Anti-Müllerian hormone: A novel biomarker for detecting bovine freemartinism.

The anti-Müllerian hormone (AMH) indicates ovarian reserve in cattle, maintaining a consistent trajectory post-puberty. In heterosexual pregnancies, the development of the Müllerian duct in female foetuses is inhibited, resulting in an anticipated minimal or absent ovarian reserve capacity. This investigation aimed to compare AMH levels in healthy Holstein heifers that had reached puberty with those of freemartin animals of the same breed and age. The study incorporated Holstein heifers reaching puberty between 11 and 15 months of age in Group 1 (G1, n = 20) and freemartin animals in Group 2 (G2, n = 19, 16). AMH measurements (AMH-1/AMH-2) were recorded at 12-day intervals for the study participants. Notably, AMH levels in three freemartin animals could not be detected, prompting statistical analysis based on measurements from the remaining 16 freemartin animals in G2. A statistically significant correlation was observed between two separate measurements in G1 and G2 (p < .001). Furthermore, AMH-1 and AMH-2 levels were statistically higher in G1 than in G2 (p < .001). In G1, AMH-1 levels ranged from 227 to 677 pg/mL, with an average of 367.3 ± 25.5 pg/mL, and AMH-2 levels ranged from 234 to 645 pg/mL, with an average of 380.8 ± 24.4 pg/mL. Conversely, in G2, AMH-1 levels ranged from 10 to 72 pg/mL, with an average of 26.8 ± 4.44 pg/mL, and AMH-2 levels ranged from 12 to 68 pg/mL, with an average of 28.75 ± 4.18 pg/mL. The mean AMH levels in G1 were approximately 14 times higher than in G2 (p < .001). Consequently, ROC analysis utilizing AMH-1 and AMH-2 data established cut-off values of ≤72 and ≤ 68 pg/mL respectively for distinguishing freemartin animals. In conclusion, AMH could be used as a reliable biomarker for identifying Holstein freemartin animals.

Evaluation of serum anti-Müllerian hormone concentrations following treatment with vitamin D in Holstein Friesian heifers.

Anti-Müllerian hormone (AMH) levels in the blood of dairy cattle are associated with the count of antral follicles (AFC), ovarian function. Numerous studies, particularly in human medicine, have shown that vitamin D3 (Vit D3) has a positive effect on AMH levels. To our knowledge, the effects of Vit D3 on serum AMH levels in heifers have not been investigated. The goal of the current study was to evaluate the concentrations of serum AMH in dairy heifers following Vit D3 treatment. The study included 20 healthy non-pregnant Holstein Friesian heifers. These animals came to estrus at least once. All heifers received a single intramuscular dosage of 5 million IU Vit D3 (Cholecalciferol). Blood samples were collected from the coccygeal veins of all animals before and after Vit D3 injection (days 7, 14, and 28) for the measurement of AMH and 25(OH)D concentrations. In our analysis, we observed that after exogenous Vit D3 administration, circular 25(OH)D increased constantly (up to day 28 after Vit D3 injection) in all animals. On day 28, AMH concentrations saw a 10% increase in comparison with those measured the day before the Vit D3 injection. However, a comparison of AMH concentrations measured across days did not reveal statistically significant differences between Day 0, 7, 14, and 28 levels (p = .10). Furthermore, no statistical correlation was observed between the pairs (age-AMH) and (weight-AMH). Considering all times, no correlation was found between 25(OH)D and blood AMH levels. These findings demonstrated that exogenous Vit D3 did not affect serum AMH in Holstein Friesian heifers.

[Comparison of different resuscitation methods for newborn calves after dystocia]. / Vergleich verschiedener Erstbehandlungsmethoden bei neugeborenen Kälbern nach Schwergeburten.

