The objective of this study was to evaluate the progression of the uterine microbiota from calving until establishment of metritis. Uterine swabs (n ‫؍‬ 72) collected at 0, 2, and 6 ؎ 2 days postpartum (dpp) from 12 metritic and 12 healthy cows were used for metagenomic sequencing of the 16S rRNA gene on the Illumina MiSeq platform. A heat map showed that uterine microbiota was established at calving. The microbiota changed rapidly from 0 to 6 ؎ 2 dpp, with a decrease in the abundance of Proteobacteria and an increase in the abundance of Bacteroidetes and Fusobacteria, which were dominant in metritic cows. Uterine microbiota composition was shared; however, metritic and healthy cows could be discriminated using relative abundance of bacterial genera at 0, 2, and 6 ؎ 2 dpp. Bacteroides was the main genus associated with metritis because it was the only genus that showed significantly greater abundance in cows with metritis. As the abundance of Bacteroides organisms increased, the uterine discharge score, a measure of uterine health, worsened. Fusobacterium was also an important genus associated with metritis because Fusobacterium abundance increased as Bacteroides abundance increased and the uterine discharge score worsened as the abundance increased. The correlation with uterine discharge score and the correlation with Bacteroides or Fusobacterium showed that other bacteria, such as Helcoccocus, Filifactor, and Porphyromonas, were also associated with metritis. There were also bacteria associated with uterine health, such as "Candidatus Blochmannia," Escherichia, Sneathia, and Pedobacter. Metritis is a huge concern for the dairy industry worldwide because it is highly prevalent (25 to 40%) and negatively affects the productivity, survival, and welfare of dairy cows (1). Diverse bacteria, including anaerobes and facultative anaerobes, were observed in the uteri of dairy cows within the first 2 weeks postpartum, but they were naturally cleared out within 60 days postpartum (dpp) (1). Culture-based studies observed that Escherichia coli, Trueperella pyogenes, Fusobacterium necrophorum, and Bacteroides spp. (e.g., Prevotella melaninogenica, formerly Bacteroides melaninogenicus) were commonly associated with endometritis or pyometra (1-3).Although culture-based studies have laid out the foundation of our understanding of the uterine microbiota, previous studies might have underestimated the microbial complexity of the intrauterine environment of cows postpartum, given that less than 1% of the microorganisms in many environments are readily cultured under standard laboratory conditions (4). In recent years, cultureindependent techniques such as clone library sequencing (5, 6) and pyrosequencing (7, 8) have been used to characterize the uterine microbiota of cows with metritis (5-7) and endometritis (7,8). Sequencing using the Illumina platform allows for deeper sequencing than has previously been feasible even with pyrosequencing (9). Indeed, evaluating the rarefaction curves from previous 16S rRNA sequencing studies ...

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Our objectives were to evaluate the efficacy of ampicillin trihydrate for the treatment of metritis in dairy cows compared with ceftiofur hydrochloride and the subsequent effects on pregnancy at first insemination (P/AI). Cows in the first 12 d in milk (DIM) with a uterine discharge score of 5 (watery, reddish or brownish discharge of foul smell) and rectal temperature <39.5°C were diagnosed with metritis based on the fetid discharge, and cows with metritis and rectal temperature ≥39.5°C were diagnosed as having puerperal metritis. Cows with metritis (n=528) were blocked by parity and type of metritis as fetid discharge or puerperal metritis and, within each block, assigned randomly to receive 11mg/kg of ampicillin (n=259) or 2.2mg/kg of ceftiofur (n=269) once daily for 5 d. Day of diagnosis of metritis was considered study d 1. A cohort of 268 cows without metritis was selected randomly at 12 DIM. Rectal temperature was measured in cows with metritis on study d 1 to 7, and 12, and vaginal discharge was scored on study d 5, 7, and 12. Metritis cure was characterized by vaginal discharge score of <5 or by vaginal discharge score of <5 and no fever. At 32±3 DIM, vaginal discharge was scored for diagnosis of purulent vaginal discharge. At 39±3 DIM, endometrial cytology was performed. At 53±3 and 67±3 DIM, ovaries were scanned to determine estrous cyclicity. Pregnancy was evaluated after the first AI. Cure of metritis based on vaginal discharge <5 was greater for ampicillin than ceftiofur on d 5 (37.1 vs. 25.2%) and 7 (57.2 vs. 46.3%), but not on d 12 (82.0 vs. 85.0%). Cure of metritis based on vaginal discharge <5 and no fever was greater for ampicillin than for ceftiofur only on d 7 (50.4 vs. 37.9%), but not on d 5 (23.1 vs. 17.6%) and 12 (66.1 vs. 67.4%). Cows with puerperal metritis had reduced cure compared with cows with fetid discharge on d 5 (30.5 vs. 12.8%), 7 (55.2 vs. 33.6%), and 12 (72.0 vs. 61.1%). The proportion of cows with fever on any day after therapy started did not differ between treatments. Fifty-three percent of cows with metritis based on fetid discharge developed fever after initiating antimicrobial therapy. Cows receiving ampicillin had less prevalence of purulent vaginal discharge than those treated with ceftiofur (57.7 vs. 67.8%), but they were both greater than cows without metritis (21.9%). Prevalence of cytological endometritis did not differ between ampicillin and ceftiofur (30.0 vs. 25.4%), but they were both greater than cows without metritis (14.5%). The proportion of estrous cyclic cows (75.0%) and P/AI did not differ among treatments (ampicillin=28.0% vs. ceftiofur=28.3% vs. without metritis=30.5%). Clinical cure was faster for ampicillin than for ceftiofur, but on study d 12 both treatments resulted in similar cure. Clinical cure was less for cows with puerperal metritis than for cows with fetid uterine discharge. Despite differences in uterine health, P/AI at the first insemination did not differ among treatments.

