Microscopic Study of Mechanoreceptors and Chemoreceptors of Anterior and Posterior Ends of Toxocara Canis Using Scanning Electron Microscopy and Light Microscope.

The present study investigated the fine structure of amphids and phasmids, cuticle, muscles, and digestive tracts of Toxocara canis using optical and electron microscopy, hematoxylin-eosin (H&E) staining, and other specific stains. A number of 38 adult T.canis worms were obtained from the animal shelter of Urmia, and their small intestines were fixated in acidified formal alcohol and 10% formalin solutions. The anterior and posterior parts of male and female T.canis worms were prepared and cut at a thickness of 4-5 μm according to the conventional method in the histological laboratory. The samples were then stained using H&E and specific periodic acid-Schiff, Masson's trichrome, and Orcein staining. The structure of amphid (anterior), phasmid (posterior), cuticle, muscles, and digestive tracts of male and female worms were studied under light microscopy. Basal, intermediate, cortex, and cuticle surface coating of the parasite were visible. Alae were also observed as the thickenings in the cuticle. The muscle layer structure consists of non-branched cylindrical cells. The intestinal tract is composed of cuticular cogs, the esophagus is of filamentous-muscular structure, and the intestine is made of columnar epithelial tissue with microvilli and glycocalyx. The amphid structure consisted of cuticular protrusions with penetrations of the cephalic framework into their inner layers. Phasmid structure also includes protrusions in the cuticle and invagination of sensory neurons. It was concluded that for the most part, the histological structure of the cuticle can be studied by optical microscopy. The muscle structure in this parasite was very similar to the skeletal muscle in mammals. Furthermore, the epithelial structure of the intestine in this parasite was largely similar to the intestinal epithelium in mammals. Finally, regarding the amphid and phasmid structures, it was observed that they were protrusions covered by cuticles where neural, filamentous, and muscular structures were the core of these protrusions.

First ultrastructural observations on gastritis caused by Physaloptera clausa (Spirurida: Physalopteridae) in hedgehogs (Erinaceus europeaus).

Ultrastructural changes of gastritis due to infection with Physaloptera clausa in 12 fresh carcasses of euthanized European hedgehogs (Erinaceus europaeus) collected from different part of Urmia, Iran, in which they were highly populated with this animal, six females and six males were subjected to detail necropsy with special reference to the stomach. Macroscopic changes of stomach were recorded and some of the worms collected. Based on number of parasites present in the stomach, they were divided into light infection, mild infection, and severe infection. Parasites were collected, and worms identification of the species was confirmed on the basis of light microscope examination with reference to keys. Tissues fixed in 3% glutaraldehyde, post-fixed in 1% osmium tetroxide and processed and plastic embedded; ultrathin sections of 60-70 nm were cut and stained with uranyl acetate and lead citrate; electron microscopic observations showed that, in light infection some changes were observed in gastric cells such as dilatation and vesiculation of the endoplasmic reticulum, large numbers of lipid granules, mitochondrial swelling, nuclear chromatin margination, and some nucleus showed washed out appearance. Other cells showed some alterations in mitochondria, dilatation of smooth endoplasmic reticulum, loss of both free and bound ribosomes, vesiculation in cytoplasm, and increase Golgi apparatus and secretory vesicles. The inflammatory cells including lymphocytes, macrophages, mast cells, and predominantly eosinophils were identified. In moderate infection, the cellular pattern of gastric mucosa replaced with inflammatory cells. The marked increase of macrophages and other inflammatory cell was observed. A particular finding in our study was the presence of globule leukocyte in the moderate stage. Moreover, scant formation and distribution of collagen fibers as well as fibroblasts were also noted. In severe infection, the most obvious observation was marked distribution of collagen fibers around the mucosal cells. The fibroblastic cells with elongated nucleus and extensive indentation were noticed. In conclusion, the result of our study revealed P. clausa could be a cause of gastritis and according to cellular pattern of inflammatory reaction, with the increase of worm burden and development of infection, chronic gastritis was stabilized. Present investigation documented the ultrastructural changes during verminous gastritis in hedgehogs.

Macroscopic and microscopic examination of pulmonary Crenosoma striatum in hedgehog.

