Ultraestructural study of effects of alkylphospholipid analogs against nematodes.

Alkylphospholipid analogs were initially developed as anticancer agents and were later found to antiparasitic activity. Miltefosine is the prototype alkylphosphocholine and is the first oral treatment against visceral leishmaniasis. Here we investigated the effects of miltefosine and two ring-substituted alkylphosphocholine derivatives, TCAN26 and TC70, on the viability, morphology, and ultrastructure of the life stages of Caenorhabditis elegans and infective larvae of the parasite Strongyloides venezuelensis. Miltefosine displayed activity against C. elegans adults at low concentrations and was more effective than TCAN26 and TC70. Miltefosine inhibited the hatching of eggs, leading to embryonic lethality, and showed larvicidal activity against C. elegans and S. venezuelensis larvae after 24 h. Mitelfosine also induced alterations in the reproductive system of hermaphrodites, causing vulvar prolapse and general effects in the body wall. Electron microscopy analysis showed that miltefosine induced selective embryonic lethality, leading to cell death. Our results suggest that alkylphospholipid analogs are a potential new alternative for anti-nematode chemotherapy.

Ultrastructural and cytochemical characterization of T1 and T2 secretory bodies from the tegument of Echinostoma paraensei.

Trematodes are lined by a syncytial layer that is named the tegument and contains small mitochondria and two different kinds of secretory inclusions. The structure and size of these bodies differs among genera and species. In a previous study, we observed many secretory bodies in the tegument of Echinostoma paraensei and named these bodies the T1 and T2 secretory bodies. No previous studies analyzed the secretory bodies of trematodes from the genus Echinostoma. Thus, the aim of this work was to use electron microscopy and cytochemistry to characterize these secretory bodies and to provide a detailed ultrastructural and morphological picture of these bodies, which are found in the tegument of E. paraensei. After ultrastructural cytochemistry analysis, we showed that both the T1 and T2 secretory bodies of E. paraensei were formed by glycoconjugates. 3D reconstruction confirmed the ovoid form of T1 secretory bodies and the biconcave and thin form of T2 secretory bodies.

Histopathological changes in the kidneys of vertebrate hosts infected naturally and experimentally with Paratanaisia bragai (Trematoda, Digenea).

Paratanaisia bragai is a trematode parasite that reaches sexual maturity in the kidney collecting ducts of domesticated and wild fowl and whose intermediate hosts are the snails Subulina octona and Leptinaria unilamellata. There are some discrepancies in descriptions of the pathology of this parasite in bird kidneys. Therefore, the purpose of this study was to analyze the kidneys of rock pigeons (Columba livia) naturally infected and of chickens (Gallus gallus) experimentally infected with Paratanaisia bragai, by means of macroscopic observation and by light and scanning electron microscopy. Both bird species showed significantly dilated collecting ducts. In addition, lymphocyte infiltration was observed in the kidneys of C. livia and metaplasia in the epithelial lining of the kidney collecting ducts of G. gallus.

Histopathological changes in the kidneys of vertebrate hosts infected naturally and experimentally withParatanaisia bragai (Trematoda, Digenea)

Paratanaisia bragai is a trematode parasite that reaches sexual maturity in the kidney collecting ducts of domesticated and wild fowl and whose intermediate hosts are the snails Subulina octona and Leptinaria unilamellata. There are some discrepancies in descriptions of the pathology of this parasite in bird kidneys. Therefore, the purpose of this study was to analyze the kidneys of rock pigeons (Columba livia) naturally infected and of chickens (Gallus gallus) experimentally infected with Paratanaisia bragai, by means of macroscopic observation and by light and scanning electron microscopy. Both bird species showed significantly dilated collecting ducts. In addition, lymphocyte infiltration was observed in the kidneys of C. livia and metaplasia in the epithelial lining of the kidney collecting ducts of G. gallus.

Paratanaisia bragai é um trematódeo que atinge sua maturidade sexual nos ductos coletores de rins de aves domésticas e silvestres, tendo os moluscos Subulina octona e Leptinaria unilamellata como hospedeiros intermediários. A patologia descrita no rim das aves apresenta uma série de divergências. Dessa forma, o presente estudo teve como objetivo analisar rins de Columba livia, naturalmente infectada, e de Gallus gallus infectados experimentalmente por Paratanaisia bragai. Através das análises, verificaram-se alterações macroscópicas, por microscopia de luz e eletrônica de varredura, sendo caracterizada significativa dilatação dos túbulos coletores. Essas alterações foram observadas nas aves infectadas naturalmente e experimentalmente. Por outro lado, foi observada infiltração linfocitária nos rins de C. livia, naturalmente infectada, e ocorrência de metaplasia no revestimento epitelial dos túbulos coletores dos rins de G. gallus, experimentalmente infectados.

