Morphological aspects of the ovaries of turtle Kinosternon scorpioides raised in captivity

The swear turtle "jurará" (Kinosternon scorpioides) is a mud turtle of the Amazon region exposed to disordering capture in the rural areas of Maranhão, Brazil. Despite its popularity in these areas, little meaningful information regarding the reproductive morphology is currently available, fact that impedes the adoption of policies for preservation of the species. To obtain more information, we studied the ovarian morphology adult jurará females kept in captivity by morphological and morphometric analysis in the dry and rainy season. The results revealed that all females were sexually mature and were in a vitellogenic period. The ovaries are two irregular structures composed by follicles in different stages of development (primary, secondary and tertiary) scattered in a stroma of loose connective highly vascularized tissue. The ovary weight was 6.25±4.23g and 2.27±1.42g, for the right and left one respectively. The gonadosomatic indexes were 2.06% for the dry season and 1.79% for the rainy season. The average of the follicles was 29.83 units per ovary. Microscopically, the mature ovaries revealed a basal layer composed by four cellular layers: the inner and outer theca, stratum granulosum with perivitelline membrane and zona radiata with vitelline membrane. No significant differences were observed in the ovaries either in the dry or wet period.

Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu)

The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu). Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05). Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89%) of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.

Morphological and morphometric evaluation of prepubertal gilt ovaries, uterine tubes and uterus at different oestrus cycle stages

Studies are performed in developing techniques/procedures that provide greater reproductive performance in farm animals, including pigs. In this sense, the study of gilts reproductive organs at different oestrus cycle stages for assessing the presence of abnormalities and/or other parameters that may affect the future animal fertility is important. In order to evaluate the morphological, morphometric and histomorphometric features of ovaries, uterus and uterine tubes (UTs) characteristics of prepubertal gilts at different oestrus cycle stages, reproductive tracts from 48 animals immediately after slaughter were obtained. After, the structures were dissected and removed, and the ovaries were used for classification of oestrus cycle stage of each gilt in follicular phase (FP) and luteal phase (FL). Then, morphometric evaluations of ovaries, UTs, uterine horns and uterine body were performed. Besides that, medial segments of UTs and uterus were fixed in Bouin solution, processed and included in paraffin, when histological sections of 5.0 micrometers (µm) were obtained and stained with Hematoxylin and Eosin. Histomorphometric analyzes using image capture system and specific software were performed. Afterwards, data were submitted to Student's t test for assessment the statistical differences (P<0.05) between the two different oestrus cycle stages (FP × LP) and between the placement of reproductive structures (right × left antimer). Among the gilts evaluated, 35 were in the FP and 13 in LP. There was no difference (P>0.05) between morphometric parameters of ovaries, UTs and uterus of gilts in FP and LP. Likewise, in respect to the placement of reproductive structures, both in the oestrus cycle stages, as in the general average, there was no difference (P>0.05). Regarding the histomorphometric variables, gilts classified in FP presented a higher (P<0.05) height of glandular and UT epithelium compared to animals in LP. On the other hand, the diameter of endometrial glands was higher (P<0.05) in gilts at LP compared to FP. Furthermore, gilts in LP presented a higher (P<0.05) proportion of endometrium occupied by glands, whereas animals in FP had a higher (P<0.05) proportion of connective tissue and blood vessels. In conclusion, in prepubertal gilts, the histomorphometric parameters as endometrial glands diameter, the height of glandular epithelium and of UT epithelium and the proportion of endometrium occupied by connective tissue, besides the glands and blood vessels varies through the oestrus cycle, possibly under the influence of ovarian steroids.

