Differential gene expression analysis reveals pathways important in early post-traumatic osteoarthritis in an equine model.

BACKGROUND: Post-traumatic osteoarthritis (PTOA) is a common and significant problem in equine athletes. It is a disease of the entire joint, with the synovium thought to be a key player in disease onset and progression due to its role in inflammation. The development of effective tools for early diagnosis and treatment of PTOA remains an elusive goal. Altered gene expression represents the earliest discernable disease-related change, and can provide valuable information about disease pathogenesis and identify potential therapeutic targets. However, there is limited work examining global gene expression changes in early disease. In this study, we quantified gene expression changes in the synovium of osteoarthritis-affected joints using an equine metacarpophalangeal joint (MCPJ) chip model of early PTOA. Synovial samples were collected arthroscopically from the MCPJ of 11 adult horses before (preOA) and after (OA) surgical induction of osteoarthritis and from sham-operated joints. After sequencing synovial RNA, Salmon was used to quasi-map reads and quantify transcript abundances. Differential expression analysis with the limma-trend method used a fold-change cutoff of log2(1.1). Functional annotation was performed with PANTHER at FDR < 0.05. Pathway and network analyses were performed in Reactome and STRING, respectively. RESULTS: RNA was sequenced from 28 samples (6 preOA, 11 OA, 11 sham). "Sham" and "preOA" were not different and were grouped. Three hundred ninety-seven genes were upregulated and 365 downregulated in OA synovium compared to unaffected. Gene ontology (GO) terms related to extracellular matrix (ECM) organization, angiogenesis, and cell signaling were overrepresented. There were 17 enriched pathways, involved in ECM turnover, protein metabolism, and growth factor signaling. Network analysis revealed clusters of differentially expressed genes involved in ECM organization, endothelial regulation, and cellular metabolism. CONCLUSIONS: Enriched pathways and overrepresented GO terms reflected a state of high metabolic activity and tissue turnover in OA-affected tissue, suggesting that the synovium may retain the capacity to support healing and homeostasis in early disease. Limitations of this study include small sample size and capture of one point post-injury. Differentially expressed genes within key pathways may represent potential diagnostic markers or therapeutic targets for PTOA. Mechanistic validation of these findings is an important next step.

MAGNETIC RESONANCE IMAGING SCORING OF AN EXPERIMENTAL MODEL OF POST-TRAUMATIC OSTEOARTHRITIS IN THE EQUINE CARPUS.

Magnetic resonance imaging (MRI) is the most sensitive imaging modality to detect the early changes of osteoarthritis. Currently, there is no quantifiable method to tract these pathological changes over time in the horse. The objective of this experimental study was to characterize the progression of MRI changes in an equine model of post-traumatic osteoarthritis using a semiquantitative scoring system for whole-organ evaluation of the middle carpal joint. On day 0, an osteochondral fragment was created in one middle carpal joint (OCI) and the contralateral joint (CON) was sham-operated in 10 horses. On day 14, study horses resumed exercise on a high-speed treadmill until the completion of the study (day 98). High-field MRI examinations were performed on days 0 (preosteochondral fragmentation), 14, and 98 and scored by three blinded observers using consensus agreement. Images were scored based on 15 independent articular features, and scores were compared between and within-groups. On days 14 and 98, OCI joints had significantly (P ≤ 0.05) higher whole-organ median scores (29.0 and 31.5, respectively), compared to CON joints (21.5 and 20.0, respectively). On day 14, OCI joints showed significant increases in high-signal bone lesion scores, and osteochondral fragment number and size. On day 98, high-signal bone lesion, low-signal bone lesion, osteophyte formation, cartilage signal abnormality, subchondral bone irregularity, joint effusion, and synovial thickening scores were significantly increased in OCI joints. Study results suggest that the MRI whole-organ scoring system reported here may be used to identify onset and progression of pathological changes following osteochondral injury.

Concentrations of stromal cell-derived factor-1 in serum, plasma, and synovial fluid of horses with osteochondral injury.

