Embryonic development of Girardia tigrina (Girard, 1850) (Platyhelminthes, Tricladida, Paludicola) / Desenvolvimento embrionário de Girardia tigrina (Girard, 1850) (Platyhelminthes, Tricladida, Paludicola)

The embryonic development of freshwater triclads is mainly known from studies of species of Dendrocoelum, Planaria, Polycelis, and, more recently, Schmidtea. The present study characterizes the development of Girardia tigrina (Girard, 1850) by means of optical microcopy using glycol methacrylate semi-thin sections. 94 cocoons were collected in the period from laying to hatching, with intervals of up to twenty-four hours. The sequence of morphological changes occurring in the embryo permitted the identification of nine embryonic stages. At the time of cocoon laying, numerous embryos were dispersed among many yolk cells, with a rigid capsule covering the entire cocoon. In the first stage (approx. up to 6 hours after cocoon laying), yolk cells and embryonic cells showed random distribution. Stage II (between 12 and 24 hours after cocoon laying) is characterized by aggregates of blastomeres, which later aggregate forming an enteroblastula. Approximately 2 days after cocoon laying (stage III), formation of the embryonic epidermis and embryonic digestive system took place, the latter degenerating during the subsequent stage. Stage V (until the fourth day) is characterized by the formation of the definitive epidermis. Between 4 and 6 days after laying, organogenesis of the definitive inner organs starts (stage VI). Approximately 14 days after laying (stage IX), formation of the nervous system is completed. At this stage, the embryo shows similar characteristics to those of newly hatched juveniles. The hatching of Girardia tigrina occurs in the period between twelve to twenty-two days after cocoon laying.

O desenvolvimento embrionário dos tricladidos é conhecido, fundamentalmente, por estudos realizados em espécies de Dendrocoelum, Planaria, Polycelis e, mais recentemente, Schmidtea. O presente estudo descreve o desenvolvimento embrionário de Girardia tigrina (Girard, 1850), a partir de análises realizadas em cortes histológicos seriados e semifinos de glicol-metacrilato, ao microscópio óptico. Noventa e quatro casulos foram coletados no período entre a postura e a eclosão, em intervalos de até vinte e quatro horas. A seqüência das modificações morfológicas no embrião permitiu a identificação de nove estágios embrionários. Na postura dos casulos, envoltos por uma cápsula rígida, observam-se numerosos embriões dispersos entre grande quantidade de células vitelinas. No estágio I (aproximadamente até 6 horas após a postura), as células vitelinas e as embrionárias mostram uma distribuição aleatória. O estágio II (entre 12 e 24 horas após a postura) caracteriza-se pela formação de agrupamentos de blastômeros, os quais posteriormente formam uma enteroblástula. Aproximadamente dois dias após a postura (estágio III), ocorre a formação da epiderme e do sistema digestivo embrionário, sendo que este último degenera no estágio seguinte. O estágio V (até o quarto dia após a postura) caracteriza-se pela formação da epiderme definitiva. Entre o quarto e o sexto dia posteriores à postura, começa a organogênese dos órgãos internos definitivos (estágio VI). Aproximadamente catorze dias após a postura (estágio IX), completa-se a formação do sistema nervoso. Neste estágio, o embrião já apresenta características similares aos espécimes juvenis. A eclosão de Girardia tigrina ocorre entre doze e vinte e dois dias após a postura dos casulos.

Embryonic development of Girardia tigrina (Girard, 1850) (Platyhelminthes, Tricladida, Paludicola).

The embryonic development of freshwater triclads is mainly known from studies of species of Dendrocoelum, Planaria, Polycelis, and, more recently, Schmidtea. The present study characterizes the development of Girardia tigrina (Girard, 1850) by means of optical microcopy using glycol methacrylate semi-thin sections. 94 cocoons were collected in the period from laying to hatching, with intervals of up to twenty-four hours. The sequence of morphological changes occurring in the embryo permitted the identification of nine embryonic stages. At the time of cocoon laying, numerous embryos were dispersed among many yolk cells, with a rigid capsule covering the entire cocoon. In the first stage (approx. up to 6 hours after cocoon laying), yolk cells and embryonic cells showed random distribution. Stage II (between 12 and 24 hours after cocoon laying) is characterized by aggregates of blastomeres, which later aggregate forming an enteroblastula. Approximately 2 days after cocoon laying (stage III), formation of the embryonic epidermis and embryonic digestive system took place, the latter degenerating during the subsequent stage. Stage V (until the fourth day) is characterized by the formation of the definitive epidermis. Between 4 and 6 days after laying, organogenesis of the definitive inner organs starts (stage VI). Approximately 14 days after laying (stage IX), formation of the nervous system is completed. At this stage, the embryo shows similar characteristics to those of newly hatched juveniles. The hatching of Girardia tigrina occurs in the period between twelve to twenty-two days after cocoon laying.

Biosynthetic capacity of rainbow trout (Oncorhynchus mykiss) interrenal tissue after cadmium exposure.