Morphological and immunohistochemical characterization of angiogenic and apoptotic factors and the expression of thyroid receptors in the ovary of tilapia Oreochromis niloticus in captivity

Morphological and immunohistochemical characterization of angiogenic and apoptotic factors and the expression of thyroid receptors in the ovary of tilapia Oreochromis niloticus in captivity were studied. The morphological evaluation of the ovaries was performed by histological paraffin embedded and stained with HE. The immunohistochemical expressions of CDC47, VEGF, Flk-1, angiopoietin, Tie-2 and thyroid receptor (TRα) were performed by the technique of streptavidein-biotin-peroxidase. Apoptosis was assessed using the TUNEL kit. The relative expression of thyroid hormone receptors (TRα and TRβ) was assessed by RT-PCR real time. The nuclear expression of CDC47 increased with the stage of maturation of the oocyte and was observed in the follicle cells. Apoptotic bodies were observed in the follicular cells of atretic follicles and postovulatory follicles from the ovaries of 150g and 350g fish. Expression of VEGF and its receptor Flk-1 was also observed in the follicular cells, and the expression of both increased with the maturity of the oocyte, with a higher intensity observed in the full-grown follicle. The expression of angiopoietin and of its receptor (Tie 2) was discrete and moderate respectively. TRα expression was independent of follicular development. However, the 350 g tilapia exhibited higher expression of TRβ compared with the 50 g tilapia. We conclude that the proliferative activity and the expression of VEGF and its receptor increase with follicular maturation and that the TRs expression increases with ovarian maturity in tilapia (Oreochromis niloticus).

Survival analysis of female dogs with mammary tumors after mastectomy: epidemiological, clinical and morphological aspects

Abstract: Mammary gland tumors are the most common type of tumors in bitches but research on survival time after diagnosis is scarce. The purpose of this study was to investigate the relationship between survival time after mastectomy and a number of clinical and morphological variables. Data was collected retrospectively on bitches with mammary tumors seen at the Small Animal Surgery Clinic Service at the University of Brasília. All subjects had undergone mastectomy. Survival analysis was conducted using Cox's proportional hazard method. Of the 139 subjects analyzed, 68 died and 71 survived until the end of the study (64 months). Mean age was 11.76 years (SD=2.71), 53.84% were small dogs. 76.92% of the tumors were malignant, and 65.73% had both thoracic and inguinal glands affected. Survival time in months was associated with age (hazard rate ratios [HRR] =1.23, p-value =1.4x10-4), animal size (HRR between giant and small animals =2.61, p-value =0.02), nodule size (HRR =1.09, p-value =0.03), histological type (HRR between solid carcinoma and carcinoma in a mixed tumor =2.40, p-value =0.02), time between diagnosis and surgery (TDS, with HRR =1.21, p-value =2.7x10-15), and the interaction TDS*follow-up time (HRR =0.98, p-value =1.6x10-11). The present study is one of the few on the subject matter. Several important covariates were evaluated and age, animal size, nodule size, histological type, TDS and TDS*follow up time were identified as significantly associated to survival time.

Morphological and physiological alterations induced by lactofen in soybean leaves are reduced with nitric oxide

Lactofen is a diphenylether herbicide recommended to control broad-leaved weeds in soybean (Glycine max) fields and its mechanism of action is the inhibition of protoporphyrinogen-IX oxidase (Protox), which acts in the chlorophyll biosynthesis. This inhibition results in an accumulation of protoporphyrin-IX, which leads to the production of reactive oxygen species (ROS) that cause oxidative stress. Consequently, spots, wrinkling and leaf burn may occur, resulting in a transitory crop growth interruption. However, nitric oxide (NO) acts as an antioxidant in direct ROS scavenging. Thus, the aim of this work was to verify, through phytometric and biochemical evaluations, the protective effect of NO in soybean plants treated with the herbicide lactofen. Soybean plants were pre-treated with different levels of sodium nitroprusside (SNP), a NO-donor substance, and then sprayed with 168 g a.i. ha-1 lactofen. Pre-treatment with SNP was beneficial because NO decreased the injury symptoms caused by lactofen in young leaflets and kept low the soluble sugar levels. Nevertheless, NO caused slower plant growth, which indicates that further studies are needed in order to elucidate the action mechanisms of NO in signaling the stress caused by lactofen in soybean crop.