OBJECTIVE: To determine whether stromal cell-derived factor-1 (SDF-1) concentrations in serum, plasma, and synovial fluid differed among untrained, race-trained, and osteochondral-injured Thoroughbred racehorses. ANIMALS: 22 racehorses without osteochondral injury and 37 racehorses with osteochondral injury. PROCEDURES: Horses without osteochondral injury were examined before and after 5 to 6 months of race training. Horses with osteochondral injury were undergoing arthroscopic surgery for removal of osteochondral fragments from carpal or metacarpophalangeal or metatarsophalangeal joints (fetlock joints). Serum, plasma, and fetlock or carpal synovial fluid samples were obtained and analyzed for SDF-1 concentration by use of an ELISA. RESULTS: In horses with fetlock or carpal joint injury, mean synovial fluid SDF-1 concentrations were significantly higher, serum SDF-1 concentrations were significantly lower, and synovial fluid-to-serum SDF-1 ratios were significantly higher than in untrained and trained horses. Synovial fluid SDF-1 concentrations were not significantly different between trained and untrained horses. Plasma SDF-1 concentrations were not different among the 3 groups. Results obtained with serum, compared with synovial fluid and plasma, had better sensitivity for differentiating between osteochondral-injured horses and uninjured horses. In horses with fetlock joint osteochondral injury, serum SDF-1 concentrations were correlated with radiographic and arthroscopic inflammation scores, but not arthroscopic cartilage scores. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that serum SDF-1 concentrations were more sensitive than plasma and synovial fluid concentrations for detection of osteochondral injury in the fetlock or carpal joint of racehorses. Analysis of serum and synovial SDF-1 concentrations in horses with experimentally induced joint injury may help define the onset and progression of post-traumatic osteoarthritis and aid in the evaluation of anti-inflammatory treatments.

Effects of yearling sale purchase price, exercise history, lameness, and athletic performance on purchase price of Thoroughbreds at 2-year-old in-training sales.

OBJECTIVE: To determine the effects of yearling sale purchase price, exercise history, lameness, and athletic performance (speed) on purchase price of 2-year-old in-training Thoroughbreds and to compare the distance exercised within 60 days prior to 2-year-old in-training sales between horses with high yearling sale purchase prices versus those with low yearling sale purchase prices and between horses with lameness during training and those without lameness during training. DESIGN: Prospective study. ANIMALS: 51 Thoroughbreds. PROCEDURES: Thoroughbreds purchased at a yearling sale were trained prior to resale at 2-year-old in-training sales. Amount of exercise and lameness status during training and speed of horses at 2-year-old in-training sales were determined. Data were analyzed via the Wilcoxon rank sum test and ANOVA. RESULTS: Median purchase price of horses at 2-year-old in-training sales was $37,000. The 2-year-old in-training sale purchase price was associated with yearling sale purchase price and distance galloped within 60 days prior to and speed recorded at 2-year-old in-training sales. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with high yearling sale purchase prices typically had high 2-year-old in-training sale purchase prices, had low distances galloped within 60 days prior to 2-year-old in-training sales, and were classified as fast at 2-year-old in-training sales. Lameness alone was not associated with 2-year-old in-training sales purchase price. However, lameness was associated with a low distance galloped before 2-year-old in-training sales, particularly for horses with a high yearling sale purchase price; this finding suggested that yearling sale purchase price can affect training management decisions for horses with lameness.

Effects of various presale radiographic findings for yearling Thoroughbreds on 2-year-old racing performance.

OBJECTIVE: To determine the effects of various presale radiographic findings for Thoroughbreds sold at a yearling sale on 2-year-old racing performance of those horses. ANIMALS: 397 Thoroughbreds. DESIGN: Cohort study. PROCEDURES: Thoroughbreds offered for sale at a Thoroughbred sales facility in Kentucky were selected via a randomization procedure. Effects of various presale radiographic findings on the following measures of 2-year-old racing performance were determined: having started a race and having placed (ie, finished in first, second, or third place) in a race at least once, total amount of money earned, and amount of money earned per start. RESULTS: Of the 397 horses, 192 (48%) started in at least 1 race during the 2-year-old racing year. The odds of failure to start a race as a 2-year-old were 1.78 times as great for horses with forelimb proximal sesamoid bone osteophytes or enthesophytes as for horses without this finding (95% confidence interval, 1.01 to 3.16). The odds of failure to start a race as a 2-year-old were 2.02 times as great for horses with hind limb proximal phalanx osteochondral fragments as for horses without this finding (95% confidence interval, 0.95 to 4.31), although this result was not significant. Radiographic findings did not have an effect on total amount of money earned, amount of money earned per start, or whether horses placed in a race. CONCLUSIONS AND CLINICAL RELEVANCE: Presale radiographic detection of forelimb proximal sesamoid bone osteophytes or enthesophytes or hind limb proximal phalanx osteochondral fragments in yearlings were associated with failure to start a race during the 2-year-old racing year in study horses.