OBJECTIVE: This study aimed to compare the use of hand-powered vacuum pumps and the suspension by the hind legs within the scope of resuscitating newborn dystocial calves. MATERIAL AND METHODS: For this study, data were collected from 60 dystocial calves of different breeds. Various reanimation methods were employed to these calves immediately after birth. In group I (n=20), the hand-powered vacuum pump from HK-Rheintechnik (Neuwied, Deutschland) was used to remove mucus and amniotic fluid from the respiratory tract. In group II (n=20), the McCulloch Medical (Elmwood, England) hand-powered vacuum pump was used to remove mucus and amniotic fluid from the respiratory tract. In group III (n=20) 10 to 12 liters of cold water were poured over the neck of 20 calves immediately after birth. In addition, the calves were suspended by the hind legs via a gate or a pole, so that the head hung downwards vertically. The calves' Time to Sternal Recumbency (T-SR) of the calves was measured following the different resuscitation procedures in order to assess their vitality. In addition, the calves were examined segmentally at one-hour post natum. Furthermore, the calves were examined clinically on the seventh day of life. In addition, we investigated whether confounding variables such as calf sex, dam parity, obstetric intervention, and the season affected T-SR. RESULTS: T-SR ranged from 1.5 to 10.5 minutes. There was no difference between the three initial treatment procedures (p=0.07). However, the T-SR of calves treated with a hand-powered vacuum pump tended to be shorter. No statistically significant difference between the pulse rates (p=0.73), respiratory rates (p=0.13), and rectal temperature (p=0.65) of the animals in the three groups on the first and seventh day of life was evident. There was no statistically significant difference in the incidence of the diseases up to the seventh day of life between the individual study groups. CONCLUSION AND CLINICAL RELEVANCE: The present study was able to show that the use of hand-powered vacuum pumps is at least equivalent to suspending calves. When using hand-powered vacuum pumps, the girth of the muzzle must be taken into account, as this can limit their utility.

Effect of uterine torsion intrapartum on concentrations of placental estrogens and progesterone in cattle.

OBJECTIVE: The current study investigates how uterine torsion influences placental oestrogens and progesterone blood concentrations in intrapartum cows. Our research tests the hypothesis that intrapartum uterine torsion impairs the ability of the placenta to synthesize steroids and may also suppress the release of synthesized steroids into the maternal circulation. METHODS: The study included a total number of 37 intrapartum dairy cows of various breeds and ages. These animals were transported to our clinic by their owners. Furthermore, general and obstetrical examinations of all these animals were performed in our clinic. The uterine torsion (UT) group consisted of 20 animals. The presence of UT was verified during clinical general examinations by vaginal and transrectal examination. The comparison (C) group included 17 animals whose birth was undisturbed or could be terminated with moderate obstetrical assistance. The clinical examination of group C animals showed no problems with their general health and genital organs. Blood samples were collected immediately after the initial obstetrical examination from 37 cows for radioimmunological measurement of estradiol-17ß (E2), free total estrogen (FTE), conjugated total estrogen (CTE), and progesterone (P4). RESULTS: In terms of P4, there was no statistical difference between the two groups. For all estrogen parameters, however, concentrations were significantly lower in the UT group than in the C group. In the correlation analysis, there was a significant correlation between the P4 and the FTE in the C group. Furthermore, the positive correlation between all estrogen parameters in the UT group was significant. In group C, significant positive correlations were found apart from the correlation between E2 and CTE. CONCLUSIONS: The results are consistent with the hypothesis and suggest that in UT animals processes dependent on estrogens or other placental hormones may be impaired during the peri- or postpartum period.

Effect of a single meloxicam administration on newborn Holstein-Friesian dystocia calves: Field results from the first 10 days of life.

Background and Aim: Calves have increased morbidity and mortality rates after dystocia. One cause is pain during birth, which reduces their colostrum intake. The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) may break this causality. This study aimed to determine the consequences of a single administration of the NSAID meloxicam to dystocia calves after birth. Materials and Methods: Fifty Holstein-Friesian calves born with dystocia from four dairy cattle farms were included in this study. The animals were randomized into two groups. The animals in Group I (n=25, treatment group) received subcutaneous 0.5 mg meloxicam/kg body weight 2-8 h after birth. The animals in Group II (n=25, control group) received a control substance (Amynin®, bovine infusion solution, Merial) with the same volume. The newborn calves were clinically examined on the 1st and 10th days of life. The information regarding the days in between was gathered by questioning the farmer. Results: There was a significant difference (p=0.04) only in calves with thin, mushy fecal consistency on the 10th day in the treatment group compared with the control group. Moreover, meloxicam had no effect on dystocia calves. Conclusion: Since NSAID administration did not produce a significant clinical effect, its necessity is questionable. Further studies should examine how modifying its application time would have an effect. The primary indicators of well-being, such as pain indicators in the blood, were not measured; however, these should be considered in subsequent studies.

The effect of GnRH on the pregnancy ratio in low-yielding local race cows: comparison of different injection times.