The disruption of endocrine system function in wildlife species, including teleosts, by contaminants such as metals is presently of major environmental concern. Recently, it has been shown that cadmium (Cd) exposure results in significant reductions in corticosteroid secretion by fish interrenal steroidogenic cells, likely through an inhibition of intracellular cortisol synthesis. In the present study, the effects of CdCl(2) on unstimulated and stimulated interrenal steroidogenesis in rainbow trout were examined with the intention of furthering an understanding of the site(s) of Cd toxic action. CdCl(2) alone reduced cortisol secretion in minced interrenal tissues to 59% and 55% of control values when exposed to 10 and 100 microM, respectively. Incubation of interrenal tissues with 0.01 IU/mL adrenocorticotropic hormone (ACTH), which activates rate-limiting steps in steroid synthesis, resulted in significant stimulation of steroidogenesis in controls. However, ACTH-stimulated steroidogenesis was reduced when tissues were previously incubated with Cd. Maximal rates of unstimulated cortisol secretion were achieved by augmentation using 5 microM 25-hydroxycholesterol (25-OHC) or 0.8 microL/mL synthetic cholesterol [SyntheChol(SC)]. Steroidogenesis augmentation by 25-OHC was significantly reduced in tissues incubated with Cd. Interestingly, cortisol secretion was significantly higher in SC-augmented tissue exposed to 1 and 10 microM Cd when compared to augmented control tissues. The results of this study show that Cd affects both stimulated and unstimulated steroidogenesis in rainbow trout, and that one major site(s) of action of Cd in the cortisol synthesis pathway is likely prior to cytochrome P450 side chain cleavage.

Histological processing techniques for the study of Dugesiidae development (Platyhelminthes, Tricladida, Paludicola).

The objective of the present study was to adapt techniques for the histological processing of Dugesiidae cocoons for the study of embryo development. The cocoons were fixed with formalin, SUSA, Bouin or paraformaldehyde/glutaraldehyde and subsequently embedded in Paraplast or glycol methacrylate (Historesin). Paraplast embedding yielded reasonable results only after the cocoon was perforated or fixed for a prolonged period of time using softening techniques with acid solutions. When the SUSA or Bouin fixative and Historesin embedding techniques were used the results were good for light microscopical analysis. Fixation with paraformaldehyde/glutaraldehyde and glycol methacrylate embedding resulted in better tissue preservation, and did not require prolonged fixation or softening techniques. Thus, we suggest this technique for light microscopical analysis of embryo development in Dugesiidae.

Histological processing techniques for the study of Dugesiidae development (Platyhelminthes, Tricladida, Paludicola)

The objective of the present study was to adapt techniques for the histological processing of Dugesiidae cocoons for the study of embryo development. The cocoons were fixed with formalin, SUSA, Bouin or paraformaldehyde/glutaraldehyde and subsequently embedded in Paraplast or glycol methacrylate (Historesin). Paraplast embedding yielded reasonable results only after the cocoon was perforated or fixed for a prolonged period of time using softening techniques with acid solutions. When the SUSA or Bouin fixative and Historesin embedding techniques were used the results were good for light microscopical analysis. Fixation with paraformaldehyde/glutaraldehyde and glycol methacrylate embedding resulted in better tissue preservation, and did not require prolonged fixation or softening techniques. Thus, we suggest this technique for light microscopical analysis of embryo development in Dugesiidae

Ultrastructural alterations in glomerulopathy associated with hydatidosis in sheep.

In the present study we investigated the ultrastructural alterations occurring in the renal glomeruli of sheep with hydatidosis. Renal samples from 39 sheep, 34 with hydatidosis and 5 without parasitosis, were examined by transmission electron microscopy. Additionally, biochemical analysis was performed by determining serum concentrations of creatinine, urea, total protein and albumin. The ultrastructural alterations identified were the presence of dense mesangial, subendothelial and intra-membranous deposits, mesangial cell proliferation with areas showing segmental sclerosis and interposition of mesangial cells with the formation of a neomembrane. Biochemical analysis revealed a significant increase in total serum protein in the experimental group compared with the control. Our results demonstrated that glomerulonephritis associated with hydatidosis in sheep can be classified into four categories: minimal lesions, mesangial glomerulonephritis, segmental and focal glomerulonephritis and membranoproliferative glomerulonephritis, being membranoproliferative and mesangial glomerulonephritis the most predominant categories.

Clastogenic effect of cisplatin on Wistar rat bone marrow cells.

The clastogenic effect of the drug cis-diamminedichloroplatinum II (cisplatin, CDDP) was investigated in Wistar rat bone marrow cells. Male rats, 3 per treatment time, aged 4 months and weighing 250-350 g were injected intraperitoneally with 6.0 mg/kg CDDP solution, and the control group received isotonic saline. The animals were sacrificed 6, 12, 18, 24 and 48 h after the injection. The chromosome preparation was obtained from bone marrow cells. Chromatid and chromosome aberrations were investigated in 300 metaphases per animal. A significant increase in number of chromosome aberrations was observed from 6 to 24 h, the majority being of the break and gap type. After 48 h a progressive reduction was observed, without differences from the negative control. These data confirm the mutagenic effect of CDDP in rats demonstrated for mice bone marrow by micronuclei assay, for murine ovary cells and mice spermatocytes.

Intraperitoneal reaction in Wistar rats after inoculation with several components of a human hydatid cyst.

Fifteen male Wistar rats were inoculated intra peritoneally with material obtained from a human hepatic hydatid cyst with 20% viability and previously inactivated with 20% NaCl. When the animals were sacrificed six months later, hydatid granulomas surrounding parasitic remains were detected in the peritoneum.

Histological preparation of the Echinococcus sp. tapeworms: staining affinity of the adult form.

Thirty specimens of Echinococcus sp. obtained by anti-helminthic treatment using arecoline hydrobromide from a dog from an Echinococcosis endemic region were fixed in 10% formalin. The material was processed histologically by a rapid manual method and stained using eleven different techniques. Green Trichrome staining as modified in the present study permitted the observation of large structures of Echinococcus sp. tapeworms.