Adult Female Horses Research Articles

Oocyte development in captive Atlantic horse mackerel Trachurus trachurus

Abstract Ndjaula, H. O. N., Hansen, T., Krüger-Johnsen, M., and Kjesbu, O. S. 2009. Oocyte development in captive Atlantic horse mackerel Trachurus trachurus. – ICES Journal of Marine Science, 66: 623–630. This 13-month experimental study focused on developing oocyte production in 101 adult female Atlantic horse mackerel. In addition, proxies of energy patterns in relation to oocyte development were investigated. The fish were captured off western Norway and fed to satiation from October 2005 to November 2006 in two replicate adjacent circular tanks. Monthly histological examination of the ovaries indicated that vitellogenic oocytes were present at all times throughout the study period, but that oocyte development did not progress past the incipient migratory nucleus stage. The oocyte diameter (OD) threshold between pre-vitellogenic and vitellogenic oocytes and the mean OD of the leading cohort were investigated, and no hiatus was observed within the OD size distribution around the 185-µm pre-vitellogenic and vitellogenic threshold. Variation in gonadosomatic index, hepatosomatic index, condition factor, and fat content (as measured by the use of a Distell Fish Fatmeter) increased with oocyte development. The observed development of oocyte recruitment and the absence of a hiatus in the oocyte distribution are characteristics of an asynchronous spawner. The species seems to have a prolonged spawning season, judging from the advanced and variety of maturity stages throughout the study period.

Comparison of pharmacokinetics of glucosamine and synovial fluid levels following administration of glucosamine sulphate or glucosamine hydrochloride

To compare the pharmacokinetics of glucosamine and the synovial fluid levels attained following treatment with glucosamine sulphate or glucosamine hydrochloride in a large animal model at clinically relevant doses. Eight adult female horses were used. Crystalline glucosamine sulphate (Dona) or glucosamine hydrochloride was administered at a dose of 20 mg/kg by either intravenous (i.v.) injection or nasogastric (n.g.) intubation. Plasma samples were collected before dosing and at 5, 15, 30, 60, 120, 360, 480 and 720 min after dosing. Synovial fluid samples were collected from the radiocarpal joints within 48 h before dosing and at 1, 6 and 12 h post-dosing. Glucosamine was assayed by Liquid Chromatography Electrospray Tandem Mass Spectrometry (LC-ESI/MS/MS). Plasma concentrations reached approximately 50 microg/mL after i.v. injection and approximately 1 microg/mL after n.g. administration of both types of glucosamine. The median oral bioavailability was 9.4% for glucosamine sulphate and 6.1% for glucosamine hydrochloride. Synovial fluid concentrations were significantly higher at 1 and 6 h following oral treatment with glucosamine sulphate compared to glucosamine hydrochloride. Twelve hours following oral administration, glucosamine levels in the plasma and the synovial fluid were still significantly higher than baseline for the glucosamine sulphate preparation, but not for the hydrochloride preparation. Following oral administration of a clinically recommended dose of glucosamine sulphate (Dona), significantly higher synovial fluid concentrations of glucosamine are attained, when compared to an equivalent dose of glucosamine hydrochloride. Whether this difference is translated into a therapeutic effect on the joint tissues remains to be elucidated.

