Sérgio Luis Pinto da Matta

Histomorphometric evaluation of the neotropical brown brocket deer Mazama gouazoubira testis, with an emphasis on cell population indexes of spermatogenic yield

Information on the reproductive biology of neotropical cervids is scarce. Therefore, the aim of this study was to perform biometric, histologic and stereologic analyses of the brown brocket deer Mazama gouazoubira testis, with an emphasis on the intrinsic yield and the Sertoli cell index. Seven adult males kept in captivity were used. The animals were immobilized; anesthetized and testicle fragments were obtained by biopsy incision. The material was fixed, processed and examined by routine histological methods for light microscopy. The average body weight was 17.2kg, from which 0.40% were allocated in gonads and 0.33% in seminiferous tubules, which represented 85.9% of the testis parenchyma. The mean albuginea width and volume were 345.7μm and 3.5mL (5.3% of the testicular weight), respectively. The mean mediastinum volume of both testicles was 1.0mL (1.5% of the testicular weight) and the testicular parenchyma volume corresponded to 93.1% of total testicular weight (64.9g). The seminiferous tubules diameter was 224.4μm, while the epithelium height was 69.6μm. On average, an adult brown brocket deer showed a total of 1418m of seminiferous tubules in both testicles (21.5m per gram of testis). Each stage I seminiferous tubular cross section contained 1.10 type A spermatogonia, 13.4 primary spermatocytes in pre-leptotene/leptotene, 13.7 spermatocytes in pachytene, 48.8 round spermatids and 3.7 Sertoli cells. The general yield of spermatogenesis was 44.7 cells and the Sertoli cell index was 13.2. The qualitative and quantitative description of testicular histology of brown brocket deer help to understand its spermatogenic process and to establish parameters for the reproductive biology of this wild species. Furthermore, the data from the present research will help further studies using other species of Brazilian cervids, especially endangered ones, making an additional effort to the species preservation.

Heteropterys aphrodisiaca infusion reduces the collateral effects of cyclosporine A on the testis.

Cyclosporine A (CsA) is known to have testicular toxicity, leading to male infertility. Stimulant and aphrodisiac properties have been attributed to the plant, Heteropterys aphrodisiaca. Thus, the present work was undertaken to evaluate the association of the drug and the medicinal herb in Wistar rats, applying testicular morphometry and ultrastructure. Twenty‐four rats were used, divided into four groups: I, control; II, CsA; III, simultaneous use of CsA and H. aphrodisiaca; IV, H. aphrodisiaca. Daily administration by gavage was carried out, during 56 days, of water (sham), CsA in a dose of 15 mg/kg per day and/or H. aphrodisiaca in a dose of 0.5 ml of the infusion prepared with 25 g of roots/100 ml of boiling water. Increased body weight was observed for all groups, but the animals that received only CsA showed the smallest body weight gain. Morphometry showed increased connective tissue volumetric proportion and decreased Leydig cell volumetric proportion in CsA‐treated rats. Using transmission electron microscopy, it was possible to ascertain that CsA caused seminiferous epithelium degeneration, resulting in Sertoli cell vacuolization, abnormal round and elongated spermatids and large accumulation of residual cytoplasm at the epithelium border next to the lumen. Expanded intercellular spaces between germ cells were still observed in H. aphrodisiaca‐treated rat testes. The administration of H. aphrodisiaca infusion to CsA‐treated rats diminished nearly all the CsA‐induced damage to the testis ultrastructure, suggesting that H. aphrodisiaca infusion may be used combined with CsA to reduce CsA‐induced injuries in the testis. Anat Rec, 291:809‐817, 2008.