Morphological cladistic analysis of Pseudobombax Dugand (Malvaceae, Bombacoideae) and allied genera

(Morphological cladistic analysis of Pseudobombax Dugand (Malvaceae, Bombacoideae) and allied genera). Pseudobombax Dugand belongs to the family Malvaceae subfamily Bombacoideae and aggregates 29 species restricted to the Neotropics. A morphological cladistic analysis of Pseudobombax and allied genera was carried out to test the monophyly of the genus and to provide hypotheses on its phylogeny. Parsimony analyses were based on 40 morphological characters and 28 species, 14 belonging to Pseudobombax and 14 to other species of Bombacoideae, Matisieae (Malvoideae) and Ochromeae. Nine most parsimonious trees (144 steps, ci 0.40, ri 0.67) were produced when 10 multistate characters were taken as ordered while only two most parsimonious trees (139 steps, ci 0.41, ri 0.67) were obtained when all characters were considered as unordered. Pseudobombax monophyly had moderate bootstrap support, appearing as sister to a clade composed of the genera Bombacopsis Pittier and Pachira Aubl., or to the genus Bombax L. according to the analysis. The petiole widened at the apex and the leaflets not jointed to the petiole are probably synapomorphies of Pseudobombax. Three main clades were found in the genus: one characterised by petiolulated leaflets and 5-angular fruits, the other by pubescent leaves and calyx, and the other by reduction of the number of leaflets. The latter includes species endemic to the Brazilian semi-arid region also characterised by the absence of phalanges in the androecium. Interspecific affinities in Pseudobombax as well as the morphological evolution in Bombacoideae are discussed.

Physiological, morphological and biochemical characteristics of the sexual propagation of Piper aduncum (Piperaceae)

In this work we attempted to characterize the diaspores and the germination process of Piper aduncum L., as well as to verify the influence of the interaction between presence and absence of light (photoperiod of 12 hours and dark) and temperature (25 °C, 30 °C and 20-30 °C) and also of gibberellin (0, 50, 100, 200 and 400 mg L-1) on the root protrusion and normal seedlings formation. The diaspores are very small with a thousand seed weight of 0.3645 g, 13% moisture and protein reserve. Diaspores are strict positively photoblastic in the tested temperature range and the optimum temperature for root protrusion was 30 °C, while for normal seedlings was 25 °C. The previous permanence in the dark led to an increase in the speed of root protrusion and percentage and speed of seedling formation. The application of gibberellic acid negatively interfered with the protrusion and growth of the radicle while favoring the elongation of hypocotyls.

Morphological, anatomical and biochemical studies on the foliar galls of Alstonia scholaris (Apocynaceae)

Morphological, anatomical and biochemical alterations in foliar galls of Alstonia scholaris R. Br. induced by the insect Pauropsylla tuberculata (Psyllidae) are described and quantified. Galls occur isolated or agglomerated on the abaxial surface of the leaf. The insect along with the egg deposits some physiologic fluid which act as a stimulant for the induction of the gall. This stimulus brings about hypertrophy followed by hyperplasia of cells next to the location of the deposited eggs. The psyllid presents three nymphal instars, from eclosion of the egg to the adult. Hyperplasia in the palisade cells is very distinctly noticed. Hypertrophy followed by hyperplasia takes place and brings about elevation of hypodermal and palisade parenchyma which undergoes repeated anticlinal divisions. Neoformation of phloematic bundles were distinctly noticed close to the site of infection. With an increase in the growth of the gall, chlorophyll content in the gall tissue decreases. A steady increase of sugar content is noticed. The immature galled tissue showed almost two fold increases in the protein content. The mature galled tissue showed a very high increase in the proline content compared to the immature galled tissue indicating a stressed condition of the galled tissue.