Surgical resolution of an avulsion fracture of the peroneus tertius origin in a giraffe (Giraffa camelopardalis reticulata).

A 4-mo-old, 185-kg male giraffe (Giraffa camelopardalis reticulata) was presented due to stifle effusion and lameness of 3-wk duration. Radiographs revealed a fracture of the extensor fossa of the lateral trochlear ridge of the femur at the origin of the peroneus tertius. Under anesthesia, dysfunction of the reciprocal apparatus was documented by flexing the stifle while the tarsus remained extended. An avulsion fracture of the origin of the peroneus tertius and extensor digitorum longus muscle was diagnosed. An exploratory arthroscopy of the femorotibial joint was followed by arthrotomy to excise the large bone fragment from its soft tissue attachments. Because of the fractious temperament of the animal, postoperative care was restricted to stall rest for 3 mo, and no postoperative complications arose. Only a mild residual lameness remained by 6 mo after surgery.

Associations of horse age, joint type, and osteochondral injury with serum and synovial fluid concentrations of type II collagen biomarkers in Thoroughbreds.

OBJECTIVE: To determine the effects of horse age, osteochondral injury, and joint type on a synthesis biomarker and 3 degradative biomarkers of type II collagen in Thoroughbreds. ANIMALS: Healthy rested adult (3- to 12-year-old) Thoroughbreds (n = 19), yearling (1- to 2-year-old) Thoroughbreds (40), and Thoroughbred racehorses (2 to 7 years old) undergoing arthroscopic surgery for removal of osteochondral fragments that resulted from training or racing (41). PROCEDURES: Samples of blood and metacarpophalangeal, metatarsophalangeal, or carpal joint synovial fluid (SF) were collected from all horses. Commercially available assays were used to analyze SF and serum concentrations of type II collagen biomarkers of synthesis (carboxy propeptide of type II collagen [CPII]) and degradation (cross-linked C-telopeptide fragments of type II collagen [CTX II], neoepitope generated by collagenase cleavage of type I and II collagen [C1,2C], and neoepitope generated by collagenase cleavage of type II collagen [C2C]). RESULTS: Osteochondral injury affected concentrations of CPII, CTX II, C1,2C, and C2C in SF, serum, or both, compared with concentrations in healthy adult horses. Compared with adult horses, yearling horses had increased SF or serum concentrations of degradative biomarkers (CTX II, C1,2C, and C2C). Concentrations were higher in carpal than metacarpophalangeal or metatarsophalangeal joints for all biomarkers in osteochondral-injured horses. Variable differences in SF concentrations between joint types were detected in healthy adult and yearling horses. CONCLUSIONS AND CLINICAL RELEVANCE: Horse age, osteochondral injury, and joint type all significantly affected type II collagen biomarker concentrations in SF and serum of Thoroughbreds.

Prevalence of various presale radiographic findings and association of findings with sales price in Thoroughbred yearlings sold in Kentucky.

OBJECTIVE: To estimate prevalences of various presale radiographic findings and of presale arthroscopy in horses offered for sale at the 2006 Keeneland September yearling sale and to compare sales prices between yearlings with and without various presale radiographic findings or a history of arthroscopy. ANIMALS: 397 Thoroughbred yearlings. DESIGN: Cross-sectional study. PROCEDURES: Presale radiographs and health records were examined to estimate prevalences of various radiographic findings and presale arthroscopy. Sales price records were used to compare sales prices between yearlings with and without various presale radiographic findings or a history of arthroscopy. RESULTS: In the forelimbs, the most common radiographic findings were vascular channels in the proximal sesamoid bones (23%), enthesophytes or osteophytes in the radiocarpal joint (22%), and osteochondritis lesions involving the sagittal ridge of the third metacarpal bone (20%). In the hind limbs, the most common radiographic findings were enthesophytes or osteophytes involving the proximal sesamoid bones (39%), abnormalities of the distodorsal aspect of the third metatarsal bone (36%), enthesophytes or osteophytes involving the distal intertarsal joint (27%), and osteochondritis lesions involving the stifle joint (8%). Thirteen percent of horses had a history of presale arthroscopy. Median sales price was significantly lower in horses with fragments of the proximal phalanx than in horses without. Median sales price was significantly higher in horses with a history of presale arthroscopy than in horses without. CONCLUSIONS AND CLINICAL RELEVANCE: Results revealed significant associations between a diagnosis of fragments of the proximal phalanx, presale arthroscopy, and sales price in Thoroughbred yearlings.

