Repositório RCAAP

CHAPTER 2: The goal of this research was to evaluate the performance of laying hens in second laying cycle, fed with minerals from organic source. The control treatment consisted of diets with inorganic mineral premix, and other treatments consisted of organic copper, iron, manganese and zinc organics in levels of 100%, 90%, 80% and 70% of inorganic minerals levels. The specific gravity and percentage of body checked eggs did not suffer influence from treatments. The egg weight had a decrease when birds were fed with organic minerals. The egg production percentage was higher when 100% of the organic minerals were included, while the feed conversion ratio better on treatments with addition of 100% and 90% organic minerals. Other addition levels provided percentage of and feed conversion ratio statistically equal to the control. Total replacement of inorganic trace minerals for equal levels of organic minerals in the diet of laying hens provided improvement in the performance. When replacement resulted in up 70% of that inclusion of inorganic minerals were maintenance of performance parameters. CHAPTER 3: The aim of this study was to evaluate the influence of the copper, iron, manganese and zinc minerals in organic form, associated to the internal and external quality parameters on hen eggs in their second production cycle. Such treatments consisted of the total replacement of this trace minerals in their inorganic form by the addition of organic mineral at levels of 100%, 90%, 80% and 70% commonly used in commercial farms. Physical and chemical analyses were performed to evaluate quality of eggs. Percentage of albumen, yolk and eggshell did not suffer influence from treatments. Percentage of protein decreased with the use of organic minerals while the mineral matter increased. The eggshell quality parameters had an improve with the addition of organic minerals on the diet of laying hens. The inclusion of minerals from organic source to laying hens diet in their second production cycle provided a highest percentage of mineral matter, in albumen and yolk, and better eggshell quality.

CHAPTER 2: The objective of this research was to evaluate the effects of fasting time on bromatology carcass, biometrics organ, water intake and performance of broiler chickens. We conducted a randomized design consisting of four days of fasting after hatching (0, 12, 24 and 36 hours) and four replications. Therefore, 640 birds were distributed in sixteen boxes containing 20 females and 20 males each. Water consumption was greater the longer the duration of fasting and was greater weight loss from hatching to the accommodation. There was also the weight loss on the yolk sac. Fasting influenced the relative weight of all organs of the digestive tract except esophagus + craw that remained unchanged. With increasing fasting time increased the relative weight of the esophagus+craw, proventriculus+gizzard, intestine+ pancreas, intestine and liver+gallbladder. However, food science and animal performance were not affected by fasting. CHAPTER 3: The objective of this research was to evaluate the effect of maltodextrin on performance and biometry of the digestive organs in chicks underwent four fasting time. Was performed a completely randomized design consisting of three treatments (1 - control, 2 - 4% sugar; 3-4% maltodextrin 20) and four times of fasting after hatching (0, 12, 24 and 36 hours) with four replications , 1920 birds in total of both sexes. The results showed that both the maltodextrin as sugar stimulated the consumption of water and after 24 hours, the group that has not been pass of fasting and was approved with maltodextrin obtained with greater weight of the small intestine and pancreas, besides a greater length of the gastrointestinal tract. The performance at 42 days was not influenced by the use of additives, however, the viability suffered positive influence after seven days of life.

In order to know more about the energy metabolic profile of crossbred cows during the peripartum, blood samples were collected from 36 animals. For each type of metabolism is dosed serum concentrations of these metabolites: total proteins, albumin and globulin to the protein profile, calcium, phosphorus and magnesium for the mineral profile, AST, ALT, GGT and alkaline phosphates for the enzyme portion and finally, non-esterifies fatty acids (NEFA), ß-hydroxybutyrate (BHB), triglycerides, cholesterol and lipoproteins (VLDL, HDL and LDL) to the energy profile. The measurements were performed in seven different times (D-7, D0, D7, D14, D21, D28 and D43). The results of protein profile indicated a hypoproteinemia, both by albumin and globulin; the parturition demonstrated influence on the mineral profile, especially in the amounts of calcium, wich can be noted subclinical hypocalcemia, but nevertheless no animal showed symptoms for this disorder on clinical trials. The enzyme concentrations, did not represent any significant change, only AST during periods close to birth, showed an increase in its concentration. Finally, there was no significant changes to indicate severe energy deficiency, since there were no losses in the BCS and milk production is not influenced by the fact these animals are of low and medium production. It is concluded that probably the diet of these animals is lower in protein than necessary, have no energy deficit and liver damage and birth is influential on the mineral concentrations.

