Ultrassonografia esplênica, hepática, biliar e renal em caprinos. Uma revisão

A utilização da ultrassonografia bidimensional é crescente na clínica de ruminantes, principalmente na avaliação abdominal destes animais. Esta é uma ferramenta segura, prática, rápida, não invasiva e sem efeito colateral conhecido tanto para o paciente quanto para o examinador. A ultrassonografia permite a avaliação de órgãos como fígado, baço, rins, abomaso, rúmen, retículo, órgãos reprodutivos e outros, fornecendo informação a respeito da localização anatômica, tamanho e características da ecotextura do parênquima, detecção de anormalidades focais ou difusamente distribuídas no parênquima, por exemplo. Estudos relacionados ao uso da ultrassonografia podem ser de grande valia no exame desses órgãos, podendo prover aos clínicos informações diagnósticas específicas, anteriormente disponíveis somente por laparotomia exploratória. Assim, este estudo teve por objetivo fornecer informações referentes à utilização da ultrassonografia para descrição da anatomia e diagnóstico de alterações morfofuncionais em órgãos como baço, fígado, vesícula biliar e rins de caprinos sadios

AVALIAÇÃO DOS EFEITOS DA PIOMETRA NA VIABILIDADE DE OÓCITOS CANINOS IMATUROS PELA UTILIZAÇÃO DO AZUL CRESIL BRILHANTE

The bitch is an experimental model of wild and even endangered canids. Therefore, the study of the factors that influence your fertility benefits scientific advances in both segments. Knowing that pyometra is one of the common uterine pathologies in bitches, this work aimed to evaluate the effects of pyometra on the morphology and competence of canine oocytes through the use of Azul CresilBrilhante (ACB). For this purpose, 1197 canine oocytes that were divided into 2 groups classified as control (healthy) and treatment (pyometra). They were morphologically classified into grade 1 (G1), grade 2 (G2) and grade 3 (G3) and according to the ACB stain as ACB (+) and ACB (–) (not stained). Bitches in the healthy group had higher amounts of total oocytes (795) and better quality (495 oocytes G1) and competence (45 ± 9.8 ACB (+)). The use of ACB was useful to distinguish the competence of the studied oocytes and can be an auxiliary tool for choosing the best oocytes.The bitch is an experimental model of wild and even endangered canids. Therefore, the study of the factors that influence your fertility benefits scientific advances in both segments. Knowing that pyometra is one of the common uterine pathologies in bitches, this work aimed to evaluate the effects of pyometra on the morphology and competence of canine oocytes through the use of Azul CresilBrilhante (ACB). For this purpose, 1197 canine oocytes that were divided into 2 groups classified as control (healthy) and treatment (pyometra). They were morphologically classified into grade 1 (G1), grade 2 (G2) and grade 3 (G3) and according to the ACB stain as ACB (+) and ACB (–) (not stained). Bitches in the healthy group had higher amounts of total oocytes (795) and better quality (495 oocytes G1) and competence (45 ± 9.8 ACB (+)). The use of ACB was useful to distinguish the competence of the studied oocytes and can be an auxiliary tool for choosing the best oocytes