The objective of this article was to investigate the efficiency of GnRH administrations at different time points after induced luteolysis on pregnancy rates in low-yielding subfertile cows. One thousand six hundred and ten healthy and subfertile dairy cows of different ages and races were used in this study. Cows were randomly divided into 4 groups. Estrus cycles were synchronized by two, with 11-day intervals, injections of the prostaglandin F2α-analogue (PG). The artificial inseminations (AIs) of all animals were achieved at the 72nd and 96th hours following the last PG injection. The animals in groups I (n 257), II (n 337), and III (n 675) were used for the administration of a single dose of GnRH at different time points. Accordingly, GnRH was applied at 48th, 64th, and 72nd hours following the last PG injection in groups I, II, and III, respectively. Group IV was accepted as a control without GnRH injection (n 341). The pregnancy rates in groups I, II, III, and IV after transrectal pregnancy examinations were found to be 89.88%, 91.09%, 83.25%, and 77.12%, respectively. In our study, maximal pregnancy rates could be obtained with GnRH injections performed at 48th and 64th hours following luteolysis induction (P < 0.001). There was a 6-8% decrease in pregnancy rates due to the injection of GnRH in the 72nd hour (P < 0.001). These dramatic losses and gains in pregnancy rates in our study emphasized the necessity of taking the time of injection into account when using GnRH to stimulate ovulation. It can be said that the success of GnRH stimulation of ovulation is directly related to the follicle wave dynamics at the time of injection point and the character of a dominant follicle.

[Administration of meloxicam to cows experiencing dystocia does not increase the incidence of retained placenta]. / Die Gabe von Meloxicam an Kühe nach Dystokie steigert nicht die Inzidenz der Retentio secundinarum.

OBJECTIVE: The post partum use of analgesics in cows has increased in consequence to society's increased awareness of animal welfare. However, it is known that the post partum administration of a subset of nonsteroidal anti-inflammatory drugs to cows may lead to an increased rate of retained placenta. Therefore, the aim of this study was clarify, whether a single administration of meloxicam to cows that experienced dystocia results in an increased rate of this disease. MATERIAL AND METHODS: The study was conducted in 63 Holstein-Friesian cows in 4 farms with random allocation into 2 groups: In animals of the treatment group (n = 27) a single subcutaneous injection of meloxicam (0.5 mg/kg) was administered within 2-8 hours after calving. The animals of the control group (n = 36) received the same volume of a bovine infusion solution (Amynin®, bovine infusion solution, Boehringer Ingelheim) subcutaneously. A clinical examination of the cows was performed on the day of parturition and on the 21st day post partum. Findings on the days in between were obtained by questioning of the farmers. The exact Fisher-test was calculated in order to test for statistical differences between the treatment and control groups. RESULTS: In cows of the treatment group, retained placenta was not observed more frequently than in cows of the control group. CONCLUSION AND CLINICAL RELEVANCE: A single administration of meloxicam to cows that experienced dystocia does not increase the incidence of retained placenta.

Effects of long-term release GnRH agonist "deslorelin" on testicular HSP expression, accessory sex glands and testicular functions in adult male rats.

The objective of the present was to determine the effect of long-term release GnRH agonists "deslorelin" on suppression and restoration of testicular and accessory sex glands functions, and expression of HSP in testes of adult male rats. A group of twenty-eight male rats and fifty-six female rats were kept for eleven months. The male rats were subdivided into treatment (n = 18; deslorelin, an analogue of GnRH, 4.7 mg, S.C; six months) and control (n = 10; untreated), and the adult female rats were introduced with either treatment or control male rats at the 2nd, 6th and 11th months post implant insertion. At 6th month of deslorelin implants insertion, six male rats from treatment and five rats from control group were sacrificed. The remaining (twelve treatment and five control) male rats were sacrificed at 11 months. The testicular dimension were measured monthly in both treatment and control rats. The blood samples were collected for testosterone and HSP70 antibody, whereas, the testes and accessory glands were isolated for histological examination at each sacrificial time. The results showed that testicular dimension were significantly lesser in treatment group until 9 months post treatment. HSP70 protein expression was negligible at 6 months in treatment group but its intensity increased in spermatids 11 months of treatment similar to control group. Significantly lower testosterone concentrations with poor semen quality, and smaller litter size were observed in treatment group. The histological picture of accessory sex glands and seminiferous tubules shown a variable integrity in treatment group than control at 6 months implant insertion. In conclusion, the subcutaneous application of 4.7 mg of the GnRH-analogue deslorelin represents a practicable, like in the female rats, method to suppress testicular, accessory sex glands functions, testicular HSP expression and fertility in male rats. Moreover, the suppressive effects of deslorelin, continued until 11th months after removal of the implant.

Postpartum uterus involution observed by real-time ultrasound scanning and vaginal cytology in Van cats.

