Fatores que influenciam a produção de embriões de vacas Nelore (Bos taurus indicus) superovuladas

Pós-graduação em Medicina Veterinária - FCAV

Obtenção de oócitos e produção in vitro de embriões em doadoras lactantes da raça Gir (Bos taurus indicus)

Pós-graduação em Medicina Veterinária - FCAV

Avaliação dos efeitos da inibição da maturação nuclear e de antioxidantes sobre a maturação oocitária, fecundação e desenvolvimento embrionário bovino in vitro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of different cryopreservation methods on post-thaw culture conditions of in vitro produced bovine embryos

The aim of this work was to evaluate the effect of cryopreservation protocols on subsequent development of in vitro produced bovine embryos under different culture conditions. Expanded in vitro produced blastocysts (n = 600) harvested on days 7-9 were submitted to controlled freezing [slow freezing group: 10% ethylene glycol (EG) for 10 min and 1.2 degrees C/min cryopreservation]; quick-freezing [rapid freezing group: 10% EG for 10 min, 20% EG + 20% glycerol (Gly) for 30 s]; or vitrification [vitrification group: 10% EG for 10 min, 25% EG + 25% Gly for 30 s] protocols. Control group embryos were not exposed to cryoprotectant or cryopreservation protocols and the hatching rate was evaluated on day 12 post-insemination. In order to evaluate development, frozen-thawed embryos were subjected to granulosa cell co-culture in TCM199 or SOFaa for 4 days. Data were analyzed by PROC MIXED model using SAS Systems for Windows (R). Values were significant at p < 0.05. The hatching rate of the control group was 46.09%. In embryos cultured in TCM199, slow freezing and vitrification group hatching rates were 44.65 +/- 5.94% and 9.43 +/- 6.77%, respectively. In embryos cultured in SOFaa, slow freezing and vitrification groups showed hatching rates of 11.65 +/- 3.37 and 8.67 +/- 4.47%, respectively. In contrast, the rapid freezing group embryos did not hatch, regardless of culture medium. The slow freezing group showed higher hatching rates than other cryopreservation groups. Under such conditions, controlled freezing (1.2 degrees C/min) can be an alternative to cryopreservation of in vitro produced bovine embryos.

Heparin and penicillamine–hypotaurine–epinephrine (PHE) solution during bovine in vitro fertilization procedures impair the quality of spermatozoa but improve normal oocyte fecundation and early embryonic development

The presence of heparin and a mixture of penicillamine, hypotaurine, and epinephrine (PHE) solution in the in vitro fertilization (IVF) media seem to be a prerequisite when bovine spermatozoa are capacitated in vitro, in order to stimulate sperm motility and acrosome reaction. The present study was designed to determine the effect of the addition of heparin and PHE during IVF on the quality and penetrability of spermatozoa into bovine oocytes and on subsequent embryo development. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes and mitochondrial function, was diminished (P<0.05) in the presence of heparin and PHE. Oocyte penetration and normal pronuclear formation rates, as well as the percentage of zygotes presenting more than two pronuclei, was higher (P<0.05) in the presence of heparin and PHE. No differences were observed in cleavage rates between treatment and control (P>0.05). However, the developmental rate to the blastocyst stage was increased in the presence of heparin and PHE (P>0.05). The quality of embryos that reached the blastocyst stage was evaluated by counting the inner cell mass (ICM) and trophectoderm (TE) cell numbers and total number of cells; the percentage of ICM and TE cells was unaffected (P>0.05) in the presence of heparin and PHE (P<0.05). In conclusion, this study demonstrated that while the supplementation of IVF media with heparin and PHE solution impairs spermatozoa quality, it plays an important role in sperm capacitation, improving pronuclear formation, and early embryonic development

Photobiological effect of low-level laser irradiation in bovine embryo production system

The objectives of this study were to evaluate the effect of low-level laser irradiation (LLLI) on bovine oocyte and granulosa cells metabolism during in vitro maturation (IVM) and further embryo development. Cumulus-oocytes complexes (COCs) were subjected (experimental group) or not (control group) to irradiation with LLLI in a 633-nm wavelength and 1 J/cm2 fluency. The COCs were evaluated after 30 min, 8, 16, and 24 h of IVM. Cumulus cells were evaluated for cell cycle status, mitochondrial activity, and viability (flow cytometry). Oocytes were assessed for meiotic progression status (nuclear staining), cell cycle genes content [real-time polymerase chain reaction (PCR)], and signal transduction status (western blot). The COCs were also in vitro fertilized, and the cleavage and blastocyst rates were assessed. Comparisons among groups were statistically performed with 5% significance level. For cumulus cells, a significant increase in mitochondrial membrane potential and the number of cells progressing through the cycle could be observed. Significant increases on cyclin B and cyclin-dependent kinase (CDK4) levels were also observed. Concerning the oocytes, a significantly higher amount of total mitogen-activated protein kinase was found after 8 h of irradiation, followed by a decrease in all cell cycle genes transcripts, exception made for the CDK4. However, no differences were observed in meiotic progression or embryo production. In conclusion, LLLI is an efficient tool to modulate the granulosa cells and oocyte metabolism

Quantificação de proteínas de fase aguda em éguas doadoras de embrião da raça quarto de milha

Pós-graduação em Ciência Animal - FMVA

Influência da resposta vacinal contra o herpesvírus bovino tipo 1 (BoHV-1) nos parâmetros reprodutivos de vacas receptoras de embrião

