Carolyn E. Arnold

A Gene Catalogue of the Euchromatic Male-Specific Region of the Horse Y Chromosome: Comparison with Human and Other Mammals

Studies of the Y chromosome in primates, rodents and carnivores provide compelling evidence that the male specific region of Y (MSY) contains functional genes, many of which have specialized roles in spermatogenesis and male-fertility. Little similarity, however, has been found between the gene content and sequence of MSY in different species. This hinders the discovery of species-specific male fertility genes and limits our understanding about MSY evolution in mammals. Here, a detailed MSY gene catalogue was developed for the horse – an odd-toed ungulate. Using direct cDNA selection from horse testis, and sequence analysis of Y-specific BAC clones, 37 horse MSY genes/transcripts were identified. The genes were mapped to the MSY BAC contig map, characterized for copy number, analyzed for transcriptional profiles by RT-PCR, examined for the presence of ORFs, and compared to other mammalian orthologs. We demonstrate that the horse MSY harbors 20 X-degenerate genes with known orthologs in other eutherian species. The remaining 17 genes are acquired or novel and have so far been identified only in the horse or donkey Y chromosomes. Notably, 3 transcripts were found in the heterochromatic part of the Y. We show that despite substantial differences between the sequence, gene content and organization of horse and other mammalian Y chromosomes, the functions of MSY genes are predominantly related to testis and spermatogenesis. Altogether, 10 multicopy genes with testis-specific expression were identified in the horse MSY, and considered likely candidate genes for stallion fertility. The findings establish an important foundation for the study of Y-linked genetic factors governing fertility in stallions, and improve our knowledge about the evolutionary processes that have shaped Y chromosomes in different mammalian lineages.

Periparturient hemorrhage in mares: 73 cases (1998–2005)

OBJECTIVE To determine signalment, physical examination and clinicopathologic abnormalities, outcome, and subsequent fertility of mares with periparturient hemorrhage (PPH) and identify factors associated with outcome (ie, survival vs death). DESIGN Retrospective case series. ANIMALS 73 mares. PROCEDURES Medical records were reviewed for information on age, breed, initial complaint, physical examination and clinicopathologic abnormalities, treatment, outcome, and subsequent fertility. RESULTS Median age was 14.0 years (range, 5 to 24 years), and median number of foals produced prior to the diagnosis of PPH was 8 (range, 1 to 16). Ten (14%) mares had prepartum hemorrhage and 63 (86%) had postpartum hemorrhage. Treatment was aimed at restoring cardiovascular volume, enhancing coagulation, controlling pain, and reducing the effects of endotoxemia. Sixty-one (84%) mares survived and 12 (16%) died or were euthanized. Common complications included fever, leukopenia, retained fetal membranes, increased digital pulses, thrombophlebitis, and cardiac arrhythmias. Of the 53 surviving mares for which subsequent breeding information was available, 26 (49%) produced 1 or more foals after recovering from PPH. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that PPH can develop in mares of any age and parity. Treatment was associated with a good prognosis for survival and a reasonable prognosis for future fertility.

Abdominal abscesses in adult horses: 61 cases (1993-2008)

OBJECTIVE To determine clinical signs, diagnostic methods, treatment, and outcome for a series of adult horses with abdominal abscesses. DESIGN Retrospective case series. ANIMALS 61 adult horses. PROCEDURES Medical records of adult horses with abdominal abscesses treated at Texas A&M Veterinary Medical Teaching Hospital (1993 to 2008) were reviewed. Information was recorded regarding signalment, history, clinical signs, diagnosis, treatment, and short- and long-term outcomes. Risk factors for survival were determined. RESULTS 61 horses met the criteria for inclusion. Clinical signs included colic (67%), fever (46%), anorexia (51%), signs of depression (57%), tachycardia (46%), and weight loss (30%). The diagnosis was made on the basis of abdominal ultrasonography, exploratory celiotomy, palpation per rectum, and necropsy. Abscesses were variable in size, location, and number. Only 15 (24.6%) horses survived to discharge. Multiple bacterial isolates were identified from aspirates of abscesses, and subsequent abdominal adhesion formation limited survival, affecting outcome. Risk factors for survival included age and heart rate at admission. CONCLUSIONS AND CLINICAL RELEVANCE Adult horses with abdominal abscesses often have severe adhesion formation. Multiple bacterial isolates are frequently identified from the abscess. Prognosis for survival is guarded.

