Equine uterine sanitizer: in vitro inhibition of endometritis-causing microorganisms and its stability to dilute antibiotics.

Endometritis is the leading cause of mare subfertility. Most mares respond to standard therapy, but alternative therapies have been developed for mares failing to respond. This study aimed to investigate a commercially available, yet unassessed, product labeled as a uterine sanitizer to determine the in vitro antimicrobial activity against microorganisms associated with endometritis and its in vitro stability to dilute antibiotics. In experiment 1, the microdilution broth technique and antimicrobial effects were assessed against Escherichia sp, Staphylococcus sp., Klebsiella sp., Pseudomonas sp., and Candida sp. Percentage inhibition was calculated by comparing the optical density. The minimum inhibitory concentration (MIC) 100% was determined using the resazurin dye technique. MIC 50% and 90% were determined using a dose-response non-linear regression. In experiment 2, the uterine sanitizer was used to dilute commonly used antibiotics achieving a final volume of 90 mL at 5°C, 21°C, and 37°C. The pH was measured at 0, 1, 3, 6, and 24 h after dilution. The uterine sanitizer had inhibitory properties against all microorganisms; Escherichia sp. being the most susceptible, and Pseudomonas sp. the most resistant. The uterine sanitizer had an acidic pH=4; however, when combined with the antibiotics, the pH of the antibiotic remained unchanged with the different temperatures and did not precipitate. In conclusion, the uterine sanitizer showed antimicrobial effects against endometritis-causing microorganisms. The dilution of antibiotics in the uterine sanitizer was stable and this association could potentiate the antimicrobial effects. Uterine sanitizer's safety and clinical efficacy in vivo remain to be tested.

Post-cooling sperm processing can rescue sperm quality of cooled-stored stallion semen.

Poor sperm quality in cooled-shipped semen has been related to subpar fertility in horses. Therefore, this study aimed to evaluate the ability of post-cooling sperm processing to improve sperm parameters of cooled-stored stallion semen for artificial insemination. For all experiments, ejaculates were collected, processed, and diluted in skimmed milk-based (SM) medium and stored at 5 °C/24h. In all experiments an aliquot of unprocessed cooled semen was used as a control. In the first experiment (Exp 1.), cooled-stored semen from 16 stallions (n = 32) was processed by SpermFilter or centrifugation (600×g/10min) and resuspended in an egg yolk-based freezing medium containing permeating cryoprotectants (EY-C) for cryopreservation. Sperm recovery and motility parameters were immediately assessed after sperm resuspension in both groups and compared with unprocessed (Unp) samples. In Exp 2., cooled semen samples from six stallions (n = 18) were processed using SpermFilter and resuspended in SM or EY-C. Motility parameters and plasma membrane integrity were assessed in all groups (Unp, SM, and EY-C). In Exp 3, cooled semen from four stallions (n = 20) was processed by SpermFilter, resuspended in SM, EY-C, or egg yolk-based medium without cryoprotectants (EY-nC); and submitted to a thermoresistance test (37 °C/3h). Motility parameters, plasma membrane integrity and stability, mitochondrial membrane potential, mitochondrial superoxide generation, and DNA fragmentation index were evaluated in all groups. Finally, in Exp 4, 39 estrous cycles of 11 mares were inseminated with unprocessed (n = 6) cooled-stored semen or semen cooled at 5 °C/24h and then processed by SpermFilter and resuspended in SM (n = 5), EY-C (n = 11), EY-nC (n = 11), or centrifuged and resuspended in EY-C (n = 6). Overall, semen processing and resuspension in EY mediums (EY-C and EY-nC) improved sperm parameters compared with those of unprocessed semen (P < 0.05). Centrifugation (91 ± 5 %) recovered more sperm than SpermFilter (84 ± 9 %; P < 0.05). Sperm resuspended in EY-nC maintained better sperm parameters throughout the thermoresistance test than those in the other groups (P < 0.05). The fertility rates were similar between all groups (P > 0.05). In conclusion, processing and resuspension in EY medium can improve sperm parameters in post-cooled-stored stallion semen.

In Vitro Antimicrobial Activity of Selected Essential Oils Against Endometritis-Causing Microorganisms in Mares.

