Superovulatory responses of cattle

Non-histological examination of superovulated ovaries of cows does not allow one to distinguish between corpora lutea and luteinized follicles. A better estimation of ovulation rate could, therefore, be made from the number of embryos recovered or from the levels of E₂-17β in the plasma 60 hours after PMSG.For comparison of different treatments, it is necessary to characterize activities of the stimulatory agents used. Administration of an FSH - LH preparation twice a day at decreasing doses gives the best mean responses, but no treatment has been found which can clearly decrease the large variation between individuals in their responses.Numerical, kinetic and endocrine ovarian factors can partly explain the variability of ovarian responses to PMSG in the heifer. Individual differences in follicular populations at the time of treatment, or in E₂-17β levels after stimulation, could be related to differences in responses in ovulation rate. Normal follicles >1.7 mm diameter before treatment would usually ovulate following PMSG injection, whereas early atretic follicles of the same size mostly luteinize.     

Superovulatory responses in dairy cows following ovulation of the dominant follicle of the first wave

In the present study we investigated the effect of hCG administration on Day 7 (Day 0 = day of standing estrus) to ovulate the dominant follicle of the first wave and the associated increase in progesterone concentration on subsequent superovulatory response in dairy cows. Twenty cyclic lactating cows were allocated at random to 2 groups: control (n = 10) and hCG-treated (n = 10). The ovaries of each cow were scanned using an ultrasound scanner on Day 7, to confirm the presence of the dominant follicle and thereafter every other day until embryo recovery. All cows received a total dose of 400 mg Folltropin-V in decreasing amounts for 5 days (Days 9 to 13) and 35 mg PGF(2alpha) on Day 12. In addition, the treated cows received 1000 IU hCG on Day 7. All cows were inseminated twice during estrus, and the embryos were collected 7 days later by a nonsurgical procedure. Blood smaples were taken at different times of the treatment period for progesterone determination. All cows possessed a dominant follicle at Day 7, and all but one of the hCG-treated cows ovulated the dominant follicle and formed an accessory corpus luteum. Plasma progesterone concentrations were significantly higher (P<0.01) in hCG-treated cows than control cows on the first day of Folltropin treatment and on the day of PGF(2alpha) injection. The mean number of follicles at estrus, the number of ovulations, the total number of embryos and the number of transferable embryos were not different (P>0.05) between control and hCG-treated cows.     

Thermoregulatory responses of Holstein cows exposed to experimentally induced heat stress

Heat stress (HS) adversely influences productivity and welfare of dairy cattle. We hypothesized that the thermoregulatory mechanisms vary depending on the exposure time to HS, with a cumulative effect on the adaptive responses and thermal strain of the cow. To identify the effect of HS on adaptive thermoregulatory mechanisms and predictors of caloric balance, Holstein cows were housed in climate chambers and randomly distributed into thermoneutral (TN; n=12) or HS (n=12) treatments for 16 days. Vaginal temperature (VT), rectal temperature (Tre), respiratory rate (RR), heart rate (HR), and dry matter intake (DMI) were measured. The temperature and humidity under TN were 25.9±0.2 °C and 73.0±0.8%, respectively, and under HS were 36.3±0.3 °C and 60.9±0.9%, respectively. The RR of the HS cows increased immediately after exposure to heat and was higher (76.02±1.70bpm, p<0.001) than in the TN (39.70±0.71bpm). An increase in Tre (39.87±0.07 °C in the HS vs. 38.56±0.03 °C in the TN, p<0.001) and in VT (39.82±0.10 °C in the HS vs. 38.26±0.03 °C in the TN, p<0.001) followed the increase in RR. A decrease (p<0.05) in HR occurred in the HS (62.13±0.99bpm) compared with the TN (66.23±0.79bpm); however, the magnitude of the differences was not the same over time. The DMI was lower in HS cows from the third day (8.27±0.33 kg d⁻¹ in the HS vs. 14.03±0.29 kg d⁻¹ in the TN, p<0.001), and the reduction of DMI was strongly affected (r=−0.65) by changes in the temperature humidity index. The effect of environmental variables from the previous day on physiological parameters and DMI was more important than the immediate effect, and ambient temperature represented the most determinant factor for heat exchange. The difference in the responses to acute and chronic exposure to HS suggests an adaptive response. Thus, intense thermal stress strongly influence thermoregulatory mechanisms and the acclimation process depend critically on heat exposure time.     