INFLUÊNCIA DA IDADE SOBRE A QUALIDADE DE OÓCITOS IMATUROS CANINOS

Among some factors that influence the quality of canine oocytes, age is well studied. Knowing that the morphological assessment of oocytes is a useful tool for this analysis, the use of new techniques such as the brilliant cresyl blue staining (BCB) increases this assessment. The objective of this work was to analyze the quantity and the oocyte quality of bitches in different age groups through the conventional morphological evaluation, in addition to the bright cresyl blue dye. For this purpose, 2.302 oocytes from 33 bitches were divided into four age groups from 1 to 3 years old; 4-6, 7-10 and over 10 years old and their oocytes were classified as G1, G2 and G3, as well as stained (BCB (+)) and non-stained (BCB (-)). The values ​​were compared using the Kruskal test -Wallis and Dunn post-test, by which it was possible to obtain higher amounts of G1 and BCB (+) oocytes in the range of 1 to 3 years, indicating that this range oocytes have a greater chance of success in fertilization processes and in vitro production, for example.Dentre alguns fatores que influenciam na qualidade dos ovócitos caninos, a idade é bem estudada. Sabendo que a avaliação morfológica dos ovócitos é uma ferramenta útil para esta análise, a utilização de novas técnicas como a coloração com azul de cresil brilhante (BCB) incrementa esta avaliação. O objetivo deste trabalho foi analisar a quantidade e a qualidade oocitária de cadelas em diferentes faixas etárias através da avaliação morfológica convencional, além do corante azul de cresil brilhante. Para tanto, 2.302 ovócitos de 33 cadelas foram divididos em quatro faixas etárias de 1 a 3 anos; 4-6, 7-10 e maiores de 10 anos e seus ovócitos foram classificados em G1, G2 e G3, corados (BCB (+)) e não corados (BCB (-)). Os valores foram comparados por meio do teste de Kruskal -Wallis e pós-teste de Dunn, pelo qual foi possível obter maiores quantidades de oócitos G1 e BCB (+) na faixa de 1 a 3 anos, indicando que nesta faixa os oócitos possuem maior chance de sucesso em processos de fertilização e produção in vitro, por exemplo

Impact Of Hypotensive Action Of Angiotensin Converting Enzyme Inhibitor On Ovarian-Intraovarian Blood Flow And Follicles Development In Goats Hormonally Stimulated With Repeated Fsh-Ecg Treatment

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductive processes, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed to use an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however, the presence of the main components of this system in reproductive tissues still need to be further investigated, since the physiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate, an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats. Materials, Methods and Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n=10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 of enalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received the same volume of saline solution. Estrus synchronization of all goats was made by intramuscular administration of PGF2α analog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight hours after device withdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3 treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetrics parameters of the ovarian artery and intraovarian blood flow, respectively, and B-mode real-time ultrasound scanner to evaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction of systolic and diastolic peak, without difference according to parity. In addition, in the third session of hormonal stimulation, only the groups (nulliparous and multiparous) not treated with enalapril maleate had a significant increase in the intraovarian blood flow (Doppler area). In treated group was also found a greater number of small follicles, a lower proportion of follicle ≥ 5 mm and a smaller follicular size. In the same group, the nulliparous females had a lower proportion of large follicles compared to the saline group, otherwise, there were no differences between the treatments in multiparous goats. In the group treated with enalapril maleate, higher number of large follicles was observed in multiparous animals than in the nulliparous ones.Discussion: In vivo and in vitro studies have shown clear effects of RAS modulation on reproductive aspects. Our results confirmed the hypotensive effect of enalapril maleate on the ovarian artery and intraovarian blood flow, also showing that the administration of the drug reduced the process of follicular depletion and the growth of large follicles. These findings suggest that the hypotensive effects observed in our experiment, result from the treatment with the ACE inhibitor, and that this alteration in hemodynamic parameters may be the main responsible for the lower follicular response observed in animals treated with enalapril-maleate. We suggest that further studies are necessary to elucidate possible effects of the administration of an ACE inhibitor on ovarian activity, especially in the expression of genes associated with follicular, oocyte and embryonic development. Finally, the study conclude that the administration of enalapril-maleate in our experimental condition exhibit a hypotensive effect on ovarian circulatory system but did not show to promote a real benefit of follicular development in goats.