This study aimed to evaluate the in vitro antimicrobial activity of essential oils (EO) from Ocimum basilicum (basil), Rosmarinus officinalis (rosemary), and Cymbopogon citratus (lemongrass) on endometritis-causing microorganisms in mares. Serial concentrations of the EO from 30.00 mg/mL to 0.47 mg/mL were tested. The major compounds of O. basilicum EO were linalyl acetate (33.32 wt.%) and citronellal (25.06 wt.%); of R. officinalis EO were borneol (26.48 wt.%), trans-ß-ocimene (16.76 wt.%), camphene (12.45 wt.%), and α-phellandrene (11.08 wt.%); and of C. citratus EO were geranial (45.96 wt.%) and neral (32.62 wt.%). Regarding antimicrobial activity, C. citratus EO has had the highest inhibition percentage (73.9%), followed by O. basilicum (67.2%) and R. officinalis (58.7%). P. aeruginosa was the only pathogen unable to establish the minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) values for the studied EO. The EOs were effective against all other microorganisms (S. equi, S. aureus, K. pneumoniae, E. coli, and C. Albicans). In conclusion, the EOs of O. basilicum, R. officinalis, and C. citratus have presented in vitro antimicrobial activity against microorganisms causing endometritis in mares.

High or Low Body Fat Deposition in the Presence of a Normal Oral Sugar Test is Not Associated With Postthaw Semen Parameters in Stallions.

This study compared the postthaw semen parameters of stallions with high and low body condition score (BCS) and evaluated associations between body morphometric parameters and postthaw semen parameters. Twenty stallions were split into Low BCS (BCS<7, n = 11) and High BCS (BCS ≥7, n = 9) groups, and underwent a complete morphometric analysis (e.g., neck scores and circumference, crest neck height, body weight, and height), and subcutaneous body fat thickness (SFT) at the tail head, withers, shoulders, and retroperitoneal space. A fasted oral sugar test (OST) was conducted on all stallions. One ejaculate from each stallion was frozen with a commercial egg yolk-based extender. Postthaw sperm motility parameters, plasma membrane integrity, mitochondrial membrane potential, hydrogen peroxide and intracellular superoxide production, and lipid peroxidation were analyzed for all stallions. The circumference at 25% and 50% of the neck's length were larger for High-BCS stallions (P < .05). There were no differences between groups for the neck crest height (P > .05). Stallions with High BCS had greater SFT at the tail head than stallions with Low BCS (P < .05); however, there were no differences between groups in the SFT at the shoulders and withers (P > .05). All stallions had resting blood glucose below the cutoff for equine metabolic syndrome. There were no differences between groups for resting glucose concentrations or for a peak at 30 or 60 minutes after initiation of the OST (P > .05). There were no differences in sperm parameters between groups (P > .05). Collectively, the findings of the present study suggest that High BCS or Low BCS in the presence of normal OST do not explain post-thaw semen parameters.

Comparative Efficacy of Histrelin Acetate and hCG for Inducing Ovulation in Brazilian Northeastern Jennies (Equus africanus asinus).

The goal of this study was to compare the efficiency of histrelin acetate (GnRH analog) and human chorionic gonadotropin (hCG) to hasten ovulation in Brazilian Northeastern jennies (Equus africanus asinus). Thirty cycles of ten jennies were randomly assigned in one of the three groups: G0 (control group), saline; G1, 250 µg of histrelin acetate; G2, 2500 IU of hCG. Jennies were evaluated by transrectal palpation and ultrasonography, and had the administration of an ovulation-inducing agent when a follicle measuring between 29 and 32 mm of diameter was diagnosed. Jennies were monitored every 6 hours by transrectal ultrasonography until ovulation. The interval between prostaglandin administration and ovulation was lower (P < .05) in jennies from the G1 (145.2 ± 34.6 hours) and G2 (147.4 ± 27.3 hours) groups compared with the control cycle (220.0 ± 41.8 hours). Both treatments (G1, 41.15 ± 3.5 hours; G2, 37.8 ± 2.5 hours) also reduced (P < .05) the interval that jennies took to ovulate after the administration of the ovulation-inducing agent compared with the control (81.8 ± 28.8 hours). All jennies from G1 and G2 ovulated up to 48 hours after ovulation induction, whereas 100% of jennies in the control cycle ovulated later (>48 hours from the administration of saline). In conclusion, both histrelin acetate and hCG at the used dose are efficient ovulation-inducing agents in jennies promoting ovulation up to 48 hours after administration.