Thermoregulatory responses of Holstein and Brown Swiss Heat-Stressed dairy cows to two different cooling systems

Thirty-seven Holstein and 26 Brown Swiss dairy cows were used to evaluate the effect of two different cooling systems on physiological and hormonal responses during the summer. A control group of cows had access only to shade (C). A second group was cooled with spray and fans (S/F) and the third group was under an evaporative cooling system called Korral Kool (KK). The maximum temperature humidity index during the trial was from 73 to 85. Rectal temperatures and respiration rates of the C group were higher (P < 0.05) than those of the S/F and KK groups in both Holstein and Brown Swiss cows. Triiodothyronine levels in milk were higher (P < 0.05) in the KK group than in the S/F and C groups, while cortisol levels were lower (P < 0.05) in the C group than in S/F and KK. There was no significant difference in the hormonal response of the two breeds. These results demonstrate that both cooling systems may be used increase the comfort of Holstein and Brown Swiss cows during summer in hot, dry climates.     

Superovulatory responses to eCG in llamas (Lama glama )

Llamas are copulation-induced single-ovulators, and multiple ovulation and embryo transfer (MOET) methods have not yet been developed for this species. Superovulatory responses to eCG given during an induced (Group A) or simulated (Group B) luteal phase were investigated using ultrasound to observe ovarian follicles and corpora lutea (CLs) and plasma progesterone was used to assess luteal function. Embryos were recovered nonsurgically. Group A (n = 19): donors were given 8 microg, im GnRH analogue (Day 0) to induce ovulation of a mature follicle, 1000 IU, im eCG (Day 7), and 250 microg PGF(2alpha) analogue (Day 9). Group B (n = 17): donors were given a subcutaneous progestagen implant (3 mg Norgestomet) at Days 0 to 7) and 1000 IU, im eCG (Day 5). When most (>65%) of the follicles in both Groups A and B had matured at 5 to 11 d post eCG, the donors were given 8 microg, im GnRH and mated once (n = 26) or twice within a 24-h interval (n = 10); embryos were recovered 6 to 9 d post ovulation. More follicles and corpora lutea were induced in Group B than in Group A, but a similar mean number of embryos were recovered (1.3 vs 1.6), and a similar proportion of donors yielded multiple embryos (35 vs 32%). The embryo recovery rate was similar for Groups A and B (39 and 37%), but it was higher (P < 0.001) with 2 (72%) rather than 1 (22%) mating, and it was negatively correlated with CL number (P < 0.05). Overall, 80% of the llamas had a precocious CL and elevated plasma progesterone concentrations when multiple follicles reached maturity. This was associated with increased subsequent superovulation and embryo recovery (P < 0.01). Peak plasma progesterone was positively correlated with the CL number (P < 0.05). From these results we conclude that superovulation may be achieved with eCG given during either an induced or a simulated luteal phase, that embryo recovery is improved following 2 matings rather than 1, and that MOET may indeed be feasible for use in the llama.     

Ovarian responses and embryo survival in recipient lactating Holstein cows treated with equine chorionic gonadotropin

