Priscila Vieira Rosa

Effect of cryopreservant combinations on the motility and morphology of curimba (Prochilodus lineatus) sperm.

This study investigated the application of intra- and extra-cellular cryoprotectant combinations on the quality of curimba Prochilodus lineatus semen subjected to cryopreservation. Semen treatments were tested with 8% DMSO or methanol as intracellular cryoprotectant, 5% egg yolk or lactose as extracellular cryoprotectant and 5% BTS. These cryoprotectant combinations are suitable for curimba but have not been tested at the lesser concentrations proposed or in combination with BTS. Semen samples collected from 19 curimbas were diluted into one of four cryoprotectant combinations: DMSO+yolk; DMSO+lactose; methanol+yolk; and methanol+lactose. After dilution, semen samples were cryopreserved in 0.5 mL straws for 10 days in a liquid nitrogen tank. Semen was thawed in a water bath at 60°C for 8s. We evaluated the quality of fresh, diluted (pre-freezing) and post-freezing semen according to sperm motility rate (%) and duration (s). Sperm morphology was also analyzed in thawed semen. Sperm motility rate decreased progressively after dilution and thawing. The motility rate in post-freezing semen was higher in the treatments using DMSO+lactose and methanol+yolk. Sperm motility duration in post-freezing sperm was greater in the treatments using methanol rather than DMSO as intracellular cryoprotectant, irrespective of the extracellular cryoprotectant used. Abnormality frequency in thawed sperm was less in semen treated with egg yolk than with lactose. Thus the use of methanol intracellular cryoprotectant is recommended along with yolk extracellular cryoprotectant in the cryopreservation process for curimba semen.

Relative distribution of gastrin-, CCK-8-, NPY- and CGRP-immunoreactive cells in the digestive tract of dorado (Salminus brasiliensis)

The endocrine cells (ECs) of the gastrointestinal mucosa form the largest endocrine system in the body, not only in terms of cell numbers but also in terms of the different produced substances. Data describing the association between the relative distributions of the peptide-specific ECs in relation to feeding habits can be useful tools that enable the creation of a general expected pattern of EC distribution. We aimed to investigate the distribution of ECs immunoreactive for the peptides gastrin (GAS), cholecystokinin (CCK-8), neuropeptide Y (NPY), and calcitonin gene-related peptide (CGRP) in different segments of the digestive tract of carnivorous fish dorado (Salminus brasiliensis) by using immunohistochemistry procedures. The distribution of endocrine cells immunoreactive for gastrin (GAS), cholecystokinin (CCK-8), neuropeptide Y (NPY), and calcitonin gene-related peptide (CGRP) in digestive tract of dorado S. brasiliensis was examined by immunohistochemistry. The results describe the association between the distribution of the peptide-specific endocrine cells and feeding habits in different carnivorous fish. The largest number of endocrine cells immunoreactive for GAS, CCK-8, and CGRP were found in the pyloric stomach region and the pyloric caeca. However, NPY-immunoreactive endocrine cells were markedly restricted to the midgut. The distribution pattern of endocrine cells identified in S. brasiliensis is similar to that found in other carnivorous fishes.

Effect of photoperiod on locomotor activity, growth, feed efficiency and gonadal development of Nile tilapia

The objective of this study was to evaluate the effect of photoperiod on locomotor activity, growth and gonadal development in Nile tilapia (Oreochromis niloticus) fingerlings. A completely randomised design was used, with five treatments (0L:24D, 6L:18D, 12L:12D, 18L:6D and 24L:0D) and four replicates, with the aquarium as the experimental unit. One hundred and sixty fingerlings of tilapia weighing 3.21±0.05 g and measuring 4.35±0.07 cm each were distributed among 20 aquaria of 20 L in a recirculation system with the temperature controlled to 27 °C, with eight fish per aquarium. Feeding was carried out twice daily for 75 days, with extruded feed containing 40% crude protein. The fingerlings subjected to a photoperiod of 12L:12D as well as those under 18L:6D and 24L:0D showed the greatest locomotor activity, whereas those under 6L:18D and 0L:24D showed the lowest activity. Fish subjected to a photoperiod of 18L:6D and 24L:0D showed the highest levels of performance. However, manipulation of the photoperiod did not influence the gonadal development, survival or the appearance of deformities in juvenile Nile tilapia. Under long photoperiods (18L:6D and 24L:0D), fish direct their energy to somatic growth and induce best feed efficiency.

