Repositório RCAAP

This study aimed to evaluate different antioxidants in embryo culture after vitrification, with or without the previous use of hydrostatic pressure (PH). Considering this, three experiments were designed to evaluate the interaction between PH and antioxidants (β-mercaptoethanol - BME, cysteamine - CYST and BME + CYST) in fresh and vitrified in vitro produced embryos. In experiment 1 hatching and degeneration rates were evaluated with 24 and 48 hours after passing through the PH and in experiment 2, the same parameters were evaluates at 12, 24, 48 and 72 hours after heating. The last step of the study evaluated the pregnancy rate of vitrified embryos, cultured for 12 hours with / without antioxidant. The first experiment showed no difference between treatments the differences, independent of the treatment, were obtained between expanded blastocyst (BX) and blastocyst (BL). The second experiment found similar results for all parameters evaluated in embryos BX. Note that the BME + cyst treatment got better hatching rate in the BL with 48 and 72 hours (76.04%) than the control group (45.83%). The same behavior was observed in degeneration 24 hours, where the BME + CYST group was 7.29 % against 32.29% in the control group. However, the pregnancy rates (55 embryo transfers) were not different between groups control fresh, control vitrified and BME (38.9%, 16.7% and 31.6%, respectively). This study showed that the use of hydrostatic pressure and antioxidant had no effect in the evaluated parameters. However. there is a significant improvement to BL embryos when combining BME + CYST in the culture medium after vitrification.

DNA methylation is one of the most studied epigenetic events and is responsible for epigenetic reprogramming which occurs during gametogenesis. Understanding how this reprogramming occurs in oogenesis is important to comprehend physiologic and genetic aspects involved in female gametogenesis in order to create parameters for oocyte competence and, consequently, to improve the in vitro embryo production, maximizing the use of gametes and improving production rates. The aim of this study was to evaluate the DNA methylation pattern in two DMRs involved in the control of XIST and IGF2 genes expression in oocytes from pre antral and antral follicles of Nellore cows. The extracted DNA from oocytes was treated with sodium bisulphite and amplified to XIST and IGF2 genes, which was cloned into DH5 cells, and then purified and sequenced. The methylation patterns found for oocytes of primordial, secondary, incompetent antral and competent antral follicles were 91.59 ± 6.4%, 85.70 ± 19.6%, 91.25 ± 7.2% and 92.58 ± ± 11.7%, respectively for XIST gene and 60.56 ± 29.1%, 59.68 ± 34.6%, 58.21 ± 33.0% and 67.47 ± 27.8%, respectively for IGF2 gene. XIST is more methylated than IGF2 gene (P<0,001). The hypermethylated pattern of XIST gene suggests that this event may be responsible for epigenetic reactivation of the X chromosome during oogenesis, which is observed in the final oocyte. The IGF2 gene was also hypermethylated, a different pattern found in matured oocytes. This suggests that the analyzed regions undergo differents epigenetic reprogramming processes during oogenenesis, which are only completed with oocyte maturation.

The objective of the studies was evaluate the effects of the animal category, season of calving, season of artificial insemination (AI), parity, numbers of previous inseminated and days in milk (DIM) at AI on pregnancy losses (PL) and the incidence of pregnancy loss before and after 60th day post-AI from females repeat breeders (FRB) and the effects of the animal category, season of calving, parturition condition and parity on the incidence of FRB. Data from Study 1 were collected from 462 pregnancies and calving of lactating dairy cows and from 118 pregnancies of heifers and the Study 2 from 997 pregnancies and calving of lactating dairy cows and from 371 pregnancies of heifers. Pregnancy check was performed by ultrasound between days 28 and 44 post-AI and re-checked between days 45 and 60 post-AI. The rates of pregnancy loss before 60th day and the incidence of RB were higher in lactating dairy cows than heifers (11.90% vs. 3.4%, P=0.01 and 24.5% vs. 6.5%, P<0.001, respectively) and pregnancy loss after day 60 was not affected by animal category (P=0.58). The season of calving affected the pregnancy loss before 60th day (which was higher in spring/summer) and there was only tendency (P=0.078) in affecting the incidence of RB cows probably due to the worst postpartum recovery. There was no relationship between season of AI, parity, number of previous inseminated and DIM at IA and pregnancy loss before and after 60th day in lactating dairy cows. The parturition condition had no effects on RB incidence, however, the effect of parity was detected. Even in crossbred lactating dairy cows the incidence of overall pregnancy loss and RB condition was higher and could be considered as a factor that compromised the reproduction efficiency.

