Detection of urinary luteinizing hormone in Japanese black cows after administration of gonadotropin-releasing hormone

The blood luteinizing hormone (LH) surge in cows is well studied. However, little is known about urinary LH in cows. This study examined urinary LH concentrations after administration of gonadotropin-releasing hormone (GnRH) in six Japanese black cows to induce LH secretion from the pituitary gland into the bloodstream. Abrupt rises in plasma and urinary LH were observed after GnRH administration. Plasma and urinary LH peaked at 2 and 5 hr, respectively. A positive correlation was observed between plasma LH concentrations and urinary LH amounts. Ovulation was confirmed in the cows after 48 hr of GnRH administration. These data strongly suggest that urinary LH is derived from plasma LH, which triggers ovulation in cows.     

Comparison of the effects of gonadotropin-releasing hormone, human chorionic gonadotropin or progesterone on pregnancy per artificial insemination in repeat-breeder dairy cows

Three different treatments were compared to improve pregnancy per artificial insemination (P/AI) in repeat-breeder (RB) dairy cows. All cows (n = 103) were assigned to one of four groups: (1) gonadotropin-releasing hormone (GnRH); (2) human chorionic gonadotropin (hCG); (3) once-used controlled internal drug release (CIDR) device; and (4) control. All treatments performed 5-6 days after artificial insemination (AI) and milk samples were collected just before treatment for progesterone assays. There were no significant differences in milk fat progesterone concentration among trial groups. Cows were observed for estrus signs thrice daily. Pregnancy per AI on day 45 in hCG and CIDR groups were significantly higher than GnRH and control groups (60.0% and 56.0% vs. 26.9% and 29.6%, respectively), but there were no differences in P/AI between GnRH and control groups. There were also no significant differences between hCG and CIDR groups. Milk fat progesterone concentrations were compared between pregnant and non-pregnant cows in each group and only in the hCG group it was significantly lower in pregnant cows. In conclusion, treating repeat-breeder cows with hCG or once-used CIDR 5-6 days after AI improved P/AI.     

Effect of subluteal concentrations of progesterone on luteinizing hormone and ovulation in lactating dairy cows

Two experiments were conducted to determine if administration of progesterone within a low, subluteal range (0.1-1.0 ng/mL) blocks the luteinizing hormone (LH) surge (experiments 1 and 2) and ovulation (experiment 2) in lactating dairy cows. In experiment 1, progesterone was administered to cycling, lactating dairy cows during the luteal phase of the estrous cycle using a controlled internal drug release (CIDR) device. CIDRs were pre-incubated in other cows for either 0 (CIDR-0), 14 (CIDR-14) or 28 days (CIDR-28). One group of cows received no CIDRs and served as controls. One day after CIDR insertion, luteolysis was induced by two injections of prostaglandin (PG) F(2alpha) (25 mg) at 12 h intervals. Two days after the first injection, estradiol cypionate (ECP; 3 mg) was injected to induce a LH surge. Concentrations of progesterone after luteolysis were 0.11, 0.45, 0.78 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14, and CIDR-0, respectively. LH surges were detected in 4/4 controls, 4/5 CIDR-28, 2/5 CIDR-14 and 0/5 CIDR-0 cows following ECP. In experiment 2, progesterone was administered to cycling, lactating, Holstein cows during the luteal phase of the estrous cycle as in experiment 1. Luteolysis was induced as in experiment 1. The occurrence of an endogenous LH surge and ovulation were monitored for 7 days. Concentrations of progesterone after luteolysis were 0.13, 0.30, 0.70 and 1.20 ng/mL for cows treated with no CIDR, CIDR-28, CIDR-14 and CIDR-0, respectively. LH surges and ovulation were detected in 5/5 controls, 3/7 CIDR-28, 0/5 CIDR-14 and 0/5 CIDR-0 cows. It was concluded that low concentrations of progesterone can reduce the ability of either endogenous or exogenous estradiol to induce a preovulatory surge of LH and ovulation.     

