ABSTRACT
ABSTRACT: The purpose of this research was to confirm the changes occurring in the foot system of the heifers challenged with lipopolysaccharides (LPS), at the clinical, serum and histological levels. We studied 16 clinically healthy heifers, 14 months of age, placed in a confinement system. All the animals were provided with an accelerometer collar to establish their activity. They were categorized into two groups: the LPS group (n=8), or those which were administered two intravenous applications of 2 mL containing 0.5 μg/kg of body weight of LPS, with a 24-hour interval and the Control group (n=8) which were given two infusions of 2 mL of saline solution in the same time interval. General clinical examination and blood collection were done at 0, 4 and 8 hours post the LPS challenges and analyses of the hemograms and paroxonese-1 were performed. The animals were then slaughtered on day 4 and the laminar tissue was collected for histological analysis. The LPS group revealed a lower total leukocyte count with heart rate and greater activity. None of the animals revealed any abnormal signs symptomatic of foot pathology after histological analysis. Hence, the challenge with LPS failed to induce any clinical and histological changes in the foot tissue compatible with laminitis.
RESUMO: O objetivo deste trabalho foi verificar alterações do sistema podal á nível clínico, hematológico e histológico de animais desafiados com lipopolissacarídeo (LPS). Foram utilizadas 16 novilhas de corte com 14 meses de idade, clinicamente saudáveis, em sistema de confinamento. Todos os animais continham uma coleira com acelerômetro para verificar atividade, sendo divididos em 2 grupos: LPS (n= 8), que recebeu duas aplicações de 2 mL contendo 0,5 μg/kg de peso corporal de LPS via intravenosa, com intervalo de 24 horas e Controle (n= 8) que recebeu duas aplicações de 2 mL de solução salina com o mesmo intervalo de tempo. Foram realizados exame clinico geral e coletas de sangue às 0, 4 e 8 horas após os desafios com LPS para análise de hemogramas e paroxonase-1. Os animais foram abatidos aos 5 dias sendo executada a coleta de tecido laminar para análise histológica. A análise estatística foi executada com a utilização do programa NCSS(2004) através de ANOVA-medidas repetidas, considerando o efeito do tratamento do período e sua interação (tratamento*horas), sendo considerados significativos valores de P<0,05. O grupo LPS apresentou superior contagem total de leucócitos com maior frequência cardíaca e atividade. Nenhum animal apresentou sinal de anormalidade compatível com laminite através da análise histológica. Entretanto, o desafio com LPS foi capaz de gerar alterações a nível clínico, constatado pelas alterações nos parâmetros de frequência cardíaca, respiratória e temperatura corporal, além de mudanças no leucograma.
ABSTRACT
The aim of this study was to evaluate the effect of exposing bovine oocytes to lipopolysaccharides (LPS) in vivo and in vitro on early embryo development. In experiment 1, cumulus oocyte complexes (COCs, n = 700/group) were challenged with 0, 0.1, 1.0 or 5.0 µg/mL of LPS during in vitro maturation (IVM). Later, in vitro fertilization (IVF) and in vitro culture (IVC) were performed. In experiment 2, COCs (n = 200/group) matured and in vitro fertilized without LPS were subjected to IVC with the same doses of LPS from experiment 1. In experiment 3, heifers received two injections of saline solution (n = 8) or 0.5 µg/kg of LPS (n = 8) 24 h apart, and 3 days later, COCs were recovered and submitted to IVM, IVF, and IVC. In experiments 1 and 3, the expression of TLR4, TNF, AREG and EREG genes in cumulus cells was evaluated. Exposure to 1 and 5 µg/mL of LPS during IVM decreased nuclear maturation (39.4 and 39.6%, respectively) compared with control (63.6%, P < 0.05). Despite that, no effect on cleavage and blastocyst rates were observed. Exposure to LPS during IVC did not affect embryonic development. In vivo exposure to LPS decreased the in vitro cleavage rate (54.3 vs 70.2%, P = 0.032), but cleaved embryos developed normally. Number of cells per embryo and gene expression were not affected by the LPS challenge in any experiment. In conclusion, although in vitro exposure to LPS did not affect early embryo development, in vivo LPS exposure reduced cleavage rate.
