Proliferative strongyloidiasis in a colony of colubrid snakes.

Strongyloides are small rhabditid nematodes primarily associated with enteric disease in a variety of animal species, including reptiles. Strongyloides spp life stages were associated with a disease outbreak in a large breeding colony of snakes. Multiple Pantherophis and Lampropeltis colubrids exhibited respiratory distress, anorexia, stomatitis, facial deformation, and waning body condition that resulted in death or necessitated euthanasia. Postmortem examinations of 13 snakes revealed epithelial hyperplasia and inflammation of the alimentary and respiratory tracts associated with varying numbers of adult and larval nematodes and embryonated or larvated ova. In a subset of snakes, aberrant nematode migration was also observed in the eye, genitourinary system, coelom, and vasculature. Histomorphology and gross examination of parasitic adult female nematodes from host tissues were consistent with a Strongyloides spp. Sedimented fecal material from 101/160 (63%) snakes housed in the affected facility was positive for nematodes and/or larvated ova. Polymerase chain reaction amplification and sequencing of portions of the 18S and 28S ribosomal ribonucleic acid (RNA) genes and the internal transcribed spacer region of adult female parasites and positive fecal samples supported the diagnosis of strongyloidiasis. Strongyloides spp possess a unique life cycle capable of alternating between parasitic (homogonic) and free-living (heterogonic) stages, resulting in the production of directly infective larvae. Commonly utilized husbandry practices in reptile collections can amplify the numbers of infective larvae generated in the captive environment, increasing the risk for rhabditid hyperinfections. This report documents morbidity, mortality, and non-enteric disease manifestations due to Strongyloides hyperinfections in a captive colubrid snake colony.

The biology of Strongyloides spp.

Strongyloides is a genus of parasitic nematodes that, unusually, has a free-living adult generation. Here we introduce the biology of this genus, especially the fascinating but complex life-cycle, together with an overview of the taxonomy, morphology, genetics, and genomics of this genus.

Morphological and molecular characterization of Strongyloides ophidiae (Nematoda, Strongyloididae).

The aim of the present study is to report morphological data from parasitic female, rhabditoid and filarioid larvae, free-living female worms and eggs of Strongyloides ophidiae (Nematoda, Strongyloididae). In addition, a molecular DNA analysis was carried out using a pool of eight S. ophidiae parasitic females. Samples were obtained from the small intestine of Oxyrhopus guibei (Serpentes, Colubridae) collected in the municipality of Lençóis Paulista, State of São Paulo, Brazil. DNA amplification by polymerase chain reaction (PCR) resulted in a 350 bp band for samples containing S. ophidiae and Strongyloides venezuelensis DNA. Strongyloides ophidiae nucleotide sequence analysis showed 98% similarity with Strongyloides procyonis and 97% with Strongyloides cebus, Strongyloides stercoralis, Strongyloides fuelleborni and Strongyloides sp. from snakes.

The stomal morphology of parasitic females of Strongyloides spp. by scanning electron microscopy.

For the discrimination of the species of the genus Strongyloides, the shape of the stoma in the apical view of parasitic females is the most important feature. Microscopical interpretations divided the shape of the stoma into four groups (simple, angular, complex, and simple but with esophageal teeth); however, the stomas classified in the latter two groups, in particular, remained unclear. In the present study, parasitic females of eight representative Strongyloides spp. with the angular stoma (S. procyonis and S. martis), complex stoma (S. robustus, S. callosciureus, S. planiceps, S. papillosus and S. myopotami), and stoma with esophageal teeth (S. ransomi) were studied by scanning electron microscopy. All species examined in this study having complex stoma or with esophageal teeth possessed at least four linguiform or verruciform projections submarginally in the stomal cavity. Namely, the number, size, shape, and the site of attachment of intra-stomal projections, along with the depth of oral cavity, determine a range of complex images of the stoma such as X-shaped or ornate stomas under light microscopy. Similarly, with the standard examination, which usually uses a lateral view of the head, erroneous interpretations of the number of lobes on the circumoral elevation of parasitic females might occur due to such intra-stomal projections in some Strongyloides spp.

A new rhabditoid nematode species in Asian sciurids, distinct from Strongyloides robustus in North American sciurids.

Strongyloides callosciureus n. sp. (Nematoda: Rhabditoidea), from Asian sciurids, is described based on morphology, morphometry, and the small and large subunit (SSU/LSU) ribosomal RNA gene (rDNA) sequences. This new species was collected from Pallas's squirrels (Callosciurus erythraeus) in the central part of mainland Japan (Honshu), which were originally introduced from Taiwan some decades ago, and plantain squirrels (Callosciurus notatus) imported from Malaysia as personal pets. For comparison, Strongyloides robustus Chandler, 1942 was collected from American red squirrels (Tamiasciurus hudsonicus) and southern flying squirrels (Glaucomys volans) imported from the United States as personal pets. The parasitic females found in North American and Asian sciurids shared some key morphological features such as the ovary running spirally around the gut, and the shapes of the stoma in the apical view and the tail. However, morphometric features of parasitic females in North American and Asian sciurids differed significantly from each other; the former was larger than the latter, and the relative position of the vulva to the whole body length from the mouth was different. The SSU/LSU rDNA sequences supported the division of sciurid Strongyloides isolates by geographical distribution of the host and morphological features, leading us to propose the erection of new species.