Morphological aspects of Angiostrongylus costaricensis by light and scanning electron microscopy.

Angiostrongylus costaricensis is a parasitic nematode that can cause severe gastrointestinal disease, known as abdominal angiostrongiliasis, in humans. This paper presents the characterization of first- and third-stage larvae and male and female adult worms of A. costaricensis by scanning electron and light microscopy. Several novel anatomical structures were identified by scanning electron microscopy, including details of the cuticular striations of the spicules in male worms and a protective flap of the cuticle covering the vulvar aperture in female worms. Other taxonomic features revealed by light microscopy include the gubernaculum and the esophageal-intestinal valve. The use of two microscopy techniques allowed a detailed characterization of the morphology of this nematode. A number of previously identified taxonomic features, such as the striated nature of the spicules and the lateral alae were confirmed; however, the use of scanning electron microscopy resulted in a reassessment of the correct number of papillae distributed around the oral opening and behind the cloacal opening. These observations, in combination with light microscopy-based characterization of the gubernaculum and esophageal valves, have allowed a more detailed description of this nematode taxonomy.

Evaluation of praziquantel effects on Echinostoma paraensei ultrastructure.

Echinostomiasis is a food-borne, intestinal, zoonotic, snail-mediated helminthiasis caused by digenean trematodes of the family Echinostomatidae with seven species of the genus Echinostoma infecting humans or domestic and wildlife animals. Echinostoma paraensei is a peristomic 37-collar-spined echinostome belonging to the "revolutum group". Praziquantel (PZQ) is the drug of choice for treatment and control of human schistosomiasis and food-borne trematodiasis. In the present study we used scanning and transmission electron microscopy to further elucidate the trematocidal effect of PZQ on adult E. paraensei and confirmed that this trematode is a suitable model to study anthelmintic drugs. Hamsters infected with E. paraensei were treated with a single dose of 30 mg kg(-1) of PZQ. The worms were recovered 15, 30, 90 and 180 min after drug administration. There was a significant decrease in worm burden in the small intestine in the hamster-E. paraensei model at the intervals of 30, 90 and 180 min after the treatment. The worms displayed damage of the peristomic collar with internalization of the spines and erosion of the tegument of the circumoral head-collar of spines. Ultrastructural analysis demonstrated an intense vacuolization of the tegument, appearance of autophagic vacuoles and swelling of the basal infolds of the tegumental syncytium. There was no change in the morphology of cells from the excretory system of adult E. paraensei, however, there was an apparent decrease of stores of glycogen particles in parenchymal cells in PZQ-treated worms. Our results demonstrated that PZQ promotes surface and ultrastructural damage of the tegument of adult E. paraensei supporting the idea that this trematode may constitute a good model to investigate drug effects mechanisms in vitro and in vivo.

New insight into the morphology of Eurytrema coelomaticum (Trematoda, Dicrocoeliidae) cercariae by light, scanning, and transmission electron microscopies.

Eurytrema coelomaticum is a digenetic trematode that parasitizes the pancreatic ducts of ruminants. In the present study, the morphology of the cercariae was analyzed using light, scanning, and transmission electron microscopies. The size of the larvae was larger than that reported in the literature. An oral sucker with many papillae and an oral aperture in its center and a ventral sucker with few discrete papillae on the edge were observed. No stylet at the anterior end of the larval body and spines on the tegument of the end of the tail were observed. The cercariae had lateral penetration glands and (central) pre-acetabular glands, from which ducts lead to the anterior region and open into a small anterior pocket. The flame cells were located laterally and communicated with fine branches, converging to two lateral excretory collecting ducts that opened into an excretory bladder, centrally located at the posterior end of the body. The tegument presented an external layer loosely attached to the larval body, below which an amorphous syncytial outer layer with many mitochondrial profiles was observed. This region exhibited many secretions and released secretory granules, indicating intense secretory activity. The circular and longitudinal muscle layers were arranged in sequence below the outer layer. The outer layer was connected by cytoplasmic bridges crossing the muscular layers to the internal region of the tegument, where the cell body with nucleus was located. In the cercarial body, it was possible to observe a typical flame cell with the barrel region, where cilia and the internal and external ribbed regions were found. The somatic cells seem to have an active metabolism, with a well-developed endoplasmic reticulum, secretory granules, and evident nuclei. The results are discussed in the light of the biology and taxonomy of this species.

Ultrastructure of the sporocysts of Eurytrema coelomaticum (Giard Et Billet, 1892) Looss, 1907.

