Systemic inflammation (SI) is increasingly studied in several species because it may be central in many metabolic disturbances and be a risk factor for clinical disease. This proof-of-concept study evaluated the effects of the anti-inflammatory drug meloxicam on markers of SI and energy metabolism, polymorphonuclear neutrophil (PMN) function, and endometritis in clinically healthy postpartum dairy cows. Cows received meloxicam (0.5 mg/kg of body weight; n = 20) once daily for 4 days (10-13 days postpartum) or were untreated (n = 22). Blood samples were collected −7,

Paraoxonase-1 (PON1) is an enzyme found in serum and follicular fluid that protects cell membrane and circulating lipids against oxidative damage. The aims of this study were to measure the direct effects of recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and to measure the carry-over effects of PON1 on pre-implantation embryo development in vitro. COCs were submitted to IVM with the addition of 0.0, 0.02, 0.04 and 0.08 mg ml(-1) of rPON1, corresponding to an average PON1 arylesterase enzyme activity of 2.2 ± 0.4, 15.5 ± 1.5, 30.2 ± 3.0 and 57.9 ± 5.0 U ml(-1) , respectively. The results indicated that addition of rPON1 during IVM improved embryo development in a dose-dependent manner as D7 embryo development was 22.2%, 29.4%, 32.2% and 37.0% for the treatment groups, respectively (p = 0.02). In conclusion, addition of PON1 enzyme during IVM exerted dose-related positive effects on embryo development rates to blastocysts.

This study evaluated the effects of treatment with meloxicam (a non-steroidal anti-inflammatory drug), parity, and blood progesterone concentration on the dynamics of the uterine microbiota of 16 clinically healthy postpartum dairy cows. Seven primiparous and 9 multiparous postpartum Holstein cows either received meloxicam (0.5 mg/kg SC, n = 7 cows) once daily for 4 days (10 to 13 days in milk (DIM)) or were untreated (n = 9 cows). Endometrial cytology samples were collected by cytobrush at 10, 21, and 35 DIM, from which the microbiota analysis was conducted using 16S rRNA gene sequence analysis. A radioimmunoassay was used to measure progesterone concentration in blood serum samples at 35 DIM and cows were classified as ˃ 1 ng/mL (n = 10) or ≤ 1 ng/mL (n = 6). Alpha diversity for bacterial genera (Chao1, Shannon-Weiner, and Camargo’s evenness indices) were not affected by DIM, meloxicam treatment, parity, or progesterone category. For beta diversity (genera level), principal coordinate analysis (Bray-Curtis) showed differences in microbiota between parity groups. At the phylum level, the relative abundance of Actinobacteria was greater in primiparous than multiparous cows. At the genus level, there was lesser relative abundance of Bifidobacterium, Lactobacillus, Neisseriaceae, Paracoccus, Staphylococcus, and Streptococcus and greater relative abundance of Bacillus and Fusobacterium in primiparous than multiparous cows. Bray-Curtis dissimilarity did not differ by DIM at sampling, meloxicam treatment, or progesterone category at 35 DIM. In conclusion, uterine bacterial composition was not different at 10, 21, or 35 DIM, and meloxicam treatment or progesterone category did not affect the uterine microbiota in clinically healthy postpartum dairy cows. Primiparous cows presented a different composition of uterine bacteria than multiparous cows. The differences in microbiota associated with parity might be attributable to changes that occur consequent to the first calving, but this hypothesis should be investigated further.

Our objective was to investigate the lipid content of uterus, blood plasma, and milk at early, mid, and late diestrus. Lactating cows (n = 30) had the estrous cycle and ovulation synchronized by administration of exogenous hormones. Cows were blocked by parity and assigned randomly to receive transcervical uterine flushing and biopsy on d 5 (early diestrus), 10 (mid diestrus) or 15 (late diestrus) of the estrous cycle. Flushing and endometrial biopsy were performed in the uterine horn ipsilateral to the corpus luteum. The recovered flushing was used for analyses of lipid composition by liquid chromatography-tandem mass spectrometry and the biopsy was used for investigation of lipid droplet abundance in endometrial cryosections using a neutral lipid fluorescent dye. In addition, blood and milk samples were collected from all cows on d 5, 10, and 15. All blood samples were used to measure the concentration of progesterone in plasma, and all milk samples were used to determine milk composition. Subsamples of blood plasma and milk were also used to evaluate the composition of fatty acids and oxylipins using the same methodology used for uterine flushing samples. The abundance of lipid droplets in the endometrium increased 1.9-fold from d 5 to 10, and 2-fold from d 10 to 15. Concentration of long-chain fatty acids and oxylipins in uterine flushing were, on average, 2.2 and 2.5 times greater in samples collected on d 15 compared with those collected on d 5 and 10. These differences were not observed in blood and milk, suggesting that accumulation of fatty acids and oxylipins in the uterus is regulated locally. In addition to concentration, the profile of individual fatty acids and oxylipins in uterine lumen changed substantially during diestrus. The main categories with increased abundance at late diestrus were mono-and polyunsaturated fatty acids, and oxylipins derived from arachidonic acid, dihomo-γlinolenic acid, and docosahexaenoic acid. In conclusion, fatty acids and oxylipins accumulate in the uterine lumen during diestrus and might work as a mechanism to supply these lipids to the developing conceptus at late diestrus, when the onset of elongation occurs and substantial synthesis of biomass and cell signaling by lipid mediators are required.

