Age-specific analysis of anti-müllerian hormone and liver biochemical parameters in freemartin holstein calves and heifers: A pilot study.

Freemartinism is the most common congenital anomaly among sexual disorders in dairy cows. This syndrome typically occurs in different-sex twin pregnancies and causes vascular anastomoses to form with the placenta in the early stages of fetal development. The study aims to determine the effectiveness of Anti-müllerian hormone (AMH) levels in calves and heifers of different age groups for diagnostic factors and to investigate the potential consequences of different hormone levels in different age groups on some liver biochemical parameters. The study involved 50 cattle from diverse age categories, divided into the freemartin group (FM Group, n=25) and the control group (C Group, n=25). Both FM and control groups were further divided into early-age (3-5 months), middle-aged (5-9 months), and older-aged groups (9-12 months). Serum AMH levels, along with total protein, albumin, and total cholesterol levels, were measured. While no statistically significant difference in AMH levels was observed in the early-age group (P:0.53), significant differences were determined in the middle (P:0.015) and older-age groups (P:0.01), where the FM group exhibited significantly decreased AMH levels compared to the control group. The evaluation of liver biochemistry revealed a statistically significant difference in total protein levels between the FM and control groups in the older age group (P:0.033). Consequently, it is reasonable to suggest that AMH levels may serve as a valid parameter for diagnosing freemartin syndrome in calves aged older than five months. Conversely, particularly in young calves, no significant differences in liver functionality were observed between freemartin-affected and healthy calves.

Antenatal pentoxifylline therapy to prevent endotoxin-induced fetal injury in the preterm goat model.

OBJECTIVE: Pentoxifylline (PTX) has immunomodulatory properties and is known to reduce sepsis-associated infant mortality. We aimed to evaluate maternal oral and intra-amniotic administration of PTX for the prevention of fetal inflammation and injury in a caprine model. MATERIALS AND METHODS: Inflammation-mediated fetal injury was induced with maternal granulocyte-colony stimulating factor and intra-amniotic endotoxin at 0.76 of gestation in date-mated pregnant goats. Eight groups were formed (n=4 each): Control, fetal injury, oral 30 mg/kg/day and 60 mg/kg/day PTX for 15 days + fetal injury, intra-amniotic 400 mg/kg and 800 mg/kg estimated fetal weight single-dose PTX with and without fetal injury. Preterm delivery by hysterotomy was performed at 0.80 of gestation to evaluate the fetal and placental effects. Immunochemistry for various markers including interleukins, caspases, cyclooxygenases, vimentin, myelin basic protein, and surfactant proteins were carried out in the fetal lungs, fetal brain, and placenta. Fetal plasma and amniotic fluid interleukins were also evaluated. Kruskal-Wallis H test and Mann-Whitney U test were used for comparisons. RESULTS: High-dose (60 mg/kg/day) maternal prophylactic oral treatment attenuated endotoxin-related histological injury and was related to low inflammatory marker expressions comparable to the controls (p>0.05 except cyclooxygenase 2). Following maternal oral administration, fetal plasma and amniotic fluid levels of the studied interleukins were also lower than the untreated endotoxin-exposed animals (p<0.05 for all comparisons). Intra-amniotic PTX was associated with inconsistent results and increased inflammatory markers in some fetuses. CONCLUSION: Oral PTX before preterm birth mitigates intrauterine inflammation with neuroprotective effects in the fetus. PTX can be considered as a candidate drug for fetal brain injury prevention in the preterm period.

Inflammation-mediated fetal injury by maternal granulocyte-colony stimulating factor and high-dose intraamniotic endotoxin in the caprine model.

OBJECTIVE: To define a novel experimental model with maternal intravenous (i.v.) granulocyte-colony stimulating factor (G-CSF) followed by a single- and high-dose of 20 mg intra-amniotic (IA) endotoxin to induce fetal brain injury in the preterm fetal goat. MATERIALS AND METHODS: Pregnant goats (n=4) were given 50 microg/day G-CSF into the maternal jugular vein through gestational days 110-115 (term, 150 days). At gestational day 115, 20 mg of IA endotoxin was administered. Following preterm delivery at day 120 by cesarean section umbilical cord, fetal lung and brain tissues were harvested for histopathology, immunohistochemistry, and electron microscopy. Inflammatory markers were evaluated in the amniotic fluid and fetal plasma. RESULTS: Necrotizing funisitis with abundant leukocyte infiltration and fetal brain injury was induced in all the fetuses in the experimental group. CONCLUSION: Maternal i.v. G-CSF for 5 days followed by 20 mg of IA endotoxin is a feasible caprine model to exacerbate intrauterine inflammation.

Ultra-high dose of intra-amniotic or direct fetal intramuscular betamethasone for lung maturation in the preterm goat model.

OBJECTIVE: To evaluate the effects of intra-amniotic (IA) and fetal injections of a single ultra-high dose of betamethasone (BM) 48 h before preterm delivery on neonatal pulmonary function, using an experimental goat model. STUDY DESIGN: Eighteen date-mated singleton pregnant Hair goats were randomized into four groups. At gestational day 118 (alveolar phase, term 150-155 days) after obtaining a sample of amniotic fluid, fetuses in group 1 (n=5) received 8 mg/kg IA BM, and in group 2 (n=5) 4 mg/kg fetal IM BM. In group 3 (n=4) (0.3mg/kg/day) maternal BM was administered at day 118 and 119 with a 24h interval; control fetuses (n=4) received 1 mL/kg of IA saline at day 118. At gestational day 120, after obtaining second sample of amniotic fluids 18 kids were delivered by preterm cesarean section, entubated, weighed, and mechanically ventilated for 15 min. Arterial blood gas samples and deflation/inflation lung pressure-volume measurements were obtained. After sacrifice, lungs were removed, weighed, gross examined and processed for further histological and immunohistochemical (IHC) evaluations. On hematoxylin and eosin (HE) stained slides, presence and severity of lung emphysema was evaluated; slides stained for surfactant proteins, and caspases were used for semi-quantitative evaluation of lung maturation. Kruskal-Wallis, Mann-Whitney, Wilcoxon signed rank, and chi-square tests were used for comparisons. RESULTS: IA BM was associated with increased number of stillbirths (60% vs. 0% in control) (p=0.06) and emphysematous changes. Bodyweight-adjusted pressure-volume measurements were improved after maternal, but not IA or fetal, BM (p=0.06). Following mechanical ventilation, arterial blood gas parameters did not significantly alter across maternal and fetal administrations. However, pH was significantly lower (p<0.05) and carbon dioxide partial pressure was higher (p<0.05) in the control group, indicating hypercapnic acidemia in non-treated pregnancies. None of the treatments induced measurable alterations in amniotic fluid lecithin/sphingomyelin (L/S) values. IA and fetal routes were associated with decreased surfactant protein expressions and increased apoptotic activity in alveolar and bronchio-alveolar epithelial cells. CONCLUSION: Ultra-high dose IA and fetal IM BM is not superior to the standard dose and maternal way of administration in our experimental design.