Short communication: Proinflammatory gene expression relative to the collection technique of endometrial samples from cows with and without subclinical endometritis.

Uterine inflammation negatively affects reproductive performance and is an important cause of infertility and subfertility in dairy cows. Several studies have investigated the use of gene expression in endometrial samples collected by biopsy or cytology to evaluate the inflammatory response of the cow uterus. This study aimed to compare the expression of the CCL5, CXCL8, IL6, and IL1B genes in the bovine endometrium according to the site of sample collection [caruncular (C) or intercaruncular (IC)], the collection method (biopsy or cytology), and the category of inflammation based on endometrial cytology (zero, medium, or high) in subclinical endometritis. The reproductive tracts of dairy cows were collected from a slaughterhouse, and punch-biopsy samples of endometrial tissues were obtained from both regions (C and IC). Endometrial cells from these regions were collected with the cytobrush technique and then used for the analysis of mRNA expression by quantitative PCR. After counting polymorphonuclear cells (PMN) by endometrial cytology, 20 uteri with an ovary at stage I (d 1-4 of estrous cycle) were categorized into 3 groups. Uteri with 0% PMN (n = 10) were assigned to group zero, uteri with 5 to 15% PMN (n = 5) to group medium (12.2 ± 1.6% PMN), and uteri with >15% PMN (n = 5) to group high (53.8 ± 32.9% PMN). All data were analyzed with 2-way ANOVA with Bonferroni multiple comparison post test. The results from gene transcripts demonstrated that the region (C or IC) of the endometrial biopsy had no influence on any of the degrees of inflammatory reaction observed. However, gene expression was more elevated in the endometrium of cows with greater inflammation compared with those without inflammation (CCL5, CXCL8, IL6, IL1B) and those with medium inflammation (CCL5, IL6). Expression of the genes evaluated did not differ between the endometrium without inflammation and with medium inflammation. However, in the high inflammation group, all genes were comparatively more expressed in samples collected by cytology relative to those derived from biopsies for both anatomical regions. In conclusion, gene expression did not differ between the C and IC tissue. Samples collected from animals with greater inflammation had greater gene expression than those with zero or medium inflammation. In addition, cytology samples had greater gene expression than biopsy samples in the high inflammation group.

Retenção de placenta e endometrite subclínica: prevalência e relação com o desempenho reprodutivo de vacas leiteiras mestiças / Retained placenta and subclinical endometritis: prevalence and relation with reproductive performance of crossbred dairy cows

Objetivou-se avaliar os efeitos do tipo de parto (eutócico vs. distócico) e do período do ano (chuvoso vs. seco) sobre a prevalência de retenção de placenta (RP) e de endometrite subclínica (ES), verificar a relação entre as duas patologias, e os efeitos dessas sobre a taxa de descarte, o período de serviço (PS) e número de IA/concepção. A RP foi avaliada no primeiro dia pós-parto (DPP). A citologia endometrial foi realizada entre 30 e 80 DPP, considerando positivos casos contendo acima de 5% de neutrófilos. Os dados foram analisados por regressão logística e análise de variância no programa Minitab (P<0,05). A prevalência de RP foi de 14,93% (69/462) e de ES de 27,49% (127/462). Foi detectada tendência de efeito (P=0,10) da RP sobre a prevalência da ES. Partos distócicos aumentaram a prevalência de RP e vacas que pariram nos meses chuvosos tiveram maior prevalência de ES (P<0,05). A RP elevou a taxa de descarte (P<0,05), a duração do período de serviço (P<0,05) e o número de IA por concepção (P<0,05), enquanto a ES não afetou essas variáveis (P>0,05). Conclui-se que a retenção de placenta tende a ser um fator de risco para endometrite subclínica, distocia predispõe à retenção de placenta e partos ocorridos no período chuvoso aumentam os casos de endometrite subclínica. Há um impacto negativo na eficiência reprodutiva de vacas leiteiras mestiças acometidas por retenção de placenta.(AU)

This study aimed to evaluate the effects of type of calving (eutocic vs. dystocic), season of the year (rainy vs. dry) on retained placenta (RP) and subclinical endometritis (SE) prevalence, to verify the relation between these diseases, as well as its effects on culling rate, days open and number of AI/conception. Retention of fetal membranes was recorded on first day postpartum. Endometrial cytology was performed between 30 and 80 days in milk (DIM) and positive cases were considered ≥5% neutrophils. Data were analysed by logistic regression and analysis of variance on Minitab program (P<0.05). The prevalence of RP was 14.93% (69/462) and of SE was 27.49% (127/462). A tendency of effect of RP on SE prevalence was detected (P=0.10). Dystocia increased RP prevalence (P<0.05). Cows that calved during rainy months had greater SE prevalence (P<0.05). RP increased culling rate (P<0.05), calving to conception interval (P<0.05) and number of AI/conception (P<0.05), although SE occurrence did not affect these variables (P>0.05). In conclusion, RP tended to be a risk factor for SE, dystocia is a predisposing factor for RP and calvings that happens during rainy period increases SE. There is a negative impact on reproductive efficiency of crossbred dairy cows that had retained placenta.(AU)

Perfil metabólico de vacas Jersey clinicamente saudáveis / Metabolic profile of clinically healthy Jersey cows