Effect of exercise and osteochondral injury on synovial fluid and serum concentrations of carboxy-terminal telopeptide fragments of type II collagen in racehorses.

OBJECTIVE: To investigate the effects of exercise and osteochondral injury on concentrations of carboxy-terminal telopeptide fragments of type II collagen (CTX-II) in synovial fluid (SF) and serum of Thoroughbred racehorses and to compare findings with radiographic and arthroscopic scores of joint injury severity. ANIMALS: 78 Thoroughbreds with (n = 38) and without (40) osteochondral injury. PROCEDURES: Serum and metacarpophalangeal or carpal joint SF samples were collected from noninjured horses before and at the end of 5 to 6 months of race training (pre- and postexercise samples, respectively) and from horses with osteochondral injury (1 joint assessed/horse). Synovial fluid and serum CTX-II concentrations were determined by use of an ELISA. Radiographic and arthroscopic scores of joint injury severity were determined for the injured horses. RESULTS: The CTX-II concentrations in SF and SF:serum CTX-II ratio were significantly higher for horses with joint injuries, compared with pre- and postexercise findings in noninjured horses. Serum CTX-II concentrations in postexercise and injured-horse samples were significantly lower than values in pre-exercise samples. On the basis of serum and SF CTX-II concentrations and SF:serum CTX-II ratio, 64% to 93% of serum and SF samples were correctly classified into their appropriate group (pre-exercise, postexercise, or injured-joint samples). In horses with joint injuries, arthroscopic scores were positively correlated with radiographic scores, but neither score correlated with SF or serum CTX-II concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that serum and SF CTX-II concentrations and SF:serum CTX-II ratio may be used to detect cartilage degradation in horses with joint injury.

High-mobility group box chromosomal protein 1 as a potential inflammatory biomarker of joint injury in Thoroughbreds.

OBJECTIVE: To investigate effects of osteochondral injury on high-mobility group box chromosomal protein 1 (HMGB-1) concentrations in synovial fluid (SF) from Thoroughbreds and to compare these results with radiographic and arthroscopic scores of severity of joint injury. ANIMALS: 40 clinically normal rested Thoroughbreds (group 1) and 45 Thoroughbreds with osteochondral injury as a result of racing. PROCEDURES: SF was obtained from the metacarpophalangeal (MCP) joints, metatarsophalangeal (MTP) joints, middle carpal joints, and radiocarpal joints. For group 2, radiographic and arthroscopic scores were determined. Concentrations of SF HMGB-1 were determined by use of an ELISA. RESULTS: SF HMGB-1 concentrations in osteochondral-injured MCP-MTP joints were significantly higher than in normal MCP-MTP joints. Similarly, SF HMGB-1 concentrations in osteochondral-injured carpal joints were significantly higher than in normal carpal joints. Radiographic and arthroscopic scores were not correlated with SF HMGB-1 concentrations. Synovial fluid HMGB-1 concentrations > or = 11 ng/mL for MCP-MTP joints and > or = 9 ng/mL for carpal joints discriminated osteochondral-injured joints from normal joints. Horses with HMGB-1 concentrations > or = 11 ng/mL for MCP-MTP joints were twice as likely to have an osteochondral injury, and horses with HMGB-1 concentrations > or = 9 ng/mL for carpal joints were 4 times as likely to have an osteochondral injury. CONCLUSIONS AND CLINICAL RELEVANCE: Osteochondral injury was associated with a significant increase in SF HMGB-1 concentrations in MCP-MTP and carpal joints, compared with results for clinically normal Thoroughbreds. Analysis of SF HMGB-1 concentrations may be useful for evaluation of joint injury in horses.

Comparison of two indirect techniques for local delivery of a high dose of an antimicrobial in the distal portion of forelimbs of horses.