12 This study evaluated the effects of treatment with meloxicam (a non-steroidal anti-inflammatory 13 drug), parity, and blood progesterone concentration on the dynamics of the uterine microbiome 14 of clinically healthy postpartum dairy cows. Seven primiparous and 9 multiparous postpartum 15 Holstein cows received meloxicam (0.5 mg/kg SC, n = 7 cows) once daily for 4 days (10 to 13 16 days in milk (DIM)) or were untreated (n = 9 cows). Endometrial cytology samples were 17 collected by cytobrush at 10, 21, and 35 DIM, from which the metagenomic analysis was done 18 using 16S rRNA gene sequence analysis. A radioimmunoassay was used to measure 19 progesterone concentration in blood serum samples at 35 DIM and cows were classified as ˃ 1 20 ng/mL (n = 10) or ≤ 1 ng/mL (n = 6). Alpha diversity for bacterial genera (Chao1, Shannon-21 Weiner, and Camargo's evenness indices) were not affected by DIM, meloxicam treatment, 22 parity, or progesterone category (P > 0.2). For beta diversity (genera level), principal coordinate 23 analysis (Bray-Curtis) showed differences in microbiome between parity groups (P = 0.01).24 There was lower overall abundance of Anaerococcus, Bifidobacterium, Corynebacterium, 25 Lactobacillus, Paracoccus, Staphylococcus, and Streptococcus and higher abundance of 26 Bacillus, Fusobacterium, and Novosphingobium in primiparous than multiparous cows (P < 27 0.05); these patterns were consistent across sampling days. Bray-Curtis dissimilarity did not 28 differ by DIM at sampling, meloxicam treatment, or progesterone category at 35 DIM (P > 0.5).29 In conclusion, uterine bacterial composition was not different at 10, 21, or 35 DIM, and 30 meloxicam treatment or progesterone category did not affect uterine microbiota in clinically 31 healthy postpartum dairy cows. Primiparous cows presented a different composition of uterine 32 bacteria than multiparous cows. The differences in microbiome associated with parity might be 33 attributable to changes that occur consequent to the first calving, but this hypothesis should be 34 investigated further.

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The aim of this study was to compare the J-Synch and conventional protocols associated with estrus detection in beef heifers and to compare pregnancy rate between non-lactating cows displaying estrus or not during the J-Synch protocol. In Experiment 1, heifers were subjected to timed artificial insemination (AI) in a conventional protocol with ECP (n=147) or J-Synch protocol plus eCG (n=149). The AI occurred 12 hours after estrus expression; or 48 (Conventional protocol) and 72 hours (J-Synch protocol) after device removal for animals not displaying estrus. The J-Synch group received 10 µg of GnRH at AI. In Experiment 2, the J-Synch was performed (n=116 cows), but without eCG injection, and estrus was monitored. Pregnancy rate was not different between protocols in Experiment 1 (Conventional: 50.68%; J-Synch: 60.4%). Heifers that displayed estrus had higher pregnancy rate only in the conventional protocol. In Experiment 2, pregnancy rate was not different between cows that displayed estrus or not. Therefore, performing AI earlier according to estrus expression increases pregnancy rate in conventional protocol, however it does not increase pregnancy rate in the J-Synch protocol.Keywords: early artificial insemination; cattle; estrus behavior; GnRH-based protocol. ResumoOs objetivos desse estudo foram comparar o protocolo J-Synch e o protocolo convencional associados com detecção de estro em novilhas de corte e comparar a taxa de prenhez entre vacas não lactantes que demonstraram ou não estro após o protocolo J-Synch. No experimento 1, as novilhas foram submetidas à inseminação artificial (IA) em tempo fixo em um protocolo convencional com ECP (n=147) ou através do protocolo J-Synch com eCG (n=149). A IA ocorreu 24 horas após o estro; ou 48 (Convencional) e 72 horas (J-Synch) após a remoção do dispositivo naqueles animais que não demonstraram estro. O grupo J-Synch recebeu 10 µg de GnRH no momento da IA. No experimento 2, foi aplicado o protocolo J-Synch (n=116 vacas) sem administração de eCG e o estro foi monitorado. No Experimento 1, a taxa de prenhez não foi diferente entre os protocolos (convencional: 50,68%; J-Synch: 60,4%). Novilhas que demonstraram estro tiveram maior taxa de prenhez apenas no protocolo convencional. No Experimento 2, a taxa de prenhez não foi diferente entre as vacas que demonstraram ou não estro. Assim, antecipar o momento da IA de acordo com o estro aumenta a taxa de prenhez no protocolo convencional, contudo não aumenta a taxa de prenhez no protocolo J-Synch.Palavras-chave: antecipação da inseminação artificial; bovinos; comportamento estral; protocolos baseados em GnRH.

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