OBJECTIVES: The objective was to investigate postpartum uterus involution by real-time ultrasonography and vaginal cytology in Van cats. METHODS: This study included 15 healthy Van cats belonging to the Van Cat Research Centre (Yuzuncu Yil University, Van, Turkey). Starting 24 h postpartum, ultrasonographic measurements were performed on the placental and interplacental uterine horn regions every day. Decreases in the diameters and uterine content were considered as criteria for uterine involution. Vaginal discharge samples were collected every day for 4 weeks postpartum. The smears were stained with Papanicolaou stain. RESULTS: The average diameters of placental and interplacental regions (IPRs) in the uterine horns were 3.12 ± 0.29 cm and 2.36 ± 0.43 cm, respectively, at 24 h postpartum. Placental regions (PRs) shrank faster than IPRs. At 48 h postpartum, it became difficult to distinguish PRs from IPRs in the uterine horns. The uterine horns could be seen in the abdominal cavity up to 5.60 ± 0.99 days postpartum. The mean of the last assessable diameter of the uterine horns from days 4 to 7 in all cats was 0.49 ± 0.07 cm. The vaginal epithelial cells appeared to be under the effect of oestrogen for 4 weeks postpartum. CONCLUSIONS AND RELEVANCE: The morphological involution of the uterus completes, to a large extent, within the first 48 h postpartum in Van cats. A more detailed hormonal analysis would contribute greatly to the understanding of the physiological processes involved in this period. Although postpartum involution appeared complete by 5.60 ± 0.99 days after parturition in Van cats, histological verification of this finding is needed.

Effect of flunixin meglumine or prostaglandin E2 treatment 15 days after breeding on fertility in Saanen does.

The objective of this study was to determine the effects of timely injections of flunixin meglumine (FM) or vaginal application of prostaglandin E2 (PgE2) on pregnancy, fertility, fecundity, and prolificacy rates in Saanen goats. One hundred and sixty-three nonlactating Saanen does were treated with a flugestone acetate (20 mg)-containing intravaginal sponge for 12 days. They also received eCG (400 IU) and a PGF2α analogue (50 µg) 10 days after progestagen priming. Does detected in estrus were mated and assigned randomly to one of three treatment groups. The PgE2 group (N = 40) received PgE2 (2.5 mg) intravaginally 15 days after mating. The FM group (N = 54) received flunixin meglumine (total dose, 100 mg) intramuscularly 15 days after mating. Flunixin meglumine was administered at 9:00 AM. Animals in the control group (N = 69) received no treatment. Pregnancy was diagnosed using transrectal ultrasonography (B-mode at 8 MHz) 30 days after mating. The pregnancy rate was significantly greater (P < 0.01) after 30 days in goats treated with PgE2 and also in the control group than in those treated with FM (67.5%, 59.4%, and 42.5%, respectively). The pregnancy rate did not differ between the PgE2 and the control group. The pregnancy and fertility rate were lowest in the FM group compared with the other groups. There was no significant difference in the prolificacy rate among experimental groups. In conclusion, our results showed that FM administration during a late luteal phase is detrimental to early pregnancy in goats.

Effects of eCG and FSH on ovarian response, recovery rate and number and quality of oocytes obtained by ovum pick-up in Holstein cows.

The goal of the present study was to compare the ovarian response, oocyte yields per animal, and the morphological quality of oocytes collected by ultrasound guided follicular aspiration from Holstein cows treated either with FSH or eCG. Twenty four normal cyclic, German Holstein cows were randomly divided into two groups. Fourteen cows received 3000 IU eCG on day-4 prior to ovum pick-up (OPU) (day 0), 2 days later (day-2), 625 microg cloprostenol was administered. On day-1 GnRH was administered i.m. and 24h later OPU (day 0) was performed. In ten cows a total dose of 500 IU follicle stimulating hormone (Pluset) was administered intramuscularly in a constant dosage for 4 days with intervals of 12h, starting on day-5. Luteolysis was induced by application of 625 microg cloprostenol on day-2. On day-1 (24h after the last FSH treatment) GnRH was administered i.m. and 24h later OPU (day 0) was performed. Ovarian follicles were visualized on the ultrasound monitor, counted and recorded. All visible antral follicles were punctured. Recovered oocytes were graded morphologically based on the cumulus investment. Average follicle number in ovaries was higher in FSH group than eCG group (p<0.05). Oocyte yields per animal did not differ between FSH and eCG groups. The proportion of grade A oocytes was higher in the FSH group in the than eCG group (p<0.05). Likewise, rate of grade C oocytes in FSH group were lower than eCG group (p<0.05). In conclusion, these results suggest that ovarian response, follicle number in ovaries and oocyte quality are affected by the type of gonadotropin and FSH is better alternative than eCG for OPU treatment.