The study of the lumbosacral plexus nerves constituents is extremely important because it relates the various evolutionary aspects of animal locomotion and posture. Considering that the femoral nerve is the largest cranial part of the lumbosacral plexus, aimed to describe the origins, distributions and ramifications of the femoral nerve in giant anteater (Myrmecophaga tridactyla), comparing with the literature described for domestic and wild animals in to establish correlations of morphological similarities and providing subsidies for similar areas. For the work three specimens were used prepared by injection of aqueous 10% formaldehyde in the femoral artery, for keeping the specimens and further dissection. The origin of the femoral nerve in the right and left sides, is the ventral brach of the lumbar spinal nerves one, two and three. As to the distributions and ramifications, the femoral nerve provides branches to the major and minor psoas muscle, lateral and medial iliac, pectineus, adductor magnus, sartorius and quadriceps. The anatomical origin of the femoral nerve in M. tridactyla shows a variation due to the difference in the number of vertebrae (L1, L2 and L3). But in most distributions and ramifications of the femoral nerve, this species has a significant degree of morphological similarities with domestic and wild animals in this study.

During the process of cloning using nuclear transfer, epigenetic marks in cells must go through a reprogramming process, so that embryonic development can occur appropriately. However, during TN this reprogramming process is not completely efficient. Analysis of cell viability and expression of genes related to pluripotency and epigenetic changes, allowed us to evaluate the action of demethylation drugs such as Procaine and SAH in somatic cell cultures. These substances are potencial inducers of epigenetic reprogramming and they could be used to improve the process of cloning by TN. The bovine fibroblasts treated with 1 mM Procaine had lower cell viability compared to the control group (non trated), while the group treated with 2 mM of SAH did not differ from the controls. OCT4 and NANOG genes were detected in control group as well as in the group treated with 1mM Procaine, while HDAC2 and DNMT1 genes were expressed in cells treated with 1 mM of Procaine as in those treated with 2 mM of SAH, showing no significant difference between the experimental groups. In this study we concluded that the OCT4 and NANOG genes are not molecular markers for cellular pluripotency in bovines and we can modify the epigenetic patterns of DNA of the nucleus donor cells for cloning by TN process, contributing to the improving of the results of this technique.

This study was performed with the objective of assessing the effects of direct inclusion of 0%, 1%, 1.5% and 2% of the alga Lithothamnium calcareum in the diet, the age hen and interaction between these variables on the production and egg shell quality of laying hens in the second cycle. A completely randomized design was used with subdivided portions (inclusion level: 0%, 1%, 1.5% and 2% x age: 112, 114, 116 and 118 weeks of age) with 6 replications, 10 hens per experimental unit. Each replications of the experiment involved use of 10 Dekalb White hens lines with an age of 110 weeks, four eggs being collected during each period. Percentage of eggs, percentage of broken eggs, percentage of cracked eggs, egg weight; yolk weight, white weight, eggshell weight, specific gravity, percentage eggshell, shell thickness ; number of shell pores; percentage of mineral matter eggshell, percentage of calcium and phosphorus in eggshell were measured and means were compared by Scott-Knott 5% test. The inclusion level of the alga increased the percentage of egg, shell thickness, percentage of mineral matter eggshell and calcium and decreased percentage of cracked eggs and number of pores in the shell. The increasing age of the laying hens decreased eggs shell quality. It was concluded that inclusion levels of Lithothamnium calcareum increased production and egg shell quality.

