Morphological, molecular and ecological characterization of a native isolate of Steinernema feltiae (Rhabditida: Steinernematidae) from southern Chile.

BACKGROUND: Steinernema feltiae is an entomopathogenic nematode used in biological control programs with a global distribution. Populations of this species show phenotypic plasticity derived from local adaptation and vary in different traits, such as location and host penetration. The aim of this work was to describe a Chilean isolate of this nematode species, using integrative approaches. METHODS: Nematode morphological and morphometric studies were conducted along with molecular analysis of nuclear genes. The symbiotic bacterium was also identified by sequencing the 16S rRNA gene. Some ecological characteristics were described, including the temperature requirements for the nematode life cycle and the effect of soil water content for optimal reproduction. RESULTS: Morphometric characterization revealed a large intra-specific variability. The isolate identity was also corroborated with the analysis of nuclear genes. Based on the 16S gene, its symbiont bacteria, Xenorhabdus bovienii, was identified. The lowest, optimal and highest temperatures found to limit the infestation and reproduction on Galleria mellonella were 10, 20 and 30 °C, respectively; the emergence from the host larvae occurred approximately 10 days after inoculation. Differences were observed in offspring, and 120 infective juveniles (IJ)/larva was the most prolific dose at 20 °C. The soil water content did not affect the number of IJ invaders, penetration efficacy and IJ emergence time or offspring per larva, but it caused a delay in achieving full mortality at the permanent wilting point with respect to saturation and field capacity. CONCLUSIONS: For the first time, a Chilean isolate of S. feltiae is described in detail considering morphological, molecular and ecological aspects. The isolate was shown to be efficient in soil containing water, with optimal temperatures ranging from 15 to 25 °C for host infestation and production of an abundant offspring; these characteristics would allow its potential use as control agents in a wide geographical area of the country.

A new species of Rhabdias (Nematoda: Rhabdiasidae), a lung parasite of Pseudopaludicola pocoto (Anura: Leptodactylidae) from north-eastern Brazil: description and phylogenetic analyses.

Rhabdias pocoto n. sp. is herein described from the lungs of the swamp frog Pseudopaludicola pocoto Magalhães, Loebmann, Nogueira, Kokubum, Baptista, Haddad & Garda, 2014, from the Caatinga biome in the state of Ceará, in north-eastern Brazil. The new species is characterized by a body that dilates posteriorly, six small lips (protuberances) and two rounded lateral expansions of cuticular inflation on the anterior end, each containing an amorphous gland-like structure inside and a short and conical tail. Additionally, molecular analysis and comparison of the partial mitochondrial cytochrome c oxidase I sequence of R. pocoto n. sp. revealed genetic divergence between the new species and the sequences of Rhabdias spp. previously deposited in GenBank. Phylogenetic analysis grouped the new taxon into the R. pseudosphaerocephala species complex + R. glaurungi clade. The new discovery represents the 19th species of Rhabdias spp. described in the Neotropical region, the ninth in Brazil and the first species of Rhabdias found parasitizing South American frogs of the genus Pseudopaludicola, as well as the first Caatinga biome species of Rhabdias.

First case of fatal equine meningoencephalitis caused by Halicephalobus gingivalis in Mexico.

Aberrant nematode larval migration in the CNS of horses is rare but frequently fatal; one of the main etiological agents involved in this illness is Halicephalobus gingivalis. This soil nematode has been associated with several fatal equine meningoencephalitis reports worldwide; however, it had never been diagnosed in horses of Mexico. A 10 year-old Andalusian horse presented dysphagia, fever, weakness, prostration and ataxia; the patient expired during the medical attention. Post mortem examination was performed and no gross alterations were found. Histopathology revealed meningoencephalitis, vasculitis and intralesional adult nematodes, larvae and eggs compatible with Halicephalobus spp. A polymerase chain reaction (PCR) for the nuclear large subunit ribosomal RNA gene (LSU rDNA) of nematodes was performed from formalin-fixed and paraffin wax-embedded sections of brain. Posterior nucleotide sequence analysis of the amplified fragment identified the agent as H. gingivalis. To our knowledge, this is the first confirmed report of Halicephalobiasis in Mexico.

Rhabdias glaurungi sp. nov. (Nematoda: Rhabdiasidae), parasite of Scinax gr. ruber (Laurenti, 1768) (Anura: Hylidae), from the Brazilian Amazon.

