Laparoscopic Application of PGE2 to the Uterine Tube Surface Enhances Fertility in Selected Subfertile Mares

Topical application of prostaglandin E₂ (PGE₂) gel to the surface of the uterine tubes via a laparoscopic procedure improved embryo recovery rates or pregnancy rates in 28 subfertile mares suspected of uterine tubal pathology. Gelatinous masses may occlude the lumen of the uterine tube and prevent sperm from reaching the site of fertilization or prevent embryos from reaching the uterus. PGE₂ is secreted by the early equine embryo, promoting passage of the embryo into the uterus; topical administration of PGE₂ onto the surface of the uterine tube has been shown to stimulate early transport of the embryo into the uterus. Embryos were produced or a pregnancy was obtained from 24 of the 28 barren mares treated with direct laparoscopic application of 0.2 mg of PGE₂ to their uterine tubes. Mares had been barren for an average duration of 1.9 ± 0.6 years and an average of 6.9 ± 3.8 estrous cycles prior to treatment, without donating an embryo or becoming pregnant. Seventeen of 20 mares bred as embryo donors produced one or more embryos with an average of 2.1 ± 1.9 embryos collected per mare (0.45 embryos per cycle) after PGE₂ treatment. Seven of 8 mares bred to carry their own pregnancy became pregnant within the first two cycles following PGE₂ treatment. These 8 mares were bred an average of 5.6 ± 1.8 cycles without a pregnancy prior to treatment. The laparoscopic PGE₂ procedure was performed during various stages of the estrous cycle; the stage varied among treated mares.     

Exploring relationships between body condition score,body fat,activity level and inflammatory biomarkers

Obesity is associated with inflammatory disorders in humans, including degenerative joint disease. While obesity is endemic in horses, its relationship to equine degenerative joint disease has not been explored. The current study sought to describe relationships between: body weight (BW), body condition score (BCS), lameness grade (AAEP), total body fat mass (kg; FM) and fat per cent (FP) [multifrequency bioelectrical impedance analysis (mfBIA)], age, gender, activity level (AL), synovial fluid (SF) and plasma (PL) PGE₂ and glycosaminoglycan (GAG) in horses. During this field investigation, the BCS (of nine) of 54 horses at multiple farms in southern Ontario, Canada, was determined. Horses were categorized as thin (BCS=3/9; n = 6), moderate (BCS=4 or 5/9; n = 18), overweight (BCS=6 or 7/9; n = 19) or obese (BCS=8 or 9/9; n = 11). Total fat mass (kg) and body fat% was measured using mfBIA, lameness was assessed (AAEP lameness scale) and synovial fluid was collected via aseptic arthrocentesis from the left intercarpal joint for assessment of inflammatory biomarkers (PGE₂, GAG). Means were compared with a one‐way ANOVA; correlation coefficients were calculated using a Spearman Rank Order Correlation to reveal correlations between variables. BCS was positively correlated with BW, FM, FP, AL and PL‐PGE₂. BW was also significantly positively correlated with PL‐PGE₂. It is concluded that BCS is significantly correlated with PL‐PGE₂, due in part to the combined effect of AL and body condition. Net inflammatory effects of body fat on risk for joint disease require further study.     

Comparative plasma and milk dispositions,faecal excretion and efficacy of per os ivermectin and pour‐on eprinomectin in horses

The horse milk gains increasing interest as a food product for sensitive consumers, such as children with food allergies or elderly people. We investigated the plasma and milk disposition, faecal excretion and efficacy of per os ivermectin (IVM) and pour‐on eprinomectin (EPM) in horses. Ten mares were divided into two groups. The equine paste formulation of IVM and bovine pour‐on formulation of EPM were administered orally and topically at dosage of 0.2 and 0.5 mg/kg bodyweight. Blood, milk and faecal samples were analysed using high‐performance liquid chromatography. The plasma concentration and persistence of IVM were significantly greater and longer compared with those of EPM. Surprisingly, EPM displayed a much higher disposition rate into milk (AUC_milk/plasma: 0.48) than IVM (AUC_milk/plasma: 0.19). IVM exhibited significantly higher faecal excretion (AUC_faeces: 7148.54 ng·d/g) but shorter faecal persistence (MRT_faeces: 1.17 days) compared with EPM (AUC_faeces: 42.43 ng·d/g and MRT_faeces: 3.29 days). Faecal strongyle egg counts (EPG) were performed before and at weekly intervals after treatment. IVM reduced the EPG by 96–100% for up to 8 weeks, whereas the reduction in the EPM group varied from 78 to 99%. In conclusion, due to the relatively low excretion in milk, EPM and IVM may be used safely in lactating mares if their milk is used for human consumption. Nevertheless, much lower plasma and faecal availabilities of EPM could result in subtherapeutic concentrations, which may increase the risk of drug resistance in nematodes after pour‐on EPM administration compared with per os IVM.     