This study described morphological aspects of the digestive tract of the giant anteater (Myrmecophaga tridactyla), five specimens were used, belonging to the collection of the Laboratório de Ensino e Pesquisa em Animais Silvestres in UFU, were processed by usual methods of macroscopic anatomical, histological and scanning electron microscope (SEM) analysis. The esophagus is a narrow tube that goes straight in the thoracic cavity. The stomach has the shape of the letter J, have the cardiac, fundic, corpus and pyloric regions. The small intestine is long, has a duodenum, jejunum and ileum. The large intestine is short, consisting of ascending colon, transverse colon, descending colon, rectum and anus. The lining epithelial of the esophagus is stratified squamous, non-glandular; and scanning electron microscopic examination of the esophagus is smooth and pleated. Stomach is simple prismatic relatively low, with shallow crypts; and rough surface. In the small intestine is simple prismatic, the duodenum has goblet cells, a huge amount of Paneth cells, the jejunum has an increase of Paneth cells, the ileum has a few Paneth cells and an increase of goblet cells; and villous surface; the large intestine is simple prismatic, an increase goblet cells; and smooth surface with openings of intestinal crypts.

The dissertation was divided into five chapters. The first was called \"General Consideration,\" which was performed a brief literature review about what was proposed in the work. In the second chapter aimed to check whether chickens immunized with Brucella abortus B19 vaccine and soluble protein extract solution, extracted from this vaccine, could produce specific polyclonal antibodies, detectable in indirect ELISA tests. Six hens with 25 week-old, White Leghorn breed, were immunized, two immunized with B19, two with a soluble protein extract solution extracted from the B19 vaccine and two were used as controls, being immunized with PBS. Blood samples were taken fortnightly and eggs daily for 13 weeks. Polyclonal antibodies IgY were purified from egg yolk using for delipidation the dilution method in acidified water (pH 5,0-5,2) and precipitation with ammonium sulfate. Was performed ELISA test to verify the specificity of IgY. Was possible to observe the production of specific polyclonal antibodies against the antigens used in the immunizations, both in serum and in egg yolks. In the third chapter aimed to check the response of IgY extracted from egg yolk of immunized chickens with B19 vaccine (IgY anti- B19) against the antigen EPS and the response of IgY extracted from egg yolk of immunized chickens with EPS (IgY anti-EPS) against the vaccine antigen B19, using the ELISA in order to verify possible antigen-antibody interactions. For this, two microtiter plates were sensitized, in one of them using B19 vaccine as antigen and another EPS antigen, were placed on each plate as primary antibodies IgY extracted from the egg yolk, for each group of immunized hens, a group immunized chickens with vaccine B19, one with EPS and one control. Both IgY anti-B19, as IgY anti-EPS were reactive to the antigens tested, so it is believed that these antibodies may serve as a tool for the identification of antigens in diagnostic tests aimed at this purpose. The objective of fourth chapter was to evaluate the humoral immune response of cattle naturally infected and newly vaccinated with B19 vaccine against EPS and B19 vaccine antigens in indirect ELISA. Was used in this study 96 samples of blood serum of cattle, 32 heifers recently vaccinated with B19 vaccine, 32 naturally infected cattle and 32 negative cattle for brucellosis. Was verified that in the indirect ELISA with antigen EPS when compared with indirect ELISA with antigen vaccine B19 and 2-ME, this test was able to distinguish newly vaccinated animals of naturally infected animals. The fifth chapter are the \"final thoughts\" on the entire dissertation, including findings of all the chapters.