Subject(s)
Embryonic Development/drug effects , Lipopolysaccharides/pharmacology , Oocytes/drug effects , Animals , Blastocyst/cytology , Blastocyst/drug effects , Blastocyst/physiology , Cattle , Cells, Cultured , Cleavage Stage, Ovum/drug effects , Cleavage Stage, Ovum/physiology , Cumulus Cells/cytology , Cumulus Cells/drug effects , Cumulus Cells/physiology , Embryo Culture Techniques/veterinary , Embryo, Mammalian , Female , Fertilization in Vitro/veterinary , Gene Expression Regulation/drug effects , In Vitro Oocyte Maturation Techniques/veterinary , Oocytes/cytology , Oocytes/physiology , PregnancyABSTRACT
The use of strategies to stimulate follicular growth are important, especially for use in timed artificial insemination (TAI) protocols, aiming to increase dairy cow's fertility. The aim of this study was to investigate the effect of insulin on follicular growth, steroid production and expression of genes related to follicular development. For this, cows were submitted to a progesterone (P4) and estradiol (E2) based synchronization protocol. In study 1, eleven primiparous lactating Holstein cows, received a single s.c. application of 0.25 IU/kg human insulin or no treatment (control) on D8 of the protocol. Blood samples were collected, and the dominant follicle diameter was assessed daily via transrectal ultrasonography, from D8 to D12. In study 2, eight multiparous non-pregnant and non-lactating Jersey cows, received a single s.c. application of 0.25 IU/kg human insulin, whereas cows from the control group received a single s.c. injection (1â¯mL) of saline solution (NaCl 0.9%). Blood samples were collected, and the dominant follicle diameter was assessed daily via transrectal ultrasonography from D6 to D9 of the protocol. Sixteen hours after insulin injection, follicular aspiration was performed. In study 1, insulin treatment decreased systemic glucose levels, but did not affect follicular growth. In study 2, the glucose decrease induced by insulin treatment was accompanied by a tendency of decreased progesterone levels in follicular fluid, along with a decrease in steroidogenic acute regulatory protein (STAR) and insulin like growth factor binding protein 2 (IGFBP2) mRNA abundance in granulosa cells. In conclusion, insulin treatment does not increase follicle growth and estradiol secretion in dairy cows, but decreases IGFBP2 and tends to increase pappalysin (PAPPA) mRNA abundance in granulosa cells, suggesting a positive effect on follicle development.
Subject(s)
Cattle/metabolism , Granulosa Cells/drug effects , Insulin/pharmacology , Ovarian Follicle/drug effects , Animals , Breeding , Female , Gene Expression/drug effects , Gene Expression Profiling , Humans , Ovarian Follicle/growth & development , Ovulation InductionABSTRACT
ABSTRACT: The objective of this study was to evaluate the effect of exogenous insulin administration on follicular growth, estrus presentation and conception rate during a protocol of ovulation synchronization. Dairy cows were subjected to the Heatsynch protocol, with the insertion of an intravaginal device containing 1.9 g of progesterone (CIDR) and an intramuscular injection containing 2.5 mg of GnRH on day 0. On day 7, the CIDR was removed and subjects were given 12.5 mg of dinoprost. Also on day 7, Insulin Group (IG, n = 21) animals received a subcutaneous injection containing 0.25 IU / kg of recombinant human insulin and the control group (CG, n = 25) received a 0.9% NaCl injection. On day 8, an injection of 0.5 mg of estradiol cypionate was administered to all cows. Animals were inseminated 12 hours after estrus presentation or at day 10 at fixed time. Follicular development was evaluated on days 7, 9, and 10 using transrectal ultrasonography, estrus presentation was observed between days 8 and 10, and conception rate was evaluated 30 days after AI. There were no differences in growth rate, follicular diameter, estrus presentation, and conception rate. Therefore, application of a single dose of insulin does not promoted an increase in follicular size, estrus presentation and conception rate in dairy cows.
RESUMO: O objetivo deste estudo foi avaliar o efeito da administração exógena de insulina sobre crescimento folicular, apresentação de cio e taxa de concepção durante um protocolo de sincronização da ovulação. As vacas holandês lactantes foram submetidas ao protocolo Heatsynch, com inserção do dispositivo intravaginal contendo 1,9 g de progesterona (CIDR) no dia 0 e uma injeção intramuscular de 2,5 mg de GnRH. No dia 7, o CIDR foi removido e foi aplicado 12,5 mg de dinoprost. Ainda no dia 7, os animais do Grupo Insulina (IG, n = 21) receberam uma injeção subcutânea de 0,25 UI / kg de insulina humana recombinante e o grupo controle (CG, n = 25) recebeu uma injeção de NaCl 0,9%. No dia 8 foi aplicado 0,5 mg de cipionato de estradiol em todas as vacas. Animais foram inseminados 12 horas após a apresentação de cio ou no dia 10 em tempo fixo. O desenvolvimento folicular foi avaliado nos dias 7, 9 e 10 por ultrassonografia transretal, a apresentação de cio foi observada entre os dias 8 e 10 e a taxa de prenhez/IA foi avaliada 30 dias após a IA. Não houve diferença quanto a taxa de crescimento e diâmetro folicular, apresentação de cio e taxa de concepção. A aplicação de uma dose única de insulina não promove o incremento no tamanho folicular, apresentação de cio e taxa de prenhez/IA em vacas de leite.