Hepatoprotective effect of Bathysa cuspidata in a murine model of severe toxic liver injury

The objective of this study was to investigate the hepatoprotective effect of a bark extract of Bathysa cuspidata extract (BCE) in a murine model of severe liver injury induced by carbon tetrachloride (CCl4). Forty‐two Wistar rats were randomized into six groups of seven animals each: Group 1(G1): CCl4; Group 2 (G2): dimethyl sulfoxide (DMSO) + CCl4; Group 3 (G3): BCE 400 mg/kg alone; Group 4 (G4): BCE 200 mg/kg + CCl4; Group 5 (G5): BCE 400 mg/kg + CCl4; Group 6 (G6): DMSO alone. The extract was administered by gavage for 18 days beginning 6 days prior to the first application of CCl4. After completing CCl4 administration, the animals were euthanized. The animals in G1, G2, G4 and G5 experienced significant body weight loss and had an increased liver somatic index compared with G3 and G6 (P < 0.05). A significant reduction in serum aspartate and alanine transaminase and gamma‐glutamyl transferase (P < 0.05) and a significant increase in the activity of the anti‐oxidant enzyme superoxide dismutase were found in G5 (P < 0.05). Lower proportions of cellular necrosis and lipid droplets were found in the livers of animals in G4 and G5 compared with G1 and G2 (P < 0.05). These results confirm the marked hepatoprotective activity of the bark extract of Bathysa cuspidata in severe injuries induced by CCl4 in rats and suggest that this effect may be associated with the inhibition of oxidative damage.

Histological alterations in gills of Astyanax aff. bimaculatus caused by acute exposition to zinc.

Increasing contamination of aquatic ecosystems by metals has caused various morphological, physiological and biochemical changes in aquatic organisms, and the gills of fish are recognized as indicators of environmental quality. In this context, the present work proposed to study the effects of different concentrations of zinc (Zn) in the histology of gills of yellow tail lambari (Astyanax aff. bimaculatus) after acute exposure. Seventy-two adult males of A. aff. bimaculatus were used, the treatments were six concentrations of Zn: 0; 3; 5; 10; 15; and 20 mg/L of water, by 96 h, and gills, muscle and bone fragments were removed. Fragments of gills were fixed and included, sectioned in a rotary microtome and stained with toluidin blue. Fragments of bone, muscle and gills were dehydrated and digested to quantify the absorption of Zn. The median lethal concentration (LC(50)) 96 h after Zn acute exposure was 10 mg/L of water. Noteworthy, Zn was highly toxic in acute exposure trials starting at the concentration 5 mg/L. The exposure of fish to the metal caused branchial histopathological changes correlated with increasing concentration, caused the death of fish at concentrations of 10, 15 and 20 mg/L. The histological alterations observed in the gills were hyperplasia, lamellar fusion, aneurysm, destruction of the lamellar epithelium, rupture of membrane, deletion of secondary lamellar high, which presented more severity in treatments exposed to the highest concentrations. In conclusion, gills of A. aff. bimaculatus presented profound histological alterations as a result of Zn exposure, and hence, proved to be excellent indicators of environmental contamination.

Long-lasting morphofunctional remodelling of liver parenchyma and stroma after a single exposure to low and moderate doses of cadmium in rats

Frequent exposure to cadmium (Cd) in low doses is common; however, the long‐lasting effects of this exposure are still poorly understood. Therefore in this study we have evaluated long‐lasting hepatic morphofunctional adaptations in rats exposed to low and moderate doses of Cd. Five experimental groups were tested: control (0.9% saline) and other four receiving single intraperitoneal doses of 0.67, 0.74, 0.86 and 1.1 mg of Cd/kg. The animals were killed after eight weeks and the following parameters were analysed: biometrics, oedema, Cd bio‐accumulation, collagen, glycogen, lipid droplets, superoxide dismutase (SOD) and catalase (CAT), serum transaminases, liver histopathology and stereology. In all groups exposed to Cd there was significant increase in SOD and CAT activities, ALP levels, proportion of binucleated hepatocytes, nuclei/cytoplasm ratio, macrophages (Kupffer cells) and collagen fibres. In these groups, glycogen accumulation by hepatocytes and the proportion of sinusoidal capillaries were significantly reduced compared with controls. The liver somatic index was increased, and liver oedema was evident in animals exposed to higher dose of Cd. Areas of necrosis were found in animals exposed to the three highest doses. These results indicate that Cd is an extremely toxic bioactive heavy metal, which even at low doses is able to disrupt liver homeostasis. At low and moderate doses, Cd exposure induces morphofunctional pathological remodelling of the hepatic stroma and parenchyma, which remain active after eight weeks. In response to injury, the liver tissue triggers a reactive process by enhancing activation of antioxidant enzymes and collagenogenesis.