OBJECTIVE: To compare isolated limb retrograde venous injection (ILRVI) and isolated limb infusion (ILI) for delivery of amikacin to the synovial fluid of the distal interphalangeal and metacarpophalangeal joints and to evaluate the efficacy of use of an Esmarch tourniquet in standing horses. ANIMALS: 6 healthy adult horses. PROCEDURES: Horses were randomly assigned in a crossover design. In ILRVI, the injection consisted of 1 g of amikacin diluted to a total volume of 60 mL administered during a 3-minute period. In ILI, the infusion consisted of 1 g of amikacin diluted to 40 mL administered during a 3-minute period followed by administration of boluses of diluent (82 mL total) to maintain vascular pressure. During ILI, the infusate and blood were circulated from the venous to the arterial circulation in 5-mL aliquots. Synovial fluid and serum samples were obtained to determine maximum amikacin concentrations and tourniquet leakage, respectively. RESULTS: Both techniques yielded synovial concentrations of amikacin > 10 times the minimum inhibitory concentration (MIC) for 90% of isolates (80 microg/mL) and > 10 times the MIC breakpoint (160 microg/mL) of amikacin-susceptible bacteria reported to cause septic arthritis in horses. These values were attained for both joints for both techniques. Esmarch tourniquets prevented detectable loss of amikacin to the systemic circulation for both techniques. CONCLUSIONS AND CLINICAL RELEVANCE: Both techniques reliably achieved synovial fluid concentrations of amikacin consistent with concentration-dependent killing for bacteria commonly encountered in horses with septic arthritis. Esmarch tourniquets were effective for both delivery techniques in standing horses.

Lameness, athletic performance, and financial returns in yearling Thoroughbreds bought for the purpose of resale for profit.

OBJECTIVE: To characterize lameness during training and compare exercise variables and financial returns among yearling Thoroughbreds that were bought for the purpose of resale for profit. ANIMALS: 40 yearling Thoroughbreds. DESIGN: Prospective study. PROCEDURES: Horses purchased at yearling sales (summer 2004) were trained prior to resale at 2-year-olds in training sales (spring 2005). Horses were monitored daily for diagnosis and treatment of lameness during training. Selected variables, including sex, age, purchase price, lameness, distance (No. of furlongs) galloped during training, and financial returns, were compared among horses that had performance speeds (assessed at 2-year-olds in training sales) classified as fast, average, or slow. RESULTS: 37 of 40 horses became lame during training, most commonly because of joint injury. Eighteen of the lame horses had hind limb injuries only; 5 horses had injuries in forelimbs and hind limbs. The frequency of new cases of lameness increased as the date of the 2-year-olds in training sales approached. At the sales, 4, 21, and 15 horses were classified as fast, average, or slow, respectively; median financial return was slightly (but significantly) different among horses classified as fast ($14,000), average ($0), or slow (-$8,000). CONCLUSIONS AND CLINICAL RELEVANCE: Incidence of lameness during training in yearling horses purchased for the purpose of resale for profit was high. Lameness more commonly affected hind limbs than forelimbs and was attributable to joint injury in most horses. Financial returns differed between horses classified as fast and average or slow at the 2-year-olds in training sales.

Pharmacokinetics of difloxacin and its concentration in body fluids and endometrial tissues of mares after repeated intragastric administration.

Pharmacokinetics of difloxacin and its distribution within the body fluids and endometrium of 6 mares were studied after intragastric (IG) administration of 5 individual doses. Difloxacin concentrations were serially measured in serum, urine, peritoneal fluid, synovial fluid, cerebrospinal fluid, and endometrium over 120 h. Bacterial susceptibility to difloxacin was determined for 174 equine pathogens over a 7-month period. Maximum serum concentration (Cmax) was 2.25 +/- 0.70 microg/mL at 3.12 +/- 2.63 h and Cmax after the 5th dose was 2.41 +/- 0.86 microg/mL at 97.86 +/- 1.45 h. The mean elimination half-life (t(1/2)) was 8.75 +/- 2.77 h and area under the serum concentration versus time curve (AUC) was 25.13 +/- 8.79 microg h/mL. Highest mean synovial fluid concentration was 1.26 +/- 0.49 microg/mL at 100 h. Highest mean peritoneal fluid concentration was 1.50 +/- 0.56 microg/mL at 98 h. Highest mean endometrial concentration was 0.78 +/- 0.48 microg/g at 97.5 h. Mean cerebrospinal fluid concentration was 0.87 +/- 0.52 microg/mL at 99 h. Highest mean urine concentration was 92.05 +/- 30.35 microg/mL at 104 h. All isolates of Salmonella spp. and Pasteurella spp. were susceptible. In general, gram-negative organisms were more susceptible than gram-positives. Difloxacin appears to be safe, adequately absorbed, and well distributed to body fluids and endometrial tissues of mares and may be useful in the treatment of susceptible bacterial infections in adult horses.

