Lidocaine administered at a continuous rate infusion does not impair left ventricular systolic and diastolic function of healthy rabbits sedated with midazolam.

Lidocaine is a versatile drug that not only provides local anesthesia, but also reduces anesthetic requirements of other agents and has antiarrhythmic, pro-kinetic, anti-inflammatory, antiendotoxemic and antioxidant effects. As it is a drug commonly used in critically ill patients, its safety from the cardiovascular system should be ensured. The aim of this study was to determine the effects of a continuous rate infusion (CRI) of lidocaine on left ventricular systolic and diastolic function of healthy rabbits sedated with midazolam by use of transthoracic echocardiography. Ten New Zealand healthy rabbits were sedated with intramuscular midazolam (1 mg/kg) and enrolled in two experimental treatments (control or lidocaine). The control treatment (CT) comprised an intravenous bolus of 0.9% sodium chloride (0.05 mL/kg) followed by CRI at 5 mL/h, whereas the lidocaine treatment (LT) comprised a bolus of 2% lidocaine without epinephrine at 1 mg/kg followed by CRI at 50 µg/kg/minute. Echocardiographic and hemodynamic variables were studied. Variables were recorded at baseline (TB) and 20, 40 and 60 minutes following start of CRI (T20, T40 and T60, respectively). No differences were found between treatments. The results of this study demonstrate that a continuous rate infusion of lidocaine at 50 µg/kg/minute does not impair echocardiographic indices of left ventricular systolic and diastolic function of healthy rabbits sedated with midazolam.

Necessidade de orientação dos tutores sobre posse responsável canina / Necessity guidance of tutors of canine responsible ownership

O objetivo dessa pesquisa foi investigar a aplicação de medidas básicas de posse responsável por tutores de cães. Para isso, foram entrevistados 50 proprietários. As questões eram referentes à procedência, detalhes ambientais e ações preventivas adotadas por eles. Apesar de alguns donos possuírem noções básicas sobre os cuidados necessários para com seus cães, existe a necessidade de se orientar a comunidade sobre posse responsável de animais de estimação, o que pode ser considerado essencial para controlar a propagação de enfermidades caninas.(AU)

This researchs goal was to investigate the application of basic measures of responsible ownership by dog tutors. For that, 50 owners were interviewed. The questions were related to origin, environment details and preventive actions adopted by them. Although some owners had basic notions about the needed care toward their dogs, there is the need to guide the community about responsible ownership of pets, that can be considered essential to control the spread of canine diseases

NECESSIDADE DE ORIENTAÇÃO DOS TUTORES SOBRE POSSE RESPONSÁVEL CANINA

O objetivo dessa pesquisa foi investigar a aplicação de medidas básicas de posse responsável por tutores de cães. Para isso, foram entrevistados 50 proprietários. As questões eram referentes à procedência, detalhes ambientais e ações preventivas adotadas por eles. Apesar de alguns donos possuírem noções básicas sobre os cuidados necessários para com seus cães, existe a necessidade de se orientar a comunidade sobre posse responsável de animais de estimação, o que pode ser considerado essencial para controlar a propagação de enfermidades caninas.

Necessidade de orientação dos tutores sobre posse responsável canina / Necessity guidance of tutors of canine responsible ownership

O objetivo dessa pesquisa foi investigar a aplicação de medidas básicas de posse responsável por tutores de cães. Para isso, foram entrevistados 50 proprietários. As questões eram referentes à procedência, detalhes ambientais e ações preventivas adotadas por eles. Apesar de alguns donos possuírem noções básicas sobre os cuidados necessários para com seus cães, existe a necessidade de se orientar a comunidade sobre posse responsável de animais de estimação, o que pode ser considerado essencial para controlar a propagação de enfermidades caninas.

This research’s goal was to investigate the application of basic measures of responsible ownership by dog tutors. For that, 50 owners were interviewed. The questions were related to origin, environment details and preventive actions adopted by them. Although some owners had basic notions about the needed care toward their dogs, there is the need to guide the community about responsible ownership of pets, that can be considered essential to control the spread of canine diseases

Effect of low oxygen tension atmosphere and maturation media supplementation on nuclear maturation, cortical granules migration and sperm penetration in swine in vitro fertilization.

