Molecular phylogeny and diagnosis of species of the family Protostrongylidae from caprine hosts in Uzbekistan.

Protostrongylids are important pulmonary parasites of ungulates, particularly caprines. Their complex life cycles involve terrestrian gastropods as intermediate hosts. Morphological discrimination of larvae in feces and snails is impossible to the species level, and molecular data are missing for many species. To improve diagnosis and epidemiology of protostrongylids, this study describes internal transcribed spacer 2 (ITS-2) sequences for five protostrongylid and one metastrongylid species from Uzbekistan. Morphological and molecular analyses identified Protostrongylus rufescens, Protostrongylus hobmaieri, Protostrongylus sp., Spiculocaulus leuckarti, and Cystocaulus ocreatus. Sequence differences between ITS-2 were sufficient to allow species identification, e.g., in the Protostrongylinae intraspecific and interspecific genetic distances ranged between 0-0.01 and 0.05 and 0.16, respectively. There was one exception in the Elaphostrongylidae with identical ITS-2 sequences in Elaphostrongylus cervi and Elaphostrongylus rangiferi questioning their status as valid species. Maximum likelihood analysis sequences largely supported the currently assumed phylogenetic relationships among Protostrongylidae as predicted using morphological characters. The monophyly of the subfamilies Varestrongylinae, Elaphostrongylinae, and Protostrongylinae was corroborated with support values in Shimodaira-Hasegawa or Bayesian modifications of the approximate likelihood ratio test (aLRT) ≥97%, but support for Muellerinae was weak (8 and 52%, respectively) since Muellerius capillaris differs significantly from the other Muellerinae. On the genus level, paraphyly of the genus Protostrongylus was confirmed since the members of the genera Spiculocaulus and Orthostrongylus were located within the Protostrongylus cluster (aLRT ≥99%). Maximum likelihood unequivocally assigned every unique sequence to the correct species confirming suitability of ITS-2 regions for diagnosis protostrongylids except of E. cervi and E. rangiferi.

Troglostrongylus brevior and Troglostrongylus subcrenatus (Strongylida: Crenosomatidae) as agents of broncho-pulmonary infestation in domestic cats.

BACKGROUND: Aelurostrongylus abstrusus is currently regarded as the main metastrongyloid infesting domestic cats, whereas the reports of Troglostrongylus spp. in domestic and wild felids largely remain anecdotic. This paper reports on pulmonary infestation caused by Troglostrongylus brevior and Troglostrongylus subcrenatus in two kittens and describes, for the first time, associated clinical presentations and pathological features. Morphometrical, molecular and phylogenetic analyses have also been conducted to differentiate here the examined Troglostrongylus species from A. abstrusus, towards a clearer delineation of metastrongyloids affecting cats. METHODS: Two kittens were referred for respiratory distress and hospitalized with a diagnosis of severe aelurostrongylosis, based on the presence of metastrongyloid larvae in the faeces. Despite prompt treatment, kittens died within 48 hours. Both kittens were submitted to necropsy to determine the cause of death. RESULTS: At necropsy, nematode specimens were found in the trachea, bronchi and bronchioles and were associated with respiratory signs (i.e., dyspnoea, polypnea, severe coughing and nasal discharge). Morphology and measurements of adult parasites found allowed the unequivocal identification of T. brevior and T. subcrenatus, even if first stage larvae were rather similar to those of A. abstrusus. Briefly, T. brevior and T. subcrenatus larvae were shorter in length and lacking the typical knob-like terminal end of A. abstrusus. Molecular and phylogenetic analyses corroborated morphological identification and provided data on mitochondrial and ribosomal DNA genes of T. brevior. CONCLUSIONS: Data presented here indicate that T. brevior and T. subcrenatus may cause major respiratory distress in domestic cats. Consequently, these two species should be included, along with A. abstrusus, in the differential diagnosis of cat bronchopulmonary affections and treatment protocols need to be evaluated. Through research on the biology, epidemiology and control of Troglostrongylus spp. infestations in domestic cats are advisable to implement current knowledge on these neglected metastrongyloids.