Intraocular pressure,specular microscopy,and prostaglandin E2 concentration in dogs with mature and hypermature cataract

This study aimed to evaluate and correlate intraocular pressure (IOP), endothelial cell density (CD), and hexagonality (HEX), and the aqueous humor prostaglandin E₂ (PGE₂) concentration in dogs with mature (MG, n = 8) and hypermature (HG, n = 8) cataracts. Eight laboratory beagles with no ocular abnormalities were included as a control group (CG). The IOP was measured using a digital applanation tonometer. Noncontact specular microscopy was used to evaluate CD and HEX. Samples of aqueous humor were used to determine prostaglandin E₂ concentration using enzyme‐linked immunoassay. Data were compared by anova and Bonferroni's multiple comparison test, and possible correlations among the PGE₂ aqueous concentration and corneal endothelium cell parameters were assessed by Person′s test (P < 0.05). Average values of IOP (P = 0.45) and CD (P = 0.39) were not significantly different between MG, HM, and CG. Average values of HEX were lower, and PGE₂ concentration was increased in the MG and HG in comparison with CG (P < 0.05); however, such parameters did not change significantly between MG and HG (P > 0.05). PGE₂ values did not correlate with IOP, CD, and HEX in any group (P > 0.05). Although there were a small number of dogs studied, our results demonstrated that cataract progression from mature to hypermature did not have a significant change in PGE2 aqueous concentration, IOP, corneal endothelial cell count, or morphology. In addition, PGE₂ concentration was not correlated with parameters of the corneal endothelium or IOP in dogs with mature or hypermature cataracts.     

Pharmacokinetics and pharmacodynamics of intravenous and transdermal flunixin meglumine in meat goats

The aim of this study was to determine the pharmacokinetics and prostaglandin E₂ (PGE₂) synthesis inhibiting effects of intravenous (IV) and transdermal (TD) flunixin meglumine in eight adult female Boer goats. A dose of 2.2 mg/kg was administered intravenously (IV) and 3.3 mg/kg administered TD using a cross‐over design. Plasma flunixin concentrations were measured by LC‐MS/MS. Prostaglandin E₂ concentrations were determined using a commercially available ELISA. Pharmacokinetic (PK) analysis was performed using noncompartmental methods. Plasma PGE₂ concentrations decreased after flunixin meglumine for both routes of administration. Mean λ_z‐HL after IV administration was 6.032 hr (range 4.735–9.244 hr) resulting from a mean V_z of 584.1 ml/kg (range, 357.1–1,092 ml/kg) and plasma clearance of 67.11 ml kg^?1 hr^?1 (range, 45.57–82.35 ml kg^?1 hr^?1). The mean C_max, T_max, and λ_z‐HL for flunixin following TD administration was 0.134 μg/ml (range, 0.050–0.188 μg/ml), 11.41 hr (range, 6.00–36.00 hr), and 43.12 hr (15.98–62.49 hr), respectively. The mean bioavailability for TD flunixin was calculated as 24.76%. The mean 80% inhibitory concentration (IC₈₀) of PGE₂ by flunixin meglumine was 0.28 μg/ml (range, 0.08–0.69 μg/ml) and was only achieved with IV formulation of flunixin in this study. The PK results support clinical studies to examine the efficacy of TD flunixin in goats. Determining the systemic effects of flunixin‐mediated PGE₂ suppression in goats is also warranted.     

