RESUMO
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Assuntos
Trematódeos , Infecções por Trematódeos , Animais , Infecções por Trematódeos/veterinária , Infecções por Trematódeos/parasitologia , Microscopia Eletrônica de Varredura , Columbidae/parasitologia , RimRESUMO
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).
Assuntos
Animais , Trematódeos/anatomia & histologia , Pesos e Medidas Corporais/veterinária , Microscopia Eletrônica de VarreduraRESUMO
Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).
Paratanaisia bragai é um trematódeo digenético que atinge a maturidade sexual nos ductos coletores de aves domésticas e silvestres, enquanto os moluscos Subulina octona e Leptinaria unilamellata atuam como seus hospedeiros intermediários no Brasil. O presente estudo analisou a morfologia e morfometria de P. bragai. Amostras adultas do parasito foram coletadas de rins de Columba livia naturalmente infectada, fixadas e preparadas para observação na microscopia de campo claro e microscopia de luz de contraste de interferência diferencial e microscopia eletrônica de varredura. O parasite possui corpo alongado e achatado, com uma ventosa oral subterminal localizada na extremidade anterior do corpo, conforme observado por todas as técnicas utilizadas. A coloração do parasito com hematoxilina-eosina permitiu observar a faringe, localizada posteriormente à ventosa oral, as glândulas vitelogênicas, que são extracecais e estendem-se anteriormente à região pré-ovariana e posteriormente à região mediana do corpo, e os cecos intestinais paralelos às glândulas vitelinas. A presença e funcionalidade do acetábulo são pontos controversos na literatura, mas foi observado em todos os espécimes analisados por microscopia eletrônica de varredura, com diâmetro maior de 38.36 ± 6.96 (28.77 - 45.39) e diâmetro menor de 31.59 ± 7,04 (21.75 - 38.16). Próximo ao acetábulo foram observadas escamas no tegumento do parasito. Escamas com (1 - 5) divisões de lâmina foram identificadas. No poro genital, foi possível visualizar o cirro evertido com formato de roseta. O poro excretor (primeiro registro morfométrico) é dorsal e subterminal, com diâmetro maior de 12.27 ± 9.16 (5.79 - 18.75) e diâmetro menor de 3.95 ± 1.49 (2.89 - 5.00).
Assuntos
Animais , Columbidae/parasitologia , Trematódeos/anatomia & histologia , Microscopia Eletrônica de VarreduraRESUMO
Toxoplasma gondii is the causative agent of toxoplasmosis, which is one of the most common parasitic diseases in the world. This pathogen causes severe damage to immunocompromised hosts, and the most frequently used therapy is the combination of pyrimethamine and sulfadiazine, which has side effects. Thus, there is a need for new therapies that target T. gondii. Herein, we present the anti-Toxoplasma effect of two new copper(II) complexes: [(H2L1) Cu (µ-Cl)2 Cu(H2L1)] Cl2·5H2O (1) and [(H2L2) Cu (µ-Cl)2 Cu(H2L2)] Cl2·6H2O (2). Complexes (1) and (2) irreversibly controlled parasite growth in vitro, with IC50 values of 0.78µM and 3.57µM, respectively, after 48h. These complexes induced part of the tachyzoite population to convert to bradyzoites, which eventually die. The cell death mechanism was unknown, but signs of apoptosis, such as membrane blebs and nuclear fragmentation, and necrosis, such as plasma membrane disruption, intense cytoplasm vesiculation and the release of cellular contents, were seen. In addition, complex (2) interfered with the correct disposition of the inner membrane complex of the parasite, affecting cell division. These results indicate that these copper complexes have potential effects against T. gondii and may be used as drugs in the future or serve as prototypes for the development of new drugs to treat toxoplasmosis.
