Adriana Cristina da Silva

Role of insulin-like growth factor-I and follicular fluid from ovarian follicles with different diameters on porcine oocyte maturation and fertilization in vitro

The objective of this study was to determine the effects of insulin-like growth factor-I (IGF-I) (0, 60, 120, 180, and 240 ng/mL) and follicular fluid (FF) derived from 2 to 5 and 6 to 10 mm diameter follicles (SpFFs and LpFFs, respectively) added during in vitro maturation (IVM) of porcine oocytes on nuclear maturation and IVF. Cumulus-oocyte complexes (COCs) were matured in NCSU-37 medium supplemented with SpFFs or LpFFs and various IGF-I concentrations. The COCs were cultured for 44 hours, and then fertilized in vitro. Maturation and IVF results were recorded 18 hours after insemination. The IVM (%) was higher (P < 0.05) in the COCs matured in LpFFs than with SpFFs when 0 (90.0 ± 6.9 vs. 76.3 ± 10.7) or 60 ng/mL IGF-I (92.0 ± 8.1 vs. 81.8 ± 10.2) was added. In SpFFs media, there was a quadratic relationship (P < 0.01) between IGF-I concentration and IVM (peak results at IGF-I = 129 ng/mL). However, when the COCs were matured with LpFFs, there was a decreasing linear effect between IGF-I concentration and IVM. At all concentrations of IGF-I, the percentage of degenerated oocytes was higher in COCs matured in SpFFs than in LpFFs. Penetration (%) did not differ (P > 0.05) between COCs matured with SpFFs or LpFFs when 60 (66.8 ± 9.4 vs. 72.7 ± 11.3) or 180 ng/mL of IGF-I (75.7 ± 10.4 vs. 73.8 ± 13.2) were used. Monospermy (%) was similar between SpFFs and LpFFs only with addition of 120 ng/mL IGF-I. The IVF performance (%) did not differ between COCs matured with SpFFs or LpFFs when IGF-I concentrations of 120 (28.5 ± 8.8 vs. 38.5 ± 8.3) and 180 ng/mL (24.3 ± 10.2 vs. 30.12 ± 8.2) were used. There was no effect of IGF-I concentration or of FF type on the number of penetrated sperm per oocyte and on male pronuclear formation. For COCs matured with SpFFs, there was a quadratic relationship between IGF-I concentration and penetration, monospermy, and IVF performance (peak results at IGF-I = 179, 122, and 135 ng/mL, respectively). Thus, on the basis of the observed quadratic relationships, we inferred that when using SpFFs, the addition of IGF-I (122-179 ng/mL) to the IVM medium produced results similar to those obtained with LpFFs without adding IGF-I. In conclusion, the addition of IGF-I to the IVM medium supplemented with SpFFs increased maturation and improved IVF results. Alternatively, IGF-I had no effect on IVM or IVF when used with LpFFs.

Comparison of two different methods for evaluating boar semen morphology

El objetivo de este estudio fue comparar el uso y la eficacia de dos tecnicas para evaluacion de la morfologia espermatica del semen porcino. La preparacion humeda con una solucion de formaldehido-citrato 2,94% y otra con el uso del colorante Rosa de Bengala. Se evaluaron 58 eyaculados de ocho verracos de alto desempeno, con edad entre 20 y 25 meses. Despues de la recogida del semen, la morfologia espermatica fue evaluada por dos tecnicas: la preparacion humeda convencional (solucion de formaldehido-citrato 2,94%) observada en microscopio de contraste de fase (1.000X de magnificacion) y la tecnica que utiliza un frotis preparado a partir del colorante Rosa de Bengala, analizado bajo un microscopio optico (400X de magnificacion). Se observo que el porcentaje de cambios de la cabeza, acrosoma, pieza intermedia y el total de cambios morfologicos no fueron diferentes (P > 0,05) entre los dos metodos. Se obtuvo un mayor numero de defectos de la cola (P < 0,05) con el uso de la tecnica de tincion con el colorante Rosa de Bengala en comparacion con la tecnica convencional. Esto se explica por el manejo en la preparacion del frotis que puede conducir a una mayor probabilidad de romper la cola, lo que conduce a la visualizacion de la anomalia cola rota. Se puede concluir que el uso del colorante Rosa de Bengala es un metodo eficaz para la evaluacion de la morfologia del semen porcino, senalando que importantes cuidados deben ser tomados, especialmente en la preparacion del frotis para que no ocurran cambios en los resultados, principalmente en los cambios de la cola.