The aim of study was to necropsy and histopathology evaluation of lung Crenosoma striatum in hedgehog. In July 2012, 10 porcupines were collected from Urmia city and transferred to parasitology lab of the veterinary faculty where they were euthanized by ketamine (over 40-90 mg/kg) intraperitoneally. In this study the lungs were assessed through naked eyes regarding parasite presence upon washing. The lung tissue was examined under loop microscope in order to finding small worms in lung parenchyma. The worms were removed by Anse forceps and kept in AFA solution, and collected for diagnosis. In order to carrying out pathological tests, some samples prepared and placed in formalin 10 % for fixation. The counted worms frequency in high severe and moderate lungs were as 86 (50 females and 36 males) and 19 (13 females and 16 males) worms respectively. But no worms were observed in healthy lungs. The infestation severity was as; low infestation (1-7 worms), moderate infestation (8-20), severe infestation (21-50) and very severe infestation (more than 50 worms). The lung examinations of non-infested lungs indicated that the lung tissues had no parasite. In addition, no inflammation reactions as inflammatory cells presence were observed, and the air spaces with alveoles' wall in some regions were observable. On histopathological examination, the observed alteration was primarily inflammatory changes, and in some cases the proliferation was also observable. Hyperemia and inflammatory cell infiltration, somehow the alveolar space was filled, representing bronchopneumonia reaction. The bronchioles had various changes as hypertrophy and hyperplastic in different parts of respiratory system. Hyperemia and hemorrhage were very severe in some cases caused hemosiderosis in the lung. In severe inflammations the pneumonia along with increasing of bronchial cells in the lumen rose as well, leading to severe verminous infestation of the lung. In regard to the obtained results, the verminous infestation of the porcupines' lung with C. striatum indicated inflammatory and proliferative alteration which was as inflammatory changes in mild cases, and inflammatory and proliferative stances in severe cases.

The phylogeny of the Schistosomatidae based on three genes with emphasis on the interrelationships of Schistosoma Weinland, 1858.

Schistosomes are digenean flukes, parasitic of birds, mammals and crocodiles. The family Schistosomatidae contains species of considerable medical and veterinary importance, which cause the disease schistosomiasis. Previous studies, both morphological and molecular, which have provided a good deal of information on the phylogenetics of this group, have been limited in the number of species investigated or the type or extent of molecular data used. This paper presents the most comprehensive phylogeny to date, based on the sequences of 3 genes, complete ribosomal small subunit rRNA and large ribosomal subunit rRNA, and mitochondrial cytochrome oxidase 1, sequenced from 30 taxa including at least 1 representative from 10 of the 13 known genera of the Schistosomatidae and 17 of the 20 recognized Schistosoma species. The phylogeny is examined using morphological characters, intermediate and definitive host associations and biogeography. Theories as to the origins and spread of Schistosoma are also explored. The principal findings are that Ornithobilharzia and Austrobilharzia form a sister group to the Schistosoma; mammalian schistosomes appear paraphyletic and 2 Trichobilharzia species, T. ocellata and T. szidati, seem to be synonymous. The position of Orientobilharzia within the Schistosoma is confirmed, as is an Asian origin for the Schistosoma, followed by subsequent dispersal through India and Africa.

Parasiticidal effects of peroxynitrite on ovine liver flukes in vitro.

Peroxynitrite (ONOO-) is a cytotoxic anion, produced by interaction between nitric oxide and superoxide in vivo in some inflammatory cells. This study investigated its effects on Fasciola hepatica and Dicrocoelium dendriticum isolated from ovine livers and kept in bile at room temperature. Peroxynitrite was synthesized using a quenched flow reactor and assayed spectrophotometrically. It was applied at different concentrations (10(-3.5) to 10(-2.3 M)) to the flukes kept in bile. The viability of the peroxynitrite-treated flukes was compared with a control group (n=5-7 per group). Control F. hepatica and D. dendriticum lived for 226+/-11and 208+/-14min, respectively. Life times were decreased by peroxynitrite at all concentrations used (P<0.001). At the highest concentration of peroxynitrite, F. hepatica and D. dendriticum lived only for 6.1+/-0.4 and 4.1+/-4.1+/-0.2min, respectively. Correlation between peroxynitrite concentration and parasite viability was significant in the case of F. hepatica (r= -0.842; P= 0.0035). A single application of peroxynitrite can decrease the life span of ovine liver flukes. A failure in the activation of hepatic macrophages in infected animals may lead to a decreased production of free radicals and, thus, peroxynitrite. Such a failure is likely to deprive the body of a defence tool against multicellular parasites.

Scanning electron microscopy of adult Gongylonema pulchrum (Nematoda: Spirurida).

Scanning electron microscopy (SEM) was used to study the surface ultrastructure of adult worms of Gongylonema pulchrum. The anterior end in both sexes was covered by numerous cuticular platelets. There was a pair of lateral cervical papillac. The buccal opening was small and extended in the dorsoventral direction. Around the mouth a cuticular elevation enclosed the labia, and eight papillae were located laterodorsally and lateroventrally. Two large lateral amphids were seen. On the lateral sides of the female's tail, phasmidal apertures were observed. The caudal end of the male was asymmetrically alate and bore 10 pairs of papillae and two phasmidal apertures.