Predictive value of prepartum serum metabolites for incidence of clinical and subclinical mastitis in grazing primiparous Holstein cows.

The aim of this study was to measure changes in biochemical markers in the peripartum period of primiparous Holstein cows diagnosed with subclinical and clinical mastitis. In this study, 37 dairy cows were monitored daily during milking until 60 days postpartum and were categorized according to the occurrence of clinical mastitis (group mastitis (GM), n = 9) or subclinical mastitis (group subclinical mastitis (GSUB), n = 10) or absence of symptoms (control group (CG), n = 18). Blood samples were collected weekly from -30 to 60 days from calving. Samples were grouped for prepartum (-30 to 0 days from calving), early postpartum (0 to 30 days from calving), and late postpartum (30 to 60 days from calving) periods. Prepartum serum non-esterified fatty acid (NEFA) concentration was higher in GM than in CG (P < 0.01). In addition, CG had higher prepartum serum glucose concentration than GM (P = 0.03). In the early postpartum period, aspartate aminotransferase (AST) activity was lower in CG than in GSUB (P < 0.05), and in the late postpartum period, AST activity was lower in CG than GSUB and GM (P = 0.01). Somatic cell count was higher during the early and late postpartum periods for GM and GSUB when compared to CG (P < 0.01). In this study, primiparous cows with low glucose and higher NEFA in the prepartum were more susceptible for mastitis in the early postpartum, probably due to low immune function associated to a more negative energy balance. In sum, increased prepartum serum NEFA concentration and decreased glucose in primiparous cows were associated with clinical mastitis incidence in the postpartum period.

Effect of recombinant bovine somatotropin (rbST) treatment on follicular population and development in non-lactating dairy cows.

The aim of this study was to evaluate the long-term effects of recombinant bovine somatotropin (rbST) on follicle population and ovulatory follicle development in non-lactating dairy cows. Twenty-one Jersey cows were allocated in rbST (n=11) or control (n=10) groups. On day -60, cows in rbST group received 500 mg of somatotropin (s.c. Lactotropin®, Elanco). On day 0, control and rbST cows received an intravaginal progesterone-releasing device (1.9 g, CIDR®, Zoetis) and GnRH (100 mg, IM, Factrel®, Zoetis). On day 8, cows received PGF2α (25 mg, IM, Lutalyse®, Zoetis) and the CIDR® was removed. Twelve hours after device removal (D8), serum, follicular fluid and granulosa cells samples were collected. Serum and follicular concentration of estradiol (E2) and progesterone (P4) were analyzed. Total RNA was extracted from granulosa cells to measure gene expression of LHCGR, STAR, HSD-3B1, CYP11A1, CYP19A1, CYP17A1, IGFR and PAPPA by real-time PCR. Ultrasonography was performed on days -60, -53, -46, -14, -7, 0 and 8 for antral follicle count. Results were analyzed by repeated measures ANOVA and t-test. There was no effect of rbST treatment on the number of follicles during the 60 days period, as well as no effect on serum and follicular fluid E2 and follicular fluid P4 at the moment of follicle aspiration. There was a reduction in PAPPA (P = 0.006), CYP11A1 (P = 0.04) and CYP19A1 (P = 0.002) mRNA levels in granulosa cells of the pre-ovulatory follicle of rbST treated cows. In conclusion, a single dose of rbST did not have long-term effects on antral follicle population, serum and follicular E2/P4 concentrations in non-lactating dairy cows. Despite that, rbST injection decreased granulosa cell expression of genes related to steroidogenesis in the pre-ovulatory follicle.

Effect of bovine somatotropin injection in late pregnant Holstein heifers on metabolic parameters and steroidogenic potential of the first postpartum dominant follicle.

The aim of this study was to determine the effect of pre-partum injections of bovine somatotropin (bST) in dairy heifers on metabolic markers and the steroidogenic potential of the first postpartum dominant follicle. Heifers were assigned to two groups: bST (ST; n = 29), that received two doses of bST (500 mg/dose) at -28 and -14 days relative to calving; and control (CTL; n = 30), that did not received bST. Follicular development was monitored via ultrasound every 3 days starting at 8 days in milk (DIM) in a subset of 20 heifers until the day the first large follicle reached a diameter of 16 mm. From these cows follicular fluid was aspirated and the follicular cells recovered (ST; n = 8 and CTL; n = 10). Blood samples were collected weekly for all heifers. Follicular fluid IGF-I concentrations of the first postpartum dominant follicle was higher (P = 0.05) in ST (87.1 ± 7.7 mg/mL) than CTL cows (64.3 ± 6.8 mg/mL). Also, E2 concentration in the follicular fluid was higher (P = 0.02) for ST (199.7 ± 55.9 ng/mL) than CTL cows (74.5 ± 37.7 ng/mL). The expression of LHCGR and STAR mRNA in follicular cells was higher (P < 0.05) in ST than CTL cows. Nonetheless, HSD3B, P450scc, P450c17, IGFr and CYP19A1 mRNA expression was not different between groups (P > 0.05). Serum IGF-I concentration was higher in ST treated heifers during the pre-partum period (P = 0.01) and no difference was observed in the postpartum period (P = 0.19). In conclusion, pre-partum bST treatment in dairy heifers increased intrafollicular IGF-I and expression of LHCGR and STAR mRNA in follicular cells of the first postpartum dominant follicle. These changes were associated to increased intrafollicular and serum E2 concentration, which can potentially increase the chance of ovulation of the first follicular wave.