Treatment of anovulatory anoestrous postpartum dairy cows with a gonadotropin-releasing hormone (GnRH), prostaglandin F2 alpha, GnRH regimen or with progesterone and oestradiol benzoate

AIM: To compare 2 treatments for anovulatory anoestrus (AA) in postpartum dairy cows. The treatments were combinations of gonadotropin-releasing hormone (GnRH) and prostaglandin F2 (PG) or progesterone (P4) and oestradiol benzoate (ODB). METHODS: Forty AA cows from each of 5 herds were blocked by age (2 or 2 years old) and randomly assigned to 1 of 2 treatments. The first group (GPG) were treated with 250 mug of a GnRH analogue, gonadorelin, followed 7 days later by 15 mg of the PG analogue, luprostiol. Two days later the cows were injected with 250 mug of gonadorelin. Cows were artificially inseminated 16-24 h after the second GnRH injection. The second group (P4+ODB) were treated with an intravaginal P4 releasing device for 6 days, followed 24 h after device removal by injection of 1 mg of ODB. Cows were pregnancy tested 35-40 days after the initial insemination and twice again at 6-8 week intervals thereafter. RESULTS: There was no significant difference between P4+ODB and GPG groups in the percentage of cows submitted for insemination in the first 7 days (94.0% vs 100% for P4+ODB vs GPG, respectively; p>0.3), in conception rate to first insemination within the first 7 days (43.6% vs 35.0% for P4+ODB vs GPG, respectively; p>0.2), in the percentage of cows conceiving in the first 28 days of the breeding period (68.0% vs 58.3%, P4+ODB vs GPG, respectively; p>0.1), or in median interval from the end of treatment to conception (20 vs 21 days; p>0.1). CONCLUSIONS: No differences in the reproductive performance of AA cows treated with either P4+ODB or GPG were detected. However, given the small number of animals enrolled, further data are required before the GPG protocol can be recommended for treatment of AA cows.     

Progesterone concentration,estradiol pretreatment,and dose of gonadotropin-releasing hormone affect gonadotropin-releasing hormone-mediated luteinizing hormone release in beef heifers

We examined whether progesterone (P₄)-induced suppression of LH release in cattle can be overcome by an increased dose of exogenous gonadotropin-releasing hormone (GnRH) or pretreatment with estradiol (E₂). In Experiment 1, postpubertal Angus-cross heifers (N = 32) had their 2 largest ovarian follicles ablated 5 d after ovulation. Concurrently, these heifers were all given a once-used, intravaginal P₄-releasing insert (CIDR), and they were randomly assigned to be given either prostaglandin F_2α (Low-P₄) or no treatment (High-P₄) at follicle ablation, and 12 h later. Six days after emergence of a new follicular wave, half of the heifers in each group (n = 8) were given either 100 or 200 μg of GnRH i.m. Plasma luteinizing hormone (LH) concentrations were higher in the Low- vs High-P₄ groups, and in heifers given 200 vs 100 μg of GnRH (mean ± SEM 15.4 ± 2.2 vs 9.1 ± 1.2, and 14.8 ± 2.1 vs 9.8 ± 1.4 ng/mL, respectively; P ≤ 0.01). Ovulation rate was higher (P = 0.002) in the Low-P₄ group (15/16) than in the High-P₄ group (6/16), but it was not affected by GnRH dose (P = 0.4). In Experiment 2, heifers (n = 22) were treated similarly, except that 5.5 d after wave emergence, half of the heifers in each group were further allocated to be given either 0.25 mg estradiol benzoate i.m. or no treatment, and 8 h later, all heifers were given 100 μg GnRH i.m. Both groups treated with E₂ (Low- and High-P₄) and the Low-P₄ group without E₂ had higher peak plasma LH concentrations compared to the group with high P₄ without E₂ (12.6 ± 1.8, 10.4 ± 1.8, 8.7 ± 1.3, and 3.9 ± 1.2 ng/mL, respectively; (P < 0.04)). However, E₂ pretreatment did not increase ovulation rates in response to GnRH (P = 0.6). In summary, the hypotheses that higher doses of GnRH will be more efficacious in inducing LH release and that exogenous E₂ will increase LH release following treatment with GnRH were supported, but neither significantly increased ovulation rate.     