The aim of this study was to evaluate the incidence of tuberculosis in a cattle herd by the cervical comparative intradermal tuberculin test (ITT), and analyze the efficiency of complementary diagnostic methods in confirming the infection. The study was conducted on a breeding dairy farm in the city of Perdizes MG. Tuberculin test was performed in 164 cattle, of which 40 were positive and slaughtered. The macroscopic post mortem inspection of carcasses was followed by the collection of nasal swabs, blood and tissue samples (liver, lung and mediastinal lymph node) for PCR tests, ELISA and histopatology with Hematoxylin-Eosin (HE) and Ziehl-Neelsen (ZN) respectively. Of the 164 cattle analysed by ITT, 41 (25%) were positive, 29 (17.68%) inconclusive and 94 (57.32%) negative. Of the 40 cattle slaughtered, 22 (55%) carcasses had macroscopic lesions suggestive of tuberculosis, 14 (35%) in the mediastinal lymph nodes, seven (17.5%) in the liver and six (15%) in the lung. The histopathology HE identified 13 (32.5%) carcasses with histological changes, six (15%) in the mediastinal lymph nodes, five (12.5%) in the liver and three (7.5%) in the lung. At ZN, the presence of acid-fast bacilli was not detected in any of samples tested. The ELISA/IDEXX identified one (2.5%) animal reagent, and the PCR test detected DNA of M. bovis in one (2.5%) cow. Being a dairy herd, the occurrence of tuberculosis may represent a zoonotic risk. The ITT should already be a routine test required for the farms that sells milk or meat, because it recognizes recent infections, often not detected by other diagnostic methods.

The aim was to evaluate ingestive behavior and establish optimum inclusion level of palm cake in sheep diet. Eighteen crossbred castrated rams were fed palm cake inclusion levels of 0%, 7.5%, 15% and 22.5% in total diet and kept in metabolic cages during the experiment that lasted 21 days. Data were subjected to regression analysis with 5% of significance for regression coefficients and equations. There was reduction in dry matter (DM), hemicellulose, neutral detergent fiber (NDF), crude protein (CP), total digestible nutrients, total carbohydrates and mineral matter intakes (P <0.01) and no significant differences in cellulose, acid detergent fiber (ADF) and non-fiber carbohydrates intakes (P>0.05). Ether extract and lignin intakes increased (P<0.01) with reduction in crude energy (CE), DM, CP, cellulose, hemicellulose and NDF digestibilities (P<0.05). The metabolizable energy (ME): CE ratio reduced because fecal CE increased (P <0.01) and nitrogen balance decreased (P<0.01). In order to compensate for unavailability of nutrients caused by palm cake, urinary excretion and feces moisture (P<0.05) reduced. There were no significant differences in total chew time, ingestion, rumination and leisure (P>0.05). Rumination and feed efficiencies (g DM / h; g NDF / h) showed no significant differences (P>0.05) and decreased (P<0,05), respectively. Palm cake inclusion levels higher than 7,5% in sheep diet reduces consumption and digestibility of nutrients with possible performance damages, especially in high-producing animals.

Reptiles are animals that attract by their diversity and among them, the snakes deserve emphasis on use commercially like pet s and biological immune production. Considering the difficulties and the risk of management, is described methods off physic contention in snakes, including equipments and cautions. Was used hook, manual contention, barrows contention, Lutz s tie, bag of fabric and fabric, contention and transport box and bedding foam in 130 animals for 4 families, Boidae, Viperidae, Elapidae and Colubridae. The snakes physics contention techniques described in these opportunity was effective, provided that the conditions imposed by each state.

This study aimed to evaluate the different types (trench silo, surface, raffia bags and pile) and times of warehousing of coproduct wet corn gluten feed (WCGF). A completely randomized split plot with the types of storage as the portion and opening days of the silage silos as the subplot. The days of collection were: 0, 1, 3, 5, 7, 14, 28, 56 days of storage. There were four treatments with three replications, totalizing 12 experimental units. The WCGF was charactirezed to temperature, pH, analysis of dry matter (DM), ether extract (EE), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber, in vitro digestibility and microbiological analysis to quantify microorganisms (yeasts and molds, lactobacilli and clostridia) of the WCGF from the experimental units. All pH values found were above the pH of ideal silage that would be of 4.2. Regarding temperature, the opening days had effect from the 7th day for the silo raffia bags and the 14th for the other trataments. Through regression it was observed that in general, the most of the values of DM, CP and DIVMS were linear behavior, while the FDA were growing and the values of NDF and EE had quadratic behavior. Treatments and days in microbiological analysis were compared, the treatment was significant only for yeasts and molds, with lower CFC counts, respectively, to the surface silo, trench silo, raffia bags and pile, for lactobacilli and clostridia had relevance only for days, the highest values found were on the 56 th day. Generally, the pH, temperature, chemical composition indicate unsuitable microbial fermentation in all treatments, it is suggested further studies about using of inoculants to enable the addition of lactic acid that could enhance product stability in conservation.