Assuntos
Apoptose/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Cobre/farmacologia , Compostos Organometálicos/farmacologia , Toxoplasma/efeitos dos fármacos , Cobre/química , Compostos Organometálicos/químicaRESUMO
Toxoplasma gondii, the causative agent of toxoplasmosis, is an obligate intracellular protozoan that can infect a wide range of vertebrate cells. Here, we describe the cytotoxic effects of the dinuclear iron compound [Fe(HPCINOL)(SO4)]2-µ-oxo, in which HPCINOL is the ligand 1-(bis-pyridin-2-ylmethyl-amino)-3-chloropropan-2-ol, on T. gondii infecting LLC-MK2 host cells. This compound was not toxic to LLC-MK2 cells at concentrations of up to 200 µM but was very active against the parasite, with a 50% inhibitory concentration (IC50) of 3.6 µM after 48 h of treatment. Cyst formation was observed after treatment, as indicated by the appearance of a cyst wall, Dolichos biflorus lectin staining, and scanning and transmission electron microscopy characteristics. Ultrastructural changes were also seen in T. gondii, including membrane blebs and clefts in the cytoplasm, with inclusions similar to amylopectin granules, which are typically found in bradyzoites. An analysis of the cell death pathways in the parasite revealed that the compound caused a combination of apoptosis and autophagy. Fluorescence assays demonstrated that the redox environment in the LLC-MK2 cells becomes oxidant in the presence of the iron compound. Furthermore, a reduction in superoxide dismutase and catalase activities in the treated parasites and the presence of reactive oxygen species within the parasitophorous vacuoles were observed, indicating an impaired protozoan response against these radicals. These findings suggest that this compound disturbs the redox equilibrium of T. gondii, inducing cystogenesis and parasite death.
Assuntos
Antioxidantes/metabolismo , Coccidiostáticos/farmacologia , Inibidores Enzimáticos/farmacologia , Compostos Férricos/farmacologia , Toxoplasma/efeitos dos fármacos , Animais , Catalase/antagonistas & inibidores , Catalase/metabolismo , Linhagem Celular , Coccidiostáticos/química , Inibidores Enzimáticos/química , Compostos Férricos/química , Macaca mulatta , Microscopia Eletrônica de Transmissão , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Toxoplasma/crescimento & desenvolvimento , Toxoplasma/metabolismoRESUMO
In the last decade ostrich farms spread throughout the world as an alternative source of investment. Although previous studies have reported hematology and biochemical values for ostriches from several regions of the world, little information is available regarding leukocyte morphology. This study reports the morphology and ultrastructure of ostrich leukocytes and hematology and biochemical values from birds raised in Brazil. Heterophils presented a lobulated nucleus, and fusiform, and acidophilic and peroxidase-negative granules. Ultrastructurally, 2 kinds of cytoplasmic granules were observed: one was large and fusiform and the other smaller with heterogeneous morphology and electrondensity; granules were peroxidase-negative. Eosinophils had a kidney-shaped eccentrically placed nucleus that was rarely lobulated and eosinophilic, round, and peroxidase-positive granules. At the ultrastructure level, 2 main kinds of granules with the same size and form but different electron density were seen; granules were peroxidase-positive. Lymphocytes and thrombocytes had the same characteristics of other avian species; monocytes presented morphological heterogeneity. Hematological and serum biochemical profiles had no sex influence and were established for ostriches raised in southeastern Brazil. These parameters will help the diagnosis of specific ostrich pathologies and serve as basic knowledge for studies in immunology and comparative avian pathology.
Assuntos
Leucócitos/ultraestrutura , Struthioniformes/sangue , AnimaisRESUMO
The use of Duddingtonia flagrans, a nematode-trapping fungus, has been investigated as a biological control method against free living larvae of gastrointestinal nematodes of livestock animals. This fungus captures and infects the nematode by cuticle penetration, immobilization and digestion of the internal contents. It has been suggested that this sequence of events occurs by a combination of physical and enzymatical activities. This report characterizes the acid phosphatase activity during the interaction of D. flagrans with the free-living nematode Panagrellus sp. The optimum pH for the hydrolysis of the acid phosphatase substrate p-nitrophenyl phosphate was 2.2, 2.8 and 5.4 from D. flagrans alone and 2.2 and 5.4 for Panagrellus sp alone, fungus-nematode interaction in liquid medium and fungus-nematode interaction in solid medium. Different acid phosphatase activity bands were detected by SDS-PAGE. Maximum acid phosphatase activity of the fungus or nematode alone and of the fungus-nematode interaction occurred within 70min of incubation in the presence of the substrate 4-methylumbelliferyl phosphate. The activity of this enzyme was significantly higher for the fungus-nematode interaction when compared to the organisms alone, indicating a synergistic response. Furthermore, structures appeared in the hyphae after 30min, nematodes were observed adhered after 40min and many were captured by the typical fungus traps after 70min of interaction. The participation of acid phosphatase activity and its importance during the interaction of the fungus with the nematode were discussed.