Treatment of anovulatory anoestrous postpartum dairy cows with a gonadotropin-releasing hormone (GnRH), prostaglandin F2𝛂, GnRH regimen or with progesterone and oestradiol benzoate

AIM: To compare 2 treatments for anovulatory anoestrus (AA) in postpartum dairy cows. The treatments were combinations of gonadotropin-releasing hormone (GnRH) and prostaglandin F_2𝛂 (PG) or progesterone (P4) and oestradiol benzoate (ODB).

METHODS: Forty AA cows from each of 5 herds were blocked by age (2 or >2 years old) and randomly assigned to 1 of 2 treatments. The first group (GPG) were treated with 250 𝛍g of a GnRH analogue, gonadorelin, followed 7 days later by 15 mg of the PG analogue, luprostiol. Two days later the cows were injected with 250 𝛍g of gonadorelin. Cows were artificially inseminated 16–24 h after the second GnRH injection. The second group (P4+ODB) were treated with an intravaginal P4 releasing device for 6 days, followed 24 h after device removal by injection of 1 mg of ODB. Cows were pregnancy tested 35–40 days after the initial insemination and twice again at 6–8 week intervals thereafter.

RESULTS: There was no significant difference between P4+ODB and GPG groups in the percentage of cows submitted for insemination in the first 7 days (94.0% vs 100% for P4+ODB vs GPG, respectively; p>0.3), in conception rate to first insemination within the first 7 days (43.6% vs 35.0% for P4+ODB vs GPG, respectively; p>0.2), in the percentage of cows conceiving in the first 28 days of the breeding period (68.0% vs 58.3%, P4+ODB vs GPG, respectively; p>0.1), or in median interval from the end of treatment to conception (20 vs 21days;p>0.1).

CONCLUSIONS: No differences in the reproductive performance of AA cows treated with either P4+ODB or GPG were detected. However, given the small number of animals enrolled, further data are required before the GPG protocol can be recommended for treatment of AA cows.     

Cervix–rectum temperature differential at the time of insemination is correlated with the potential for pregnancy in dairy cows

This study sought to establish whether temperature gradients between the cervix, vagina, and rectum at and 7 days post-artificial insemination (AI) were associated with the incidence of pregnancy in lactating dairy cows (Experiment I; n = 90 ovulating cows) and to evaluate temperature gradient dynamics from the time of insemination to 7 days post-AI under heat stress conditions (Experiment II; n = 16 ovulating and 4 non-ovulating cows). In Experiment I, 39 cows (43.3%) became pregnant. The odds ratio for pregnancy was 2.5 for each one-tenth of a degree drop in cervical temperature with reference to the control rectal temperature at the time of AI (P = 0.01), whereas the same decrease in the cervix–rectum temperature differential 7 days post-AI resulted in an odds ratio of 0.44 (P = 0.02). In Experiment II, 5 of the ovulating cows (31.3%) became pregnant. The mean values of the vagina–rectum, vagina–cervix, and cervix–rectum temperature differentials at AI (day 0), 8 h, 24 h, and 7 days post-AI changed significantly from day 0 to day 7 (within-subject effect; P < 0.02) in ovulating cows but not in non-ovulating cows. Temperature differentials on days 0 and 7 were similar between ovulating cows and cows of Experiment I. Overall, our findings support the notion that a temperature differential between the caudal cervical canal and rectum at AI may be an indicator of the likelihood of pregnancy. Possible prospects of confirming estrus at the herd-level are also suggested.     