Achatina Fulica como hospedeiro intermediário de nematódeos de interesse médico-veterinário em Goiás, Brasil / Achatina fulica as an intermediate host of nematoda of medical-veterinary interest in Goiás, Brazil

Achatina fulica, também conhecido como caramujo africano, é um molusco terrestre que pode atuar como hospedeiro intermediário de vários helmintos, entre eles alguns com importância médica e veterinária como Angiostrongylus cantonensis e Angiostrongylus costaricensis, nematódeos responsáveis pela meningoencefalite eosinofílica e pela angiostrongilose abdominal, respectivamente. Este estudo objetivou conhecer a distribuição deste molusco no estado de Goiás e pesquisar a ocorrência de larvas de nematódeos de interesse parasitológico. Detectado, inicialmente, em 2003 no município de Morrinhos, A. fulica teve sua distribuição geográfica ampliada e encontra-se atualmente presente em 39,5por cento, dos municípios de Goiás. A pesquisa da helmintofauna,realizada pela técnica de digestão artificial das amostras obtidas em Caldas Novas, Morrinhos e BelaVista de Goiás, resultou no encontro de larvas de Aelurostrongylus abstrusus (prevalência de 35por cento), Rhabditis sp. (47,5por cento), Strongyluris sp. (15por cento) e de outros metastrongilídeos (2,5por cento). Este estudo ampliou o conhecimento da distribuição geográfica de A. fulica em Goiás e reforçou a participação deste molusco em ciclos biológicos de helmintos, indicando a necessidade de controle e vigilância epidemiológica em áreas urbanas com grande densidade deste molusco em razão da facilidade de contato com as populações humanas e de animais domésticos, o que propicia a transmissão de zoonoses.

Phylogenetic analysis of the Metastrongyloidea (Nematoda: Strongylida) inferred from ribosomal RNA gene sequences.

Phylogenetic relationships among nematodes of the strongylid superfamily Metastrongyloidea were analyzed using partial sequences from the large-subunit ribosomal RNA (LSU rRNA) and small-subunit ribosomal RNA (SSU rRNA) genes. Regions of nuclear ribosomal DNA (rDNA) were amplified by polymerase chain reaction, directly sequenced, aligned, and phylogenies inferred using maximum parsimony. Phylogenetic hypotheses inferred from the SSU rRNA gene supported the monophyly of representative taxa from each of the 7 currently accepted metastrongyloid families. Metastrongyloid taxa formed the sister group to representative trichostrongyloid sequences based on SSU data. Sequences from either the SSU or LSU RNA regions alone provided poor resolution for relationships within the Metastrongyloidea. However, a combined analysis using sequences from all rDNA regions yielded 3 equally parsimonious trees that represented the abursate Filaroididae as polyphyletic, Parafilaroides decorus as the sister species to the monophyletic Pseudaliidae, and a sister group relationship between Oslerus osleri and Metastrongylus salmi. Relationships among 3 members of the Crenosomatidae, and 1 representative of the Skrjabingylidae (Skrjabingylus chitwoodorum) were not resolved by these combined data. However, members of both these groups were consistently resolved as the sister group to the other metastrongyloid families. These relationships are inconsistent with traditional classifications of the Metastrongyloidea and existing hypotheses for their evolution.

A retrospective study of 11 cases of lungworm (Didelphostrongylus hayesi) infection in opossums (Didelphis virginiana).

A juvenile, female North American opossum (Didelphis virginiana) died of verminous pneumonia caused by Didelphostrongylus haysei despite aggressive treatment with oral fenbendazole, corticosteroids, and antibiotics. This prompted a retrospective study of lungworm infection in opossums, during which 19 additional necropsy reports from opossums were reviewed. Including the subject of this report, a total of 11 (55%) of these cases included a diagnosis of lungworm infection. This diagnosis was considered to have contributed to death in eight out of the 11 cases (73%). Histologically, 10 of the 11 (91%) opossums had granulomatous bronchopneumonia with small to moderate numbers of adult nematodes in the airways and parenchyma. Four of the 11 (36%) opossums had free larvae within the parenchyma or terminal airways. Inflammation was usually associated with larvae, degenerating parasites, and nonintact adult nematodes. Superimposed bacterial pneumonia was evident in three animals, and sections of lung examined from all the opossums were characterized by moderate to severe smooth-muscle hyperplasia in airways, including terminal respiratory bronchioles and alveolar ducts. Nine animals had prominent medial smooth-muscle hyperplasia in small- and medium-sized arterioles. Lesions in other organs, particularly in liver, heart, and gastrointestinal tract, were frequently identified. Three animals had concomitant septicemia or bacterial bronchopneumonia (or both), which contributed to the cause of death. Seven animals had gastric nematodosis (Physaloptera sp.), although three of them had been treated with a 14-day course of fenbendazole.

[The occurrence of broncho-pulmonary nematodes in sheep from Emilia-Romagna]. / Indagini sulla diffusione dei nematodi bronco-polmonari in ovini dell'Emilia-Romagna.

In the present investigations 510 sheep have been examined for the presence of lungworm larvae: 342 (67,05%) of them proved positive. The identification of the parasites demonstrated the presence of Dyctiocaulus filaria in 18,62% of the examined animals, Muellerius capillaris in 50%, Cystocaulus ocreatus in 26,86% and Protostrongylus spp. in 12,35%. All positive flocks were positive for two or more genera of lungworms.