Objective: To investigate effects of 1% hyaluronic acid–chondroitin sulfate–N-acetyl glucosamine (HCNAG) on the damage repair response in equine articular cartilage. Sample: Articular cartilage from 9 clinically normal adult horses. Procedures: Full-thickness cartilage disks were harvested from the third metacarpal bone. Cartilage was single-impact loaded (SIL) with 0.175 J at 0.7 m/s and cultured in DMEM plus 1 % (vol/vol) HCNAG or fibroblastic growth factor (FGF)-2 (50 ng/mL). Histologic and immunohistochemical techniques were used to identify tissue architecture and apoptotic cells and to immunolocalize type I and II collagen and proliferating nuclear cell antigen (PCNA). Results: Type II collagen immunoreactivity increased in SIL cartilage, compared with control samples. At days 14 and 28 (day 0 = initiation of culture), control samples had significantly fewer repair cells than did other treatment groups. In control samples and SIL + HCNAG, there was a significant decrease in apoptotic cell number, compared with results for SIL and SIL + FGF-2 samples. At days 14 and 28, there was a significant increase in chondrocytes stained positive for PCNA in the control samples. Conclusions and Clinical Relevance: 1% HCNAG significantly affected apoptotic and repair cell numbers in an SIL damage-repair technique in adult equine articular cartilage. However, HCNAG had no effect on the number of PCNA-positive chondrocytes or on type II collagen immunohistochemical results. The inclusion of 1% HCNAG in lavage solutions administered after arthroscopy may be beneficial to cartilage health by increasing the number of repair cells and decreasing the number of apoptotic cells.

Objective: To determine whether increased gene expression of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) in laminae of horses with starch gruel–induced laminitis was accompanied by increased enzyme activity and substrate degradation. Sample: Laminae from the forelimb hooves of 8 healthy horses and 17 horses with starch gruel–induced laminitis (6 at onset of fever, 6 at onset of Obel grade 1 lameness, and 5 at onset of Obel grade 3 lameness). Procedures: Gene expression was determined by use of cDNA and real-time quantitative PCR assay. Protein expression and processing were determined via SDS-PAGE and quantitative western blotting. Protein distribution and abundance were determined via quantitative immunofluorescent staining. Results: ADAMTS-4 gene expression was increased and that of versican decreased in laminitic laminae, compared with expression in healthy laminae. Catalytically active ADAMTS-4 also was increased in the tissue, as were ADAMTS-4–cleavage fragments of versican. Immunofluorescent analyses indicated that versican was depleted from the basal epithelia of laminae of horses at onset of Obel grade 3 lameness, compared with results for healthy laminae, and this was accompanied by regional separation of basal epithelial cells from the basement membrane. Aggrecan gene and protein expression were not significantly affected. Conclusions and Clinical Relevance: Changes in gene and protein expression of ADAMTS-4 and versican in the basal epithelium of laminitic laminae indicated a fundamental change in the physiology of basal epithelial cells. This was accompanied by and may have caused detachment of these cells from the basement membrane.

Objective: To determine the expression and distribution of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), its substrates aggrecan and versican, and their binding partner hyaluronan in laminae of healthy horses. Sample: Laminae from the forelimb hooves of 8 healthy horses. Procedures: Real-time quantitative PCR assay was used for gene expression analysis. Hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts combined with SDS-PAGE and western blotting were used for protein and proteoglycan analysis. Immunofluorescent and immunohistochemical staining of tissue sections were used for protein and hyaluronan localization. Results: Genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were expressed in laminae. The ADAMTS-4 was predominantly evident as a 51-kDa protein bearing a catalytic site neoepitope indicative of active enzyme and in situ activity, which was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts. Aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae. The ADAMTS-4 localized to these cells but was also present in some cells in the dermal laminae. Conclusions and Clinical Relevance: Within digital laminae, versican exclusively and aggrecan primarily localized within basal epithelial cells and both were constitutively cleaved by ADAMTS-4, which therefore contributed to their turnover. On the basis of known properties of these proteoglycans, it is possible that they can protect the basal epithelial cells of horses from biomechanical and concussive stress.

Objective: To determine effects of syringe type and storage conditions on blood gas and acid-base values for equine blood samples. Sample: Blood samples obtained from 8 healthy horses. Procedures: Heparinized jugular venous blood was equilibrated via a tonometer at 37°C with 12% O2 and 5% CO2. Aliquots (3 mL) of tonometer-equilibrated blood were collected in random order by use of a glass syringe (GS), general-purpose polypropylene syringe (GPPS), or polypropylene syringe designed for blood gas analysis (PSBGA) and stored in ice water (0°C) or at room temperature (22°C) for 0, 5, 15, 30, 60, or 120 minutes. Blood pH was measured, and blood gas analysis was performed; data were analyzed by use of multivariable regression analysis. Results: Blood Po2 remained constant for the reference method (GS stored at 0°C) but decreased linearly at a rate of 7.3 mm Hg/h when stored in a GS at 22°C. In contrast, Po2 increased when blood was stored at 0°C in a GPPS and PSBGA or at 22°C in a GPPS; however, Po2 did not change when blood was stored at 22°C in a PSBGA. Calculated values for plasma concentration of HCO3 and total CO2 concentration remained constant in the 3 syringe types when blood was stored at 22°C for 2 hours but increased when blood was stored in a GS or GPPS at 0°C. Conclusions and Clinical Relevance: Blood samples for blood gas and acid-base analysis should be collected into a GS and stored at 0°C or collected into a PSBGA and stored at room temperature.