65 CLONED EMBRYOS FROM HORSE SOMATIC CELLS AND ENUCLEATED BOVINE AND OVINE OOCYTES AND THEIR DEVELOPMENTAL COMPETENCE

The objective of this study was to investigate the reprogramming potential of equine somatic cell donor nuclei in either bovine or ovine recipient oocyte cytoplasmic environments. Heterogeneous embryos were reconstructed by somatic cell nuclear transfer (NT). The percentage of fusion and developmental competence, assessed by rates of cleavage and morula and blastocyst formation, were determined. Skin fibroblast cells, obtained from the ear of an adult female Mongolian horse, were dissociated using 0.25% trypsin and cultured in vitro in a humidified atmosphere of 5% CO2 in air at 37°C. Donor somatic cells were serum-starved before NT and used between passages 4 and 6. Bovine and ovine oocytes derived from slaughterhouse ovaries were matured in vitro for 17–19 and 22–24 h, respectively, in a humidified atmosphere of 5% CO2 in air at 38.5°C, before they were enucleated and used as recipient cytoplasts. The fibroblasts were injected under the zona pellucida of the cytoplasts and electrically fused by 2 DC electrical pulses of 1.58 kV cm–1 for 10 μs, with an interval of 0.13 s. The reconstructed embryos were then activated with 5 μm ionomycin in H-M199 for 5 min and then in 2 mm 6-DMAP for 4 h. The equine-bovine and equine-ovine reconstructed embryos were co-cultured, respectively, with bovine and ovine cumulus cells in synthetic oviduct fluid supplemented with amino acids (SOFaa) and 10% fetal calf serum (FCS) for 168 h. The data were analyzed with ANOVA and differences among the groups were evaluated with t-test. The results of the percentages of fusion, cleavage, and development to morula (8 to 64 cells) and blastocyst stages of equine-bovine and equine-ovine heterogeneous embryos are shown in Table 1. This study demonstrates that heterogeneous embryos can undergo early embryonic divisions and that reprogramming of equine fibroblast nuclei can be initiated in foreign cytoplasts. It appears that embryos reconstructed with equine somatic nuclei and ovine cytoplasts have a higher developmental potential than those using bovine cytoplasts. Table 1. Developmental competence of equine-bovine and equine-ovine reconstructed embryos

Fatal Myocardial Degeneration in an Adult Quarter Horse with Vitamin E Deficiency

Abstract A carcass of an adult female Quarter Horse that spontaneously died was received for general investigation at the veterinary diagnostic laboratory of North Dakota State University. Routine necropsy examination demonstrated multiple pale and firm foci in the heart that were histologically consistent with severe myocardial degeneration/necrosis and fibrosis. Analysis of a liver sample confirmed a suboptimal vitamin E level and a selenium value that was within reference range. Because there was no anamnestic indication for ingestion of cardiotoxic plants or evidence of other etiologic factors, vitamin E deficiency was concluded to have led to the development of the myocardial lesions, which precipitated acute heart failure resulting in sudden death.

Effects of continuous oral administration of phenylbutazone on biomarkers of cartilage and bone metabolism in horses

To evaluate the effects of continuous oral administration of phenylbutazone on serum and synovial fluid biomarkers of skeletal matrix metabolism in horses. 11 adult female horses without clinical or radiographic evidence of joint disease. Horses were randomly assigned to control or treatment groups. Phenylbutazone was administered orally twice daily at a dose of 4.4 mg/kg for 3 days to the treatment group and subsequently at a dose of 2.2 mg/kg for 7 days. Serum and radiocarpal synovial fluid samples were obtained at baseline and thereafter at regular intervals for 4 weeks. Biomarkers of cartilage aggrecan synthesis (chondroitin sulfate 846) and type II collagen synthesis (procollagen type II C-propeptide) and degradation (collagen type II cleavage) were assayed. Biomarkers of bone synthesis (osteocalcin) and resorption (C-terminal telopeptide of type I collagen) were also measured. No significant differences were found between control and treatment groups or temporally for the biomarkers chondroitin sulfate 846, procollagen type II C-propeptide, collagen type II cleavage, and C-terminal telopeptide of type I collagen in serum or synovial fluid. A significant increase in osteocalcin concentration occurred in synovial fluid during treatment in the treated group. No treatment effect was detected for serum osteocalcin concentration. Results suggested that continuous phenylbutazone administration at recommended doses altered some biomarkers in healthy equine joints after short periods of administration. Increased osteocalcin concentration may indicate an undetermined anabolic effect of phenylbutazone administration on periarticular bone or transient induction of osteogenesis in articular chondrocytes or a mesenchymal subpopulation of synoviocytes.