The genus Rhabdias Stiles & Hassal, 1905 includes about 83 species of nematodes parasitic in amphibians and reptiles worldwide. Herein, we describe Rhabdias glaurungi sp. nov. from the hylid frog Scinax gr. ruber (Laurenti, 1768) in the Gunma Ecological Park, Santa Bárbara municipality, state of Pará, Brazil. This species has six small lips, an inflated cuticle along the entire body and a cup-shaped buccal capsule with smooth internal surface of its anterior part and irregularly folded internal surface of its posterior part in apical view. From the 17 valid species recognized in the Neotropical realm, the new species can be distinguished by the number of lips, the morphology and size of its buccal capsule, as well as the extent and shape of its cuticular inflation; in addition, there are molecular differences. Sequences of the mitochondrial Cytochrome c Oxidase subunit I gene strongly support the status of this form as a separate species. Molecular phylogenetic analysis shows R. glaurungi sp. nov. nested within the R. pseudosphaerocephala Kuzmin, Tkach & Brooks, 2007 species complex. Rhabdias glaurungi sp. nov. is the second species of the genus described from hosts of the family Hylidae in the Neotropical realm. We conclude that the diversity of Rhabdias within the Neotropics is likely largely underestimated.

Description, biology and molecular characterisation of Serpentirhabdias moi n. sp. (Nematoda: Rhabdiasidae) from Chironius exoletus (Serpentes: Colubridae) in Brazil.

A new species of the genus Serpentirhabdias Tkach, Kuzmin et Snyder, 2014, S. moi n. sp., is described from a colubroid snake Chironius exoletus from Caxiuanã National Forest, State of Pará, Brazil. The species is characterised by having a triangular oral opening, absence of the buccal capsule, presence of six minute onchia in the oesophastome, and excretory glands of approximately the same length as the oesophagus. These qualitative morphological characters, as well as some measurements, differentiate the new species from other Neotropical and Nearctic Serpentirhabdias spp. The morphological description of parasitic adults of S. moi n. sp. is complemented by the description of free-living stages including males, females, and infective larvae. Comparative analysis of partial sequences of cox1 and 12S mitochondrial genes strongly supported the status of S. moi n. sp. as a new species. Molecular phylogeny based on sequences of the nuclear DNA region spanning the 3' end of the 18S nuclear rRNA gene, ITS region (ITS1 + 5.8S + ITS2) and 5' end of the 28S gene supported monophyly of all rhabdiasid genera included in the analysis and placed the new species into the Serpentirhabdias clade as sister taxon to S. fuscovenosa.

A new species of Diomedenema (Nematoda, Rhabditida, Spiruromorpha) from Spheniscus magellanicus (Aves, Sphenisciformes) found on the southern coast of Brazil.

A new species of the genus Diomedenema, a spiruromorph nematode, collected from the lung of Spheniscus magellanicus (Sphenisciformes) found on the southern coast of the state of Rio Grande do Sul, Brazil, is described. The new species is differentiated from the only previously described species of the genus, D. diomedeae Johston & Mawson, 1952, by males possessing a set of caudal papillae with three pairs of precloacal, two pairs of adcloacal and one pair of postcloacal papillae; precloacal papillae with the papillae of the first two pairs being closer to each other than those of the third pair; a longer and pointed tail in males; and females with the vulva at mid-body. This is the first report of a nematode infecting the lung of a sphenisciforme host.

Morphometric and phylogenetic analyses of Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) from the lancehead snake Bothrops moojeni Hoge, 1966 (Reptilia: Serpentes: Viperidae) in Brazil.

Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) is described from the lungs of the 'Brazilian lancehead' Bothrops moojeni (Hoge, 1966) from the savannah in São Paulo State, Brazil. The new species is the eighth species of Serpentirhabdias described in the Neotropical region, and differs from other species mainly by a combination of characters: lips slightly notable, presence of fine striations at posterior ends, presence of two parallel lines with intercalated pores, a pore-shaped phasmid situated at the level of the anal aperture and another two in the posterior half of the tail. It is the first species of Serpentirhabdias reported in this snake host and the second species of this genus found parasitizing South American viperidian snakes. Molecular phylogenetic analysis using ribosomal (ITS and 28S partial) genes confirms Serpentirhabdias viperidicus n. sp. as a new species that clustered in the Serpentirhabdias clade, sister taxon to Serpentirhabdias fuscovenosa and Serpentirhabdias elaphe. This is the first description of Serpentirhabdias species from Brazil using molecular approaches and morphological characters to confirm the monophyly of this recent genus.

