Determination of anti-Müllerian hormone levels in blood and urine in fertile cats.

In recent years, blood anti-Müllerian hormone (AMH) levels have been investigated in female animals to diagnose many conditions, such as the presence of ovarian tissue, follicle reserve, and granulosa cell tumors. Since blood collection is an invasive method, diagnosis with a non-invasive method is important in terms of practicality and animal welfare. This study aimed to investigate the presence of AMH in cat urine and determine whether a correlation exists between blood and urine AMH levels. In addition, it was aimed at revealing whether there was a change in blood and urine AMH levels according to ovarian follicle distribution. Twenty-seven healthy, fertile female cats in the follicular phase were included. Following blood and urine sample collection, a routine ovariohysterectomy was performed. Histological analysis of the removed ovarian tissue was used to determine ovarian follicle types. While both AMH and estrogen levels were determined in blood samples, only AMH levels were investigated in urine samples. Blood AMH levels averaged 10.61 ± 0.75 ng/mL (range: 5-16 ng/mL), while urine AMH levels averaged 5.67 ± 0.91 ng/mL (range: 0.2-13 ng/mL). While urinary AMH level was <1 ng/mL in 7 cats, urinary AMH was >1 ng/mL in all remaining cats. While the study demonstrated AMH excretion in urine, no correlation was found between blood and urine AMH values. However, a significant positive correlation was observed between blood AMH levels and serum estrogen levels (P < 0.001). These findings suggest that urinary AMH may be a product of proteolytic degradation, potentially leading to inaccurate estimations of ovarian activity based solely on urine AMH levels.

Determination of natural antibodies, beta-hydroxybutyric acid, and non-esterified fatty acid levels in the serum of peripartum Tuj and Hemsin sheep.

BACKGROUND AND AIM: Many metabolic and immunological changes occur during the transition period. Innate immunity plays an important role against to infections and natural antibodies (NAb) are important in immunity. This study aims to determine a connection between serum NAb titers, beta-hydroxybutyric acid (BHBA), and non-esterified fatty acid (NEFA) concentrations in Tuj and Hemsin sheep during the peripartum period. MATERIALS AND METHODS: Serum NAb, BHBA, and NEFA levels were determined from the blood samples collected from Tuj and Hemsin sheep on days 30 and 15 before birth, on the day of birth (day 0), and on days 15 and 30 after birth. RESULTS: NAb titers were found to be higher in Tuj than in Hemsin sheep (p<0.001). No statistically significant difference was found in serum BHBA concentrations of both breeds on all sampling days (p>0.05). The serum NEFA level was lower in Tuj sheep in the last 15 days of pregnancy compared to Hemsin sheep (p<0.05), while no difference was found in samples collected at the other time points. CONCLUSION: This study indicated that serum NAb titers significantly changed in Tuj and Hemsin sheep during the transition period. Serum BHBA and NEFA concentrations increased during the last stages of pregnancy and decreased after birth. Based on these findings, it is suggested that the immunological status could vary by the breed of sheep or various factors that affect the sheep's metabolic state.

Efficacy of a commercial test kit to determine early pregnancy in cows using whole blood and blood serum.

The objective of this study was to determine the efficacy of the Fassisi®BoviPreg visual test kit (Fassisi®BoviPreg) in determining early pregnancies in cows by measuring the pregnancy-associated glycoprotein (PAG) in whole blood and blood serum. The study was conducted on 50 cows, the artificial insemination (AI) dates of which were designated as day 0. Pregnancy diagnosis was performed with transrectal ultrasonography (USG), and serum samples were simultaneously collected and used with Fassisi®BoviPreg to determine pregnancies on 30 days after AI. The results of the Fassisi®BoviPreg test on serum and whole blood samples, respectively, on 30 days after AI were as follows: sensitivity, 61.54% and 50.0%; specificity, 79.17% and 75%; accuracy, 70.0% and 62.0%; positive predictive values, 76.2% and 68.4%; negative predictive values, 65.5% and 58.1%; false-positive diagnoses, 23.8% and 31.6%; and false-negative diagnoses, 34.5% and 41.9%. On day 50 after AI, sensitivities were 63.64% and 50.0%, specificities were 100.0% and 100.0%, and accuracies were 75.0% and 65.62% in serum and whole blood, respectively. Higher pregnancy rates were obtained using Fassisi®BoviPreg in cows with ≥ 5 ng/mL P4 (P < 0.001). In conclusion, the results from the Fassisi®BoviPreg tests on cows on 30 and 50 days after AI showed that the use of blood serum is more accurate and suitable than that of whole blood. The results also showed a higher confidence level in specificity 50 days after AI.

Pre-pubertal treatment with a GnRH agonist in bitches-Effect on the uterus and hormone receptor expression.

