host and/or geographic area.

Helminth parasites of Mabuya arajara Rebouças-Spieker, 1981 (Lacertilia: Mabuyidae) from Chapada do Araripe, northeastern Brazil.

We provide data on the helminth fauna from the digestive tract of the lizard Mabuya arajara Rebouças-Spieker, 1981 from Chapada do Araripe, northeastern Brazil. Seventy one of the 127 lizards examined (56%) were infected with four nematode species: Physalopteroides venancioi and Physaloptera sp. (Physalopteridae), Strongyluris oscari (Heterakidae), and Parapharyngodon alvarengai (Pharyngodonidae), the latter being the component species (prevalence 53.5%; mean intensity of infection 3.37 ± 2.0; discrepancy index D = 0.69). The helminth P. alvarengai infected M. arajara throughout the year and showed increased infection rates in July, at the beginning of the dry season. In addition to the relationship with seasonality, lizards with greater body length and/or body mass were more infected. Relationships between number of parasites and body mass and with the sexes of lizards, on the other hand, were not found. Mabuya arajara represents a new host for these nematodes. This study contributes to the knowledge of the helminth fauna associated with the digestive tract of lizards from South America and the Caatinga domain.

Two nematode species from freshwater and marine fishes in Thailand, including Ascarophis scatophagi sp. nov. (Cystidicolidae) from Scatophagus argus (Scatophagidae).

One new and one known species of parasitic nematodes are reported from Thailand for the first time: Falcaustra kaverii (Karve et Naik,1951) (Kathlaniidae) from the intestine of the freshwater fish Poropuntius deauratus (Valenciennes) (Cyprinidae) (new host record) and Ascarophis scatophagi sp. nov. (Cystidicolidae) from the intestine of the marine fish Scatophagus argus (Linnaeus) (Scatophagidae). The morphology of both these species was studied with the use of light (LM) and scanning electron microscopy (SEM) (the latter method used for F. kaverii for the first time). The new species (A. scatophagi) is mainly characterized by nonfilamented, markedly elongate-oval eggs (size 42-45 × 18 µm), spicules 555-642 µm and 105-150 µm long, the muscular/glandular oesophagus length ratio (1:13-34), bifurcate deirids, the length of the female tail (159-249 µm) and cephalic structures typical of the nominotypical subgenus Ascarophis. Representatives of Falcaustra Lane, 1915 and Ascarophis van Beneden, 1871 were not previously reported from fishes in Thai waters.

Redescription and genetic characterization of Cystidicoloides vaucheri, including first description of male and current status on the phylogeny of Cystidicolidae (Nematoda: Habronematoidea).

Cystidicoloides vaucheri collected in the stomach of the redtail catfish Phractocephalus hemioliopterus from River Acre, State of Acre, Brazil is redescribed, including the first description of males and the first genetic characterization based upon 18S and 28S genes of the rRNA. Newly collected females were biometrically smaller than those reported in the original description, but similar morphology shared by the two samples revealed that they belong to the same species. Scanning electron micrographs showed the accurate structure of the cephalic region, described here in detail. Furthermore, the morphology of males completed the specific diagnosis, strengthening the validity of the species. The three other congeners differ from C. vaucheri mainly as follows: in C. dlouhyi the area rugosa is absent, the cephalic structures in C. fischeri are completely distinct, and in both species the spicules have membranous outgrowths, absent in C. vaucheri. Despite the dubious generic assignment of C. izecksohni, it differs from C. vaucheri in several biometrical and morphological features. Because of data availability, only sequences of the 18S were used for phylogenetic reconstructions. Results showed that the genus Ascarophis and the families Cystidicolidae and Physalopteridae are not monophyletic. Cystidicoloides vaucheri formed an independent branch clustering with representatives of Cystidicolidae, confirming its validity. The inclusion of Salmonema and Spinitectus within Cystidicolidae should be reviewed, since they formed an assemblage with species from Rhabdochonidae. In fact, current classification of some taxa belonging to Habronematoidea, Physalopteroidea and Thelazioidea need to be re-evaluated, mainly based on molecular data from different genes.

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.