Assuntos
Fosfatase Ácida/metabolismo , Ascomicetos/enzimologia , Rabditídios/microbiologia , Fosfatase Ácida/fisiologia , Animais , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Cinética , Rabditídios/enzimologia , Fatores de TempoRESUMO
This study investigated the effects of Photorhabdus temperata infection on the activities of digestive enzymes of the sugarcane stalk borer Diatraea saccharalis. Non-infected D. saccharalis larvae present a major alpha-amylase, several proteinases, three sucrose hydrolases and two alpha-glucosidases in their midgut. Analysis of these hydrolases by electrophoresis and "in gel" assays showed that the activities of all enzymes decreased following infection, with an initial decline observed 12 h after infection. The activities of alpha-glucosidases decreased by 50% twelve hours after infection, whereas, at this time, the alpha-galactosidase activities decreased by 70%. Interestingly, the animals died 48 h after infection, but approximately 5% of all the enzymes tested remained active in the midgut following host death. At this time, most of the cultivable native intestinal bacteria had died.
Assuntos
Lepidópteros/enzimologia , Lepidópteros/microbiologia , Photorhabdus/patogenicidade , Animais , Sistema Digestório/enzimologia , Sistema Digestório/microbiologia , Glicosídeo Hidrolases/metabolismo , Interações Hospedeiro-Patógeno , Larva/enzimologia , Larva/microbiologia , Peptídeo Hidrolases/metabolismo , Saccharum/parasitologia , alfa-Amilases/metabolismo , alfa-Glucosidases/metabolismoRESUMO
Ric c 1 and Ric c 3 are the major castor bean allergens. In order to identify continuous IgE-epitopes in Ric c 1 and Ric c 3, pools of sera from rats immunized with a pool of 2S albumin from these seeds, Ric c 1 and Ric c 3 overlapping synthetic peptides, were used to screen for IgE-binding epitopes. The allergenic properties were monitored by mast cell degranulation assays, histamine quantification and human-IgE binding. Large and small chains isolated from these proteins present allergenic properties. Four continuous epitopes were identified in Ric c 3 and two in Ric c 1. This knowledge may allow the induction of protective antibody responses to antagonize the IgE recognition.
Assuntos
Antígenos de Plantas/imunologia , Epitopos/imunologia , Imunoglobulina E/imunologia , Mastócitos/imunologia , Proteínas de Plantas/imunologia , Ricinus/imunologia , Albuminas 2S de Plantas , Alérgenos , Sequência de Aminoácidos , Animais , Antígenos de Plantas/isolamento & purificação , Antígenos de Plantas/metabolismo , Degranulação Celular , Mapeamento de Epitopos , Feminino , Humanos , Mastócitos/citologia , Dados de Sequência Molecular , Peptídeos/síntese química , Peptídeos/química , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Ratos , Ratos Endogâmicos , Alinhamento de SequênciaRESUMO
Snake venoms are toxic to a variety of cell types. However, the intracellular damages and the cell death fate induced by venom are unclear. In the present work, the action of the South American rattlesnake Crotalus durissus terrificus venom on CHO-K1 cell line was analyzed. The cells CHO-K1 were incubated with C. d. terrificus venom (10, 50 and 100g/ml) for 1 and 24 hours, and structural alterations of actin filaments, endoplasmic reticulum and nucleus were assessed using specific fluorescent probes and agarose gel electrophoresis for DNA fragmentation. Significant structural changes were observed in all analyzed structures. DNA fragmentation was detected suggesting that, at the concentrations used, the venom induced apoptosis.