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito das características uterinas e dia do ciclo na taxa de prenhez e níveis séricos de progesterona em éguas candidatas à receptora de embrião

Pós-graduação em Medicina Veterinária - FMVZ

Soroprevalência, isolamento e diagnóstico molecular para leptospirose em bovinos de fazendas de cria e de centrais de receptoras de embrião da região de Bauru- SP: Diagnóstico de leptospirose bovina em Bauru - SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biological performance of a polycaprolactone-based scaffold used as fusion cage device in a large animal model of spinal reconstructive surgery

A bioactive and bioresorbable scaffold fabricated from medical grade poly (epsilon-caprolactone) and incorporating 20% beta-tricalcium phosphate (mPCL–TCP) was recently developed for bone regeneration at load bearing sites. In the present study, we aimed to evaluate bone ingrowth into mPCL–TCP in a large animal model of lumbar interbody fusion. Six pigs underwent a 2-level (L3/4; L5/6) anterior lumbar interbody fusion (ALIF) implanted with mPCL–TCP þ 0.6 mg rhBMP-2 as treatment group while four other pigs implanted with autogenous bone graft served as control. Computed tomographic scanning and histology revealed complete defect bridging in all (100%) specimen from the treatment group as early as 3 months. Histological evidence of continuing bone remodeling and maturation was observed at 6 months. In the control group, only partial bridging was observed at 3 months and only 50% of segments in this group showed complete defect bridging at 6 months. Furthermore, 25% of segments in the control group showed evidence of graft fracture, resorption and pseudoarthrosis. In contrast, no evidence of graft fractures, pseudoarthrosis or foreign body reaction was observed in the treatment group. These results reveal that mPCL–TCP scaffolds could act as bone graft substitutes by providing a suitable environment for bone regeneration in a dynamic load bearing setting such as in a porcine model of interbody spine fusion.

The evaluation of a biphasic osteochondral implant coupled with an electrospun membrane in a large animal model

Conventional clinical therapies are unable to resolve osteochondral defects adequately, hence tissue engineering solutions are sought to address the challenge. A biphasic implant which was seeded with Mesenchymal Stem Cells (MSC) and coupled with an electrospun membrane was evaluated as an alternative. This dual phase construct comprised of a Polycaprolactone (PCL) cartilage scaffold and a Polycaprolactone - Tri Calcium Phosphate (PCL - TCP) osseous matrix. Autologous MSC was seeded into the entire implant via fibrin and the construct was inserted into critically sized osteochondral defects located at the medial condyle and patellar groove of pigs. The defect was resurfaced with a PCL - collagen electrospun mesh that served as a substitute for periosteal flap in preventing cell leakage. Controls either without implanted MSC or resurfacing membrane were included. After 6 months, cartilaginous repair was observed with a low occurrence of fibrocartilage at the medial condyle. Osteochondral repair was promoted and host cartilage degeneration was arrested as shown by the superior Glycosaminoglycan (GAG) maintenance. This positive morphological outcome was supported by a higher relative Young's modulus which indicated functional cartilage restoration. Bone in growth and remodeling occurred in all groups with a higher degree of mineralization in the experimental group. Tissue repair was compromised in the absence of the implanted cells or the resurfacing membrane. Moreover healing was inferior at the patellar groove as compared to the medial condyle and this was attributed to the native biomechanical features.

Monitoring animal behaviour and environmental interactions : GPS collars and satellite remote sensing within a satellite remote sensing context

Remote monitoring of animal behaviour in the environment can assist in managing both the animal and its environmental impact. GPS collars which record animal locations with high temporal frequency allow researchers to monitor both animal behaviour and interactions with the environment. These ground-based sensors can be combined with remotely-sensed satellite images to understand animal-landscape interactions. The key to combining these technologies is communication methods such as wireless sensor networks (WSNs). We explore this concept using a case-study from an extensive cattle enterprise in northern Australia and demonstrate the potential for combining GPS collars and satellite images in a WSN to monitor behavioural preferences and social behaviour of cattle.

Monitoring animal behaviour and environmental interactions using wireless sensor networks, GPS collars and satellite remote sensing

Remote monitoring of animal behaviour in the environment can assist in managing both the animal and its environmental impact. GPS collars which record animal locations with high temporal frequency allow researchers to monitor both animal behaviour and interactions with the environment. These ground-based sensors can be combined with remotely-sensed satellite images to understand animal-landscape interactions. The key to combining these technologies is communication methods such as wireless sensor networks (WSNs). We explore this concept using a case-study from an extensive cattle enterprise in northern Australia and demonstrate the potential for combining GPS collars and satellite images in a WSN to monitor behavioural preferences and social behaviour of cattle.

The design and evaluation of a mobile sensor/actuator network for autonomous animal control

This paper investigates a mobile, wireless sensor/actuator network application for use in the cattle breeding industry. Our goal is to prevent fighting between bulls in on-farm breeding paddocks by autonomously applying appropriate stimuli when one bull approaches another bull. This is an important application because fighting between high-value animals such as bulls during breeding seasons causes significant financial loss to producers. Furthermore, there are significant challenges in this type of application because it requires dynamic animal state estimation, real-time actuation and efficient mobile wireless transmissions. We designed and implemented an animal state estimation algorithm based on a state-machine mechanism for each animal. Autonomous actuation is performed based on the estimated states of an animal relative to other animals. A simple, yet effective, wireless communication model has been proposed and implemented to achieve high delivery rates in mobile environments. We evaluated the performance of our design by both simulations and field experiments, which demonstrated the effectiveness of our autonomous animal control system.