Effect of benzocaine concentration and fish size on anesthesia and recovery in Nile tilapia

To evaluate the influence of benzocaine concentration on the induction and recovery from anesthesia in different sized Nile tilapia (Oreochromis niloticus), it was developed a random block experiment design in 4 × 4 factorial consisted of four fish lengths (6; 10; 14 and 18 cm total lenght) and four benzocaine concentrations (60; 120; 180 and 240 mg of benzocaine per liter of water). The induction and the recovery from anesthesia were divided in three stages based on the behavior of the fishes under anesthetic effect. The time spent in each stage was recorded. Differences in length did not affect significantly the anesthesia and recovery time. Linear plateu regressions were generated to estimate the point which increases on anesthetic doses does not imply in further effects. During the anesthesia induction, the plateu occurred in concentration time of 132 mg/L, and estimated time of 86 seconds would be necessary to achieve the complete sedation (stage 3). For permanence of the fish under total sedation, it is needed 190 g/L benzocaine and estimated time of permanence in this stage is 91.1 seconds. Benzocaine concentrations above 190 mg/L do not produce any additional effects that justify its use.

Daily rhythms of the expression of genes from the somatotropic axis: The influence on tilapia (Oreochromis niloticus) of feeding and growth hormone administration at different times.

The aim of this research was to investigate the presence of daily rhythms in the somatotropic axis of tilapia fed at two times (mid-light, ML or mid-dark, MD) and the influence of the time of day of growth hormone (GH) administration on the response of this axis. Two different GH injection times were tested: ZT 3 (3h after lights on) and ZT 15 (3h after lights off). In both experiments, the mRNA expression levels of hypothalamic pituitary adenylate cyclase-activating polypeptide (pacap), pituitary growth hormone (gh), liver insulin-like growth factors (igf1 and igf2a), and liver and muscle growth hormone receptors (ghr1 and ghr2) and IGF receptors (igf1ra and igf2r) were evaluated by means of qPCR. Daily rhythms were observed in the liver for ghr1, ghr2 and igf2r but only in fish fed at ML, with the acrophases located in the light phase (ZT 3:30, 3:31 and 7:38 h, respectively). In the muscle, ghr1 displayed a significant rhythm in both groups and ghr2 in ML fed fish (acrophases at ZT 5:29, 7:14 and 9:23h). The time of both GH administration and feeding influenced the response to GH injection: ML fed fish injected with GH at ZT 15 h showed a significant increase in liver igf1, igf2a and ghr2; and muscle ghr2 expression. This is the first report that describes the existence of daily rhythms in the somatotropic axis of tilapia and its time-dependent responses of GH administration. Our results should be considered when investigating the elements of the somatotropic axis in tilapia and GH administration.

Activity of glutamate dehydrogenase and protein content in the breast of broilers fed diets containing different sources and levels of glycerine

According to scientific literature, glycerol in the diet can spare glucogenic amino acids by inhibiting the activity of enzymes, such as glutamate dehydrogenase, thereby promoting protein deposition in muscle tissues. Therefore, the objective of this study was to evaluate the effect of three sources of glycerine (crude glycerine from soybean oil--CGSO, mixed crude glycerine from frying oil and lard--MCG and a semipurified glycerine from soybean oil--SPGSO) in four concentrations in the diet (17.5, 35.0, 52.5 and 70.0 g of each type of glycerine/kg of feed) on the activity of hepatic glutamate dehydrogenase, performance and protein content in the breast of broilers, during 22-35 days of age (experiment I) and 33-43 days of age (experiment II). In both experiments, an increase in MCG induced a linear decline in glutamate dehydrogenase activity (p < 0.05). In contrast, increasing the concentration of SPGSO in the diet caused a linear increase in enzyme activity (p < 0.05). There was no (p > 0.05) isolated effect of glycerine on the enzyme activity in either evaluated phase; however, during 33-42 days of age, MCG inhibited (p < 0.05) the glutamate dehydrogenase activity by up to 34.43%. During 22-35 days of age, the diet containing SPGSO induced a higher protein content (p < 0.05) in the breast, and regardless of the source utilized, the maximum protein deposition was estimated (p < 0.05) when broilers were fed with 55.08 g glycerine/kg of diet. There was no (p > 0.05) interaction or isolated effects of the sources and levels of glycerine on the protein content in the breast of broilers at 33-42 days of age, and moreover, all diets containing glycerine promoted a similar protein deposition in the breast compared with birds that received the diet without glycerine. The bird age also showed to influence the feed intake and weight gain of broilers fed diet containing glycerine. It is concluded that for both rearing phases, an increase in glycerine in the diet did not necessarily reduce the glutamate dehydrogenase activity, and the protein deposition in the breast of broilers may not be strictly correlated with the activity of this enzyme.