Disposition of orally administered cefpodoxime proxetil in foals and adult horses and minimum inhibitory concentration of the drug against common bacterial pathogens of horses.

OBJECTIVES: To determine the disposition of orally administered cefpodoxime proxetil in foals and adult horses and measure the minimum inhibitory concentrations (MICs) of the drug against common bacterial pathogens of horses. ANIMALS: 6 healthy adult horses and 6 healthy foals at 7 to 14 days of age and again at 3 to 4 months of age. PROCEDURE: A single dose of cefpodoxime proxetil oral suspension was administered (10 mg/kg) to each horse by use of a nasogastric tube. In 7- to 14-day-old foals, 5 additional doses were administered intragastrically at 12-hour intervals. The MIC of cefpodoxime for each of 173 bacterial isolates was determined by use of a commercially available test. RESULTS: In 7- to 14-day-old foals, mean +/- SD time to peak serum concentration (Tmax) was 1.7 +/- 0.7 hours, maximum serum concentration (Cmax) was 0.81 +/- 0.22 microg/mL, and elimination half-life (harmonic mean) was 7.2 hours. Disposition of cefpodoxime in 3- to 4-month-old foals was not significantly different from that of neonates. Adult horses had significantly higher Cmax and significantly lower Tmax, compared with values for foals. The MIC of cefpodoxime required to inhibit growth of 90% of isolates for Salmonella enterica, Escherichia coli, Pasteurella spp, Klebsiella spp, and beta-hemolytic streptococci was 0.38, 1.00, 0.16, 0.19, and 0.09 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration at a dosage of 10 mg/kg every 6 to 12 hours would appear appropriate for the treatment of equine neonates with bacterial infections.

Risk factors associated with fecal Salmonella shedding among hospitalized horses with signs of gastrointestinal tract disease.

OBJECTIVE: To estimate prevalence of and identify risk factors for fecal Salmonella shedding among hospitalized horses with signs of gastrointestinal tract disease. DESIGN: Cross-sectional study. ANIMALS: 465 hospitalized horses with gastrointestinal tract disease. PROCEDURE: Horses were classified as positive or negative for fecal Salmonella shedding during hospitalization by means of standard aerobic bacteriologic methods. The relationship between investigated exposure factors and fecal Salmonella shedding was examined by means of logistic regression. RESULTS: The overall prevalence of fecal Salmonella shedding was 13%. Salmonella serotype Newport was the most commonly isolated serotype (12/60 [20%]), followed by Anatum (8/60 [13%]), Java (13%), and Saint-paul (13%). Foals with gastrointestinal tract disease were 3.27 times as likely to be shedding Salmonella organisms as were adult horses with gastrointestinal tract disease. Adult horses that had been treated with antimicrobial drugs prior to hospitalization were 3.09 times as likely to be shedding Salmonella organisms as were adult horses that had not been treated with antimicrobial drugs prior to hospitalization. Adult horses that underwent abdominal surgery were 2.09 times as likely to be shedding Salmonella organisms as were adult horses that did not undergo abdominal surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a history of exposure to antimicrobial drugs prior to hospitalization and abdominal surgery during hospitalization were associated with Salmonella shedding in adult horses with gastrointestinal tract disease. Foals with gastrointestinal tract disease were more likely to shed Salmonella organisms than were adult horses with gastrointestinal tract disease.

Pharmacokinetics and plasma concentrations of acetylsalicylic acid after intravenous, rectal, and intragastric administration to horses.