IFN‐τ Acts in a Dose‐Dependent Manner on Prostaglandin Production by Buffalo Endometrial Stromal Cells Cultured in vitro

Interferon‐τ (IFN‐τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F_2α (PGF_2α) and prostaglandin E₂ (PGE₂). PGF_2α is responsible for the luteolysis; however, PGE₂ favours establishment of pregnancy by its luteoprotective action. In this study, the dose‐response effect of recombinant bovine IFN‐τ (rbIFN‐τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN‐τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN‐τ treatment enhanced the secretion of both PGE₂ and PGF_2α, and also its ratio in a strict dose‐dependent manner with a significant increase (p < 0.01) in PGE₂ production at 1 μg/ml dose of rbIFN‐τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time‐dependent fashion in all the groups at 6, 12 and 24 h post‐treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE₂ remained higher than PGF_2α indicating PGE₂ as the major PG produced by endometrial stromal cells. The dose‐dependent response of rbIFN‐τ signifies the importance of optimum concentration of IFN‐τ for the embryonic development especially during the critical period to establish successful pregnancy.     

Pharmacokinetics and pharmacodynamics of intravenous and transdermal flunixin meglumine in alpacas

The aim of this study was to determine the pharmacokinetics and prostaglandin E₂ (PGE₂) synthesis inhibiting effects of intravenous (IV) and transdermal (TD) flunixin meglumine in eight, adult, female, Huacaya alpacas. A dose of 2.2 mg/kg administered IV and 3.3 mg/kg administered TD using a cross‐over design. Plasma flunixin concentrations were measured by LC‐MS/MS. Prostaglandin E₂ concentrations were determined using a commercially available ELISA. Pharmacokinetic (PK) analysis was performed using noncompartmental methods. Plasma PGE₂ concentrations decreased after IV flunixin meglumine administration but there was minimal change after TD application. Mean t_1/2λz after IV administration was 4.531 hr (range 3.355 to 5.571 hr) resulting from a mean Vz of 570.6 ml/kg (range, 387.3 to 1,142 ml/kg) and plasma clearance of 87.26 ml kg^?1 hr^?1 (range, 55.45–179.3 ml kg^?1 hr^?1). The mean C_max, T_max and t_1/2λz for flunixin following TD administration were 106.4 ng/ml (range, 56.98 to 168.6 ng/ml), 13.57 hr (range, 6.000–34.00 hr) and 24.06 hr (18.63 to 39.5 hr), respectively. The mean bioavailability for TD flunixin was calculated as 25.05%. The mean 80% inhibitory concentration (IC₈₀) of PGE₂ by flunixin meglumine was 0.23 µg/ml (range, 0.01 to 1.38 µg/ml). Poor bioavailability and poor suppression of PGE₂ identified in this study indicate that TD flunixin meglumine administered at 3.3 mg/kg is not recommended for use in alpacas.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.     

Anti‐inflammatory effects of four potential anti‐endotoxaemic drugs assessed in vitro using equine whole blood assays

Despite the severity and common occurrence of equine endotoxaemia, the available anti‐endotoxic treatments do not effectively target key inflammatory mechanisms such as leucocyte activation and cytokine production. In this study, four compounds with potential anti‐endotoxic effects, namely rolipram, azithromycin, ethyl pyruvate and metformin, were investigated in vitro using equine whole blood stimulated with bacterial lipopolysaccharide. TNF‐α and IL‐1β production were measured in plasma. Rolipram was the most potent inhibitor of cytokine production (IC₅₀ 0.84 and 4.68 μm for TNF‐α and IL‐1β, respectively) with almost complete inhibition of TNF‐α, but inhibited IL‐1β by only 39.46%. Azithromycin produced almost complete inhibition of both cytokines, but tended to be less potent than rolipram (IC₅₀ 10.66 and 17.4 μm for TNF‐α and IL‐1β, respectively). Metformin inhibited TNF‐α production with similar potency to rolipram and azithromycin (IC₅₀ 3.35 μm ) but showed significantly lower efficacy (45.93%; P < 0.05), and had no inhibitory effect on IL‐1β. Ethyl pyruvate was the least potent (IC₅₀ 68.35 μm and >10 mm for TNF‐α and IL‐1β production, respectively). Further work is required to investigate whether these or related compounds may have potential use in the treatment of equine endotoxaemia in vivo.     