Nowadays there are no more discussions related to the efficience use of sorghum in feed of broilers as well as anatomo-physiological changes triggered by the larger particle size of the ration particle. The success of the combination of these factors can lead to great economic advantage in the modern poultry industry. In this sense, three experiments were carried out in order to evaluate the effect of whole grain sorghum compared to maize and milled sorghum on the gastrointestinal tract and on carcass characteristics of broilers at 7 and 42 day old. The whole sorghum grain: (1) In broilers at 7 and 42 day old it provides greater body weight, due to its higher digestibility and increased gizzard weight by exercising greater mechanical work because of the larger particle size; (2) It increases duodenal crypt depth at 7 and 42 day old and jejunum at 42 day old without, however, affecting the characteristic of the villi of the small intestine and absorption surface area of intestinal mucosa in both evaluated age, (3) It does not affect the relative weight and length of the intestine, (4) It can replace corn without harming the commercial cuts yield as well as the chemical characteristics of the carcass; (5) It increases the visceral content and it is not advantageous for the carcass yield; for this purpose it is recommended to use the milled grain sorghum; (6) It excludes costs with grinding reducing the production cost of poultry.

The infection of humans with Salmonella, particularly by serovars Enteritidis and Typhimurium is a worldwide public health concern. The analysis of this pathogen in birds feces and environment is a way to monitoring the infection in batches and to verify the need of introduction of controls, with recommendation of European Commission Regulation (EC) (No. 646/2007) that lots of birds are being analyzed for the presence of these serovars in phases prior to slaughter. Different methodologies can be used to research these microrganisms in environmental samples, however, most of them demands a long time to obtain the results. Thus, it is desirable that other methods, faster and more practical have their performance verified for possible deployment in routine. The aim was to evaluate the equivalence between the results obtained by the methods: BAX ®, Half Semi-solid Rappaport- Vassiliadis Modified (MSRV) (ISO 6579) method and the traditional official reference in Brazil (Ordinance 126, MAPA) for Salmonella Typhimurium and S. Enteritidis in poultry environmental samples. Prope and feces samples fortified with an average of 100 to 1000 CFU/g of each serovar, and the same samples were evaluated without fortification by the three methods. Were obtained 504 diagnoses that were analyzed for the same repeatability, reproducibility, sensitivity, specificity and accuracy. These indices were conducted individually and in relation to the traditional method (relative rates). The results indicated that the methods analyzed showed satisfactory performance and compliance testing verified the correspondence between the alternative methods and official method, which allow us to affirm that the methods have equivalent performances.

We analyzed the distribution of the nervus fibularis communis and tibialis in three giant anteaters (Myrmecophaga tridactyla) from the collection of Animal Anatomy Laboratories at Universidade Federal de Uberlândia and Universidade Federal de Goiás, which were injected and preserved in an aqueous solution of 10% formaldehyde. It was noted that those nerves derive from the ischiadicus, and the fibularis communis divided into nervus fibularis profundus and superficialis, distributing to the musculis abductor cruris caudalis, biceps femoris, extensor digitorum lateralis, longus and brevis, fibularis longus and brevis, and tibialis cranialis; The nervus tibialis innerved the musculis flexor digitorum superficialis and profundi, gastrocnemius and popliteus. Drawing an imadinary line in the mid-lateral region of the tuberositatis tibia, the nervus fibularis communis may be blocked in its proximal portion, the caudal third, between the tendon of insertion of the biceps femoris and lateral side of musculus gastrocnemius (middle third); and on the lateral side of the tibiotarsal joint, between the tendons of insertion of the musculi extensor digitorum lateralis and longus. Considering another imaginary line on the medial side of the leg, originating in the caudal region of the condylus medialis tibia, the nervus tibialis may be blocked in its proximal third, between the lateral and medial portions of the musculus gastrocnemius, and distally, between the tuber calcanei and the tibia caudodistal face.

