Effects of Intrauterine Devices in Mares: A Histomorphological and Immunohistochemical Evaluation of the Endometrium

Oestrous suppression by intrauterine devices (IUDs) is caused by prolongation of luteal function, but the biological mechanism is unknown. The aim of the study was to investigate mechanisms which could explain the action of IUDs. Thirty mares were age‐matched and either inseminated (AI, n = 15) or fitted with an IUD (IUD, n = 15) and subsequently divided into four groups: AI‐P, pregnant (n = 8); AI‐N, non‐pregnant (n = 7); IUD‐P, prolonged luteal phase (n = 7); and IUD‐N, normal luteal phase (n = 8). The median ages were 5.5 and 7 years in AI‐P and IUD‐P groups and 14 and 11 years in AI‐N and IUD‐N groups, respectively. On Day 15 after ovulation, an endometrial biopsy was obtained to study histomorphological and immunohistochemical expression patterns of uterine proteins (uteroferrin, UF; uterocalin, UC; uteroglobin, UG), oestrogen and progesterone receptors (ER, PR), proliferation marker Ki‐67 and content of inflammatory cells. Expression of UF was higher in IUD mares; the difference between pregnant and IUD‐P mares was significant. Mares exhibiting a prolonged luteal phase (AI‐P, IUD‐P) showed only mild angiosclerosis and lower expression of both ER and PR than mares with a normal luteal phase (AI‐N, IUD‐N). No significant differences were detected in the numbers of inflammatory cells, with the exception of macrophages, which were more numerous in AI‐P than AI‐N mares. Although inflammatory cells were not detected in IUD mares, increased UF levels may indicate chronic inflammation. Young age and normality of the endometrial blood vessels may improve the efficacy of IUDs.     

Effects of crocin on frozen-thawed sperm apoptosis,protamine expression and membrane lipid oxidation in Yanbian yellow cattle

Glycerol is used as a bovine semen osmotic cryoprotectant that greatly improves the quality of frozen and thawed bovine sperm. However, high glycerol concentrations can have a toxic effect on frozen and thawed bovine sperm. Therefore, this experiment investigated the effect of replacing a portion of the glycerol in a cryoprotectant solution with crocin on the sperm apoptosis, protamine deficiency and membrane lipid oxidation of frozen and thawed Yanbian yellow cattle sperm. The experiment included a control group (6% glycerol) and four treatment groups: I (3% glycerol), II (3% glycerol +0.5 mM crocin), III (3% glycerol + 1 mM crocin) and IV (3% glycerol + 2 mM crocin). Computer assisted semen analysis was used to detect sperm motility, Hoechst 33,342, propidium iodide, and JC-1 staining were used to analyse sperm viability and mitochondrial membrane potential, chromomycin A3 staining was used to detect protamine deficiency and DNA damage, flow cytometry was used for sperm membrane lipid disorder detection and analysis, and real-time quantitative RT-qPCR was used to detect the mRNA expression levels of protamine-related genes (PRM2, PRM3), sperm acrosome-associated genes (SPACA3), oxidative stress-related genes (ROMO1) and apoptosis-related genes (BCL2, BAX). Compared to the control group, replacing a portion of glycerol with 1 mM crocin significantly improved sperm motility, plasma membrane integrity, membrane lipid disorders (p < .05) and viability, mitochondrial membrane potential, protamine deficiency (p < .01). The expression level of PRM2, PRM3, SPACA3 and BCL2 significantly increased (p < .05), while the expression levels of ROMO1 and BAX significantly decreased (p < .05). Accordingly, the BCL2/BAX ratio significantly increased (p < .05). In summary, the substitution of a portion of glycerol with crocin in cryoprotective solution improved the quality of Yanbian yellow cattle sperm after freezing and thawing.     

Luteolytic Effect of Prostaglandin F2α on Bovine Corpus Luteum Depends on Cell Composition and Contact

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.     

IMAGING DIAGNOSIS—TEMPOROMANDIBULAR JOINT DYSPLASIA IN A BASSET HOUND

A 5‐month‐old intact male Basset Hound presented for evaluation of pain and crepitation during manipulation of the temporomandibular joint, worse on the right side. A computed tomography (CT) scan of the head was performed. The CT images demonstrated the osseous features of temporomandibular joint dysplasia and facilitated a 3D reconstruction, which allowed better visualization of the dysplastic features. The patient responded to conservative management with a tape muzzle with no recurrence reported by the owner 6 months after presentation.     

Feline paraneoplastic syndrome associated with thymoma

CASE HISTORY: A 6-year-old, spayed, female, domestic short-haired cat presented with severe erythroderma and scaling skin. She showed disturbed gait and mild behavioural changes.

CLINICAL FINDINGS: The cat had a generalised, erythematous, scurfy dermatitis with marked, multifocal crusting and skin thickening. The skin was painful and contracted, which appeared to prevent normal freedom of movement.