Use of Cholesterol‐Loaded Cyclodextrin in Donkey Semen Cryopreservation Improves Sperm Viability but Results in Low Fertility in Mares

The use of cholesterol‐loaded cyclodextrin (CLC) on semen cryopreservation has been related with better sperm viability in several species; however, the effect on fertility is not known in donkey semen. Ejaculates (n = 25) from five donkeys were diluted in S‐MEDIUM with 0, 1, 2 or 3 mg of CLC/120 × 10⁶ spermatozoa. Semen was frozen, and thawed samples were evaluated by computer‐assisted sperm analyser system (CASA), supravital test, hyposmotic swelling test and fluorescent dyes to assess the integrity of sperm membranes. Mares (n = 60) were inseminated with frozen‐thawed semen treated with the doses of 0 or 1 mg CLC. Percentages of sperm with progressive motility and with functional plasma membrane were greater (p < 0.05) in the CLC‐treated groups than in the control. Percentages of intact plasma membrane and intact plasma membrane and acrosome detected by fluorescent dyes were also greater (p < 0.05) in CLC‐treated groups. Although no difference (p > 0.05) in conception rates was detected between groups (control, 3/30, 10%; CLC‐treated, 1/30, 3.3%), fertility was low for artificial insemination programs in mares. Therefore, we firstly demonstrated that frozen semen treated with CLC in S‐MEDIA extender before freezing improves the in vitro sperm viability, but semen treated or not with CLC in S‐MEDIUM extender results in a very low conception rate in mares inseminated with thawed donkey semen.     

Case series: Persistent dorsal displacement of the soft palate in eight horses

Persistent dorsal displacement of the soft palate (pDDSP) is a relatively rare equine disorder. This case series reports histories and clinical findings in 8 cases of pDDSP, and outcome after treatment of concurrent epiglottic entrapment (n = 3), laryngeal tie‐forward (n = 6) ± laser staphylectomy (n = 2), and/or sectioning of mucosa ventral to the epiglottis and the hyoepiglotticus muscles (n = 2). Four of 6 horses that underwent tie‐forward ± laser staphylectomy had complete resolution of pDDSP and returned to their previous level of work, as did one horse that had a subepiglottic releasing incision. Correction of concurrent epiglottic entrapment in 3 horses with pDDSP did not result in resolution of pDDSP in any case. Previous laryngeal surgery including laryngoplasty and ventriculocordectomy appear to be risk factors for the development of pDDSP.     

Collagen,glycosaminoglycans and matrix metalloproteinase‐2 and metalloproteinase‐9 in the cervix of the ewe during prepubertal development

The tortuous nature of the ovine cervix restricts the transcervical passage of the cannula, and many studies have aimed to understand the endocrine mechanism of the remodelling of cervical tissue in adult ewe. However, little is known about the remodelling of the cervical tissue during the prepubertal development of the lambs. To obtain histochemical and biochemical evidence about the nature of the prepubertal development of the cervix of the ewe, cervices of Corriedale lambs obtained at 0, 1, 2, 4, 6 and 8 months of age (n = 5 to 6 in each) were processed. Neutral and acidic glycosaminoglycans (by PAS‐Alcian stain) were weakly in the cervical stroma and not shown change during the development, whereas the percentage volume of fibrillar collagen (by van Gieson stain) increases throughout the experimental period in the superficial fold stroma and deep wall stroma (p < 0.05). The relative cervical weight (g/kg of body weight) and the collagen concentration (by spectrophotometry, mg/mg wet tissue) showed an early decreasing phase from months 0 to 4 and a later increasing phase from months 4 to 8 (p < 0.05). The latent form of matrix metalloproteinase‐2 (MMP‐2) detected by gelatin zymography (ng/mg protein) decreased from months 0 to 2 and increased from months 4 to 8, whereas the activated form decreased from months 0 to 2, remained low until month 6 and then recovered on month 8 (p < 0.0001). Data suggest that the relative cervical weight biphasic pattern during the development is related to MMP‐2‐dependent changes in the collagen content.     

Efficacy of a mephentermine‐based product as a vasopressor and a cardiac performance enhancer when given intramuscularly to cattle