PRIMEIRA AVALIAÇÃO DA OBTENÇÃO E QUALIDADE DE OÓCITOS POST MORTEM DE LHAMAS PELO AZUL CRESIL BRILHANTE

This study aimed to demonstrate the applicability of the ovarian slicing technique in the recovery of post mortem oocytes in a llama (lama glama) victim of acute bronchopneumonia and the effectiveness of the brilliant cresyl blue (ACB) technique to select quality oocytes for a subsequent in vitro culture. The ovary-salpinge-hysterectomy (OSH) was performed 12 hours after the animal's death. The ovaries were transported refrigerated immediately to the laboratory in Leibovitz medium (L-15). The oocytes were identified, quantified under a stereomicroscopic and classified according to two methods: conventional morphological classification and ACB assay. 26 oocytes classified by conventional morphology were obtained: 10 oocytes grade 1, 12 oocytes grade 2 and 4 oocytes grade 3. The use of the ACB dye demonstrated that 88% of the evaluated oocytes maintained their quality and were competent for the conservation of genetic or future material application in reproductive biotechnologies in this species. Further studies are suggested aiming to use selected oocytes with ACB to enhance the success of in vitro culture in this species.Este estudo teve como objetivo demonstrar a aplicabilidade da técnica de fatiamento ovariano na recuperação de oócitos post mortem em uma lhama (lama glama) vítima de broncopneumonia aguda e a eficácia da técnica de azul de cresil brilhante (ACB) para selecionar oócitos de qualidade para posterior incubação cultura in vitro. A ovário-salpinge-histerectomia (OSH) foi realizada 12 horas após a morte do animal. Os ovários foram transportados refrigerados imediatamente ao laboratório em meio Leibovitz (L-15). Os oócitos foram identificados, quantificados em estereomicroscópio e classificados de acordo com dois métodos: classificação morfológica convencional e ensaio ACB. Obtiveram-se 26 oócitos classificados pela morfologia convencional: 10 oócitos grau 1, 12 oócitos grau 2 e 4 oócitos grau 3. A utilização do corante ACB demonstrou que 88% dos oócitos avaliados mantiveram sua qualidade e foram competentes para a conservação do patrimônio genético ou futura aplicação de material em biotecnologias reprodutivas nesta espécie. Novos estudos são sugeridos com o objetivo de utilizar oócitos selecionados com ACB para aumentar o sucesso do cultivo in vitro nesta espécie

Equine ovarian tissue xenografting: impacts of cooling, vitrification, and VEGF.

Ovarian tissue transplantation methods using cooled and cryopreserved samples have been attractive options for fertility preservation in animal models and humans. The aim of this study was to evaluate the impact of previous exposure to cooling, cryopreservation, and VEGF on the overall efficiency of equine ovarian tissue after heterotopic xenotransplantation in mice. The end points evaluated were follicular morphology and development, follicular and stromal cell densities, angiogenesis (i.e. the density of new and mature blood vessels), collagen types I and III fiber densities, and total fibrosis. Ovaries of adult mares were harvested after ovariectomy, and ovarian fragments were xenografted in the i.p. wall of BALB nude mice. Ten types of treatments involving different combinations of cooling, cryopreservation, xenografting procedures, and VEGF exposure were compared. The novel aspect of this study was the use of equine ovarian tissue xenotransplantation in mice, challenging the fragments with different combinations of treatments. The main findings were (i) cooling but not cryopreservation was effective in preserving the follicular morphology, (ii) a greater percentage of developing follicles but lower follicular and stromal cell densities were observed after ovarian tissue engraftment, (iii) exposure to VEGF increased new and mature vessels in cryopreserved-transplanted tissue, and (iv) an appropriate balance in the collagen types I and III fiber ratio in cooling-transplanted tissue was observed after exposure to VEGF. This study contributes to advancing knowledge in the preservation of ovarian tissue after cooling-cryopreservation and transplantation aiming to be applied to genetically superior/valuable horses, livestock, endangered animals, and, possibly, humans. LAY SUMMARY: Due to ethical limitations involving humans, the female horse (mare) has recently emerged as an alternative model for reproductive comparisons with women to optimize fertility restoration using ovarian tissue transplantation techniques. This study determined if ovarian tissue from donor mares (n = 3), exposed or not to vascular endothelial growth factor (VEGF) before transplantation, better survives for 7 days after transplantation into mouse hosts (n = 12). Tissues submitted to different combinations of cooling, freezing, and transplanting treatments, along with control groups, were evaluated using the parameters morphology, development, the density of immature eggs (follicles), the density of supportive (stromal) cells, collagen protein proportions, and density of blood vessels. Frozen-thawed treatments had lower percentages of normal follicles. Exposure to VEGF increased blood vessel densities in frozen tissue and favored adequate collagen levels in cooled-transplanted treatments. In conclusion, VEGF exposure seems to be beneficial for mare ovarian tissue transplantation and warrants further investigation