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.

Description of Serpentirhabdias atroxi n. sp. (Nematoda: Rhabdiasidae), a parasite of Bothrops atrox (Linnaeus) (Reptilia: Serpentes: Viperidae) in Brazilian Amazonia.

A new lung-dwelling nematode species is described from the common lancehead Bothrops atrox (Linnaeus) in the Brazilian Amazon Region. The species is assigned to the genus Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on the presence of six lips arranged in two lateral groups, the absence of prominent cuticular inflations, and lung parasitism in snakes. Serpentirhabdias atroxi n. sp. differs from other species of the genus mainly by details of the morphology of the anterior end: cuticularised ring surrounding the anterior part of the buccal cavity and six minute onchia present in the oesophastome. Serpentirhabdias atroxi n. sp. is the seventh species of the genus known from the Neotropical Realm and the second species described from viperid snakes.

Sicarius uncinipenis and Deletrocephalus cesarpintoi in captive greater rheas of Minas Gerais State, Brazil.

The present study details the pathological and parasitological findings of parasitic ventriculitis and nematode infections in the large intestines of two female Rhea americana americana birds. The birds were housed in captivity, and both exhibited poor body condition and lethargy. The rheas were sent to the Veterinary Hospital of the Veterinary School, Universidade Federal de Minas Gerais (UFMG) and, despite medical care, the clinical condition of the birds did not improve. The birds died two days after admission, and were submitted to necropsy. Gross, histopathology and parasitological analysis resulted in the identification of Sicarius uncinipenis, which is associated with parasitic ventriculitis, while Deletrocephalus cesarpintoi was identified in the large intestine of both rheas. The apparent clinical indications, including loss of appetite and death, combined with the discovery of numerous parasites and other pathology changes, supported the conclusion that the death of the birds was caused by the parasitic infection. Further investigations of these infections in free-living and captive rheas are required, such that accurate data on the incidence and pathogenicity of these parasites can be obtained.

Sicarius uncinipenis and Deletrocephalus cesarpintoi in captive greater rheas of Minas Gerais State, Brazil / Sicarius uncinipenis e Deletrocephalus cesarpintoi em emas de cativeiro no Estado de Minas Gerais, Brazil

The present study details the pathological and parasitological findings of parasitic ventriculitis and nematode infections in the large intestines of two female Rhea americana americana birds. The birds were housed in captivity, and both exhibited poor body condition and lethargy. The rheas were sent to the Veterinary Hospital of the Veterinary School, Universidade Federal de Minas Gerais (UFMG) and, despite medical care, the clinical condition of the birds did not improve. The birds died two days after admission, and were submitted to necropsy. Gross, histopathology and parasitological analysis resulted in the identification of Sicarius uncinipenis, which is associated with parasitic ventriculitis, while Deletrocephalus cesarpintoi was identified in the large intestine of both rheas. The apparent clinical indications, including loss of appetite and death, combined with the discovery of numerous parasites and other pathology changes, supported the conclusion that the death of the birds was caused by the parasitic infection. Further investigations of these infections in free-living and captive rheas are required, such that accurate data on the incidence and pathogenicity of these parasites can be obtained.

O presente estudo relata os achados patológicos e parasitológicos de ventriculite parasitária e da infecção por nematódeo no intestino grosso em duas fêmeas Rhea americana americana. As aves eram mantidas em cativeiro e ambas apresentaram condição corporal ruim e inapetência. As emas foram encaminhadas para o Hospital Veterinário da Escola de Veterinária da UFMG e, apesar dos cuidados médicos, não houve melhora na condição clínica. As aves morreram dois dias após a internação e foram encaminhadas para a necropsia. Ao exame macroscópico, histopatológico e parasitológico, Sicarius uncinipenis foi identificado e associado com ventriculite parasitária, enquanto Deletrocephalus cesarpintoi foi identificado no intestino grosso. Possivelmente, o quadro de inapetência e morte foi causado pela infecção parasitária, pois os parasitos eram numerosos. Mais investigações dessa infecção são necessárias em emas de vida livre e cativeiro, para a obtenção de informações mais precisas da incidência e patogenicidade desses parasitos.