The goal of this study was to confirm the vasopressor and cardiac effects of POTENAY^® INJETÁVEL (POT), a mephentermine‐based product, given to cattle with induced vascular/cardiac depression. Ten healthy Holstein cattle (206 ± 13 kg) followed a randomized‐complete‐block design (RCBD) utilizing crossover study design. Each animal randomly received (1 ml/25 kg, IM) of either POT (n = 10) or volume‐matched placebo control (0.9%NaCl, CP, n = 10). A subset of animals (n = 5) received POT first (day 0) while the remaining (n = 5) received CP; after a six‐day washout period, cattle received the opposite compound. Animals were anesthetized and catheterized for systemic/left ventricular hemodynamic monitoring. Myocardial dysfunction/hypotension was induced by increasing the end‐tidal isoflurane concentration until arterial blood pressure was 20% lower than at baseline and remained stable. Once the animal was determined to be hypotensive and hemodynamically stable, steady‐state hypotensive baseline data (BL2) were acquired, and treatment with either POT or CP was given. Data were acquired post‐treatment at every 15 min for 90 min. POT improved cardiac output (+68 L/min, ±14%, p < 0.05), MAP (+14 mmHg, ±4%, p < 0.05), HR (+22 bpm, ±8%, p < 0.05), and peak rates of ventricular pressure change during both systole (dP/dt_max: +37 mmHg/s ±13%, p < 0.05) and diastole (dP/dt_min: +31 mmHg/s, ±7%, p < 0.05). No improvements were noted following placebo‐control administration. Results indicate that POT improves cardiac performance and systemic hemodynamics in cattle with induced cardiovascular depression when given as single intramuscular injection.     

Cervical disc width index is a reliable parameter and consistent in young growing Dutch Warmblood horses

Intervertebral disc disease, as well as the associated alteration of the radiographic intervertebral disc space width, has been reported in horses. Disc height index (DHI) has proven to be an accurate and objective parameter in other species but data related to this parameter are lacking in horses. Therefore, the aims of this retrospective longitudinal diagnostic accuracy study were (a) to evaluate the reliability of measurements within and between observers of the equine Disc Width Index (EDWI) as a parameter for radiographic equine cervical intervertebral disc space width, and (b) to evaluate the sequential development of the EDWI over time. For this, EDWI from all intervertebral disc spaces between second cervical (C) to first thoracic (Th) vertebrae were obtained in a group of 39 Dutch Warmblood horses at 1, 5, and 18 months of age, by one European College of Veterinary Diagnostic Imaging (ECVDI) board‐certified veterinary radiologist (S.V.) and two veterinary students. Bland‐Altmann plots and intraclass Correlation Coefficient revealed a good intra‐ and interobserver agreement. A linear mixed‐effect model did reveal that mean EDWI increases significantly toward the caudal cervical spine, but did not differ significantly for a certain location over time or between sexes. Spearman's rank test did show a significant correlation between the vertebral alignment angle induced by different head‐neck positions and a normalized EDWI (ρ = 0.33, P < .0001). Student's t‐test revealed that the presence of C6‐C7 transposition of the transverse processes did not influence EDWI significantly. It was concluded that EDWI represents a reliable parameter for equine cervical radiographic intervertebral disc space width. Practical implementation of EDWI warrants monitoring in a group of adult horses while maintaining a standardized head‐neck position.     

Two successful embryo transfers of mini-donkey embryos in Brazilian Northeastern jennies using an alternative method: Case report

Pregnancy rates after embryo transfer (ET) are disappointing in donkey species. This study aims to report two successful ET of mini-donkey embryos using Brazilian Northeastern jennies as recipients. Eighteen embryo flushes were performed 9 days post-ovulation in two non-pregnant mini-donkeys jennies (11 and 7 cycles per jenny). Eleven embryos (61%, 11/18) were collected and transferred to Brazilian Northeastern jennies 4–6 days post-ovulation by conventional (n = 6) or an alternative (n = 5) technique. The alternative method consisted of inserting a Polansky equine vaginal speculum smeared with lubricant in the vagina of the recipient jenny. The arms of the speculum were extended to allow the visualization of the cervix. Then, using an adapted crafted, elongated, toothed tissue grasping forceps, the external cervical os was held, and the cervix was gently pulled backward, aiming to straight the cervical canal. The ET gun was inserted through the vagina and cervix by visual inspection, and the embryo was released into the uterine lumen. All embryos collected were Grade 1 and classified as Expanded Blastocysts. No jennies become pregnant after conventional ET (0/6), whereas two recipient jennies (40%, 2/5) become pregnant and delivered offspring in the following year after ET using the alternative technique. In conclusion, Brazilian Northeastern jennies can be used as embryo recipients using the alternative method proposed in the present study. However, further investigations are needed to improve the knowledge and results of ET in donkey species.     