Aerobic exercise and not a diet supplemented with jussara açaí ( Euterpe edulis Martius) alters hepatic oxidative and inflammatory biomarkers in ApoE-deficient mice

The pulp of jussara açaí (Euterpe edulis Martius) fruit is rich in anthocyanins that exert antioxidant and anti-inflammatory effects similar to those exerted by aerobic exercise. In the present study, we investigated the effects of jussara açaí fruit pulp consumption, either alone or in combination with aerobic exercise, on the hepatic oxidative and inflammatory status of ApoE-deficient (ApoE - / - ) mice. Male mice were divided into four groups (control (C), control plus açaí, exercise plus açaí (EXA) and exercise (EX)) and fed the AIN-93M diet or the AIN-93M diet formulated to contain 2 % freeze-dried açaí pulp. Mice in the EX and EXA groups were subjected to a progressive running programme (5 d/week, 60 min/d, 16 m/min) for 12 weeks. Mice that were made to exercise exhibited reduced (40·85 %; P< 0·05) hepatic superoxide dismutase activity when compared with the C mice, independent of the açaí diet. Mice in the EX group exhibited a lower (42 %; P< 0·05) mRNA expression of monocyte chemotactic protein-1 in the liver compared with the C mice. Mice in the EXA and EX groups had lower percentages of hepatic lipid droplets (70 % and 56 %, respectively; P< 0·05) when compared with the C mice. Mice in the EX group had smaller (58 %; P< 0·05) area of lesions in the aorta when compared with the C mice. Serum lipid profile was not affected (P>0·05). In conclusion, aerobic exercise training rather than açaí fruit pulp consumption or a combination of both enhances the hepatic oxidative and inflammatory status of ApoE - / - mice.

Morphometry of testis and seminiferous tubules of the adult crab-eating fox (Cerdocyon thous, Linnaeus, 1766)

Body and testicular biometric parameters are very important for establishing reproductive patterns and, consequently, the development of protocols for assisted reproduction in different species. A direct correlation between the testis weight and the sperm population was observed in other studied species, because the testis size primarily reflects the total volume of the seminiferous tubule, its main component. The objective of this study was to determine the testicular volume parameters and correlate data from morphometry of testis and seminiferous tubules with body mass in six adult crab-eating foxes. The mean body weight of the crab-eating foxes in this study was 6.53 kg, with approximately 0.068% allocated to the testicular mass and 0.042% specifically to seminiferous tubules, which represented 87.5% of the testicular parenchyma. The albuginea comprised 12.5% of the testicular mass. The mean diameter of seminiferous tubules was 236 μm, and the mean thickness of the seminiferous epithelium was 62.9 μm. Values of tubular parameters indicate a sperm productivity close to those observed in previously studied carnivores.

Evaluation of the Cell Population of the Seminiferous Epithelium and Spermatic Indexes of the Bat Sturnira lilium (Chiroptera: Phyllostomidae)