Six healthy adult horses (5 mares and 1 stallion) were given a single dose of acetylsalicylic acid (ASA), 20 mg/kg of body weight, by intravenous (IV), rectal, and intragastric (IG) routes. Serial blood samples were collected via jugular venipuncture over a 36-h period, and plasma ASA and salicylic acid (SA) concentrations were determined by high-performance liquid chromatography. After IV administration, the mean elimination rate constant of ASA (+/- the standard error of the mean) was 1.32 +/- 0.09 h(-1), the mean elimination half-life was 0.53 +/- 0.04 h, the area under the plasma concentration-versus-time curve (AUC) was 2555 +/- 98 microg x min/mL, the plasma clearance was 472 +/- 18.9 mL/h/kg, and the volume of distribution at steady state was 0.22 +/- 0.01 L/kg. After rectal administration, the plasma concentration of ASA peaked at 5.05 +/- 0.80 microg/mL at 0.33 h, then decreased to undetectable levels by 4 h; the plasma concentration of SA peaked at 17.39 +/- 5.46 microg/mL at 2 h, then decreased to 1.92 +/- 0.25 microg/mL by 36 h. After rectal administration, the AUC for ASA was 439.4 +/- 94.55 microg x min/mL and the bioavailability was 0.17 +/- 0.037. After IG administration, the plasma concentration of ASA peaked at 1.26 +/- 0.10 microg/mL at 0.67 h, then declined to 0.37 +/- 0.37 microg/mL by 36 h; the plasma concentration of SA peaked at 23.90 +/- 4.94 microg/mL at 4 h and decreased to 0.85 +/- 0.31 microg/mL by 36 h. After IG administration, the AUC for ASA was 146.70 +/- 24.90 microg x min/mL and the bioavailability was 0.059 +/- 0.013. Administration of a single rectal dose of ASA of 20 mg/kg to horses results in higher peak plasma ASA concentrations and greater bioavailability than the same dose given IG. Plasma ASA concentrations after rectal administration should be sufficient to inhibit platelet thromboxane production, and doses lower than those suggested for IG administration may be adequate.

Hand-assisted laparoscopic left nephrectomy in standing horses.

OBJECTIVE: To describe a hand-assisted, laparoscopic technique to remove the left kidney in standing horses. STUDY DESIGN: Prospective evaluation. ANIMALS: Eight horses. METHODS: Food was withheld for a minimum of 12 hours. Horses were sedated with detomidine hydrochloride (0.01-0.02 mg/kg, intravenously) and restrained in standing stocks. The left paralumbar fossa was prepared for surgery, and the surgical site was infiltrated with 2% mepivacaine. Hand-assisted, laparoscopic removal of the left kidney was performed through an incision in the center of the paralumbar fossa; the surgeon's hand was used to isolate the left kidney and associated vasculature. The renal artery and vein were isolated and individually ligated. After vessel transection distal to the ligatures, the left kidney was exteriorized, the ureter ligated and transected, and the incision closed. RESULTS: Laparoscopic removal of the left kidney was successfully performed in all horses. Retroperitoneal infiltration of local anesthesia provided adequate anesthesia. Intraoperative hemorrhage occurred in 3 horses. Surgical duration (initial skin incision to transection of the left kidney) ranged from 20 to 90 minutes. In 2 horses, no signs of pain were noted for 48 hours postoperatively. CONCLUSION: Hand-assisted laparoscopic surgery can be used for removal of the left kidney in horses. Clinical Relevance-Hand-assisted laparoscopic nephrectomy can be safely performed in standing horses; however, care should be taken to identify accessory branches of the renal artery to limit potential complications with hemorrhage.

Regional and zonal variations in the sulfation patterns of chondroitin sulfate in normal equine corneal stroma.

OBJECTIVE: To determine regional and zonal variation in sulfation patterns of chondroitin sulfate in normal equine corneal stroma. SAMPLE POPULATION: 22 normal eyes from 11 horses. PROCEDURE: Corneas were collected within 24 hours of death from equine necropsy specimens. After papain-chondroitinase digestion of corneal tissue, disaccharides deltaDi4S and deltaDi6S were quantified by use of capillary zone electrophoresis in the superficial, middle, and deep zones of central and peripheral regions of the cornea. RESULTS: For the 2 regions combined, deltaDi6S/deltaDi4S values were significantly lower in the deep and middle zones, compared with that of the superficial zone. In the central region, deep and middle zones had significantly lower deltaDi6S/deltaDi4S values than the superficial zone did. In the peripheral region, the deep zone had significantly lower deltaDi6S/deltaDi4S values, compared with superficial and middle zones. In the deep zone, the peripheral region had significantly lower deltaDi6S/deltaDi4S values than the central region did. CONCLUSIONS AND CLINICAL RELEVANCE: Distribution of deltaDi6S/deltaDi4S values follows a gradient across the healthy equine cornea, being smallest in the deep and middle zones of the central region and the deep zone of the peripheral region. Regional and zonal differences in the distribution of stromal deltaDi6S and deltaDi4S may influence the role of glycosaminoglycans in health, disease, and wound repair of the equine cornea.