Methylome Patterns of Cattle Adaptation to Heat Stress.

Heat stress has a detrimental impact on cattle health, welfare and productivity by affecting gene expression, metabolism and immune response, but little is known on the epigenetic mechanisms mediating the effect of temperature at the cellular and organism level. In this study, we investigated genome-wide DNA methylation in blood samples collected from 5 bulls of the heat stress resilient Nellore breed and 5 bulls of the Angus that are more heat stress susceptible, exposed to the sun and high temperature-high humidity during the summer season of the Brazilian South-East region. The methylomes were analyzed during and after the exposure by Reduced Representation Bisulfite Sequencing, which provided genome-wide single-base resolution methylation profiles. Significant methylation changes between stressful and recovery periods were observed in 819 genes. Among these, 351 were only seen in Angus, 366 were specific to Nellore, and 102 showed significant changes in methylation patterns in both breeds. KEGG and Gene Ontology (GO) enrichment analyses showed that responses were breed-specific. Interestingly, in Nellore significant genes and pathways were mainly involved in stress responses and cellular defense and were under methylated during heat stress, whereas in Angus the response was less focused. These preliminary results suggest that heat challenge induces changes in methylation patterns in specific loci, which should be further scrutinized to assess their role in heat tolerance.

Functions of the GABAergic system on serum LH concentrations in pre-pubertal Nellore heifers.

This study was conducted to evaluate the luteinizing hormone (LH) secretion pattern after gamma-aminobutyric acid (GABAA) antagonist to determine the effects of the GABAergic system on LH secretion during reproductive maturation in pre-pubertal Nellore heifers. Nellore heifers (n = 10) were administered a picrotoxin injection of 0.18 mg/kg, i.v. Blood samples were collected every 15 min for 3 h at different developmental stages (8, 10, 14 and 17 mo of age). Plasma concentrations of LH were quantified using an RIA (sensitivity of 0.04 ng/mL and CV of 15 %). There was an interaction between treatment and age (P = 0.034). Picrotoxin-treated heifers had lesser (P ≤  0.05) LH mean concentrations during a 3 h period at 10 and 17 mo of age compared to control heifers (P ≤  0.05). Comparing the period before and after Picrotoxin injection in the same animals, there was a 33 % decrease in LH concentration at 8 mo of age (P = 0.0165). These results indicate the GABAergic system has a stimulatory function in inducing LH secretion in pre-pubertal Nellore heifers. These findings corroborate previous results that GABA increases GnRH/LH secretion in other species during the pre-pubertal period.

Early resynchronization of follicular wave emergence among Nelore cattle using injectable and intravaginal progesterone for three timed artificial inseminations.

Follicular dynamics and pregnancy per AI (P/AI) of cattle submitted to resynchronization 13 d after TAI using a progesterone (P4) insert and supplementary injectable progesterone (iP4) were determined. There was synchronization of ovulation timing imposed (Day 0 = expected estrus; Experiment 1). On Day 13, animals were assigned to: control (only P4 insert; 15 cows and 13 heifers) and iP4 (P4 insert+100 mg iP4; 13 cows and 12 heifers) groups, and submitted to daily ovarian ultrasonography and blood collection (Day 13 to Day 22). In Experiment 2, 760 suckled cows and 498 heifers were submitted to a TAI on Day 0 and assigned into control and iP4 groups on Day 13. In animals with luteolysis on Day 22, there was a second TAI on Day 24, and on Day 37 were subjected to the opposite treatments as imposed in first resynchronization procedure. On Day 37, there was pregnancy diagnosis in animals with functional CL. The third TAI was performed on Day 48. Day of follicular wave emergence did not differ between groups and parities. Dominant follicles were larger in cows than heifers, and in animals of the control group on Day 24. Greater P4 concentrations were detected on Day 14 and Day 15 in the iP4-treated animals. Luteolysis occurred earlier in cows than heifers. Overall P/AI percentage as a result of second and third TAIs, regardless of parity, was greater in iP4-treated animals. In conclusion, females treated with a supplementary iP4 had a greater plasma P4 concentration and P/AI, but there was no effect of iP4 treatment on synchrony of timing of follicular wave.

Serum Progesterone and Conception Rates in Acyclic Embryo Recipient Mares Using a Bovine Progesterone-Releasing Intravaginal Device.

The objective of this study was to quantify serum progesterone levels, uterine features, and pregnancy rates in acyclic, embryo recipient mares using a bovine progesterone-releasing intravaginal device in a commercial embryo transfer (ET) program. The study included 73 recipient mares of unknown breed, aged 3-10 years, weighing 350-500 kg, and kept under an intensive management system on Tifton 85 (Cynodon spp.) pastures with water and mineral salt ad libitum. The horses were divided into two groups: a group with a progesterone-releasing intravaginal device (1 g progesterone, G-IVP4, n = 24) and a control group (G-iP4, n = 49) receiving an injection of 1,500 mg long-acting progesterone. Jugular blood was collected for the G-IVP4 group for subsequent progesterone measurement by radioimmunoassay on three occasions: Day 0 (D0), intravaginal device was placed; Day 5 (D5), day of the ET; and Day 9 (D9), day of pregnancy diagnosis. There was an increase (P < .0001) in serum progesterone levels on D5 and D9 compared with D0 (4.09 ± 0.81 and 6.45 ± 1.03 ng/mL vs. 0.71 ± 0.14 ng/mL). There were no differences among groups in the pregnancy rate (P > .05), with rates of 83.33% and 73.46% for G-IVP4 and G-iP4, respectively. In conclusion, the intravaginal route for absorption of 1 g of progesterone device increased the serum level of progesterone sufficiently to prepare the uterus of acyclic recipient mares for ET, and the conception rate was similar to the standard protocol using long-acting injectable progesterone.