Exposure of medical staff to Strongyloides stercolaris from a patient with disseminated strongyloidiasis.

We examined whether medical staff were infected with Strongyloides stercolaris through exposure to the body substances of a patient with disseminated strongyloidiasis. The patient excreted a large number of S. stercolaris organisms in respiratory secretions and stool-like excretions from a nasogastric tube. Blood tests in six physicians and three nurses, who were highly suspected of having had contact with the substances without appropriate protection during medical care of the patient for about 1 week, showed no increase of eosinophiles or IgG antibodies against S. stercolaris. We conclude that adherence to the standard precautions is sufficient for preventing the nosocomial transmission of this organism.

Identification and characterization of the threadworm, Strongyloides procyonis, from feral raccoons (Procyon lotor) in Japan.

Strongyloides procyonis Little, 1966 was detected about 45 years ago in raccoons (Procyon lotor) of southern Louisiana, U.S.A., and was demonstrated experimentally to cause creeping eruption and a short-lived intestinal infection in a healthy human volunteer. After its description and demonstration of its pathogenicity in humans, S. procyonis has not been found in raccoons in North America despite repeated surveys. During a survey on feral raccoons in Japan, S. procyonis parasitic females were identified in 66 (28.3%) of 233 raccoons collected between May 2004 and January 2005. The number of parasitic females recovered from individual raccoons was 1-197 (geomean, 3.2). Both the morphological features and the nucleotide sequences of the small and large subunit ribosomal RNA genes (SSU/LSU rDNA) of S. procyonis closely resembled those of zoonotic Strongyloides stercoralis. The sequences of internal transcribed spacer (ITS)1 and 28S rDNA could differentiate clearly these 2 species. Awareness of S. procyonis in raccoons in North America and other places worldwide where raccoons are introduced and naturalized is important to assess the epidemiological significance of this potentially zoonotic helminth species.

Characterization of the protective response against a homologous challenge infection with Strongyloides venezuelensis in rats.

The protective response in rats against a homologous challenge infection with Strongyloides venezuelensis was characterized. In an initial infection with 1000 filariform larvae and migrating larvae (L(3)) of S. venezuelensis, the population of L(3) in the lungs on day 3 postinfection (PI), and that of adult worms in the small intestine on day 7 PI, were 180.8+/-14.5 and 336.8+/-70.7, respectively. The latter were gradually expelled towards day 42 PI. After the initial infection, the rats developed strong immunity against a homologous challenge infection as manifested by a marked reduction in worm populations, stunted body length and width, damage to reproductive organs, impaired egg production and rapid expulsion of the worms by day 14 after challenge. Expulsion of the worms was preceded by a significantly elevated (P<0.05) peripheral blood eosinophil (PBE) count, both in the initial (200.0+/-26.5 x 10(3)ml) and the challenge infection (400.9+/-165.4 x 10(3)ml). These findings suggest that rats acquire strong homologous immunity following initial exposure to S. venezuelensis. It is suggested that PBEs are involved in worm expulsion. A major target of these effector mechanisms is the reproductive system of S. venezuelensis.

Secreted adhesion molecules of Strongyloides venezuelensis are produced by oesophageal glands and are components of the wall of tunnels constructed by adult worms in the host intestinal mucosa.

The parasitic female of Strongyloides venezuelensis keeps invading the epithelial layer of the host intestinal mucosa. Upon invasion, it adheres to the surface of the intestinal epithelial cells with adhesion molecules secreted from the mouth. It has been demonstrated that S. venezuelensis are expelled from the intestine because mucosal mast cells inhibit the attachment of adult worms to the mucosal surface. In the present study, we generated specific antibodies against secreted adhesion molecules to investigate their function in vivo, because these molecules have been demonstrated only in vitro in spite of the importance in the infection processes. A mouse monoclonal antibody specific to S. venezuelensis adhesion molecules inhibited the attachment of adult worms to plastic dishes and the binding of adhesion molecules to rat intestinal epithelial cells. Immunohistochemical study revealed that adhesion molecules were produced by oesophageal glands and were continuously secreted in vivo to line the wall of the tunnels formed by adult worms in the intestinal mucosa. Our findings indicate that adhesion molecules play essential roles in the infection processes of S. venezuelensis in the host intestine.

Strongyloidiasis in a Cope's grey tree frog (Hyla chrysoscelis).

A male Cope's grey tree frog (Hyla chrysoscelis) died spontaneously with ventral subcutaneous edema and was necropsied. Thickening of the intestinal mucosa was observed histopathologically, with villous atrophy and intraepithelial nematodes present. Adult female Strongyloides sp. nematodes were isolated from the fixed intestinal tract, the first time this nematode genus has been recovered from this frog genus. Intestinal strongyloidiasis should be considered as a cause for protein-losing enteropathy and death in frogs.