The purposes of this study were to describe macro and microscopic patterns of cutaneous wound healing on Blue-fronted parrots and to evaluate the influence of the polihexanide compound in this process. An experiment has been taken place within 20 animals sorted into 5 groups. After the anesthetic induction, two small pieces of skin within 8,0 mm diameter were taken from their chest. A histological evaluation were carried out in 4 groups and in the 5th one an macroscopic and morfometric analyses. The wounds of the right hand side were treated with sodium chloric solution at 0,9% and the left hand side applied polihexanide gel solution at 0,1%. For histological evaluation, the cuts were collected in the following days: 3, 7, 14 and 21. After macro and microscopic analyses no significant pattern were observed among the treatments. In the first day, the wounds were reddish and bleeding though no caseous secretion was seen. By the 4th day tissue granulation started to develop. The cutaneous regeneration were observed in the 21st day and the hauling a bit later, in the 25th. The microscopic results of the 3rd and 7th days befitted the healing sore process phase. In the 14th day lymphocytes have been seen mainly. The blood vessels were maintained congested and the neovascularization were observed intensely in the 3rd and 7th days. Along with healing process evolution, epidermal ulceration was not observed. The re-epithelization process has started on the 21st day within epithelial tissue recovering half of the wound area. The inflammation and healing process of the Amazona aestiva came together and its recovery patterns were similar to the ones reported on mammals. Daily wound antisepsis prevented microorganism proliferation. The use of polyhexamethylene biguanide hydrochloride (PHMB) has not improved the cutaneous recovery process significantly on Blue-fronted parrots.

The aim of the present study was to determine the influence of transport distance of cattle in winter and summer on pH 24 hours post mortem and occurrence of bruises. 420 cows were slaughtered in Uberlândia - MG and come from Triângulo Mineiro. The experimental design was completely randomized in a 2x2 factorial, with two distances (15-85 km and 140-200 km) and two seasons (summer and winter), with four replicates each, except trucks that traveled for 15-85 km in summer (nine replications). The research was developed in July 2011 and January 2012. The distance on unpaved roads in the route 15-85 km was 10-18 km (winter) and 4-10 km (summer). On the other side, in the route 140-200 km, the animals traveled for 13-30 km (winter) and 12-18 km (summer). The occurrence, location, amount and degree of recent lesions, and pH 24h were assessed. Quantitative variables were subjected to analysis of variance followed by medium test, while the qualitative ones were evaluated through chi-square test. The pH value of 5.908 in carcasses of cattle transported in winter was higher than in summer - 5.711, not varying in the measured distances. The frequency of injuries in animals transported in winter - 97.78% - was higher than in summer - 88.26%. 77.39% of them were injured in rear and 94.02% presented grade I. The amount of injuries ranged from 1.9 to 2.36 kg by injured carcass, with a total weight of 168.575 kg. There was a greater presence of bruising in cattle transported over greater distances - 97.45% - than in short distances - 85.95%. Cows transported by road for up to 200 km have no change in pH 24h, but longer distances cause more bruising, especially grade I in the rear. In winter, there is a higher frequency of injuries than in summer.

The objective was to evaluate the internal and external quality of hens eggs, classified in eggs with normal eggshell and eggs with vitreous eggshell, produced by hens in three different ages. Forty-five eggs of each eggshell type, normal and vitreous, were sampled at age of 30, 50 and 70 weeks. After the execution of physical and chemical analysis, it was observed that specific gravity, for both eggs with normal eggshell and eggs with vitreous eggshell, had a decrease with the advancing age of the hen. The weight of the egg, in both types of eggshell increased with the advancing age of the hen. Higher concentration of calcium was found on eggs with normal eggshell comparing to eggs with vitreous eggshell. The phosphorus percentage wasn t affected by the eggshell type. The mineral deposition of the eggshell was constant, the thickness of the eggshell in both eggshell types increased throughout the reproductive life of the hen and decreased when the hen got older. The albumen weight increased, showing its relation with the egg size. The percentage of albumen moisture with the advancing age of the hen didn t show any relation with the egg weight. Protein concentration and pH of the albumen decreased along with hen ages. The weight of egg yolk, the moisture percentage and the yolk pH decreased with the advancing age of the hen. Crude protein and ether extract increased with the advancing age of the hen. Eggs with vitreous eggshell showed eggshell with lower quality compared to those with normal eggshell, without, however, compromise the internal quality of albumen and yolk.