AVALIAÇÃO DA EFICIÊNCIA DE REUTILIZAÇÃO DO CIDR® EM PROTOCOLOS DE SUPEROVULAÇÃO DE OVELHAS

Sheep farming is an area of veterinary medicine of great importance at regional and national level, among the easily found breeds of economic relevance in northeastern Brazil are Morada Nova and Santa Inês. In addition, ultrasonography has been used systematically in animals superovulatory response to the sheep supervoulation protocols. In order to assess, based on the number of yellow bodies and number of viable embryos produced, the efficiency of the reuse of CIDR® in superovulation protocols in sheep using ultrasonography in color Doppler mode, a superovulatory protocol was performed in a total 40 sheep, with G-USED when 10 Morada Nova ewes and 10 Santa Inês ewes received reused CIDR® and G-NOVO when 10 Morada Nova ewes and 10 Santa Inês ewes received new CIDR®. The mean and standard error of yellow bodies was 10.35±1.33 for G-USED and 9.25±1.65 for G-NOVO, while the number of viable embryos was 6.50±2, 54 for G-USED and 8.25±1.49 for G-NEW. Thus, the reuse of CIDR® demonstrated efficiency in superovulation and did not interfere with the development of corpus luteum and the production of quality transferable embryos in sheep.A ovinocultura é uma área da medicina veterinária de grande importância a nível regional e nacional, dentre as raças de relevância econômica facilmente encontradas no nordeste do Brasil estão a Morada Nova e a Santa Inês. Além disso, a ultrassonografia tem sido utilizada sistematicamente na resposta superovulatória de animais aos protocolos de supervolução de ovinos. Com o objetivo de avaliar, com base no número de corpos amarelos e no número de embriões viáveis produzidos, a eficiência do reaproveitamento do CIDR® em protocolos de superovulação em ovelhas por ultrassonografia no modo Doppler colorido, foi realizado um protocolo de superovulação em um total de 40 ovelhas, com G-USED quando 10 ovelhas Morada Nova e 10 ovelhas Santa Inês receberam CIDR® reutilizado e G-NOVO quando 10 ovelhas Morada Nova e 10 ovelhas Santa Inês receberam CIDR® novo. A média e erro padrão dos corpos amarelos foi de 10,35±1,33 para G-USED e 9,25±1,65 para G-NOVO, enquanto o número de embriões viáveis foi de 6,50±2, 54 para G-USED e 8,25±1,49 para G-NEW. Assim, a reutilização do CIDR® demonstrou eficiência na superovulação e não interferiu no desenvolvimento do corpo lúteo e na produção de embriões transferíveis de qualidade em ovinos

AVALIAÇÃO DO USO DO CIDR NOVO E REUTILIZADO NO INÍCIO E DURAÇÃO DO ESTRO EM OVINOS

Study evaluated the use of new and re-utilized CIDR® to increase the reproductivebreeding efficiency of the Santa Inês (SI) and Morada Nova (MN). In the synchronization protocol, 40 sheep was divided into four groups: MN with re-utilized CIDR®; MN with new CIDR®; SI with re-utilized CIDR® and SI with new CIDR®. The protocol lasted for 15 days. At D0, females received CIDR®. Between D7 and D9, ovarian superstimulation was induced by two doses application with a 12-hour break of p-FSH. Estrus confirmation was evaluated with ruffians, and the duration determined with the first and last mounts. The ovulation was determined by B-mode ultrasound and color Doppler. All animals of the experiment showed signs of estrus after removal of the device and there was no difference between treatments (p>0.05). In Treatment 1 (T1, CIDR® used), the beginning of estrus was 22.3±1.53h and in Treatment 2 (T2, new CIDR®) was 22.9±1.64h. In estrus duration, T1 and T2 was similar (21.6±1.81 and 22.2±0.63, respectively). Conclusion: the treatments are efficient to induce and synchronize estrus, with satisfactory results in their duration.Estudo avaliou o uso de CIDR® novo e reutilizado para aumentar a eficiência reprodutiva de Santa Inês (SI) e Morada Nova (MN). No protocolo de sincronização, 40 ovinos foram divididos em quatro grupos: MN com CIDR® reutilizado; MN com o novo CIDR®; SI com CIDR® reutilizado e SI com novo CIDR®. O protocolo durou 15 dias. No D0, as fêmeas receberam CIDR®. Entre D7 e D9, a superestimulação ovariana foi induzida pela aplicação de duas doses com intervalo de 12 horas de p-FSH. A confirmação do estro foi avaliada com rufiões, e a duração determinada com a primeira e a última monta. A ovulação foi determinada por ultrassom modo B e Doppler colorido. Todos os animais do experimento apresentaram sinais de estro após a retirada do dispositivo e não houve diferença entre os tratamentos (p>0,05). No Tratamento 1 (T1, CIDR® utilizado), o início do estro foi de 22,3±1,53h e no Tratamento 2 (T2, novo CIDR®) foi de 22,9±1,64h. Na duração do estro, T1 e T2 foram semelhantes (21,6±1,81 e 22,2±0,63, respectivamente). Conclusão: os tratamentos são eficientes para induzir e sincronizar o estro, com resultados satisfatórios na duração dos mesmos