Chitosan (CHI), a non-toxic and biodegradable biopolymer has been successfully used in the food, human and veterinary medicine industries, especially because of its antimicrobial properties. This study aimed to investigate the effects of CHI on dry matter intake (DMI), nutrient digestibility, blood metabolites, milk yield and composition, and milk fatty acids profile of lactating dairy cows. Sixteen Holstein cows (91.3715.1 days in milk, 29.8 74.1 kg/d of milk, 610 755 kg of body weight [BW]) were used in a 4 Â 4 Latin square design, and each experimental period consisted of 14 days of diet adaptation and 7 days for data collection. Animals were allocated to receive one of treatments: C0: basal diet, without CHI addition; C50, C100 and C150 received 50, 100 and 150 mg/kg BW of CHI, respectively. Chitosan addition did not affect DMI, but increased (Pr 0.05) the digestibility of dry matter (DM), organic matter (OM), crude protein (CP) and neutral detergent fiber (NDF). Blood urea nitrogen concentration was increased (P ¼0.01) with CHI addition. Chitosan decreased (P ¼0.02) nitrogen (N) fecal excretion without affect nitrogen balance. There was no effect of CHI on milk yield, fat corrected milk, and milk composition. Chitosan decreased (P r0.05) C6:0 and C18:1 c9, and quadratically affected C14:1 fatty acids (FA) concentration in milk fat. Chitosan had no effect on total saturated and unsaturated FA of milk fat. Chitosan improved nutrient digestibility without affect productive performance and milk fatty acid profile of midlactation dairy cows.

The main objective was to evaluate the association between vulvovaginal laceration and uterine diseases in dairy cows. The secondary objectives were to evaluate the association between vulvovaginal laceration and cyclicity, and reproductive performance. The vulvovaginal region of 660 Holstein cows from a 5,000 lactating-cows herd was inspected at 4d in milk (DIM) for the presence of lacerations, and scored (VLS) as follows: 0=no laceration; 1=laceration <2cm at the dorsal commissure of the vulva or lateral walls of the vulva/vagina; 2=laceration ≥2cm at the dorsal commissure of the vulva or at the lateral walls of the vulva/vagina, or both. Vaginal discharge was scored at 4, 6, and 8 DIM for diagnosis of metritis, and then at 32 DIM for diagnosis of purulent vaginal discharge (PVD). Data were analyzed using LOGISTIC and PHREG procedures of SAS. Cows with VLS 2 had greater incidence of metritis than cows with VLS 0 (69.1 vs. 42.4%), and cows with VLS 1 tended to have greater incidence of metritis than cows with VLS 0 (52.0 vs. 42.4%). Cows with VLS 2 had greater incidence of PVD than cows with VLS 0 (56.5 vs. 43.1%). A lower proportion of cows with VLS 2 than VLS 0 were cyclic by 64 DIM (70.0 vs. 86.8%). A lower proportion of cows with VLS 2 than VLS 0 were pregnant at 60 d after first AI (28.7 vs. 33.6%). Proportion of pregnant cows at 60d after AI tended to be lower for VLS 1 than VLS 0 (28.4 vs. 33.6%). Hazard of pregnancy by 300 DIM was not affected by VLS. Hazard of pregnancy was decreased for cows with metritis, PVD, and anovular cows. In summary, vulvovaginal laceration was associated with uterine disease and cyclicity, which were negatively associated with reproductive performance. Vulvovaginal laceration was recognized as a risk factor for postpartum uterine disease.