Spectral analysis of femoral artery blood flow waveforms of conscious domestic cats.

The qualitative and quantitative aspects of femoral artery blood flow waveform spectra were evaluated in 15 male and 15 female Persian and mixed breed domestic cats (Felis catus), which were healthy and not sedated, using duplex Doppler ultrasonography (DDU). Spectral Doppler demonstrated a biphasic characteristic in 16 (53.34%) of the animals evaluated, and a triphasic characteristic in the 14 (46.66%) remaining animals. The systolic blood pressure and heart rate values were within the normal range for the species. The quantitative parameters evaluated, based on the spectral Doppler, were as follows: systolic velocity peak (SVP), recent diastolic velocity peak (RDVP), end diastolic velocity peak (EDVP), mean velocity (MV), integral velocity time (ITV), artery diameter (AD), femoral flow volume (FFV), pulsatility index (PI), resistive index (RI), systolic peak acceleration time (AT) and deceleration time (DT). The respective mean values were: 36.41 ± 7.33 cm/s, 4.69 ± 0.90 cm/s, 10.74 ± 2.74 cm/s, 23.06 ± 4.86 cm/s, 3.91 ± 1.05 cm, 0.17 ± 0.04 cm, 0.11 ± 0.08 cm3, 3.85 ± 0.19, 1.40 ± 0.20, 39.84 ± 7.38 ms, and 114.0 ± 22.15 ms. No significant differences were found between males and females. The analyses carried out on the femoral artery flow spectrum obtained by DDU showed that it is easy to use and highly tolerated in non-sedated, healthy cats. It appears that DDU may be a useful diagnostic technique, but further studies are needed to evaluate how it compares with invasive telemetric methodology or high-definition oscillometric waveform analytic techniques.

Avaliação morfológica e morfométrica de ovários, tubas uterinas e útero de fêmeas suínas pré-púberes em diferentes fases do ciclo estral

Studies are performed in developing techniques/procedures that provide greater reproductive performance in farm animals, including pigs. In this sense, the study of gilts reproductive organs at different oestrus cycle stages for assessing the presence of abnormalities and/or other parameters that may affect the future animal fertility is important. In order to evaluate the morphological, morphometric and histomorphometric features of ovaries, uterus and uterine tubes (UTs) characteristics of prepubertal gilts at different oestrus cycle stages, reproductive tracts from 48 animals immediately after slaughter were obtained. After, the structures were dissected and removed, and the ovaries were used for classification of oestrus cycle stage of each gilt in follicular phase (FP) and luteal phase (FL). Then, morphometric evaluations of ovaries, UTs, uterine horns and uterine body were performed. Besides that, medial segments of UTs and uterus were fixed in Bouin solution, processed and included in paraffin, when histological sections of 5.0 micrometers (µm) were obtained and stained with Hematoxylin and Eosin. Histomorphometric analyzes using image capture system and specific software were performed. Afterwards, data were submitted to Students t test for assessment the statistical differences (P 0.05) between morphometric parameters of ovaries, UTs and uterus of gilts in FP and LP. Likewise, in respect to the placement of reproductive structures, both in the oestrus cycle stages, as in the general average, there was no difference (P>0.05). Regarding the histomorphometric variables, gilts classified in FP presented a higher (P<0.05) height of glandular and UT epithelium compared to animals in LP. On the other hand, the diameter of endometrial glands was higher (P<0.05) in gilts at LP compared to FP. Furthermore, gilts in LP presented a higher (P<0.05) proportion of endometrium occupied by glands, whereas animals in FP had a higher (P<0.05) proportion of connective tissue and blood vessels. In conclusion, in prepubertal gilts, the histomorphometric parameters as endometrial glands diameter, the height of glandular epithelium and of UT epithelium and the proportion of endometrium occupied by connective tissue, besides the glands and blood vessels varies through the oestrus cycle, possibly under the influence of ovarian steroids.