The objectives of Experiment 1 were to determine a dose of eCG that would increase total luteal volume and plasma progesterone (P4) concentration on estrous cycle Day 7 in cows. The objectives of Experiment 2 were to determine the effects of treating embryo recipient lactating Holstein cows with eCG on pregnancy per embryo transfer (P/ET). In Experiment 1, lactating dairy cows at 63 ± 3 d postpartum (DIM) received no treatment (control, n = 10), or 600 (eCG6, n = 19), or 800 (eCG8, n = 19) IU of eCG 2 d after the start of the ovulation-synchronization protocol, Day -8 (Day -10 GnRH, Day -3 PGF_2α, Day 0 GnRH). Blood was sampled on Days -10, -8, -3, 0, 7, and 14 for P4 concentration. Ovaries were examined by ultrasound on Days -10, -3, 0, and 7. In Experiment 2, lactating dairy cows were paired according to parity and previous insemination (0 or > 1 insemination) and assigned to receive 800 IU of eCG (eCG8, n = 152) 2 d after the start of the ovulation-synchronization protocol (Day -10 GnRH, Day -3 PGF_2α, Day 0 GnRH) or to receive no treatment (control, n = 162). Blood was sampled on Days -10, -3, 0, 7, and 14 for determination of P4 concentration. Ovaries were examined by ultrasound on Days -10, -3, and 7, and cows with a CL > 20 mm in diameter on Day 7 received an embryo. In Experiment 1, P4 concentration on Day 7 was higher (P < 0.05) for eCG8 cows (2.3 ± 0.3 ng/mL) compared with control (1.2 ± 0.3 ng/mL) and eCG6 (1.1 ± 0.3 ng/mL) cows. In Experiment 2, eCG8 primiparous cows had more (P < 0.01) follicles > 10 mm on Day -3 compared with control primiparous cows (2.5 ± 0.9 vs 1.7 ± 0.5 mm), but multiparous control and eCG8 cows did not differ. A larger (P = 0.03) percentage of control cows received an embryo (87.5 vs 79.1%) compared with eCG8 cows. Among cows that received an embryo, total luteal volume on Day 7 was affected (P = 0.05) by treatment (eCG8 = 8.3 ± 0.4 cm³, control = 6.2 ± 0.4 cm³), but P4 concentration on Day 7 did not differ significantly between treatments. The percentage of cows pregnant 53 d after ET (overall, 24.2%) was not significantly different between control and eCG8 cows. In the current study, no differences in P/ET were observed between control and eCG8 cows and treatment with eCG increased the percentage of cows with asynchronous estrous cycle.     

Estrous and related behaviour in postpartum Holstein cows

Over a period of 1 year, three consecutive groups, each consisting of 12 postpartum Holstein cows aged from 3 to 9 years, were observed continually for 80 days. Animals were housed in a windowless free stall environment under husbandry and nutrition practices similar to those employed in commercial dairy units. A time lapse videorecorder, and two low light intensity television cameras were used to monitor behavioural activities of the cows. The stall was continuously illuminated with an intensity of 32 lux at the eye level of a standing cow. Each day the entire videotape was viewed and the number of mounts, the feeding, drinking, resting and walking activities of each cow were recorded.The distribution of mounts showed evidence of circadian rhythm with the highest frequency occurring during the nocturnal period. The total number of mounts and mounting/mounted ratios per estrus varied among individuals and were affected by estrus synchronisation. The number of mounts per cow increased from an average 11.2 with one cow in estrus to 52.6 with three cows in estrus at the same time. The sequence of postpartum estrus indicated a gradual increase both in mounted and mounting activities and mounting/mounted ratios. The data indicate that on the average, more mounted cows were in estrus (90%) than mounting ones (79%). The length of mounting between cows in synchronized estrus lasted on the average longer (7.4±0.41 sec) than between nonsynchronized cows with only one estrous partner (4.6±1.12 sec). Detected duration of true estrus was 7.5, 7.8 and 10.1 h in one, two and three cow estral groups respectively. Behavioural symptoms of the estrus onset also differed among individuals and revealed a dependency on social factors.Estrus stage caused changes in the proportional composition of daily activities. A comparison with 4 pre- and post estral days showed a high increase of walking time at the cost of both resting time and time spent at the feeder. The frequency distribution of agonistic interactions was approximately doubled during the estrous period while the distribution of instigator/victim ratios reached the highest value (1.44) one day before estrus and the lowest value (0.60) on the first post estrus day. The most frequent physical contact in agonistic interactions was in the head to head position which accounted for 64% of all fights. The head-neck position was second (16%), followed by the head-shoulder (9%), head-flank (6%), head-rear (3%) and head-side (2%). Further analysis of agonistic interactions revealed detectable motivation in about 25% of all fights. The number of mounts per estrus indicated a positive relationship to the rank of the victimizing. The mounting/mounted ratio was positively related to instigation order and negatively to age.     