The giant anteater, Myrmecophagidae representative of the family, has a wide geographic distribution and is threatened with extinction due to their habits of life and human occupation, which is restricting its area of life. This study aimed to describe the origins and distributions of nerves suprascapular, subscapular and thoracodorsal cranial and caudal in giant anteater (Myrmecophaga tridactyla). We used three adult specimens, one female and two males, weighing approximately 40 kg, fixed in formaldehyde solution 10%. In the three specimens studied the suprascapular nerve originated from the ventral rami of the sixth (C6) and seventh (C7) cervical spinal nerves and sent branches to the muscles supraspinatus and infraspinatus. Subscapular nerves cranial and caudal originated from the ventral rami of the sixth (C6) and seventh (C7) cervical spinal nerves and distributed to the subscapularis muscle and teres major muscle. The thoracodorsal nerve originated in the ventral rami of the sixth (C6), seventh (C7) and eighth (C8) and first cervical spinal nerves (T1) thoracic spinal nerve, and sent branches to the latissimus dorsi muscle.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

This study aimed to determine the minimum time after the contact of commercial white and red eggs with material contaminated by Salmonella Heidelberg and its penetration to the inner side of the eggs. White and red eggs recently collected from among layer hens aged between 45 50 weeks, proceeding from a commercial farm in Uberlândia MG, were artificially contaminated by contact with wettish ribbands in inoculated liquid at a stagnant phase of growth (103-104 UFC.g-1 of Salmonella Heidelberg). In the study of each kind of egg, the eggs were subdivided into the following groups: the negative control (without artificial contamination), the positive control (analyzed externally after contamination and internally after the maximum period of maintenance of the test group) and the test group, four repetitions were conducted. The eggs of the test group were maintained at temperature between 25ºC and 30ºC and its contents were analyzed after 60, 90, 120, 150, 180, 210 and 240 minutes, as for the presence of Salmonella in 25g of sample. Each sample unit was compounded of five eggs. The analysis protocol used was the traditional one with pre-enrichment, selective enrichment, plateled in selective agar, biochemical and serological tests. The results obtained were submitted to logistics regression that indicated that the presence of Salmonella Heidelberg in the inner side of the red eggs happens after 168 minutes of contact and in the white eggs after 141 minutes of contact.

The morphological study, in particular the anatomical, contributes significantly to the applied areas. Hence the importance of studying the facial arteries in fetuses zebul-crossed cattle, which were obtained from slaughterhouses in Uberlândia MG, 20 males and 10 famales. The arterial system received injenção neoprene latex 450 50% (Du Pont do Brazil SA Chemical Industries) via the right common carotid arteries and left. The pieces were fixed in aqueous 10% formaldehyde by subcutaneous injection, intracavitary and intramuscular, as well as his immersion in this solution for a minimum of 48 hours. And subsequently dissected. It was observed that the right facial artery originated from the trunk linguofacial by 90%, and the remaining 10% emerged directly from the external carotid artery and the left originated from the trunk linguofacial in 90% and the remaining 10% emerged from the carotid artery both external and irrigate the digastric, irrigate the digastric, buccinator, depressor upper and lower lip, the zygomatic, nasobabial elevator, lift the upper lip, canine and masseter.