Germination and morphological characterization of the fruits, seeds, and seedlings of Pilosocereus gounellei

The present work examined the germination and morphology of the fruits and seeds of the cactus Pilosocereus gounellei (xique-xique) and describeb the different stages of seedling growth. Germination tests examined the effects of combinations of two illumination regimes (a 12 hour photoperiod and total darkness) and three temperature regimes (25, 30, and 20-30 ºC) on the seeds of this species. Germination counts were made daily until the 16th day after sowing and the germination percentage and velocity index, and average germination time were evaluated. Descriptions of seed morphology considered both their external and internal aspects. The processes of germination and seedling growth were evaluated during 150 days and eight developmental stages were identified based on morphological changes. The fruits of the xique-xique cactus are 40.67 ± 4.40 mm long and 48.09 ± 3.23 mm in diameter, weigh 53.85 ± 10.03 g, and contain 3713 ± 689.50 seeds. Alternating temperatures of 20-30 ºC together with a photoperiod of 12 hours of light resulted in the highest seed germination rate. The seeds are positively photoblastic and germination is epigeal and fanerocotyledonary. Seedling growth is slow, but at 150 days after sowing the plants had developed epicotyls, large numbers of thorns, and the whole plant averaged 59.56 mm in height.

Ecological distribution of stream macroalgae in different spatial scales using taxonomic and morphological groups

We examined the ecological distribution of macroalgal communities in streams using species groups (taxonomic units = algal phyla, and morphological = morphological types) with similar structures and functions instead of the species themselves. The study was conducted from June to July/2007 in two drainage basins located in mid-southern region of Paraná State , Brazil. Evaluations of macroalgal communities took into consideration the following spatial scales: the drainage basin (the Pedras river and Marrecas river basins), shading regime (open and shaded stream segments), mesohabitats (riffles and pools), and microhabitats (sampling units of 0.05m2). A total of 29 taxa (23 subgeneric, one generic, and five vegetative groups) were identified. On these, 12 taxa belong to Chlorophyta, 11 to Cyanobacteria, four to Heterokontophyta, and two to Rhodophyta. The proportions of morphological types were: 24% free filaments, 17.25% mats, tufts, gelatinous colonies, and gelatinous filaments, 7% crusts. In terms of spatial scales, we observed a predominance of Chlorophyta in open stream segments and Cyanobacteria in shaded stream segments, reflecting the loss of competitive advantage of green algae in sites with low energy availability. In the mesohabitats, the morphological types recorded in pools were predominantly poorly adapted to fast currents (free filaments), while those found in riffles (mats, tufts and gelatinous filaments) were highly resistant to fast water flows. As such, the use of species groupings based on algal taxonomy associated with morphological characteristics proved to be useful to understanding the distributions of these organisms in lotic environments.

Control of the phosphorylation of the astrocyte marker glial fibrillary acidic protein (GFAP) in the immature rat hippocampus by glutamate and calcium ions: possible key factor in astrocytic plasticity

The present review describes recent research on the regulation by glutamate and Ca2+ of the phosphorylation state of the intermediate filament protein of the astrocytic cytoskeleton, glial fibrillary acidic protein (GFAP), in immature hippocampal slices. The results of this research are discussed against a background of modern knowledge of the functional importance of astrocytes in the brain and of the structure and dynamic properties of intermediate filament proteins. Astrocytes are now recognized as partners with neurons in many aspects of brain function with important roles in neural plasticity. Site-specific phosphorylation of intermediate filament proteins, including GFAP, has been shown to regulate the dynamic equilibrium between the polymerized and depolymerized state of the filaments and to play a fundamental role in mitosis. Glutamate was found to increase the phosphorylation state of GFAP in hippocampal slices from rats in the post-natal age range of 12-16 days in a reaction that was dependent on external Ca2+. The lack of external Ca2+ in the absence of glutamate also increased GFAP phosphorylation to the same extent. These effects of glutamate and Ca2+ were absent in adult hippocampal slices, where the phosphorylation of GFAP was completely Ca2+-dependent. Studies using specific agonists of glutamate receptors showed that the glutamate response was mediated by a G protein-linked group II metabotropic glutamate receptor (mGluR). Since group II mGluRs do not act by liberating Ca2+ from internal stores, it is proposed that activation of the receptor by glutamate inhibits Ca2+ entry into the astrocytes and consequently down-regulates a Ca2+-dependent dephosphorylation cascade regulating the phosphorylation state of GFAP. The functional significance of these results may be related to the narrow developmental window when the glutamate response is present. In the rat brain this window corresponds to the period of massive synaptogenesis during which astrocytes are known to proliferate. Possibly, glutamate liberated from developing synapses during this period may signal an increase in the phosphorylation