Resumption of postpartum ovarian cyclicity in high-producing Holstein cows

The objective of this study was to investigate the resumption of ovarian cyclicity postpartum in high-producing dairy cows in commercial dairy farms under subtropical conditions. The cows were kept in a free-stall or tie-stall barn. Milk samples were collected from cows twice weekly, and progesterone in the skim milk was assayed by double-antibody ELISA. Cows were examined rectally and vaginoscopically at 2-week intervals after calving. Body condition score (BCS) and body weights were taken before and after calving. A cow was considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Thirty seven percent (n=20/54) of the cows had normal resumption of ovarian cyclicity (resumption within 45 days after calving), and 63% (n=34/54) had delayed resumption (resumption did not occur until >45 days after calving). Delayed resumption Type I (one or more ovarian cycles with luteal phase >20 days, i.e. prolonged luteal phase; 31.5%) and delayed resumption Type II (first ovulation did not occur until > or =45 days after calving, i.e. delayed first ovulation; 24.1%) were the most common types of delayed resumptions. Almost half (46.3%) of the cows did not resume their ovarian cyclicity until >65 days postpartum. Cows with delayed resumption Type I had a higher incidence of abnormal cervico-vaginal discharge (64.7%) and incomplete uterine involution (94.1%) compared to cows with normal resumption (P<0.01). The BCS of cows with delayed resumption Type II were lower than those of normal resumption cows at 5 weeks and later in the postpartum period (P<0.05). Approximately two-thirds of high-producing cows had delayed resumption of ovarian cyclicity postpartum. Prolonged luteal phase and delayed first ovulation were two important ovarian dysfunctions that delayed postpartum resumption of cyclicity in high-producing dairy cows.     

Genetic analysis of reproductive disorders in Israeli Holstein dairy cows

The heritability of common reproductive disorders was asessed in Israeli Holstein cows. Data were collected from 76,000 calvings on 102 collective farms (kibbuts) over a 36-mo period. The disorders considered were those of retained placenta, abnormal lochia, endometritis, anestrus, ovarian cysts with anestrus and ovarian cysts with nymphomania. The statistical model included fixed effects of herd-year, month of calving, duration of previous pregnancy, interaction between calving difficulty and sex of offspring, and the random effect of the sire. Herd-year effects consituted 0.68 of the explained variance of the incidence of retained placenta and approximately 0.89 of all the other disorders. Sire and calving-month effects were significant (P<0.05) for all the disorders. Heritability estimates of the disorders were 0.01 for retained placenta, abnormal lochia, and endometritis; 0.02 for anestrus; and 0.03 for all disorders. Correlations between any two disorders did not exceed 0.20. High genetic correlations were found between retained placenta and either abnormal lochia (0.90) or endometritis (0.70), while moderate genetic correlations were found among each of these and anestrus (0.30). Reproductive disorders occur as part of a complex which may be efficiently controlled by management measures. Nevertheless, breeding for resistance to reproductive disorders may also be feasible by using appropriate sires evaluated on a large number of daughters.     

Evaporative cooling for Holstein dairy cows under grazing conditions

Twenty-four grazing Holstein cows in mid and late lactation were randomly assigned to two treatment groups: control and cooled. The trial was performed at the Experimental Dairy Unit, Rafaela Agricultural Experimental Station (INTA), Argentina. The objective was to evaluate the effects of sprinkler and fan cooling before milkings on milk production and composition. The effects of the cooling system on rectal temperature and respiration rate were also evaluated. Cooled cows showed higher milk production (1.04 l cow^–1 day^–1). The concentration and yield of milk fat and protein increased in response to cooling treatment. The cooling system also reduced rectal temperature and respiration rate. No effects were observed on body condition. It was concluded that evaporative cooling, which is efficient for housed animals, is also appropriate to improve yields and animal well-being under grazing systems. These results are impressive since the cooling system was utilized only before milkings, in a system where environmental control is very difficult to achieve. This trial was performed during a mild summer. The results would probably be magnified during hotter weather.     