Due to the scarcity of information about patterns of spermatogenesis in bats, this study aimed to provide information on the testicular activity of the bat Sturnira lilium along the annual seasons. Thus, a series of morphometrical and stereological analyses were made using the testes of adult S. lilium in order to achieve a better understanding of the sperm production dynamics. Light and transmission electron microscopy analyses were performed in testicular fragments of animals captured during dry and rainy seasons. The testes followed the pattern of organization described for other mammals, and there were no morphological differences between organs collected either in dry or in rainy seasons. Each tubular cross-section in stage 1 was made of 0.5 type-A spermatogonia, 4.4 primary spermatocytes in preleptotene/leptotene, 3.7 in zygotene, 11.9 in pachytene, 35.6 round spermatids and 8.5 Sertoli cells. The mitotic and meiotic indexes were 15.4 and 2.9 cells, respectively, while the spermatogenesis yield was 68.7 cells. The testicular sperm reserves was 37.61×106 cells, and daily sperm production per gram of testis averaged 209.68×106 cells, both highest averages occurring in the rainy season. S. lilium male bats have a continuous reproductive pattern, high spermatogenesis yield and low support capacity by the Sertoli cells.

The energy density of laser light differentially modulates the skin morphological reorganization in a murine model of healing by secondary intention

This study investigates the influence of gallium–arsenide (GaAs) laser photobiostimulation applied with different energy densities on skin wound healing by secondary intention in rats. Three circular wounds, 10 mm in diameter, were made on the dorsolateral region of 21 Wistar rats weighting 282.12 ± 36.08 g. The animals were equally randomized into three groups: Group SAL, saline solution 0.9%; Group L3, laser GaAs 3 J/cm2; Group L30, laser GaAs 30 J/cm2. Analyses of cells, blood vessels, collagen and elastic fibres, glycosaminoglycans and wound contraction were performed on the scar tissue from different wounds every 7 days for 21 days. On day 7, 14 and 21, L3 and L30 showed higher collagen and glycosaminoglycan levels compared to SAL (P < 0.05). At day 21, elastic fibres were predominant in L3 and L30 compared to SAL (P < 0.05). Type‐III collagen fibres were predominant at day 7 in both groups. There was gradual reduction in these fibres and accumulation of type‐I collagen over time, especially in L3 and L30 compared with SAL. Elevated density of blood vessels was seen in L30 on days 7 and 14 compared to the other groups (P < 0.05). On these same days, there was higher tissue cellularity in L3 compared with SAL (P < 0.05). The progression of wound closure during all time points investigated was higher in the L30 group (P < 0.05). Both energy densities investigated increased the tissue cellularity, vascular density, collagen and elastic fibres, and glycosaminoglycan synthesis, with the greater benefits for wound closure being found at the density of 30 J/cm2.

Histomorphometric evaluation of the Molossus molossus (Chiroptera, Molossidae) testis: The tubular compartment and indices of sperm production

Insectivorous bats play a very important role in the regulation of tropical ecosystems, but information about their reproductive cycle is lacking. Thus, male Molossus molossus were captured over the four seasons, and morphometric analyses of their testes were conducted to infer on the gonadal dynamics and the reproductive capacity of the species. Testes were immersed in Karnovsky fixative, and fragments were embedded in methacrylate and paraplast for morphometric and TUNEL assay respectively. The least gonadosomatic index (0.3%), tubulesomatic index (0.2%) and tubular diameter (133.2μm) occurred in summer. An adult M. molossus showed a total average of 48.9m of seminiferous tubules per gram of testis. Primary spermatocytes were observed in the zygotene at Stage 1 of the seminiferous epithelium cycle. The greatest meiotic index was obtained in winter (3.8 cells), and the general yield of spermatogenesis was higher in winter (64.5 cells) than in summer (19.1 cells). There was no difference in the apoptotic cells count among seasons. The Sertoli cell index was less in summer (5.9) than in fall (11.6), while the number of Sertoli cells per gram of testis did not vary significantly among the seasons (28.0×10(7)). The spermatic reserve per gram of testis was greater in the fall (63.9×10(7)) and winter (69.8×10(7)) than summer (37.1×10(7)). We conclude that M. molossus males show a continuous reproductive cycle, featuring greater spermatogenic activity during the fall and winter, a tubular length above the average of other mammals and a less support capacity of the Sertoli cells.