The digenetic trematode Eurytrema coelomaticum is a parasite of pancreatic ducts of ruminants. The ultrastructure of the mother and daughter sporocysts of E. coelomaticum was analyzed. The mother sporocyst was attached to the coelome of the intestine wall of intermediate snail host Bradybaena similaris, intimately adhered in some regions. It presents a highly folded tegument with granules and the body wall was composed by an outer syncitial layer, basal lamina, and circular and longitudinal muscle layer. Below was the cell body (cyton) with the nucleus. The daughter sporocysts obtained by dissection exhibited many granules and secretory vesicle in the outer layer indicating an intense secretory activity. The body wall presented the same layers of the mother sporocysts, but the outer syncitial layer invaginated and an amorphous layer was present between the syncitial and circular muscle layers. The protonephridial excretory system was viewed. The anterior and posterior end of the expelled sporocyst exhibit a degenerated structure, but biological activity still occurred in these regions. The swollen middle of the body was filled by a lamellar structure formed by degenerating membranes, but the excretory system was preserved. The endocyst wall was fibrilar and filled by cercariae and amorphous, membranous and secretory material inside it. These results were discussed.

Light and scanning electron microscopy of sporocysts of Eurytrema coelomaticum (Giard et Billet, 1892) Looss, 1907.

Eurytrema coelomaticum is a fluke that infects ruminants in South America, Europe and Asia. The morphology of the mother and daughter sporocysts of E. coelomaticum obtained from Bradybaena similaris, the first intermediate host, is described for the first time by light and scanning electron microscopy. The intermediate host was exposed to E. coelomaticum eggs and after 30 days the mother sporocyst was found in the coelom adhered to the intestine wall. This sporocyst was a rounded or elongated mass (0.1078 mm), with numerous germinal balls in it, and a folded tegument with no specializations. The daughter sporocysts obtained following dissection of infected snails have varied shape, one hollow tapered region with many transversal and longitudinal striations, named anterior end. The expelled daughter sporocyst presented an oval sac-like central region with a small anterior and a posterior longer filament-like prolongation. The measures of the expelled sporocysts are presented and compared to previous descriptions.

Effects of albendazole on Litomosoides chagasfilhoi (Nematoda: Filarioidea) females in vivo.

Gerbils (Meriones unguiculatus) experimentally infected with Litomosoides chagasfilhoi were treated with a single oral dose of 40 or 80 mg of albendazole, respectively. Observation of the microfilaremia after the treatment showed that both single oral doses of albendazole decreased the microfilaremia in L. chagasfilhoi infection. The body wall was composed of a cuticle, a hypodermis, and a muscular layer, and treated nematodes showed no morphological alterations. The ultrastructural alterations produced by treatment with 40 mg of albendazole included a higher number of membrane invaginations in the basal labyrinth of the uterine epithelium and the presence of myelin figures in this region. Inside the uterus, most embryos and microfilariae were disintegrated. The treatment with 80 mg of albendazole did not produce alterations in the uterine wall, and the number of vesicles near the microfilariae sheath was smaller than that observed in the untreated and in the 40-mg treatment groups. However, all the microfilariae observed in the uterus were extensively damaged with cytoplasmic vacuolization and cellular degeneration. No alterations in the intestinal cells were observed after treatment with 40 or 80 mg of albendazole. The present study contributes to the knowledge of albendazole's effects in filariids and demonstrates the potential embryotoxic and microfilaricidal consequences of this drug.

Preliminary observations on fluids incubated with Wuchereria bancrofti using the immunochromatographic test.

To assess the performance of the immunochromatographic test for filariasis, adult Wuchereria bancrofti worms were incubated under different conditions. The tests were strongly positive with incubation fluids from both living and mechanically damaged females. Negative results were observed with incubation fluids from all male worms and from intact dead females.

Preliminary observations on fluids incubated with Wuchereria bancrofti using the immunochromatographic test / Observações preliminares de fluidos incubados com Wuchereria bancrofti usando o teste imunocromatográfico

To assess the performance of the immunochromatographic test for filariasis, adult Wuchereria bancrofti worms were incubated under different conditions. The tests were strongly positive with incubation fluids from both living and mechanically damaged females. Negative results were observed with incubation fluids from all male worms and from intact dead females.

Para a valiar o desempenho do teste imunocromatográfico para filariose, vermes adultos de Wuchereria bancrofti foram incubados em diferentes condições. Os testes foram fortemente positivos com fluidos de incubação de fêmeas vivas e danificadas mecanicamente. Resultados negativos foram obtidos com fluidos de todos os machos e de fêmeas mortas intactas.