Probability of pregnancy to artificial insemination either after detected oestrus or at a fixed time in dairy cows: Influence of intrinsic and extrinsic factors in a large-scale,on-farm study

The first aim of this study was to determine the influence of the procedures [hormonal treatments for fixed time artificial insemination (FTAI) versus insemination at spontaneous oestrus (SEAI)] on several sequential inseminations (AI). A second aim was to determine the influence of some intrinsic and extrinsic factors and their interactions, including characteristics of the animals such as age, season, farm, sire, and AI technician on the response to both procedures. A retrospective analysis was performed from a data base of 120.807 AIs of healthy cows with at least 40–70 days post-partum at first service. Overall, FTAI achieved slighter greater pregnancy rates than insemination after detected oestrus. The second AI seems to be a key insemination as effects of sire and technician were greater than in the following ones. The use of FTAI or SEAI in one AI did not affect the results of the following AIs, regardless if FTAI or SEAI procedures were used in that AI. Technician had greater variation than sire or farm on final pregnancy rate. The results of each sire for pregnancy rate varied according to the type of insemination, with sires achieving greater results with one or other procedure. Pregnancy rate was positively related to the days in milk in the first two AIs. Results were greater in autumn than in spring services.     

Use of two cloprostenol administrations 11.5 days apart efficiently synchronizes oestrus in photostimulated multiparous dairy goats in the non-breeding season

This study assessed the efficiency of synchronous oestrous induction by light programme followed by two doses of cloprostenol in acyclic Saanen goats of different parity orders. Primiparous (n = 22) and multiparous (n = 33) goats were subjected to 16 hr of light and 8 hr of darkness for 60 days (D0-D60), starting 10 days after the winter solstice. All goats received 120 µg cloprostenol doses on D130 (morning) and D141.5 (afternoon) (11.5 days apart). Oestrus behaviour, ovarian follicular dynamics and serum progesterone (P4) analyses were recorded from D0 to D174 at different intervals. Animals in oestrus after D141.5 were randomly assigned into two groups: assisted natural mating (NM) or artificial insemination (AI; 10–24 hr after oestrus onset with frozen-thawed semen). From D57 to D120, 89.0% of goats presented large follicles (5–8 mm) and P4 concentrations were subluteal from D0 to D120. More multiparous compared to primiparous goats (54.5%, 18/33 vs. 18.2%, 4/22) exhibited oestrus after both injections. More primiparous compared to multiparous goats (54.5%, 12/22 vs. 12.1%, 4/33) did not exhibit oestrus after any injection. A total of 35 goats (64%) were in oestrus after the second prostaglandin injection and were subjected to NM or AI. The conception rate was similar among primiparous (70.0%, 7/10) and multiparous (68.0%, 17/25) goats but the pregnancy rate differed, being 31.8% (7/22) and 51.5% (17/33), respectively. No interaction was found between parity order and P4 concentrations in does that became pregnant or not. Thus, the association between light programme (60 days, starting at the beginning of winter) and two cloprostenol administrations 11.5 days apart (starting 70 days after the end of the light treatment) resulted in sufficient synchronous oestrous response in multiparous acyclic Saanen goats to reach satisfactory fertility levels after both NM and AI.     

Relationship between the milk yield response to short-term bovine somatotropin treatment and the lipolytic response to adrenaline in dairy cows

The aim of this experiment was to determine if the milk yield response of dairy cows to short-term treatment with bovine somatotropin (bST) was correlated with the non-esterified fatty-acid (NEFA) response to an adrenaline challenge. Twenty-six multiparous Holstein cows (58+/-5.4 days postpartum) received daily sub-cutaneous injections of saline for 7 days followed by sub-cutaneous injections of 20mg/day of bST for 14 days. On day 7 of the saline treatment and day 14 of the bST treatment the cows were given an intravenous injection of adrenaline (1.4 microg/kg body weight). Blood samples were taken before and after the adrenaline challenge. The difference in milk yield between the saline and the second week of bST treatment (MYR) varied considerably between animals (from -0.4 to +8.0 kg/day). MYR was positively correlated with the change in the basal concentration of NEFA between the saline and second week of bST treatment, as well as with the change in the area under the profile of NEFA above basal values following the adrenaline challenge. It remains to be established whether the greater lipolytic responses to adrenaline of the cows with the greater MYR reflects the deeper negative energy that these animals also experienced or a fundamental difference in the physiology of their adipose tissue.     