Pharmacokinetics of an orally administered methylcellulose formulation of gallium maltolate in neonatal foals

Gallium is a trivalent semi-metal with anti-microbial effects because of its incorporation into crucial iron-dependent reproductive enzyme systems. Gallium maltolate (GaM) provides significant gallium bioavailability to people and mice following oral administration and to neonatal foals following intragastric administration. To study the prophylactic and therapeutic effects of GaM against Rhodococcus equi pneumonia in foals, we developed a methylcellulose formulation of GaM (GaM-MCF) for oral administration to neonatal foals. Normal neonatal foals were studied. Six foals received 20 mg/kg and another six foals received 40 mg/kg of GaM-MCF orally. Serial serum samples were collected and serum gallium concentrations were determined using inductively coupled plasma mass spectroscopy. Gallium was rapidly absorbed (T(max) of 4 h), and a mean C(max) of 0.90 or 1.8 microg/mL was achieved in foals receiving 20 or 40 mg/kg respectively. Marked variability existed in C(max) among foals: only half of the foals receiving 20 mg/kg attained serum concentrations of >0.7 microg/mL, a level suggested to be therapeutic against R. equi by previous studies. Mean elimination half-life was 32.8 or 32.4 h for foals receiving 20 or 40 mg/kg respectively. The results of this study suggest that at least 30 mg/kg orally every 24 h should be considered in future pharmacodynamic and efficacy studies.

Laparoscopic evaluation of oviductal patency in the standing mare.

Intraluminal masses in the oviduct might cause infertility and/or subfertility in mares by preventing transport of ova, sperm, or both, to the site of fertilization or the embryo into the uterus. Currently, there is no clinical test for oviductal patency in mares. The objective was to determine if the passage of microspheres from the oviduct to the uterus was associated with the presence or absence of intraluminal masses. In this trial, a standing laparoscopic technique was used to cannulate the oviducts and instill 15-μm fluorescent beads in 16 light-breed mares. At 48 hours after surgery, uterine lavage was performed to collect and quantify the number of beads transported to the uterus. Mares were immediately euthanized, and their reproductive tracts recovered. The presence of intraluminal masses was determined from postmortem evaluation and compared with the number beads recovered in the uterine lavage fluid. A test was considered positive for intraluminal masses if no beads were transported to the uterus. The sensitivity and specificity of the test were 71.4% and 85.7%, respectively.

Influence of an intra-articular lipopolysaccharide challenge on markers of inflammation and cartilage metabolism in young horses

Nineteen weanling Quarter Horses (225 to 380 kg) were used in a randomized complete block design to investigate the effects of intra-articular lipopolysaccharide (LPS) to induce acute joint inflammation in young horses. Horses were blocked by age, BW, and sex and were randomly assigned to 1 of 3 treatments for a 35-d experiment. Treatments included intra-articular injection of 0.25 ng (n = 7) or 0.50 ng (n = 6) of LPS obtained from Escherichia coli O55:B5 or sterile lactated Ringers solution (n = 6; control) into the radial carpal joint. Synovial fluid was obtained at preinjection h 0 and 2, 6, 12, 24, 168, and 336 h postinjection and was analyzed for PGE2, carboxypeptide of type II collagen (CPII), and collagenase cleavage neopeptide (C2C) biomarkers via commercial ELISA kits. Rectal temperature (RT), heart rate (HR), respiratory rate (RR), and carpal circumference were recorded before each sample. Lameness scores on a 0 to 5 scale were conducted after arthrocentesis. Data were analyzed using PROC MIXED procedure of SAS. Linear and cubic effects were tested in the form of contrasts. Clinical assessment of HR, RR, and RT were not influenced by treatment (P ≤ 0.16). All horses exhibited increased lameness scores over time (P ≤ 0.01), and horses receiving LPS, regardless of dose, had greater recorded lameness scores at 12 and 24 h postinjection (P ≤ 0.05). Joint circumference increased (P ≤ 0.01) across treatments in response to repeated arthrocentesis. Mean synovial fluid PGE2 concentrations increased linearly with increasing levels of LPS administration (P ≤ 0.01). Additionally, regardless of treatment, PGE2 increased over time and peaked at 12 h postinjection (P ≤ 0.01) and remained elevated above baseline at 336 h postinduction. Synovial concentrations of anabolic CPII increased linearly (P ≤ 0.01) with increasing dosage of LPS and increased (P ≤ 0.01) over 24 h in all horses, beginning at 6 h and peaking at 24 h postinjection. Concentrations of C2C in synovial fluid were not influenced by treatment and decreased from 0 to 6 h and steadily increased to 24 h in all horses (P ≤ 0.01). These results indicate that intra-articular LPS induced intra-articular inflammation and collagen synthesis in young horses and that the response is dose dependent. The use of this model to induce predictable joint inflammation may provide insight to the efficacy of preventative strategies relating to joint disease in the young horse.

Antimicrobial activity of gallium maltolate against Staphylococcus aureus and methicillin-resistant S. aureus and Staphylococcus pseudintermedius: An in vitro study

Gallium is a trivalent semi-metallic element that has shown antimicrobial activity against several important human pathogens. This antimicrobial activity is likely related to its substitution in important iron-dependent pathways of bacteria. The genus Staphylococcus, which includes human and animal pathogens that cause significant morbidity and mortality, requires iron for growth and colonization. In this study, gallium maltolate, at various concentrations between 50 and 200μM, inhibited the in vitro growth of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) at time-points between 8 and 36h after inoculation. The inhibitory activity of gallium maltolate against clinical isolates of MRSA and methicillin-resistant Staphylococcus pseudintermedius (MRSP) from a veterinary teaching hospital was determined.