The acute effect of intravenous lipopolysaccharide injection on serum and intrafollicular HDL components and gene expression in granulosa cells of the bovine dominant follicle.

The aim of this study was to evaluate the effect of an acute systemic inflammatory response induced by lipopolysaccharide (LPS) in the serum and follicular fluid (FF) high-density lipoprotein (HDL) components, hormone concentrations and granulosa cell gene expression. For this purpose, twenty non-lactating Jersey dairy cows were submitted to a progesterone (P4) - estradiol (E2) based synchronization protocol. Cows received a single i.v. dose of LPS (2.5 µg/kg of body weight) or saline solution (CTL Group) 2 h after P4 insert removal. Blood, granulosa cells and FF samples were collected six hours after LPS injection. Five hours after LPS injection rectal temperature was increased in LPS (P < 0.0001, 40.4 ± 0.1 °C) compared to the CTL cows (38.8 ± 0.1 °C). Serum PON1 activity was reduced by LPS injection (130.2 ± 5.1 vs. 99.6 ± 3.3 U/mL; P < 0.001), as well as HDL-cholesterol concentrations (70.3 ± 5.3 vs. 50.1 ± 6.2 mg/dL; P < 0.05). The FF E2 and P4 concentrations were not different between groups (P > 0.05). The PON1 activity in the FF was also decreased by LPS injection (P = 0.01). In comparison to CTL group, cows injected with LPS had a ten fold reduction in STAR, TLR4 and TNF mRNA expression (P < 0.05). In conclusion, an intravenous LPS challenge in cows induced an acute systemic inflammatory response reducing HDL and its components in serum but not in the FF. Only PON1 activity serum reduction was reflected in the FF in the short term. Additionally, steroidogenic and inflammatory genes had reduced expression in the granulosa cells.

Variações metabólicas e endócrinas de cachaços híbridos (Landrace x Large White) durante o primeiro ano de vida e sua relação com o estabelecimento da puberdade / Metabolic and endocrine changes of hybrid boars (Landrace x Large White) during the first year of life and its relation to the establishment of puberty

O objetivo deste estudo foi determinar variações em parâmetros metabólicos e endócrinos de suínos machos durante o primeiro ano de vida e sua relação com a puberdade. Foram utilizados seis suínos acompanhados, de 22 a 360 dias deidade, com pesagens corporais e coletas de sangue a cada sete dias para avaliação dos níveis séricos de glicose, insulina, colesterol, albumina, ureia, cálcio, fósforo, aspartato aminotransferase (AST), gama glutariltransferase (GGT), ácidos graxos não esterificados (NEFA) e testosterona. A partir do ganho de peso e níveis de testosterona foram estabelecidos três períodos: pré-puberdade (1-5 meses de idade); puberdade (6-9 meses de idade); pós-puberdade (10-12 meses de idade), em que os níveis de testosterona foram de 0,6 ± 0,1 ng/mL, 1,8 ± 0,2 ng/mL e 1,8 ± 0,2 ng/mL, respectivamen- te (P < 0,0001). Foi observada correlação positiva entre a testosterona e níveis de albumina, idade e peso corporal, e correlação negativa entre testosterona e os níveis de cálcio, GGT e glicose. Os níveis de glicose diminuíram ao longo do desenvolvimento (P < 0,0001), e os níveis de NEFA foram maiores em pré-púberes (P = 0,01). A enzima GGT teve diminuição dos seus níveis a partir da puberdade (P < 0,0001), e a AST teve seus menores níveis na puberdade (P = 0,0003). A albumina apresentou maiores níveis durante a puberdade (P < 0,0001) e a ureia no período pós-puberdade (P < 0,0001). Os níveis de cálcio e fósforo apresentaram menores níveis no período pós-puberdade (P<0,0001). Assim, a puberdade representa um período de flutuação nos níveis de marcadores metabólicos de suínos machos híbridos, devido ao efeito da testosterona sobre o metabolismo energético, proteico e mineral.

The present study aimed to determine changes in endocrine and metabolic parameters in male pigs during the first year of life and its relation with the puberty. Six male pigs of 22 at 360 days of age were used in this study. Body weights and blood collection were performed every seven days for evaluation of serum glucose, insulin, cholesterol, albumin, urea,calcium, phosphorus, aspartate aminotransferase (AST), gamma glutamyltransferase (GGT), non-esterified fatty acids (NEFA) and testosterone. From the weight gain and testosterone levels were established three periods: prior to puberty (1-5 months old); puberty (6-9 months old); postpubertal (10-12 months old). During these periods testosterone levels were 0.6 ± 0.1 ng/mL, 1.8 ± 0.2 ng/mL and 1.8 ± 0.2 ng/mL, respectively (P < 0.0001). Positive correlation was observed between testosterone levels and albumin levels, age and body weight, and a negative correlation between testosterone levels and calcium, glucose and GGT. Glucose levels decreased throughout development (P < 0.0001) whereas NEFA levels were higher in pre-pubertal (P = 0.01). The enzyme GGT levels had decreased from puberty (P < 0.0001), and AST had their lowest levels at puberty (P = 0.0003). Albumin showed higher levels during puberty (P < 0.0001). The urea values were stable until puberty, and increased post-puberty (P < 0.0001). The levels of calcium and phosphorus levels had lower post-puberty (P < 0.0001). Thus, puberty is a period of fluctuation in the levels of metabolic markers of hybrid male pigs due to the effect of testosterone on energy, protein and mineral metabolism.