Synovial fluid levels and serum pharmacokinetics in a large animal model following treatment with oral glucosamine at clinically relevant doses

To examine the concentration of glucosamine in the synovial fluid and its pharmacokinetics in serum in a large animal model following dosing with glucosamine HCl at clinically relevant levels. Eight adult female horses were studied. After an overnight fast, glucosamine HCl (20 mg/kg of body weight) was administered by either nasogastric (NG) intubation or intravenous (IV) injection. Blood samples were collected before dosing and at 5, 15, 30, 60, 120, 180, 240, 360, 480, and 720 minutes after dosing. Synovial fluid samples were collected from the radiocarpal joints 48 hours before dosing and at 1 and 12 hours after dosing. Glucosamine was assayed by fluorophore-assisted carbohydrate electrophoresis. The maximum concentration of glucosamine in serum reached approximately 300 muM ( approximately 50 microg/ml) following IV dosing and approximately 6 microM (approximately 1 microg/ml) following NG dosing. Synovial fluid concentrations reached 9-15 microM with IV dosing and 0.3-0.7 microM with NG dosing, and remained elevated (range 0.1-0.7 microM) in most animals even at 12 hours after dosing. Following NG dosing, the median serum maximal concentration of 6.1 microM (range 4.38-7.58) was attained between 30 minutes and 4 hours postdose. The mean apparent volume of distribution was 15.4 liters/kg, the mean bioavailability was 5.9%, and the mean elimination half-life was 2.82 hours. Clinically relevant dosing of glucosamine HCl in this large monogastric animal model results in serum and synovial fluid concentrations that are at least 500-fold lower than those reported to modify chondrocyte anabolic and catabolic activities in tissue and cell culture experiments. We conclude that the apparent therapeutic benefit of dietary glucosamine on pain and joint space width in humans and animals may be secondary to its effects on nonarticular tissues, such as the intestinal lining, liver, or kidney, since these may be exposed to much high levels of glucosamine following ingestion.

Muscle fiber types in Crioulo horses

SUMMARY Sixty-three Crioulo horses were used for this work. The horses belong to 7 different establishments in South America (six located in the South of Brazil and one in Uruguay) and were divided into 4 categories or groups consisting of 19, 17, 14, and 13 animals, respectively. Group A consisted of adult female horses of lineage selected by endurance tests, and that had rested for at least a year. These animals were winners or descendents of the winners of endurance tests. Their pedigrees were analyzed to check the absence of the blood of Chilean horses in their lineage. Group B consisted of adult female horses whose lineage was selected by “Freio de Ouro” tests, and that had been kept out of training for at least a year. They were winners, or were classified or descendents of animals that had won the “Freio de Ouro” test. Their pedigrees were analyzed and the blood of horses of Chilean origin was found to be present, or of pure Chileans found in their pedigree. Group C consisted of female horses from 2 to 2.5 years old, whose lineage was selected by “Freio de Ouro” testing. These were raised in paddocks and had received no training. They were the descendents of “Freio de Ouro” test champions, the pedigrees were analyzed, and the presence of Chilean blood was found in their crossbreeding. Group D consisted of adult animals of both sexes and gelded male horses that had been kept in training through daily work at a cattle ranch until one week before the collection of the material. These animals had no Chilean blood in their cross-breeding.