Reproductive performance in dairy cows following postpartum treatment with gonadotrophin releasing hormone and/or prostaglandin: a field trial.

Three hundred and five Holstein Friesian cows were given either 250 micrograms gonadotrophin releasing hormone (GnRH) or saline on day 15 postpartum followed by 500 micrograms cloprostenol or saline on day 24 postpartum. Four treatment groups were formed using random allocation: Group I -- placebo (Day 15)/placebo (Day 24), Group II -- GnRH (Day 15)/placebo (Day 24), Group III -- placebo (Day 15)/cloprostenol (Day 24), Group IV -- GnRH (Day 15)/cloprostenol (Day 24). Double blind techniques were used during the follow-up period. Rectal palpation, to assess uterine involution and ovarian activity was performed just prior to each treatment and again at 28 days postpartum. In addition blood samples were collected at 15, 24 and 28 days postpartum for measurement of plasma progesterone. There were no significant differences among treatment groups with respect to services per conception, number of heats detected before first service and culling for infertility. Cows treated only with GnRH had an increased calving to first estrus and calving to first breeding interval, and tended to have an increased calving to conception interval. Treatment with cloprostenol significantly decreased calving to conception and calving to first observed estrus intervals. Treatment with GnRH on day 15 postpartum resulted in a significant increase in the subsequent incidence of pyometra and prebreeding anestrus. On the other hand, cloprostenol treatment on day 24 postpartum resulted in a decreased incidence of pyometra, regardless of GnRH treatment and a decreased incidence of prebreeding anestrus in GnRH treated cows compared to cows receiving only GnRH at day 15 postpartum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Clinical prospects proposing an increase in the luteolytic dose of prostaglandin F2α in dairy cattle

Over the past few decades, the luteolytic dose of prostaglandin F_2α (PGF_2α) and its analogs, used to synchronize estrus for fixed-time insemination in dairy cattle, have remained unchanged. Given the beneficial effects of PGF_2α on a young corpus luteum and on multiple ovulations in a fixed-time insemination protocol, and its therapeutic abortive effects on multiple ovulations in pregnant cows, we propose the use of a double PGF_2α dose or two PGF_2α treatments 24 hours apart. Ultrasonography procedures serve to identify luteal structures and may therefore help to determine the best PGF_2α dose to improve the fertility of high-producing dairy cows.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.     

Presence of multiple corpora lutea affects the luteolytic response to prostaglandin F2α in lactating dairy cows

Since the 1970s, luteolytic doses used for synchronizing estrus in dairy cattle have remained unchanged. This study aimed to evaluate the dose-response effect of prostaglandin F_2α (PGF_2α), which is used for synchronizing estrus, and subsequent fertility in cows with two or more corpora lutea (CL). The study population consisted of 1,683 cows with a single CL (1CL), 501 cows with multiple CL receiving a single dose of PGF_2α (2CL1), and 252 cows with multiple CL receiving a 1.5 × PGF_2α dose (2CL1.5). Cows with a single CL (n = 1,245) showed estrus significantly (P < 0.01) earlier (3.01 ± 1.23 days; mean ± SD) than cows with multiple CL (n = 287; 3.33 ± 1.69 days). Using 1CL cows as reference, the odds ratio (OR) for the estrus response in 2CL1 cows was 0.13 (P < 0.0001), whereas the ORs for estrus response and pregnancy of 2CL1.5 cows were 1.8 (P = 0.0001) and 1.7 (P = 0.001), respectively. Based on the results for only the 2CL1 cows, the OR for the estrus response was 0.7 (P = 0.01) for cows producing ≥ 45 kg of milk at treatment, compared to the remaining cows producing < 45 kg of milk. Our results showed that the presence of multiple CL reduced the estrus response to that induced by a single PGF_2α dose and milk production was inversely associated with this response, whereas an increased PGF_2α dose improved the estrus response. Therefore, an increase in the standard PGF_2α dose is recommended.     