Sicarius uncinipenis and Deletrocephalus cesarpintoi in captive greater rheas of Minas Gerais State, Brazil / Sicarius uncinipenis e Deletrocephalus cesarpintoi em emas de cativeiro no Estado de Minas Gerais, Brazil

The present study details the pathological and parasitological findings of parasitic ventriculitis and nematode infections in the large intestines of two female Rhea americana americana birds. The birds were housed in captivity, and both exhibited poor body condition and lethargy. The rheas were sent to the Veterinary Hospital of the Veterinary School, Universidade Federal de Minas Gerais (UFMG) and, despite medical care, the clinical condition of the birds did not improve. The birds died two days after admission, and were submitted to necropsy. Gross, histopathology and parasitological analysis resulted in the identification of Sicarius uncinipenis, which is associated with parasitic ventriculitis, while Deletrocephalus cesarpintoi was identified in the large intestine of both rheas. The apparent clinical indications, including loss of appetite and death, combined with the discovery of numerous parasites and other pathology changes, supported the conclusion that the death of the birds was caused by the parasitic infection. Further investigations of these infections in free-living and captive rheas are required, such that accurate data on the incidence and pathogenicity of these parasites can be obtained.

O presente estudo relata os achados patológicos e parasitológicos de ventriculite parasitária e da infecção por nematódeo no intestino grosso em duas fêmeas Rhea americana americana. As aves eram mantidas em cativeiro e ambas apresentaram condição corporal ruim e inapetência. As emas foram encaminhadas para o Hospital Veterinário da Escola de Veterinária da UFMG e, apesar dos cuidados médicos, não houve melhora na condição clínica. As aves morreram dois dias após a internação e foram encaminhadas para a necropsia. Ao exame macroscópico, histopatológico e parasitológico, Sicarius uncinipenis foi identificado e associado com ventriculite parasitária, enquanto Deletrocephalus cesarpintoi foi identificado no intestino grosso. Possivelmente, o quadro de inapetência e morte foi causado pela infecção parasitária, pois os parasitos eram numerosos. Mais investigações dessa infecção são necessárias em emas de vida livre e cativeiro, para a obtenção de informações mais precisas da incidência e patogenicidade desses parasitos.

Description of Rhabdias breviensis n. sp. (Rhabditoidea: Rhabdiasidae) in two Neotropical frog species.

Nematodes of the genus Rhabdias Stiles & Hassall, 1905 (Rhabditoidea: Rhabdiasidae) have a dioecious free-living stage and a hermaphroditic stage that parasitises the lungs of amphibians and reptiles. Approximately 94 species of Rhabdias have been described. Because the similar morphological characteristics such as the labial structures, the location of the vulva and the shape of the tail of Rhabdias spp. hinder their identification, molecular biology techniques and scanning electron microscopy have been employed to diagnose species of this genus. This study describes Rhabdias breviensis n. sp., parasitic in the lungs of two Neotropical frog species Leptodactylus petersii Steindachner and Leptodactylus macrosternum Miranda-Ribeiro. The description of this species integrates classical taxonomy, scanning electron microscopy and a molecular analysis of the mitochondrial COI gene. The new species differs from all other Rhabdias species parasitic in Neotropical hosts in certain morphometric parameters, the position of the vulva, the host group and the cephalic characters.

Helminth parasites of Pseudacris hypochondriaca (Anura: Hylidae) from Baja California, Mexico, with the description of two new species of nematodes.

The helminth parasite fauna of the hylid frog Pseudacris hypochondriaca in several localities along the Baja California Peninsula in northwestern Mexico is presented. The helminth fauna consists of 4 species of nematodes (Oswaldocruzia pipiens, a larval form of an Ascaridid, 2 new species belonging to the genera Rhabdias and Cosmocercoides), and 1 species of digenean ( Gorgoderina sp.). The new species of Rhabdias represents the 88th species assigned to the genus and the third species described from Mexican anurans. Also, the species of Cosmocercoides represents the 20th species assigned to the genus and the first representative of this genus described from Mexico.

Rhabdias filicaudalis n. sp. (Nematoda: Rhabdiasidae) from the snake Spilotes pullatus (Serpentes: Colubridae) in Brazil.