Cardiopulmonary and sedative effects of intravenous administration of low doses of medetomidine and xylazine to adult horses

Abstract Objective To determine the cardiopulmonary and sedative effects of medetomidine hydrochloride in adult horses and to compare those effects with effects of an equipotent dose of xylazine hydrochloride. Animals 10 healthy adult female horses. Procedure 5 horses were given medetomidine (4 μg/kg of body weight, IV), and the other 5 were given xylazine (0.4 mg/kg, IV). Heart rate, respiratory rate, arterial blood pressures, pulmonary arterial blood pressures, and cardiac output were recorded, and sedation and ataxia scores were assigned before and every 5 minutes after drug administration for 60 minutes. Rectal temperature and blood gas partial pressures were measured every 15 minutes after drug administration. Results Arterial blood pressure was significantly decreased throughout the study among horses given medetomidine and was significantly decreased for 40 minutes among horses given xylazine. Compared with baseline values, cardiac output was significantly decreased 10, 20, and 40 minutes after administration of medetomidine and significantly increased 40 and 60 minutes after administration of xylazine. Despite the significant decrease in respiratory rate in both groups, results of blood gas analyses were not significantly changed over time. Ataxia and sedation scores were of similar magnitude for the 2 groups, but ataxia persisted slightly longer among horses given medetomidine. Horses resumed eating hay 10 to 55 minutes after drug administration. Conclusions and Clinical Relevance Results suggest that equipotent low doses of medetomidine and xylazine induce comparable levels of ataxia and sedation and similar cardiopulmonary changes in adult horses. (Am J Vet Res 1999;60:1371–1376)

Hemodynamic and metabolic alterations associated with intravenous infusion of a combination of adenosine triphosphate and magnesium chloride in conscious horses

Abstract Objective To determine hemodynamic and metabolic effects of IV infusion of ATP-MgCl2 combination and maximal safe IV infusion rate in conscious horses. Animals 6 adult female horses. Procedure All horses received an IV infusion of ATP-MgCl2 combination, beginning at a rate of 0.05 mg of ATP/kg of body weight/min, which was increased by 0.05 mg/kg/min increments at 10-minute intervals until a rate of 1.0 mg/kg/min was achieved. Data were collected prior to the start of the infusion, at the end of each infusion rate, and at 15-minute intervals for the next hour after discontinuation of the infusion. Measured or calculated hemodynamic variables included cardiac output, cardiac index, heart rate, stroke volume, systemic and pulmonary arterial pressures, and systemic and pulmonary vascular resistances. Arterial blood gas tensions, CBC, plasma biochemical profiles, urine volume and specific gravity, and selected clinical signs of disease also were evaluated. Results Intravenous infusion of ATP-MgCl2 significantly increased cardiac output, decreased systemic vascular resistance, and caused mild pulmonary hypertension. Magnitude of the hemodynamic alterations was dependent on rate of infusion. Maximal safe infusion rate for these horses was 0.3 mg/kg/min. All horses became lethargic, and their appetites diminished during the infusion; 5 horses had mild signs of abdominal discomfort. Flank sweating was observed in all horses as infusion rate increased. Urine volume and specific gravity and hematologic, biochemical, and arterial blood gas alterations were detected during and after infusion. Conclusions and Clinical Relevance Intravenous administration of ATP-MgCl2 in healthy, conscious, adult horses caused various metabolic and hemodynamic alterations that were without appreciable detrimental effects. (Am J Vet Res 1999;60:1140–1147)

Plasma and urine nitric oxide concentrations in horses given a low dose of endotoxin