Negative energy balance increases periprandial ghrelin and growth hormone concentrations in lactating dairy cows

The reported effects of feeding on growth hormone (GH) secretion in ruminants have been inconsistent, and are likely influenced by energy status of animals. High-producing dairy cows in early lactation and late lactation were used to assess the effects of energy balance on temporal variation of plasma metabolites and hormones. Cows were fed a single diet once daily, and feed was withdrawn for 90 min prior to feeding. Beginning at the time of feed withdrawal, plasma samples were collected via jugular catheters hourly for 24h. Concentrations of non-esterified fatty acids and GH were measured for all samples, while insulin, glucose, and acylated (active) ghrelin were quantified for four sample times around feeding. As expected, calculated energy balance was significantly lower in early lactation than late lactation cows (-43.5 MJ retained/day versus 7.2 MJ retained/day). Following the primary meal of the day, a GH surge was observed in early lactation but not in late lactation cows. This difference was not explained by temporal patterns in non-esterified fatty acid, insulin, or glucose concentrations. However, a preprandial ghrelin surge was observed in early lactation only, suggesting that ghrelin was responsible for the prandial GH surge in this group. Results of a stepwise regression statistical analysis showed that both preprandial ghrelin concentration and energy balance were significant predictors of prandial GH increase over baseline. Adaptations to negative energy balance in lactating dairy cattle likely include enhanced ghrelin secretion and greater GH response to ghrelin.     

Changes in circulating adiponectin and metabolic hormone concentrations during periparturient and lactation periods in Holstein dairy cows

Although our previous report demonstrated that adiponectin and AdipoR1 gene expressions changed among different lactation stages in the bovine mammary gland, its in vivo kinetics remain unclear in ruminant animals. In this study, we investigated the changes in circulating concentrations of adiponectin, as well as other metabolic hormones and metabolites, (i) during the periparturient period and (ii) among different lactation stages, in Holstein dairy cows. In experiment 1, serum adiponectin concentrations increased after parturition. Serum insulin concentrations were lower in the postpartum than prepartum period, whereas serum growth hormone (GH) concentrations increased in the postpartum period. Serum nonesterified fatty acids (NEFA) levels were increased during the postpartum period and were dependent on the parity. In experiment 2, there was no significant difference in plasma adiponectin concentrations among lactational stages. Plasma insulin concentrations tended to be lower in early lactation while plasma GH levels tended to be higher. Plasma NEFA concentrations were significantly lower in mid‐ and late‐lactation stages than non‐lactation stages. These findings indicate that elevation of serum adiponectin might be involved in energy metabolism just around parturition, and might exert its action through regulation of receptor expression levels in target tissues in each lactational stage in Holstein dairy cows.     

Efficacy of two antibiotic treatments in curing clinical and subclinical mastitis in lactating dairy cows

Aim. To compare the bacteriological and clinical cure rates for clinical and subclinical mastitis in New Zealand dairy cows following treatment with either an intramammary penicillin-dihydrosptreptomycin preparation or a subcutaneous injection of penethamate hydriodide.

Methods. Milk samples were collected from clinical and subclinical cases of mastitis before and 14 and 21 days after initiation of treatment for bacteriological culture, somatic cell count determination and conductivity testing.

Results. No significant differences in the bacteriological cure rate of major Gram-positive pathogens, clinical cure rate, somatic cell count or conductivity were found between treatments. However, the bacteriological cure ralte of coagulase-negative staphylococcus infections and the overall bacteriological cure rate was lower for quarters treated with penethamate than with penicillin-dihydrosptreptomycin.

Conclusions. The bacteriological cure rate of mastitis caused by major Gram-positive pathogens, the clinical cure rate, somatic cell count and conductivity did not differ between the two antibiotic treatments.