Aim of the study was to examine the effect of deslorelin on uterine tissues of eleven pre-pubertal bitches aged 4.2 ± 0.6 m. Implants containing placebo (sodium chloride 0.9%; n = 4, G I), 4.7 mg (n = 3, GII) or 9.4 mg (n = 4, GIII) deslorelin acetate (Suprelorin® ; Virbac, France), were administered subcutaneously. Signs of oestrus, vaginal cytology, serum progesterone (P4) and estradiol-17ß (E2) concentrations were monitored until the occurrence of oestrus. Bitches were ovariohysterectomized and sections from the uterine tissue were subjected to immunohistochemistry (IHC) for detection of GnRH receptor (R), Kisspeptin (KP)10, Kisspeptin receptor (GPR54), androgen receptor (AR), oestrogen receptor (ER) α,ß, and progesterone receptor (PR). Tissue sections were scored semi-quantitatively using an immunoreactivity score (IRS) ranging from 0 to 300 (3). Since some animals were ovariohysterectomized before puberty (n = 1 from GII and n = 2 from GIII), and some in metestrus (all controls and 2 from GII and GIII each), results from these animals were separately evaluated and compared to the controls. Results: No abnormalities were seen in uterine tissues. Kisspeptin 10 expression was low in all cell types, highest IRS were seen in the vascular endothelial cells. The GPR54 was mainly detected in the luminal epithelial cells, superficial and deep uterine glands. The expression of GPR54 and ERα,ß was especially high in bitches operated prepubertally. No difference was observed between the controls and experimental bitches operated in their first metestrus. The PR and ERα,ß were exclusively expressed in superficial and deep uterine glands and luminal surface epithelial cells. The AR and GnRH-R expression was negative in all cells of all groups. We conclude that application of 4.7 or 9.4 mg deslorelin at the age of 4 months did not cause uterine disturbances. GPR54 expression might be influenced by pre-pubertal deslorelin treatment or the changings related to approaching puberty; the latter is supposed in case of ERα,ß.

Association of luteal blood flow with follicular size, serum estrogen and progesterone concentrations, and the inducibility of luteolysis by PGF2α in dairy cows.

The aim of this study was to investigate the compatibility of the visual evaluation result of the blood flow characteristics and the blood flow measurements of the CL and the predictability of the responses given by corpora lutea with varying levels of blood flow to an induction of luteolysis by a PGF2α injection and to determine the possibility of increase in serum estrogen and progesterone concentrations in parallel with increased luteal blood flow (LBF). The cows, bearing a CL (n = 60; postpartum 35 days), were injected with PGF2α and were monitored for signs of estrous following the first injection. The cows, which did not show estrous signs, were examined for the presence of a CL on Day 14, whereas those that showed signs of estrous were examined on Day 10 following the onset of estrous. The level of LBF was visually graded as + (low; GI), ++ (medium; GII), +++ (high; GIII), and ++++ (very high; GIV). Immediately after the examination of LBFs, a second intramuscular injection of PGF2α was injected. In the cows, which were determined to be in estrous, the diameter of the Graafian follicles was measured by B-mode ultrasonography. Subsequently, these animals were artificially inseminated. The animals, which did not show estrous after the second injection, were examined as previously described and monitored for signs of estrous. A strong correlation (r = 0.654; P < 0.001) was determined to exist between the results of the visual examination of the images and the results obtained for the LBF area with the use of the Pixel Flux software. GIII (0.83 ± 0.15 cm2) and GIV (1.03 ± 0.48 cm2) were found to differ from GI (0.47 ± 0.23 cm2) and GII (0.51 ± 0.12 cm2) for the size of the LBF (P < 0.001). Serum progesterone levels in groups (GI, GII, GIII, and GIV) were determined to be 4.44 ± 2.42 ng/mL, 6.03 ± 2.37 ng/mL, 7.01 ± 2.94 ng/mL, and 7.17 ± 1.69 ng/mL, respectively. The comparative evaluation of the study groups showed that the groups did not statistically differ for the period between PGF2α injection and the onset of estrous, mean Graafian follicle size and estrogen levels. No direct correlation existed between these reproductive parameters and LBF.

Subclinical mastitis causes alterations in nitric oxide, total oxidant and antioxidant capacity in cow milk.

The aim of this study was to investigate total antioxidant (TAC), and oxidant capacity (TOC) and nitric oxide (NO) levels in milk of cows with subclinical mastitis. Brown Swiss and Holstein breed cows were screened with California Mastitis Test (CMT) to determine mammary glands with subclinical mastitis. Moreover, somatic cell counts (SCC) were determined electronically in all milk samples. Mammary quarters were classified as healthy (n=25) or subclinical mastitis (n=35) based on CMT scores and somatic cell count (SCC: < or =200,000/ml or >200,000/ml) in milk. Nitric oxide, TOC and SCC levels were significantly higher (p<0.001, p<0.005 and p<0.001, respectively) in milk from mammary quarters with subclinical mastitis compared to those from healthy mammary quarters. In conclusion, subclinical mastitis results in higher NO concentrations, TOC and SCC, and NO and TOC were positively correlated with SCC. Moreover, alterations in NO levels and TOC in milk could be used as an alternative diagnostic tool to screen for subclinical mastitis.