Testicular Morphological and Ultrasonographic Characterization of Male Gray Brocket Deers (Mazama gouazoubira) in Different Reproductive Status

Background: Gray brocket deer (Mazama gouazoubira) populations have been declining due to human intervention. Yet, only a few studies have assessed ultrasonographic testicular characteristics in cervids. Considering the relevance of monitoring testicular size, blood flow, and parenchyma, the present study aims to establish baseline information on scrotal circumference, testicular volume, and spectral Doppler parameters, to describe differences among adult male gray brocket deer in different reproductive status, and to correlate ultrasound parameters with testes size measurements.Materials, Methods & Results: Six adult male gray brocket deers were used in the study. Scrotal circumference and testicular volume were measured. B mode ultrasound images of testes (longitudinal and cross-sectional views) and epididymes were subjected to computer-assisted analysis, obtaining the numerical pixel values (NPV) and pixel standard deviation (PSD). Using spectral Doppler, supratesticular artery blood flow velocities (peak systolic velocity - PSV, end diastolic velocity - EDV, time-average maximum velocity - TAMAX, resistivity - RI and pulsatility indices - PI) were obtained. Semen was analyzed through total motility, vigor, and concentration tests. Three animals were normospermic (F+ group) and three were oligo/azoospermic (F- group). Groups were compared using were compared using a one-way ANOVA or Kruskal-Wallis followed by Student-Newman-Keuls (SNK) test. Ultrasound parameters were correlated to testes size parameters using Pearson’s correlation for parametric variables and Spearman’s correlation for non-parametric variables. F+ group presented significantly higher scrotal circumference (14.57 ± 1.19 cm), testicular volume (26.18 ± 4.94 cm3), and testes cross-sectional NPV (69.88 ± 24.00) and PSD (10.78 ± 3.42) than group F- (NPV: 28.26 ± 13.75, PSD: 6.70 ± 1.84). No significant differences were observed between the groups regarding the spectral Doppler ultrasound parameters. Significant correlations were observed between scrotal circumference and longitudinal (r = 0.76) and cross-sectional testes NPV (r = 0.89), and testicular volume was correlated with longitudinal (r = 0.78) and cross-sectional testes NPV (r = 0.91) and with cross-sectional testes PSD (ρ = 0.82).Discussion: Increased testicular echogenicity (higher NPV) has been positively associated with improved testicular growth, cell population expansion, inner and outer seminiferous tubules diameter, spermatids percentages and testis weight. In addition, more heterogenous testes (higher PSD) were associated with higher sperm output. It was suggested that the animals in group F- had compromised testicular development and spermatogenesis. The correlation observed between testes NPV and scrotal circumference was proposed to be associated with seminiferous tubules impairment. The F- group showed lower testicular volume, NPVs and PSDs in cross-sectional testicular images, suggesting higher protein levels and lower lipid contents were present in their parenchyma, influencing in testicular echogenicity and echotexture. No differences in spectral Doppler parameters were observed between the two groups. Also observed in stallions. However, PSV, EDV, TAMAX could be potential infertility indicators in other mammalians. These different results may be due to different locations of the evaluated vessel, species and techniques, age, ambient temperature, pathological conditions, and anaesthesia. Thus, it is suggested that scrotal circumference, testicular volume, and testes NPV are good indicators of male reproductive health in gray brocket deer and may help with better male selection in the species