Morphology, homogonic development, and lack of a free-living generation in Strongyloides robustus (Nematoda, Rhabditoidea), a parasite of North American sciurids.

Adult females of Strongyloides robustus Chandler, 1942, a parasite of sciurids in North America, were found in the duodenal mucosa of 30 of 32 red squirrels (Tamiasciurus hudsonicus (Erxleben)) collected in Cape Breton Island, Nova Scotia, Canada. The parasitic female is illustrated and redescribed; characteristics include: body 3.8-8.0 mm long, cephalic extremity with X-shaped mouth and 8 circumoral lobes, ovaries spiralling around intestine, and tail bluntly rounded. Eggs in fresh feces contained tadpole-stage larvae. In fecal cultures, eggs hatched and larvae invariably developed to the filariform infective third stage; i.e. a free-living generation did not occur and is probably absent in S. robustus in Cape Breton and possibly other parts of North America. It is hypothesized that homogonically developing S. robustus might be more fecund or more efficiently transmitted than species of Strongyloides that exhibit both homogonic and heterogonic development. Larvae of S. robustus in fecal cultures, i.e. homogonic larvae, are described in detail. Intestinal walls of second- and third-stage larvae, as well as the lateral chords of young third-stage larvae, contained numerous round bodies, likely nutrient stores. Third-stage larvae were present within 2 days in cultures maintained at 30 degrees C, 4 days at 20 degrees C, and 7 days at 15 degrees C. They lived for at least 33 and 30 days at 15 degrees C and 20 degrees C, respectively. Third-stage larvae probably die when their nutrient stores are exhausted.

Toxocara vitulorum in the milk of buffalo (Bubalus bubalis) cows.

The duration of excretion of Toxocara vitulorum larvae in the milk of buffalo cows determines the optimum time for treating calves. Studies on 10 cows showed that a few larvae occur in the colostrum of some cows before the calf has suckled, but most are present from the day after calving and for a further five days. From day 9 onwards, very few larvae were found in the milk. The total number of larvae found was comparable with the number of adult parasites collected from the calves of cows with similar histories. The larvae were 1254 +/- 60 microns long and 36 +/- 6.7 microns in diameter at the ventriculus, figures which are substantially different from some published results.

Strongyloidiasis in cotton rats (Sigmodon hispidus) from central Oklahoma.

Thirty-one of 40 cotton rats (Sigmodon hispidus) collected from central Oklahoma were infected with Strongyloides sp. (78% prevalence). Larvae of Strongyloides sp. (rhabditiform or filariform) were not demonstrable in intestinal contents and scrapings. Female nematodes recovered from intestinal contents and scrapings had morphological similarities with Strongyloides sigmodontis. Cotton rats infected with Strongyloides sp. were indistinguishable clinically from non-infected hosts. Infected animals had no significant gross lesions, but the presence of Strongyloides sp. in the intestinal mucosa was associated with villus atrophy and mild to moderate infiltration of the lamina propria by lymphocytes, plasma cells and occasional eosinophils. Other organs or tissues examined were free from lesions induced by Strongyloides sp.

Intestinal nematode infections.

This article discusses strongyloidiasis, hookworm infection, trichostrongyliasis, ascariasis, trichuriasis (whipworm infection), and enterobiasis (pinworm infection). For each infection, the author describes the organism, the epidemiology and geographic distribution, symptomatology and pathogenesis, and diagnosis and treatment.

Scanning electron microscopy of the penetration of newborn mouse skin by Strongyloides ratti and Ancylostoma caninum larvae.

The scanning electron microscopical appearances of infective larvae of Strongyloides ratti and Ancylostoma caninum have been described. Particularly noteworthy was the tail of S. ratti which was found to have a distal aperture surrounded by a row of eight projections. The penetration of larvae through newborn mouse skin was investigated. S. ratti larvae forced its way rapidly and directly through the stratum corneum. No larvae of A. caninum were observed in stages of partial penetration but the occasional empty sheath was seen.

Nematode parasites of Puerto Rican tree frogs, Eleutherodactylus spp: two new species and a proposal of Poekilostrongylus gen. nov. (Trichostrongylidae).

Poekilostrongylus puertoricensis gen.nov., sp.nov. is proposed for nematodes recovered from Eleutherodactylus coqui, in Puerto Rico. The new genus is similar to Oswaldocruzia Travassos, 1917, but lacks longitudinal ridges on the cuticle. Oswaldocruzia lenteixierai Viqueras, 1938, is partially redescribed, and a key to the genera of the Oswaldocruziinae is given. Thelandros (Parathelandros) garciai sp.nov. is described from E. antillensis and E. portoricensis. Strongyloides sp. was found in E. antillensis and Aplectana spp. (females) were recovered from E. locustus, E. richmondi and E. coqui.