Abstract Objective To quantify plasma and urine nitric oxide (NO) concentrations before and after low-dose endotoxin infusion in horses. Animals 11 healthy adult female horses. Procedure Eight horses were given endotoxin (35 ng/kg of body weight, IV) over 30 minutes. Three sentinel horses received an equivalent volume of saline (0.9% NaCl) solution over the same time. Clinical signs of disease and hemodynamic variables were recorded, and urine and plasma samples were obtained to measure NO concentrations prior to endotoxin infusion (t = 0) and every hour until postinfusion hour (PIH) 6, then every 2 hours until PIH 24. Blood for hematologic and metabolic analyses and for serum cytokine bioassays were collected at 0 hour, every hour until PIH 6, every 2 hours through PIH 12, and finally, every 6 hours until PIH 24. Results Differences in plasma NO concentrations across time were not apparent, but urine NO concentrations significantly decreased at 4 and 20 to 24 hours in endotoxin-treated horses. Also in endotoxin-treated horses, alterations in clinical signs of disease, and hemodymanic, metabolic, and hematologic variables were significant and characteristic of endotoxemia. Serum interleukin-6 (IL-6) activity and tumor necrosis factor (TNF) concentrations were increased above baseline values from 1 to 8 hours and 1 to 2 hours, respectively. Conclusions and Clinical Relevance Plasma and urine NO concentrations did not increase in horses after administration of a low dose of endotoxin, despite induction of an inflammatory response, which was confirmed by increased TNF and IL-6 values characteristic alterations in clinical signs of disease, and hematologic, hemodynamic and metabolic variables. (Am J Vet Res 1999:60:969-976)

Ticarcillin administration to the equine: Intrauterine and intramuscular

Serum levels of ticarcillin disodium, a semi-synthetic penicillin (Beecham Laboratories, Bristol, Tennessee, 37620), were measured at various time intervals up to and including 24 h after intrauterine and intramuscular administration in adult female horses. Three separate studies were conducted in Part I: in the first and second studies, serum levels were measured after intrauterine administration of 1 and 3 g of ticarcillin, respectively, and in the third study, levels were measured after intramuscular administration of 6 g of ticarcillin. In Part II, serum levels of ticarcillin were measured after intramuscular administration of ticarcillin at dosages based on body weight: 10 mg/lb (22 mg/kg) of body weight, 15 mg/lb (33 mg/kg) of body weight and 20 mg/lb (44 mg/kg) of body weight. Results showed little if any absorption of the antibiotics into the systemic circulation after intrauterine administration. All serum levels measured after administration of both intrauterine dosages of 1 and 3 g were less than 4.0 ug/ml. Peak serum concentrations, after intramuscular administration of 6 g of ticarcillin, occurred between 20 and 45 min after administration and averaged 15.97 ± 5.0 ug/ml. Average peak serum concentrations, after intramuscular administration of 10 mg/lb (22 mg/kg) of body weight, 15 mg/lb (33 mg/kg) of body weight, 15 mg/lb (33 mg/kg) and 20 mg/lb (44 mg/kg) of body weight, were 16.12 ug/ml, 25.75 ug/ml and 50.25 ug/ml, respectively. After intramuscular administration of 6 g of ticaracilin, average serum half-life was determined to be 2.9 h.

Some Influences of Meteorological Factors on Flight Activity of Female Horse Flies (Diptera: Tabanidae)

Adult female horse flies ( Tabanus spp. L.) were collected hourly in modified Malaise traps. Regression analyses of numbers of flies collected and the values of various meteorological factors in the ranges observed indicated which factors influenced activity. From greater to lesser influence, these factors were: barometric pressure, temperature, evaporation and evaporation change per hour, total sky radiation change per hour, total sky radiation and wind velocity change per hour, and relative humidity and temperature change per hour. The influence of these factors upon activity of the 6 most abundant species was also determined. Prediction equations are presented and the traps described.

Herpestype Virus and Type C Particles in Spontaneous Equine Lymphoma

Herpestype and type C virus particles are known to produce tumors in animals of certain species, and to be present in a variety of human tissues. Similar particles were found in a horse lymphoma.Electron microscope study was carried out on a tumorous prescapular lymph node from an adult female horse with spontaneous lymphoma. Lymph node cells grown in vitro for a total of 105 days (6th passage) were also studied. The lymph node was infiltrated by blastic cells of reticulum cell type and of lymphoblast type. No virus particles were found in the original lymph node. The cultured cells have a large cytoplasm with well developed rough endoplasmic reticulum and Golgi's apparatus, and an indented nucleus with marginated chromatin. Occasionally, intranuclear fibrous structures were observed. Elongated nuclear protrusion were frequently seen.