The aim of this study was to evaluate the efficiency of low oxygen tension (5% CO(2) , 5% O(2) and 90% N(2) ) on in vitro oocyte maturation using defined media (0.1% polyvinyl alcohol - PVA) or 10% porcine follicular fluid (PFF)-supplemented media. To achieve this goal, oocytes were evaluated regarding cortical granules (GCs) migration, nuclear maturation and sperm penetration. Oocytes were in vitro matured under different conditions: 5% or 20% O(2) atmosphere and 0.1% PVA- or 10% PFF-supplemented media and evaluated at 0 and 44 h of maturation. To evaluate the migration of CGs and nuclear maturation, by confocal microscopy, oocytes were incubated with 100 µg of FITC-PNA/ml and 10 µg/ml of propidium iodide. To address sperm penetration, after maturation, in vitro fertilization for 6 h and in vitro culture for 18 h, zygotes were incubated with 10 mg/ml Hoechst 33342. Pronuclei and polar bodies were quantified using an epifluorescence microscope. Atmosphere conditions did not affect the CGs migration, but media supplementation did. Oocytes matured in 10% PFF media had a higher percentage of CGs in the oocyte periphery than oocytes matured in PVA-supplemented media. However, this fact did not have effect on in vitro sperm penetration levels. No effect of atmosphere conditions and media supplementation was observed on the rates of metaphase II oocytes. Therefore, the use of low oxygen tension in association with PVA maturation media does not improve the in vitro maturation system of porcine oocytes, because its use did not improve nuclear maturation, CGs migration and zygotes monospermic rates.

Use of chromomycin A3 staining in bovine sperm cells for detection of protamine deficiency.

Sperm chromatin integrity is essential for accurate transmission of male genetic information, and normal sperm chromatin structure is important for fertilization. Protamine is a nuclear protein that plays a key role in sperm DNA integrity, because it is responsible for sperm DNA stability and packing until the paternal genome is delivered into the oocyte during fertilization. Our aim was to investigate protamine deficiency in sperm cells of Bos indicus bulls (Nelore) using chromomycin A3 (CMA3) staining. Frozen semen from 14 bulls were thawed, then fixed in Carnoy's solution. Smears were prepared and analyzed by microscopy. As a positive control of CMA3 staining, sperm from one bull was subjected to deprotamination of nuclei. The percentage of CMA3-positive bovine sperm did not vary among batches. Only two bulls showed a higher percentage of CMA3-positive sperm cells compared to the others. CMA3 is a simple and useful tool for detecting sperm protamine deficiency in bulls.

Effects of serum deprivation and cycloheximide on cell cycle of low and high passage porcine fetal fibroblasts.

Arrest of cells in G0/G1 cell cycle phase is desired for nuclear transfer procedures. Serum starvation and cell cycle inhibitors are different ways to induce synchronization of the cell cycle. This study investigated the effects of serum starvation and cycloheximide (CHX) on the cell cycle of low (5th) and high (15th) passages fetal porcine fibroblasts. Cell cycle phases were determined using fluorescent activated cell sorting. Fifth passage fibroblast cultures had higher (p < 0.05) proportion of cells in G0/G1 only after 72 h of serum starvation (77.60 +/- 0.65) when compared with non-starved cells (71.44 +/- 1.88). Serum starvation for all periods tested induced an increase (p < 0.05) on proportion of cells in G0/G1 on the 15th passage. No significant differences were observed on the 5th passage cultures exposed to CHX, although, on the 15th passage an increase on proportion of cells was observed after all periods of exposure (p < 0.05). These data indicates that high passage cells in vitro are more susceptible to serum starvation and CHX G0/G1 synchronization.

Production of a cloned calf from a fetal fibroblast cell line

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5 percent) were successfully fused and 24 (28.9 percent) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8 percent) of which were pregnant on day 35 and 3 (16.7 percent) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

Production of a cloned calf from a fetal fibroblast cell line.

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).