Effects crude protein levels on female Nile tilapia (Oreochromis niloticus) reproductive performance parameters

The goal of the present work was to study the reproductive performance of Nile Tilapia (Oreochromis niloticus) female broodstock fed diets containing different levels of crude protein (CP). Two hundred and forty Nile tilapia (O. niloticus) were used at an average age of 30 months, with 180 females and 60 males. The broodstock were lodged separately in masonry tanks with continuous water flow. The females were stocked in thirty tanks with dimensions of 8 m(3) in a completely randomized design consisting of five treatments and six replications. The treatments consisted of five diets with different levels of CP (32, 34, 36, 38 and 40%) and with digestible energy per gram of protein of 9.5 kg of feed. The crude protein (CP) levels positively influenced (p<0.05) reproductive parameters (female relative and absolute fecundity, egg diameter, fasting larvae survival capacity), the somatic indexes (gonadosomatic (GSI), hepatosomatic (HIS), viscerosomatic (VSI)), total plasma protein, albumin and triglycerides. There were no significant differences (p>0.05) observed with regard to spawning weight and female weight. The reproductive parameters studied in the present research indicate that diets formulated with 38% CP with digestible energy per gram of CP of 9.5 were the best diets for tilapia females during the reproductive period.

Fotoperíodo sobre parâmetros fisiológicos relacionados ao estresse em alevinos de tilápia-do-nilo

The objective of this study was to evaluate the influence of different photoperiods on physiological parameters related to stress in Nile tilapia fingerlings (Oreochromis niloticus). A completely randomized design with five treatments (0L, 6L, 12L, 18L, 24L) and four replications was used, and the aquarium as the experimental unit. One hundred and sixty tilapia fingerlings, 3.21±0.05g and 4.35±0.07cm were distributed in 20 aquariums of 20L in a recirculation system with temperature controlled at 27 ° C and density of eight fish per aquarium. Feeding was performed twice daily with extruded feed containing 40% crude protein, for a period of 75 days. At the end of experimental period, the data of survival, cortisol, glucose and hematocrit were subjected to analysis of variance and, subsequently, in case of significance (P 0.05).

Adaptation to physical training in rats orally supplemented with glycerol.

We evaluated training adaptation and physical performance parameters in rats orally supplemented with glycerol, glucose, or saline, and submitted to moderate aerobic exercise. Thirty male rats were trained for 6 weeks and administered the supplements during the last 4 weeks of the experiment. Animals were distributed in a completely randomized factorial 2 × 3 design (with or without exercise and 3 substrates). Data were subjected to analysis of variance (ANOVA) and means were compared using the Student-Newmann-Keuls test at 5%. Among the trained animals, none of the substances caused differences in the percentages of protein, fat, or water content in the carcass. Compared with the sedentary animals, the trained animals supplemented with saline and glucose showed a higher protein percentage in the carcass. The relative mass of the heart and adrenal glands was higher in the trained animals. Glycerol improved the protein content in non-trained animals and increased the relative adrenal mass in both groups. Glycerol reduced the variation in levels of lactate and aspartate aminotransferase (AST) during the last exercise session. There was no difference between groups regarding the relative mass of the thymus and gastrocnemius or with the diameter of muscle fibers or the neutrophil-lymphocyte ratio. Supplementation with glycerol was efficient at attenuating variation in AST and lactate levels during exercise.

Content of plasmatic glycerol and activity of hepatic glycerol kinase in broiler chickens fed diets containing different sources and concentrations of glycerine

The objective of this study was to evaluate the effect of three sources of glycerine (crude glycerine from soya bean oil - CGSO, mixed crude glycerine from frying oil and lard--MCG, and a semipurified glycerine from soya bean oil--SPGSO) in four concentrations in the diet (17.5, 35.0, 52.5 and 70.0 g of each type of glycerine per kg of feed) on the levels of plasmatic glycerol and the activity of the hepatic enzyme glycerol kinase in broilers of 22-35 days old (experiment I) and 33-42 days old (experiment II). The highest (p < 0.05) plasmatic glycerol level was detected in broilers fed diet containing CGSO. Independent of the source, increasing the concentration of glycerine led to a linear increase (p < 0.05) in the plasmatic glycerol concentration. In experiment I, all the diets containing glycerine resulted in increased concentrations of plasmatic glycerol, in relation to the control diet without glycerine. However, in experiment II, only the diet containing 17.5 g of CGSO per kg of feed and the diets formulated with any of the three types of glycerine in the inclusion concentrations of 35, 52.5 and 70 g/kg of feed resulted in higher plasmatic glycerol levels than those observed in the control broilers. The source of glycerine influenced the glycerol kinase activity only in experiment II, where the use of CGSO in the diet increased the enzyme activity. For both experiments, the glycerol kinase activity increased with the inclusion of glycerine in the diet. In conclusion, based on the absence of saturation of the glycerol kinase activity for the three glycerine sources and for both rearing periods evaluated, the broilers can metabolize the glycerol (at the level of the phosphorylation of the glycerol to glycerol-3-phosphate in the liver) present in the glycerine when the diet is supplemented with up to 70 g of glycerine per kg of feed.