DIAGNOSIS: The cat was suspected to have a paraneoplastic syndrome. A mediastinal mass was located and histologically confirmed as thymoma. The cat was diagnosed with a thymoma-associated cutaneous paraneoplastic syndrome.

CLINICAL RELEVANCE: This is a rare condition with few reports in the literature. The skin changes, both grossly and histologically, were considered to be different from those described in cases of paraneoplastic dermatosis associated with pancreatic neoplasia. The clinical presentation was characteristic and more cases may occur in practice than are recognised. In this case, as in previous reports, the tumour was grossly resectable, which could lead to cure of the clinical condition.     

Enhanced bioavailability of oral docetaxel by co-administration of cyclosporin A in dogs and rats

Pharmacokinetic and pharmacodynamic endpoints of intravenously and orally administered docetaxel (DT) with or without oral cyclosporine were characterized in rats and dogs. Plasma samples were analysed for DT using liquid chromatography–mass spectrometry. DT area‐under‐the‐concentration–time curve, plasma clearance and maximum serum clearance were significantly affected by cyclosporine in rats (P ≤ 0.0005). Bioavailability of DT alone and DT combined with cyclosporine was 1.3 and 82%, respectively, in rats. In dogs, cyclosporine resulted in a 14–17‐fold increase in peak DT concentrations (P < 0.0005) and area‐under‐the‐curve (P < 0.0005), and a 17‐fold reduction in DT clearance (P < 0.005). Bioavailability of DT alone was 18 ± 16% and statistically exceeded 100% (251 ± 104%) when combined with cyclosporine in dogs. Two of the six dogs receiving DT plus cyclosporine developed neutropenia, and one of the six dogs experienced vomiting and diarrhoea. Cyclosporine significantly alters the disposition of oral DT in dogs and rats, such that clinically relevant serum concentrations are achievable.     

Cardiovascular effects of butorphanol in isoflurane-anesthetized alpacas

Butorphanol has been used clinically to provide analgesia in alpacas, but cardiovascular effects have not been reported. Using a randomized cross‐over design, eight healthy, young adult female alpacas (3 ± 1 SD years) weighing 64 ± 9 SD kg were anesthetized with isoflurane by mask followed by tracheal intubation and maintenance of anesthesia with 1.75% et (isoflurane) in oxygen. Two treatments, butorphanol (0.1 mg kg^–1 IV) and control (saline, IV) were assigned to the animals in a randomized manner allowing a minimum of two weeks between treatments. While anesthetized, animals were instrumented for measurement of cardiovascular variables including systolic, diastolic, and mean arterial blood pressure, pulmonary arterial pressure, pulmonary capillary wedge pressure, central venous pressure, cardiac output (CO) and pulmonary temperature (TEMP). CO was measured via thermodilution using 5 mL of iced 5% dextrose and recording the average of three replicate measurements. Cardiac index, systemic vascular resistance (SVR) and pulmonary vascular resistance were also calculated. Arterial and mixed venous blood samples were collected for blood gas analysis [pH, pO₂, pCO₂, (HCO₃^?), BE, Hb_sat]. Variables were collected at baseline (time 0) and at 5, 10, 15, 30, 45, and 60 minutes following injection. Variables were analyzed by anova for repeated measures with post‐hoc differences between means identified using the Bonferroni comparison (p < 0.05). SVR decreased five minutes after administration of butorphanol (Huynh Feldt corrected p = 0.045) and remained decreased for 60 minutes. TEMP decreased with time in both groups (Huynh Feldt corrected p = 0.000027), but groups were not different between each other. Other cardiovascular and blood gas variables were not different between groups. We conclude that butorphanol (0.1 mg kg^–1 IV) had minimal effects on the cardiovascular system of these alpacas, causing a mild decrease in SVR.     

Effects of Constant, 9 and 16-h Light Cycles on Sperm Quality, Semen Storage Ability and Motile Sperm Subpopulations Structure of Boar Semen

This study was performed to test the effect that two separate, daily, constant-light regimes of both 9 and 16 h could have on the main parameters of boar-semen quality analysis, as well as on the motile sperm subpopulations structure and the ability of its conservation at 16 degrees C. Results show that both luminous regimes have slight, specific effects on the main parameters of boar-semen quality analysis, as well as on the motile sperm subpopulations structure. Furthermore, the conservation ability at 16 degrees C of boar semen was not significantly different between both photoperiods. When a temporal study was performed, results showed that semen quality and motility parameter changes were stabilized at nearly constant values from the second month of the study to the last month in both luminous regimes, indicating a rapid light-related effect on testicular function. Our results indicate that light regimes oscillating from 9 h daily to 16 h daily are of little importance in the control of boar-semen quality in a farming environment.