Porcine follicular fluid concentration of free insulin-like growth factor-I collected from different diameter ovarian follicles

The study aimed to quantify the concentrations of free IGF-I in serum and fluid of ovarian follicles in pre-pubertal gilts and describe the ovarian morphology by measuring the size of the ovaries and counting the number of surface follicles. Ovaries (n=1,000) from pre-pubertal gilts were obtained immediately after slaughter. A total of 10 samplings were performed, with ovaries obtained from 50 females for each collection. The follicles situated on the surface of each ovary were classified as small (SFs, 2 to 5mm in diameter) or large (LFs 6 to 10mm in diameter) and the follicular fluid was obtained by follicle aspiration. The collection of serum samples was performed after the gilts exsanguination using sterile tubes. From the pool of serum and follicular fluid obtained from 50 females, the concentration of free IGF-I was determined in each sample using an enzyme immunoassay kit (ELISA). The description of ovarian morphometry was performed in 100 ovaries from randomly selected gilts. The larger and smaller lengths of ovaries were measured, and the total number of SFs and LFs present on the surface of each ovary were also counted. The IGF-I concentration was greater (P<0.05) in LFs (170.92±88.29 ng/mL) compared with SFs (67.39±49.90ng/mL) and serum (73.48±34.63ng/mL). The largest and smallest length of the ovaries was 26.0±3.0 and 19.0mm ±2.0mm, respectively. The number of SFs (70.86±25.76) was greater (P<0.01) than LFs (6.54±5.26). The study concluded that LFs present greater levels of IGF-I when compared with SFs and blood, which is related to increased activity of the LFs and its differentiation to ovulation. In addition, ovaries of pre-pubertal gilts have a higher number of SFs compared to LFs. Therefore, our study demonstrated unique data regarding the physiological concentration of free IGF-I in ovarian follicles, that can be used in future research to evaluate the addition of this hormone in the in vitro production media of porcine embryos with the goal to improve the technique efficiency.

Feline hypertrophic cardiomyopathy: an echocardiographic approach

RESUMEN La cardiomiopatia hipertrofica felina es una enfermedad primaria del miocardio que se caracteriza por una leve a severa hipertrofia concentrica primaria del miocardio ventricular. Los factores hereditarios y mutaciones causales se han atribuido al desarrollo de la enfermedad en algunas razas, como los gatos Maine Coon y Ragdoll. Sin embargo, esta enfermedad sigue siendo un reto para los veterinarios debido a la dificultad del diagnostico precoz y el riesgo de muerte subita de los animales afectados. La ecocardiografia es una herramienta no invasiva de eleccion para el diagnostico de las enfermedades cardiacas en los gatos. Nuevas tecnicas ecocardiograficas como el Doppler Tesidual (el color bidimensional, el modo M a color y el modo de onda pulsada), Strain y Strain rate , han demostrado ser eficaces en el diagnostico temprano de la disfuncion del miocardio, incluso en ausencia de hipertrofia en el examen ecocardiografico convencional. Esta revision pretende acercar la informacion mas reciente sobre el diagnostico ecocardiografico de la cardiomiopatia hipertrofica felina.

Sperm concentration on the intrauterine artificial insemination in swine

Conduziu-se este estudo, com o objetivo de avaliar a eficiencia da inseminacao intra-uterina (IIU) em suinos, considerando as taxas de retorno ao estro, aborto, parto, alem do tamanho da leitegada (numero de leitoes nascidos e nascidos vivos). Na IIU, as femeas foram inseminadas nos tempos de 24 e 48 horas apos a deteccao do estro, utilizando-se as concentracoes de 500 milhoes, 1 bilhao, 1,5 bilhao e 2 bilhoes de espermatozoides, em 20mL de diluente. A passagem do cateter de IIU atraves da cervix foi possivel em 97,9% das femeas. Foi realizado diagnostico de retorno ao estro a partir do 18o dia e diagnostico de gestacao por ultrassonografia transcutânea entre o 28o e 30o dias apos a inseminacao. A taxa de retorno ao estro foi de 6,3% na IIU. A taxa de parto na IIU foi de 87,2%, sendo a taxa de parto para a concentracao de 500 milhoes de 100%. Com relacao ao numero de leitoes nascidos totais e nascidos vivos, nao houve diferencas, entre as diferentes concentracoes espermaticas (P>0,05). A utilizacao da concentracao de 500 x 106 espermatozoides em 20mL de diluente, com inseminacao intra-uterina, obteve-se um bom desempenho reprodutivo.