O conhecimento do metabolismo dos animais é de suma importância para se obter sucesso em qualquer atividade que envolva rebanhos. Objetivou-se determinar o perfil bioquímico sérico de vacas Jersey clinicamente saudáveis no pré e pós-parto, mantidas em sistema de criação semi-intensivo na região de Uberlândia, Minas Gerais, Brasil. Realizou-se o exame clínico e coleta de sangue de 40 vacas da raça Jersey multíparas e lactantes em 15 momentos, entre -150 dias pré-parto até 60 dias pós-parto (DPP). No laboratório, foram analisadas as concentrações séricas de proteínas totais, albumina, globulinas, ureia, creatinina, ácidos graxos não esterificados (NEFA), ß-hidroxibutirato (BHBA), triglicerídeos, colesterol, lipoproteínas de alta densidade (HDL), lipoproteínas de muita baixa densidade (VLDL), lipoproteínas de baixa densi­dade (LDL), aspartato aminotransferase (AST), gama-glutamil transferase (GGT), creatina quinase (CK), cálcio, fósforo e magnésio. Dos 40 animais, apenas 21 vacas atenderam aos critérios de inclusão e seus dados foram utilizados. Procedeu-se o teste de Tukey para os dados paramétricos e para os dados não paramétricos realizou-se o teste de Kruskal Wallis na comparação entre os momentos avaliados. Não houve interferência do balanço energético negativo e da hipocalcemia subclínica na ocorrência de doenças uterinas e no desempenho reprodutivo. Concluiu-se que vacas da raça Jersey apresentam perfil bioquímico caracterizado por hiperglobulinemia durante o periparto, além de níveis elevados de triglicérides, entretanto, sem comprometimento da função hepática e desempenho reprodutivo.(AU)

The understanding of animal metabolism is important for the success of livestock activity. The aim of this study was to determine the serum biochemical profile of clinically healthy Jersey cows in pre and postpartum, housed in semi-intensive production system in Uberlândia, Minas Gerais, Brazil. Clinical examination and blood collection from 40 multiparous lactating dairy cows, between -150 days prepartum up to 60 days postpartum (DPP) were performed. At the laboratory, serum parameters of total proteins, albumin, globulins, urea, creatinine, non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BHBA), triglycerides, cholesterol, high density lipoprotein (HDL), very low density lipoproteins (VLDL), low density lipoproteins (LDL), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), creatine kinase (CK), calcium, phosphorus and magnesium were analyzed. To the total of cows (n=40) enrolled, only 21 cows were complied with the inclusion criterias. It was proceeded the Tukey test for parametric data, and to the nonparametric data it was held the Kruskal Wallis test for the comparison between days evaluated. There was no interference of negative energy balance and subclinical hypocalcemia in the occurrence of uterine diseases and reproductive performance. In conclusion, Jersey cows have biochemical profile characterized by hyperglobulinemia during peripartum, and high levels of triglycerides, but without impaired liver function and reproductive performance.(AU)

Perfil metabólico de vacas mestiças leiteiras com baixo escore de condição corporal no periparto / Metabolic profile in crossbred dairy cows with low body condition score in the peripartum period

Objetivou-se avaliar o perfil metabólico energético, proteico e enzimático de vacas mestiças leiteiras com baixo escore de condição corporal (ECC) no periparto. Foram colhidas amostras sanguíneas uma semana antes do parto, no dia do parto, e aos sete, 14, 21, 28 e 43 dias pós-parto (DPP) de 36 animais, com média de ECC de 2,6±0,5, com eutocia e pós-parto fisiológico e sem tratamentos nesta fase. Analisaram-se as concentrações séricas de proteínas totais, albumina e globulinas para o perfil protéico; AST, ALT, GGT e fosfatase alcalina para o perfil enzimático; ácidos graxos não-esterificados (NEFA), β-hidroxibutirato (BHBA), triglicerídeos, colesterol e lipoproteínas (VLDL, HDL e LDL) para o perfil energético. As vacas apresentaram no pré-parto hipoproteinemia, hipoalbuminemia, hipocolesterolemia e aumento das enzimas GGT e AST. No dia do parto houve lipólise e hipoglobulinemia. Concluiu-se que vacas mestiças leiteiras com baixo ECC apresentam balanço energético negativo, hipoproteinemia com hipoalbuminemia e lesão hepática no periparto, com restabelecimento aos 30 DPP, mas não recuperam sua condição corporal até o final do puerpério.

The aim of this study was to evaluate the metabolic profile of protein, energy and enzyme in crossbred dairy cows with low body condition score (BCS) in the peripartum period. Blood samples were collected from 36 animals with 2.6±0.5 BCS, eutocia, physiological postpartum and without any treatment, on following days: one week before calving, calving and 7, 14, 21, 28 and 43 days in milk (DIM). It was evaluated serum total protein, albumin and globulins for protein profile; AST, ALT, GGT and alkaline phosphatase for mineral profile; nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), triglycerides, cholesterol and lipoproteins (VLDL, HDL and LDL) for energy profile. Crossbred dairy cows had hypoproteinemia, hypoalbuminemia, hypocholesterolemia and increased that both enzymes AST and GGT at the precalving. There were lipolysis and hypoglobulinemia at parturition. It was concluded that crossbred dairy cows with low BCS have negative energy balance, hypoproteinemia with hypoalbuminemia and hepatic injury in the peripartum. This condition is restored at 30 DIM, but there is no recovery of the body condition by the end of puerperium.

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa.

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.