With the intensification of the agricultural sector, the need to develop new cultivars of tropical forage increases, which have better yield capacity and nutritional quality that suits Brazil s soil and climate conditions. The purpose of this study was to evaluate livestock, nutritional and morphological aspects of the Brachiaria hybrid cv. Mulato II and Brachiaria brizantha cv. Piata. The experiment was conducted on the Capim Branco Farm in Uberlândia Federal University. The experimental design used for the evaluation was done in randomized blocks (DRB). For the forages evaluation it was used a split-plot in time system. It was admitted two treatments - the cultivar Mulato II and Piata. We have evaluated the tiller density, leaf proportion, stem and dead material, crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin content, in vitro degradability of dry matter (IVDDM), weight gain and average grazing pressure. The Cultivar Mulato II had greater (p<0.05) tiller density and leaf: stem in relation to Piata. Regarding the FDA, FDN and lignin content, the Piata was significantly (p<0.05) superior than cultivar Mulato II. As for the PB content and DIVMS, the Cultivar Mulato II showed superiority (p<0.05) compared to Piata. There was no significant difference (p>0.05) among cultivars in relation to the average grazing pressure, however, the weight gain was higher (p<0.05) to the animals that underwent the cultivar Mulato II.

The dissertation was divided in two chapters. The aim was to determine whether hens immunized with an inactivated suspension of Leptospira and a solution of outer membrane proteins extracted from the serovar Hardjo, could produce specific polyclonal antibodies to Leptospira, detected in ELISA assay. Eight hens White Leghorn race with 25-weeks-old were immunized, three with an inactivated suspension of Leptospira, three with a solution of outer membrane proteins (OMP) extracted from the serovar Hardjo and two controls immunized with saline. Blood samples were collected fortnightly and eggs daily. The IgY was purified from the egg yolk using the method for the delipidation of dilution with water acidic and ammonium sulfate precipitation. The ELISA assay was performed to verify the specificity of the IgY, these was possible to observe the production of specific antibody to Leptospira both in serum and purified egg yolk. The specific antibody titers peaked in the fifth week post immunization. The production of polyclonal IgY was effective for producing high titers of specific antibodies. The aim of the second chapter was search to epitopes or mimetops (mimetic sequence of the true epitopes) through technique of phage display using IgY specific to Leptospira, outer membrane proteins the serovar Hardjo and IgY control. To realize the biopanning was used a commercial library of random peptides Ph.D. 12 mer, New England BioLabs®Inc Phage Display Library was first incubated with IgY control, performing a process called negative selection, and then with IgY specific to Leptospira and to OMP Leptospira Hardjo, selected phages were eluted competitively and acid. After three rounds of selection, 288 clones were isolated and sequenced, generating 132 valid sequences. The selected clones were tested in diagnosis by ELISA assay with the antibody used as a biopanning target in order to verify the specificity of these. The results of ELISA testing it was found that nine phage (PMEcomp17, PMEac10, PMEac12, PMEac19, PMEac29, PMEac35, Lep6, Lep7 e Lep20) showed better results. These were aligned with the primary sequence of proteins of Leptospira spp. and Leptospira interrogans serovar Hardjo and insilico analyzes showed that nine peptides has similarity with immunogenic proteins of Leptospira. In this work we selected nine peptides with similarity to proteins of Leptospira interrogans that were considered immunogenic. The peptides were similar to outer membrane proteins, lipoprotein, LipL32, LipL41, LigA, SecY, LenE.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Fundação de Amparo a Pesquisa do Estado de Minas Gerais

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.