Correlation of discocyte frequency and ATP concentration in preserved blood: A morphological indicator of red blood cell viability

Red blood cells (RBC) are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP) concentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echinocytes and stomatocytes and, at final stages, by spherocytes, the last step before splenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4oC. We now report a significant correlation (r2 = 0.98) between the percentage of discocytes (89 to 7%) and ATP (100 to 10%) concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator of ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilities.

Role of mast cell- and non-mast cell-derived inflammatory mediators in immunologic induction of synaptic plasticity

We have previously discovered a long-lasting enhancement of synaptic transmission in mammal autonomic ganglia caused by immunological activation of ganglionic mast cells. Subsequent to mast cell activation, lipid and peptide mediators are released which may modulate synaptic function. In this study we determined whether some mast cell-derived mediators, prostaglandin D2 (PGD2; 1.0 µM), platelet aggregating factor (PAF; 0.3 µM) and U44619 (a thromboxane analogue; 1.0 µM), and also endothelin-1 (ET-1; 0.5 µM) induce synaptic potentiation in the guinea pig superior cervical ganglion (SCG), and compared their effects on synaptic transmission with those induced by a sensitizing antigen, ovalbumin (OVA; 10 µg/ml). The experiments were carried out on SCGs isolated from adult male guinea pigs (200-250 g) actively sensitized to OVA, maintained in oxygenated Locke solution at 37oC. Synaptic potentiation was measured through alterations of the integral of the post-ganglionic compound action potential (CAP). All agents tested caused long-term (LTP; duration ³30 min) or short-term (STP; <30 min) potentiation of synaptic efficacy, as measured by the increase in the integral of the post-ganglionic CAP. The magnitude of mediator-induced potentiation was never the same as the antigen-induced long-term potentiation (A-LTP). The agent that best mimicked the antigen was PGD2, which induced a 75% increase in CAP integral for LTP (antigen: 94%) and a 34% increase for STP (antigen: 91%). PAF-, U44619-, and ET-1-induced increases in CAP integral ranged for LTP from 34 to 47%, and for STP from 0 to 26%. These results suggest that the agents investigated may participate in the induction of A-LTP

Morphological changes of the jejunal mucosa in protracted diarrhea and their correlation with disease duration, weight loss and serum albumin levels

The pathogenesis of protracted diarrhea is multifactorial. In developing countries, intestinal infectious processes seem to play an important role in triggering the syndrome. Thirty-four children aged 1 to 14 months, mean 6.5 months, with protracted diarrhea were studied clinically and in terms of small intestinal mucosal morphology. Mild, moderate or severe hypotrophy of the jejunal mucosa was detected in 82% of cases, and mucosal atrophy was observed in 12%. The intensity of the morphological changes of the jejunal mucosa correlated negatively with serum albumin levels. No correlation was detected between mucosal grading and duration of diarrhea or between mucosal grading and weight reported as percentile. After nutritional support was instituted, serial jejunal biopsies were obtained from 12 patients: five patients submitted to parenteral nutrition for 7 to 38 days, mean 17 days, and 7 patients receiving a hypoallergenic oral diet (semi-elemental formula, 3; chicken formula, 3; human milk, 1). In seven cases (58%) a progressive increase in villus height and a decrease in the number of inflammatory cells were noted. Recovery of the morphologic pattern was accompanied by clinical improvement in all patients