A preliminary study on the suitability of Cervidil to induce cervical dilation for artificial insemination in ewes

The aim of this study was to evaluate the efficacy of Cervidil^®, a prostaglandin E₂ (PgE₂)-releasing vaginal insert used for induction of cervical ripening and labour in women, to enhance the ease of transcervical artificial insemination (TCAI) in anestrous ewes (June). It was hypothesized that the use of Cervidil^® prior to AI would cause dilation of the cervix, and thus alleviate the difficulty associated with traversing the cervix for semen deposition in sheep. Cervidil^® was inserted 12 h before insemination in six Rideau-Arcott ewes; six ewes served as untreated controls.Semen was deposited into the uterus of all six treated ewes but TCAI was possible only in four of the six control ewes. It can be concluded that Cervidil^® facilitated transcervical semen deposition in anestrous ewes. The treatment with Cervidil^® has the promise of a technique to improve transcervical AI and to enable non-invasive embryo transfer procedures in sheep.     

Measurements of canine aqueous humor inflammatory mediators and the effect of carprofen following anterior chamber paracentesis

Objectives Phase I: To evaluate levels of prostaglandin E₂ (PGE₂), nitrites and nitrates (NO_x), tumor necrosis factor‐alpha (TNF‐α) and expression of inducible cyclo‐oxygenase (COX‐2), nitric oxide synthase (NOS‐2), and matrix metalloproteinases (MMP‐3 and ‐9) in canine aqueous humor following repeated anterior chamber paracenteses (ACP). Phase II: to evaluate the effect of carprofen on PGE₂, NO_x, and TNF‐α in canine aqueous humor following ACP. Animals studied Four beagles in phase I and 8 beagles in phase II. Procedures Phase I: ACP was performed at time (T) 0, 4 and 8 h. Phase II: A randomized, placebo‐controlled cross‐over design with four dogs per group where carprofen was given 4.4 mg/kg/day on day (D) 1, 2 and 3. ACP was performed at T0 and T1.5 on D3. Statistical analysis was performed with repeated measures anova and post hoc Tukey‐Kramer multiple‐comparison procedure. In phase II, TNF‐α level was analyzed with a Wilcoxon signed‐rank test. Results Phase I: PGE₂ significantly increased (P < 0.0001) to plateau at T4. NO_X was decreased at T4 (P < 0.06), but increased at T8 (P < 0.0001). COX‐2 showed detectable expression only at T8. TNF‐α, NOS‐2, MMP‐3 and ‐9 were undetectable at all time points. Phase II: At T1.5, PGE₂ was significantly elevated in both groups but was lower in the carprofen group (P = 0.037). NO_x and TNF‐α did not statistically increase in either group. Conclusions Following ACP, significant increases in PGE₂ levels confirmed inflammation characterized by a rise of COX‐2. The NO_x pathway took longer to induce as compared with PGE₂. Carprofen decreased PGE₂ levels and could help control intraocular inflammation.     

Distribution of mast cells in vaginal, cervical and uterine tissue of non-pregnant mares: investigations on correlations with ovarian steroids

Mast cells constitute part of the cellular immune system of the genital tract. They play a potential role in cervical remodelling during parturition and are subject to the influence of ovarian steroids. In this study, the influence of oestradiol‐17β and progesterone on the distribution of mast cells in equine vaginal, cervical and uterine tissue was evaluated. Genital tracts were retrieved from healthy mares at a local slaughterhouse. The cervix was divided along the longitudinal axis into five equivalent regions from cranial to caudal (R1 to R5). Toluidine blue staining was used for the detection of mast cells. Hormone values were determined via radioimmunoassay. In cervical tissue, mast cells were present at higher frequency and higher density than in vaginal and uterine tissue. Statistically significant differences were obtained between cervical R1 (cranial) and uterus as well as cervical R2 and uterus (p < 0.05). In cervical tissue, an association between the presence of mast cells and peripheral oestrogen concentrations was observed, whereas such correlation could not be established for vaginal and uterine tissue. With increasing oestradiol‐17β concentrations, the density of mast cells in cervical tissue increased to a statistically significant degree in the cranial (r = 0.46; p = 0.05) and caudal (r = 0.5; p = 0.03) regions. We propose that mast cells play a role in the cyclic remodelling of the equine cervix during the course of the oestrous cycle. Elucidation of mechanisms controlling cervical remodelling could contribute to development of therapies for mares showing a lack in cervical opening during oestrus.