Rhabdias filicaudalis n. sp. (Nematoda: Rhabdiasidae) from the lung of Spilotes pullatus (Serpentes: Colubridae) is described. The host snake was captured in the municipality of Avaré, São Paulo State, Brazil. Rhabdias filicaudalis n. sp. differs from all other species by the combination of the following characters: straight body, truncated anterior end, six weakly developed lips arranged in two opposite groups of three, pre-equatorial vulva, oesophagus length/body length ratio (%) 4.9-7.5 (5.8 +/- 0.6), nerve ring distance from anterior end/oesophagus length ratio (%) 36.9-61.1 (49.8 +/- 6.4), tail length/body length ratio (%) 3.0-5.5 (4.0 +/- 0.5), vulva distance from anterior end/body length ratio (%) 39.9-51.7 (45.9 +/- 3.5), and a cuticular filiform tail tip.

Origin of the parasites of an invading species, the Australian cane toad (Bufo marinus): are the lungworms Australian or American?

Phylogeographical analyses that identify the geographical origin of parasites in invading species can clarify the parasites' potential for biological control of the invader and the risks posed by the parasite to native species. Our data on nuclear and mitochondrial genetic sequences show that the nematode lungworms (Rhabdias spp.) in invasive Australian populations of cane toads (Bufo marinus) are Rhabdias pseudosphaerocephala, a South American species. We did not find this lungworm species in any Australian frogs sympatric with cane toads, suggesting that the parasite does not attack Australian frogs and hence may offer potential as a biocontrol agent of the toad.

New species of Rhabdias (Nematoda: Rhabdiasidae) and other helminths from Norops capito (Sauria: Polychrotidae) from Nicaragua.

Rhabdias nicaraguensis n. sp. (Rhabditida: Rhabditidae) from the lungs of Norops capito (Sauria: Polychrotidae) is described and illustrated. Rhabdias nicaraguensis n. sp. represents the 54th species assigned to the genus and the 12th from the Neotropical realm. Of the 12 Neotropical Rhabdias species, nicaraguensis is most similar to tobagoensis and vellardi. These 3 species have equatorial placement of the vulva, inflated cuticle, and 6 small circumoral lips. Rhabdias nicaraguensis is easily separated from R. tobagoensis by the shape of the buccal cavity and from R. vellardi by body size and shape of the tail. Rhabdias nicaraguensis differs from both species by host preference, the amount of inflated cuticle covering the body, and the phasmids situated posterior to the midpoint of the tail.

Two new species of Rhabdias (Nematoda: Rhabdiasidae) from the marine toad, Bufo marinus (L.) (Lissamphibia: Anura: Bufonidae), in Central America.

Two new Rhabdias species are described from the lungs of the cane toad Bufo marinus (L.) from Costa Rica and Nicaragua. Rhabdias alabialis n. sp. differs from other known species of the genus by the remarkable morphology of its head end, i.e., the absence of lips or pseudolabia, the slitlike oral opening, and the triangular shape of the buccal capsule in apical view. Rhabdias pseudosphaerocephala n. sp. is identified as a form previously known in Central and South America as Rhabdias sphaerocephala Goodey, 1924, a species initially described from toads in Europe. The new species is differentiated from R. sphaerocephala based on head-end morphology and sequences of nuclear rDNA.

New species of Rhabdias (Nematoda: Rhabdiasidae) from Bufo occidentalis (Anura: Bufonidae) from Sierra Madre del Sur, Mexico.

Rhabdias kuzmini n. sp., a parasite of the lungs of Bufo occidentalis, is described and illustrated. This Mexican taxon differs from the related species in the genus by the possession of 4 lips (2 subdorsal and 2 subventral) and 2 lateral pseudolabia, corpus not inflated, a larger barrel-shaped buccal capsule, equatorial vulva, and the presence of a slightly swollen cuticle in the anterior and posterior ends of the body. This is the 16th species described in the Neotropical Realm and the first species of Rhabdias described from endemic anurans in México.

Helminths in Mesaspis monticola (Squamata: Anguidae) from Costa Rica, with the description of a new species of Entomelas (Nematoda: Rhabdiasidae) and a new species of Skrjabinodon (Nematoda: Pharyngodonidae).

Entomelas duellmani n. sp. (Rhabditida: Rhabdiasidae) from the lungs and Skrjabinodon cortagoensis n. sp. (Oxyurida: Pharyngodonidae) from the intestines of Mesaspis monticola (Sauria: Anguidae) are described and illustrated. E. duellmani is the sixth species assigned to the genus and is the third species described from the Western Hemisphere. It is easily separated from other neotropical species in the genus by pre-equatorial position of its vulva. Skrjabinodon cartagoensis is the 24th species assigned to the genus and differs from other neotropical species in the genus by female tail morphology.