Our expanding knowledge of ovarian function during the buffalo estrous cycle has given new approaches for the precise synchronization of follicular development and ovulation to apply consistently assisted reproductive technologies (ART). Recent synchronization protocols are designed to control both luteal and follicular function and permit fixed-time AI with high pregnancy rates during the breeding (autumn-winter) and nonbreeding (springsummer) seasons. Additionally, allow the initiation of superstimulatory treatments at a self-appointed time and provide opportunities to do fixed-time AI in donors and fixed-time embryo transfer in recipients. However, due the scarce results of in vivo embryo recovery in superovulated buffaloes, the association of ovum pick-up (OPU) with in vitro embryo production (IVEP) represents an alternative method of exploiting the genetics of high yeld buffaloes. Nevertheless, several factors appear to be critical to OPU/IVEP efficiency, including antral follicle population, follicular diameter, environment, farm and category of donor. This review discusses a number of key points related to the manipulation of ovarian follicular growth to improve assisted reproductive technologies in buffalo.

A regionalização de vazões é uma técnica que permite transferir informação entre bacias hidrográficas semelhantes, a fim de calcular, em sítios que não dispõem de dados, as variáveis fluviométricas de interesse; de tal modo, a mesma caracteriza-se por ser uma ferramenta útil na obtenção de outorga de direitos de uso de recursos hídricos, instrumento previsto na Lei 9433/97. O Departamento de Águas e Energia Elétrica (DAEE) é o órgão responsável pela gestão dos recursos hídricos no Estado de São Paulo;utiliza um modelo de regionalização de vazões formulado na década de 80. Diante de algumas limitações do modelo utilizado pelo DAEE, este estudo tem como objetivo geral formular um modelo, por intermédio de uma nova metodologia, e assim testar a precisão dos dois modelos, no que diz respeito à predição de vazões utilizadas como critério de outorga. A avaliação dos modelos fez-se pela validação cruzada,conjunta ao índice de confiança (c), utilizando como valor de estimativa as vazões geradas pelos modelos, e como valor padrão as calculadas por intermédio das séries históricas de estações fluviométricas. O novo modelo rejeitou a hipótese de ter capacidade preditiva inferior e desvantagens de uso, confrontado ao antigo; portanto, a nova metodologia apresenta potencial, quando comparada às comumente utilizadas neste tipo de estudo.

The objective of the current study was to evaluate the effects of supplemental n-3 and n-6 fatty acid (FA) sources on cellular immune function of transition dairy cows. Animals were randomly assigned to receive 1 of 4 diets: control (n=11); whole flaxseed (n-3 FA source; n=11), 60 and 80g/kg of whole flaxseed [diet dry matter (DM) basis] during pre- and postpartum, respectively; whole raw soybeans (n-6 FA source; n=10), 120 and 160g/kg of whole raw soybeans (diet DM basis) during pre- and postpartum, respectively; and calcium salts of unsaturated FA (Megalac-E, n-6 FA source; n=10), 24 and 32g/kg of calcium salts of unsaturated FA (diet DM basis) during pre- and postpartum, respectively. Supplemental FA did not alter DM intake and milk yield but increased energy balance during the postpartum period. Diets containing n-3 and n-6 FA sources increased phagocytosis capacity of leukocytes and monocytes and phagocytosis activity of monocytes. Furthermore, n-3 FA source increased phagocytic capacity of leukocytes and neutrophils and increased phagocytic activity in monocytes and neutrophils when compared with n-6 FA sources. Supplemental FA effects on adaptive immune system included increased percentage of T-helper cells, T-cytotoxic cells, cells that expressed IL-2 receptors, and CD62 adhesion molecules. The results of this study suggest that unsaturated FA can modulate innate and adaptive cellular immunity and trigger a proinflammatory response. The n-3 FA seems to have a greater effect on phagocytic capacity and activity of leukocytes when compared with n-6 FA.