Repeatability and heritability of response to superovulation in Holstein cows

The objective of this study was to estimate the relative effects of genetic and phenotypic factors on the efficacy and efficiency of superovulation for Holstein-Friesian cows reared in Brazil. A database, established by the Associacao Brasileira de Criadores de Bovinos da Raca Holandesa, consisting of a total of 5387 superovulations of 2941 cows distributed over 473 herds and sired by 690 bulls was used for the analysis. The records were analyzed by MTDFREML (Multiple Trait Derivative-Free Restricted Maximum Likelihood), using a repeatability animal model. The fixed effects included in the model were contemporaneous group (veterinarian, herd, year and season of the superovulation); number of semen doses; cow age; and superovulation order. The estimated repeatability of the number of the transferable embryos was low (0.13), and the estimated heritability was 0.03. These results indicate that environmental factors play a critical role in the response of a cow to a superovulation treatment. There is little evidence that future responses to superovulation by individual females can be predicted by previous treatment(s) or that superovulation response is an heritable trait.     

Respiratory heat loss of Holstein cows in a tropical environment

In order to develop statistical models to predict respiratory heat loss in dairy cattle using simple physiological and environmental measurements, 15 Holstein cows were observed under field conditions in a tropical environment, in which the air temperature reached up to 40°C. The measurements of latent and sensible heat loss from the respiratory tract of the animals were made by using a respiratory mask. The results showed that under air temperatures between 10 and 35°C sensible heat loss by convection decreased from 8.24 to 1.09 W m^–2, while the latent heat loss by evaporation increased from 1.03 to 56.51 W m^–2. The evaporation increased together with the air temperature in almost a linear fashion until 20°C, but it became increasingly high as the air temperature rose above 25°C. Convection was a mechanism of minor importance for respiratory heat transfer. In contrast, respiratory evaporation was an effective means of thermoregulation for Holsteins in a hot environment. Mathematical models were developed to predict both the sensible and latent heat loss from the respiratory tract in Holstein cows under field conditions, based on measurements of the ambient temperature, and other models were developed to predict respiration rate, tidal volume, mass flow rate and expired air temperature as functions of the ambient temperature and other variables.This paper forms part of A. S. Campos Maias doctoral thesis.     

Field trial on glucose-induced insulin and metabolite responses in Estonian Holstein and Estonian Red dairy cows in two herds

Background  

Insulin secretion and tissue sensitivity to insulin is considered to be one of the factors controlling lipid metabolism post partum. The objective of this study was to compare glucose-induced blood insulin and metabolite responses in Estonian Holstein (EH, n = 14) and Estonian Red (ER, n = 14) cows.     

IgG1 variations in the colostrum of Holstein dairy cows

High-immune quality colostrum (IgG1 concentration ⩾50 g/l) is crucial for the health and development of the young calf. Studies on colostrum quality tend to focus on external factors such as breed, parity or dry period length, but few have focused on within-cow variations. Here we ran experiments to gain a deeper insight into within-cow variation in IgG1 concentrations in dairy cow colostrum. Trials were performed in an experimental farm, located in the Western part of France. Colostrum from each quarter and a composite sample (mix of four quarters) were concomitantly collected on 77 Holstein dairy cows just after calving to assess the influence of sample type on IgG1 concentrations. Variation in IgG1 concentrations during the first milking was studied on samples from nine cows collected every minute from the start of milking. Repeatability of colostral IgG1 concentration was estimated from 2009 and 2010 data on 16 healthy cows. IgG1 concentrations were tested using a radial immunodiffusion method. Sensitivity and specificity were similar regardless of sample type tested (individual quarter or composite milk). Mean average IgG1 concentration was 54.1 g/l in composite colostrum, and was significantly higher in hind quarter teats (56.2 g/l) than front quarter teats (53.1 g/l). Average IgG1 concentration did not change significantly during colostrum milking, and the variations observed (15% or less) were likely due to the laboratory method (CV 15%). IgG1 concentrations in dam colostrum increased slightly from 2009 to 2010 due to BW and parity effects. In 56% of cases, colostrum quality could have been assessed on either individual or composite colostrum samples collected at any time during the first milking without affecting the reliability of the measurement. However, in other cases, differences were significant enough to mean that estimates of average IgG1 concentration in colostrum from any one quarter would not be reliable. It is concluded that colostrum quality, from an IgG1 concentration point of view, could be assessed with a composite sample taken at any time during the first milking.     