Pharmacokinetics of gallium maltolate after intragastric administration in adult horses.

OBJECTIVE To determine the pharmacokinetics of gallium maltolate (GaM) after intragastric administration in adult horses. ANIMALS 6 adult horses. PROCEDURES Feed was withheld for 12 hours prior to intragastric administration of GaM (20 mg/kg). A single dose of GaM was administered to each horse via a nasogastric tube (time 0). Blood samples were collected at various time points from 0 to 120 hours. Serum was used to determine gallium concentrations by use of inductively coupled plasma-mass spectroscopy. Noncompartmental and compartmental analyses of serum gallium concentrations were performed. Pharmacokinetic models were selected on the basis of the Akaike information criterion and visual analysis of plots of residuals. RESULTS Serum concentration data for 1 horse were such that this horse was considered an outlier and excluded from noncompartmental and compartmental analyses. Noncompartmental analysis was used to determine individual pharmacokinetic parameters. A 1-compartment model with first-order input and output and lag time was selected as the best-fit model for the data and used to determine mean ± SD values for maximum observed serum concentration (0.28 ± 0.09 μg/mL), time of maximum concentration (3.09 ± 0.43 hours), time to the first measurable concentration (0.26 ± 0.11 hours), apparent elimination half-life (48.82 ± 5.63 hours), area under the time-concentration curve (20.68 ± 757 h•μg/mL), and apparent volume of distribution (73,493 ± 18,899 mL/kg). CONCLUSION AND CLINICAL RELEVANCE Further studies are necessary to determine the bioavailability of GaM after intragastric administration in adult horses.

Thermoregulation of the testicle in response to exercise and subsequent effects on semen characteristics of stallions

Stallions (n = 8) were implanted with a thermal sensory device in the muscle of the neck and the subcutaneous tissue of the scrotum and then assigned to either a nonexercise (Non-EX; n = 4) or exercise (EX; n = 4) group. A motorized equine exerciser was used to work EX stallions 30 min/d for 4 d/wk during a 12-wk period from July through October 2010. Temperatures (subcutaneous scrotal, intramuscular neck, and rectal) were recorded at 0, 22, and 30 min after the start of exercise, as well as 60 and 120 min post-exercise. Hourly ambient temperature and relative humidity data were also obtained. Semen was collected at 0, 4, 8, and 12 wk and analyzed for volume, sperm concentration, total sperm numbers, percentages of total and progressively motile sperm, sperm morphologic characteristics, and sperm DNA quality. No effect (P > 0.05) of exercise was observed on any of the measured semen variables. Implantation of thermal sensory devices had no demonstrable acute or chronic effects on the scrotal or neck tissue, indicating that the thermal sensory devices are a safe and effective way to measure subcutaneous scrotal and neck temperatures. At 22 and 30 min of exercise, rectal and neck temperatures increased (P < 0.0001) approximately 1.9 and 2.4°C, respectively, and scrotal temperatures simultaneously increased, although not significantly (P = 0.33), approximately 0.8°C. Correlations existed between scrotal, neck, rectal, and ambient temperatures, with the correlation between scrotal and rectal temperatures being greatest (r(s) = 0.76; P < 0.0001). Although moderate exercise for a short duration in extreme heat and humidity did significantly increase core body temperatures in stallions, scrotal temperatures did not significantly increase, and sperm parameters were unaffected.

Use of a modified Vinsot technique for partial phallectomy in 11 standing horses.

CASE DESCRIPTION 6 geldings and 5 stallions were evaluated from January 2007 through April 2009 for the following conditions requiring phallectomy: chronic paraphimosis (n = 7), squamous cell carcinoma of the penis (3), and priapism (1). CLINICAL FINDINGS None of the 7 horses with paraphimosis was able to retract the penis. Chronicity of the paraphimosis in 6 horses ranged from 2 weeks to 2 months and was unknown in the seventh horse. Horses with paraphimosis had been medically treated without success. The horse with priapism had developed the condition secondary to acepromazine administration 2 days prior to referral and was unsuccessfully treated once by intracavernosal administration of phenylephrine and irrigation of the cavernosal tissues prior to surgery. The 3 horses with squamous cell carcinoma of the penis had had the condition for 2 years and had been treated by repeated application of a cryogen or chemotherapeutic agent to the lesions. TREATMENT AND OUTCOME All 11 horses underwent a partial phallectomy by means of a modified Vinsot technique. Modifications to the original technique included creation of a linear urethrostomy, alteration of the location and shape of the urethrostomy, application of a latex tourniquet, concurrent castration of stallions, and use of the procedure in standing horses. The procedure was technically easy to perform, well tolerated by the horses, and cosmetically acceptable to the owners, and had minimal postoperative complications. Long-term follow-up information was obtained from owners of 10 horses a median of 454 days after surgery; 2 owners reported mild urine scalding as the only adverse effect. CONCLUSIONS AND CLINICAL RELEVANCE The modified Vinsot technique of partial phallectomy was effective and may be useful for horses that are unsuitable candidates for general anesthesia because of medical or owner financial constraints.