Effect of dietary supplementation of palm kernel cake on ovarian and hepatic function in buffalo (Bubalus bubalis).

To determine the optimal inclusion amount of palm kernel cake (PKC) in a buffalo diet, in the present study there was evaluation of the ovarian activity, metabolism and hepatic function of females that were treated to synchronize the time of ovulation. Twenty-four estrous-cyclic and non-lactating Murrah buffalo with a mean age of 5.7 years were supplemented with 0%, 0.25%, 0.5% and 1% of their body weight (BW) with PKC. Animals were subjected to the Ovsynch protocol (beginning of protocol = D0). The ovaries were examined and the blood was collected on D10 (follicular phase) and D17 (luteal phase). Follicular and luteal development and serum progesterone concentrations were not affected by diet (P > 0.05). Serum concentrations of cholesterol were greater in animals supplemented with PKC in amounts at 0.5% of BW or less with PKC, regardless of the phase of the estrous cycles when evaluations occurred (P < 0.05). Concentrations of HDL-cholesterol were similar (P > 0.05) during the follicular and luteal phases. Triglyceride concentrations increased linearly (P = 0.03) as percentage of PKC inclusion diets increased during the follicular phase, but were similar in the luteal phase (60.0 mg/dL; P = 0.51). Amount of PKC supplementation did not affect the concentrations of alanine aminotransferase, but there was a greater amount of aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT) during both phases of the estrous cycle (P < 0.05). Animals supplemented at 1.0% of BW with PKC had greater AST and GGT concentrations than what is recommended for buffalo. The results of the present study indicate PKC supplementation of buffalo diets does not affect the development of the ovarian follicle and corpus luteum nor the peripheral concentration of progesterone, even though there are greater serum concentrations of total cholesterol and triglycerides. Because the amount of PKC supplementation in the present study does not result in hepatic dysfunction when fed at the 0.5% of BW amount, it is suggested that this agro-industrial byproduct of high nutritional value may be a new alternative for dietary supplementation of grazing buffalo.

Characterization of the LH peak after short and long fixed-time artificial insemination protocols in sheep raised in the tropics.

The aim of this study was to evaluate the peak in luteinizing hormone (LH) and the pregnancy rate of sheep (Texel × Santa Inês) in the tropics using short- (6 days) and long-term (12 days) progesterone protocols followed by artificial insemination (AI) both in and out of the breeding season. Experiment 1 was conducted within (IN) the breeding season (autumn, n = 36), and experiment 2 was conducted outside (OUT) of the breeding season (spring, n = 43). In each experiment, the sheep were divided into two groups (6 or 12 days) according to the duration of treatment with a single-use progesterone release vaginal device (CIDR® , Pfizer, São Paulo, SP, Brazil), and blood samples were collected from 10 animals per group every 4 hr to measure the LH and progesterone concentrations. In the spring, the characteristics of the LH peak did not differ between groups; but in the autumn, there were differences between groups at the beginning (G-6 IN: 36.44 ± 5.46 hr; G-12 IN: 26.57 ± 4.99 hr) and end of the LH peak (G-6 IN: 46.22 ± 7.51 hr; G-12 IN: 34.86 ± 8.86 hr). The results showed alterations in the LH peak during the breeding season only in the sheep undergoing the short-term protocol.

Effect of intravenous propofol and remifentanil on heart rate, blood pressure and nociceptive response in acepromazine premedicated dogs.

OBJECTIVE: To investigate the cardiorespiratory, nociceptive and endocrine effects of the combination of propofol and remifentanil, in dogs sedated with acepromazine. STUDY DESIGN: Prospective randomized, blinded, cross-over experimental trial. ANIMALS: Twelve healthy adult female cross-breed dogs, mean weight 18.4 ± 2.3 kg. METHODS: Dogs were sedated with intravenous (IV) acepromazine (0.05 mg kg(-1) ) followed by induction of anesthesia with IV propofol (5 mg kg(-1) ). Anesthesia was maintained with IV propofol (0.2 mg kg(-1) minute(-1) ) and remifentanil, infused as follows: R1, 0.125 µg kg(-1) minute(-1) ; R2, 0.25 µg kg(-1) minute(-1) ; and R3, 0.5 µg kg(-1) minute(-1) . The same dogs were administered each dose of remifentanil at 1-week intervals. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (f(R) ), end tidal CO(2) (Pe'CO(2) ), arterial hemoglobin O(2) saturation, blood gases, and rectal temperature were measured before induction, and 5, 15, 30, 45, 60, 75, 90, and 120 minutes after beginning the infusion. Nociceptive response was investigated by electrical stimulus (50 V, 5 Hz and 10 ms). Blood samples were collected for plasma cortisol measurements. Statistical analysis was performed by anova (p<0.05). RESULTS: In all treatments, HR decreased during anesthesia with increasing doses of remifentanil, and increased significantly immediately after the end of infusion. MAP remained stable during anesthesia (72-98 mmHg). Antinociception was proportional to the remifentanil infusion dose, and was considered satisfactory only with R2 and R3. Plasma cortisol concentration decreased during anesthesia in all treatments. Recovery was smooth and fast in all dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Infusion of 0.25-0.5 µg kg(-1) minute(-1) remifentanil combined with 0.2 mg kg(-1) minute(-1) propofol produced little effect on arterial blood pressure and led to a good recovery. The analgesia produced was sufficient to control the nociceptive response applied by electrical stimulation, suggesting that it may be appropriate for performing surgery.