Use of discriminant analysis based on echocardiography for classification of congestive heart failure in dogs with myxomatous mitral valve disease

Mixomatous mitral valve disease (MMVD) is one of the most common cardiac abnormalities in dogs and humans that can lead to cardiac heart failure (CHF). Its diagnosis remains based on echocardiography and clinical signs. However, the early diagnose of MMVD can contribute to a better prognosis and avoid CHF. The aim of this study was to evaluate the clinical, radiographic and echocardiographic presence of CHF in dogs with MMVD in combination with a statistical model as a mathematical tool. For this purpose, dogs were divided into three groups (healthy; MMVD without CHF; and MMVD with CHF), according the clinical, radiographic and echocardiographic evaluation findings. Thus, linear discriminant functions were obtained by analyzing the variables weight, body surface area, aortic diameter, the ratio of the left atrium/aortic diameter, the ratio between the mitral regurgitation jet area and the left atrial area, vena contracta diameter and mitral valve proximal isovelocity surface area. Then, mathematical equations were established for each group of dogs. Statistical functions obtained in this study enabled to classify the dogs, regarding the presence of CHF with a probability of correct classification of 90.4%. Thus the statistical model demonstrated that it could be used as an auxiliary method for identifying CHF in dogs with MMVD.

Efficacy of sildenafil therapy for pulmonary hypertension in dogs: a systematic review

This systematic review aimed to assess the efficacy of sildenafil in the treatment of pulmonary hypertension (PH) in dogs and, when possible, to propose recommendations for or against its use. Electronic searches were carried out using six international databases. The inclusion criterion was in vivo clinical trials that assessed the efficacy of sildenafil in the treatment of naturally occurring PH. The scoring system was designed to take into account aspects such as the form of distribution and masking of experimental treatments, the number of animals, association with other drugs, diagnostic methods for PH, evaluations of the clinical response to treatment and adverse effects. A total of five papers were found. In general, the clinical condition and quality of life of the animals improved after sildenafil therapy, and the occurrence of adverse effects was uncommon. Echocardiographic findings provided evidence for changes in pulmonary arterial pressure which could be associated with alterations in physical activity and survival time. Still, the methodological quality of the studies was low, only one of them was a randomised, placebo-controlled and double-blind clinical trial. Furthermore, with modest evidence, this study allowed the recommendation that sildenafil is an important therapy which can play a positive therapeutic role in the treatment of canine PH. Additionally, prospective, randomised, blinded and controlled studies with a larger number of animals assessing physical activity and the adverse effects of sildenafil and other drugs in association are recommended to evaluate the efficacy of this drug in dogs.

Strain and strain rate by two-dimensional speckle tracking echocardiography in a maned wolf

The measurement of cardiovascular features of wild animals is important, as is the measurement in pets, for the assessment of myocardial function and the early detection of cardiac abnormalities, which could progress to heart failure. Speckle tracking echocardiography (2D STE) is a new tool that has been used in veterinary medicine, which demonstrates several advantages, such as angle independence and the possibility to provide the early diagnosis of myocardial alterations. The aim of this study was to evaluate the left myocardial function in a maned wolf by 2D STE. Thus, the longitudinal, circumferential and radial strain and strain rate were obtained, as well as, the radial and longitudinal velocity and displacement values, from the right parasternal long axis four-chamber view, the left parasternal apical four chamber view and the parasternal short axis at the level of the papillary muscles. The results of the longitudinal variables were -13.52±7.88, -1.60±1.05, 4.34±2.52 and 3.86±3.04 for strain (%), strain rate (1/s), displacement (mm) and velocity (cm/s), respectively. In addition, the radial and circumferential Strain and Strain rate were 24.39±14.23, 1.86±0.95 and -13.69±6.53, -1.01±0.48, respectively. Thus, the present study provides the first data regarding the use of this tool in maned wolves, allowing a more complete quantification of myocardial function in this species.