Campylobacter is considered a fragile microorganism ans sensitive to environmental conditions, but demostrate strategies to survive in unfavorable environmental conditions. This study evaluated the viability and production of transcipts of the genes sodB, p19 ciaB and dnaJ in strains ATCC 33291, NCTC 11351 and IAL 2383 stored in UHT milk and neopeptona + 12% glycerol,whether or not subject to the pre-treatment temperature of 4°C or 10°C for 30 minutes.Analyses were performed immediately after freezing in liquid nitrogen (day 0) and after maintenance for 30, 60 and 90 days at -20°C.The viability was evaluated by traditional method of cultivation and production of transcripts by RT-PCR technique. The quantification was only possible on the first day of analysis (day 0) and had a mean of 3.0x107UFC and in the remaining periods of storage strains showed confluent growth not allowing their enumeration. The set of results has shown that the UHT milk was the most appropriate for cryopreservation than the use of neopeptona +12% glycerol. The pretreatment at 4°C for 30 minutes favored the production of transcripts for ciaB and dnaJ genes. For the strains ATCC 33291 and NCTC 11351 was verified a possible interconnection of sodB genes and p19, however, this link was not observed for the strain IAL 2383, which also showed different behavior from other strains for viability in both cryoprotectants and production of transcripts. The results of this study show that when the maintenance of viability of the strains is essential, it is necessary to use different combinations of cryoprotectants / treatments to increase the chances of recovery and, when the primary purpose is the production of transcripts, the option to maintain the reliability of the results is the immediate extraction of DNA of isolated strains.

It is proposed to evaluate the induction period, muscle relaxation and recovery, as well as adverse effects of propofol in jacaré-tinga (Caiman crocodilus) using the cervical sinus vertebral as a means of application. Ten specimens were used of caiman who received a dose of 5 mg / kg of propofol intravenously in vertebral venous sinus in the neck. Evaluated the physiological parameters of cloacal temperature and heart and respiratory rates, as well as the reaction postural righting, muscle relaxation, head support and response to nociceptive stimulation. Evaluated the periods of sensory and motor block, and calculated the latency periods, skilled anesthesia and recovery. In this study don t occurred sensory block in the animals, motor block was 88.5 ± 55.35 minutes, the latency period was 57 ± 9.77 seconds, there was no reasonable period of anesthesia and recovery was 34.1 ± 0.2 minutes. Apnea occurred in 90% of the animals and decreased respiratory rate. It is concluded that the use of propofol at a dose of 5 mg / kg applied to the cervical sinus vertebral in jacaré-tinga Caiman crocodilus produces a rapid period of latency with good muscle relaxation and rapid recovery does not cause sensory block and its adverse effect cause was apnea.

The weight´s effect on egg´s quality, performance characteristics and incubation of chicks were observed in two experiments. Three treatments were used, defined by the weight of the eggs, breeding Leghorn 38 weeks old: small (S) 53 to 57g, medium (M) 58 to 62g and large (L) 63 to 67g. We have evaluated: a) the eggs - strength and shell thickness, Haugh units (HU), albumen height and percentage of egg compounds b) in incubation - eggs weight loss , hatching and embryo diagnosis, c) in one day old chicks in the yolk sac: weight and length of the chick, absolute and relative weight of the yolk sac, d) dry material (DM), crude protein (CP) albumen´s energy , and the yolk sac. Eggs range of different weights produced by Leghorn breeders of the same age (38 weeks) did not show differences among them for shelll strength, relative weight of albumen and yolk, albumen and yolk in DM and gross energy in the yolk. There are differences in the shell thickness, UH, albumen height, absolute weight of egg components, the relative weight of shell, the yolk and albumen protein and gross energy albumen. In the incubation, the weight loss of eggs and the hatch decreases with increasing egg weight range. Unhatched eggs increase with the increase of initial mortality and pecked alive and dead. The chick weight, absolute and relative weight of the yolk sac are directly proportional to the weight of the egg. The proportion of protein in the yolk sac decreases with increasing egg weight and gross energy content is higher in the yolk sac of chicks hatched from medium eggs.