Sequence variation of the alpha-lactalbumin gene in Holstein and Nellore cows

The alpha-lactalbumin is a subunit of lactose-synthase, an enzyme responsible for lactose production, a disaccharide that influences milk production. Sequence variations of bovine alpha -lactalbumin have been associated with differences in milk yield. This study aimed to analyze allelic frequency differences at position - 1689 (g. A > G) and + 15 (g. A > G) of the alpha-lactalbumin gene in Holstein (Bos taurus) and Nellore (Bos indicus) cows. Blood samples were analyzed from 34 Holstein, 104 Nellore, and 99 Dairy Nellore cows using PCR-RFLP. The different RFLP patterns were sequenced and a novel sequence variation on nucleotide - 46 was identified. An adenine at this position was designated as the A allele and a guanine was designated B allele. The frequencies of alleles A - 1689, A - 46, and A + 15 differed between Holstein and both Nellore breeds. The results show that differences in alpha-lactalbumin allelic variants in the 5'-flanking and the 5'-UTR region might be associated with differences in milk production between Holstein cows and cows from Nellore breeds. However, the lack of difference between Nellore and Dairy Nellore suggests that other sequence variantions that regulate milk production might be responsible for the selection of Dairy Nellore cows with superior milk production.     

Superovulatory and endocrine responses in heifers treated with FSH-P at different stages of the estrous cycle

Forty-two Holstein heifers were superovulated with FSH-P (total dose, 30 mg) and cloprostenol. Treatment was initiated on Day 3 (Group D3, n = 11), Day 6 (Group D6, n = 11), Day 9 (Group D9, n = 10) or Day 12 (Group D12, n = 10) of the estrous cycle. Heifers were bled daily for serum progesterone and estradiol-17beta determinations and every 6 h for a 48-h duration at the expected time of estrus for luteinizing hormone (LH) assay. Ova and embryos were flushed from the reproductive tracts and the number of corpora lutea (CL) were recorded after slaughter on Day 7 post-estrus. Mean (+/- SEM) numbers of observed CL were higher (P < 0.05) in Group D9 (33.3 +/- 4.8) than in Group D3 (15.3 +/- 3.8), with Group D6 (17.0 +/- 2.9) and Group D12 (23.9 +/- 7.3) being intermediate. Similarly, mean (+/- SEM) numbers of fertilized embryos were highest (P < 0.05) in Group D9 (13.3 +/- 2.2). There was also a nonsignificant trend for the number of transferable embryos to be greatest in Group D9. Neither serum progesterone concentrations 3 d after the LH peak nor peak serum estradiol 17beta concentrations differed among groups, but both were significantly correlated with numbers of observed CL and total ova and embryos.     

Ovarian function in Holstein cows immunized against pregnant mare serum gonadotrophin

Six Holstein-Friesian cows were immunized against pregnant mare serum gonadotrophin (PMSG) using Freunds' adjuvant during the mid-luteal phase of the estrous cycle. Antibody response was maintained by five booster immunizations at 2- to 3-wk intervals. Four cows were treated with a single intramuscular injection of PMSG (2350 I U) 107 d after primary immunization. Cloprostenol (500 ug) was administered at 56 h and 72 h after the treatment with PMSG; the cows were inseminated three times at 12-h intervals starting 56 h after cloprostenol treatment. Five days after insemination, the animals were slaughtered and their reproductive organs were recovered to quantify the population of corpora lutea and unovulated follicles (>10 mm dia). Antibody titres and progesterone concentrations were determined from blood samples collected either on alternate days or twice a week. Initially, progesterone concentrations were measured in milk samples. All cows produced antibodies, and titres were elevated within 6 to 9 d following each booster immunization. After each boost, however, the antibody titres declined rapidly. Progesterone concentrations declined to below 1 ng/ml after two weeks of initial immunization and remained low throughout the study, except in one cow that ovulated on Day 75. All animals were observed to have large follicular cysts during this period. Treatment with PMSG induced a single ovulation in one cow. Ovulations were neither induced by PMSG nor observed in any of the other animals. In PMSG-treated animals, the mean number of large follicles (5.0) was greater than in those which were not treated (2.0). The results of this study suggest that low titres of antibodies against PMSG are sufficient to disturb ovarian activity, result in follicular cysts and block multiple ovulations in response to exogenous PMSG.