Objective: To evaluate effects of the arytenoid lateralization technique and suture tension on airway pressure in the canine larynx. Sample: 7 canine cadaver larynges. Procedures: Negative pressure was elicited aboral to the larynx. Airway pressure was measured at airflows of 15 to 120 L/min before and after thyroarytenoid lateralization (TAL), cricoarytenoid lateralization (CAL), and combined TAL and CAL (cricothyroarytenoid lateralization [CTAL]) at 100 and 500 g of suture tension and with sectioning of the sesamoid cartilage (SSC) and disarticulation of the cricothyroid joint (DCTJ). Rima glottidis area (RGA) was measured. Effects of technique, modification, and suture tension on pressure and RGA were evaluated statistically. Results: Increased suture tension significantly reduced airway pressure for TAL at 30 L/min, CAL at 45 to 120 L/min, and CAL after SSC and DCTJ at 60, 75, and 105 to 120 L/min. The CAL and CTAL caused significantly lower airway pressures than did TAL > 30 L/min, but SSC and DCTJ did not significantly reduce pressure. All procedures, except TAL at 100 g of tension, resulted in a significant RGA increase from baseline. The CAL and CTAL caused a significantly greater RGA than did TAL. For TAL at 100 g of tension, SSC significantly increased RGA. Conclusions and Clinical Relevance: CAL and CTAL caused lower airway pressures than did TAL. No significant pressure differences were detected between CAL and CTAL; SSC and DCTJ had little effect on pressure. Pressure may be a more sensitive indicator of airflow than is RGA in the larynx of canine cadavers.

Objective: To determine MRI characteristics of the skulls and brains of sheep with chronic cerebral coenurosis (CC) caused by naturally acquired Taenia multiceps infection. Animals: 33 sheep with CC and 10 healthy control sheep. Procedures: Sheep underwent MRI of the head. Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were determined. For CC-affected sheep, the number, location, and volume of T multiceps cysts were determined and the percentage volumes of cysts in the cranial cavity and rostral and caudal fossas of the cranial cavity were calculated. Focal and diffuse abnormalities of cranial bones in CC-affected sheep were identified. Brain edema and hemorrhage and signs of increased cranial pressure (ICP) in MRI images were determined. Results: Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were significantly larger for CC-affected sheep versus healthy control sheep. Total volumes of cysts ranged from 4.40% to 46.93% in cranial cavities of sheep, 4.12% to 51.53% in rostral fossas of cranial cavities of sheep, and 15.24% to 68.30% in caudal fossas of cranial cavities of sheep. Moderate to severe diffuse cranial bone abnormalities and signs of increased ICP in MRI images were detected in 21 and 24 sheep, respectively, and were positively correlated with cyst volumes. Conclusions and Clinical Relevance: Results suggested that cranial cavity volume and morphological abnormalities can be detected in sheep with CC. These changes may reflect abnormalities in ossification of the cranial bones secondary to chronically increased ICP caused by development of T multiceps cysts.

Objective: To determine the tissue depletion profile of tulathromycin and determine an appropriate slaughter withdrawal interval in meat goats after multiple SC injections of the drug. Animals: 16 healthy Boer goats. Procedures: All goats were administered tulathromycin (2.5 mg/kg, SC) twice, with a 7-day interval between doses. Blood samples were collected throughout the study, and goats were euthanized at 2, 5, 10, and 20 days after the second tulathromycin dose. Lung, liver, kidney, fat, and muscle tissues were collected. Concentrations of tulathromycin in plasma and the hydrolytic tulathromycin fragment CP-60,300 in tissue samples were determined with ultrahigh-pressure liquid chromatography–tandem mass spectrometry. Results: The plasma profile of tulathromycin was biphasic. Absorption was very rapid, with maximum drug concentrations (1.00 ± 0.42 μg/mL and 2.09 ± 1.77 μg/mL following the first and second doses, respectively) detected within approximately 1 hour after injection. Plasma terminal elimination half-life of tulathromycin was 61.4 ± 14.1 hours after the second dose. Half-lives in tissue ranged from 2.4 days for muscle to 9.0 days for lung tissue; kidney tissue was used to determine the withdrawal interval for tulathromycin in goats because it is considered an edible tissue. Conclusions and Clinical Relevance: On the basis of the tissue tolerance limit in cattle of 5 ppm (μg/g), the calculated withdrawal interval for tulathromycin would be 19 days following SC administration in goats. On the basis of the more stringent guidelines recommended by the FDA, the calculated meat withdrawal interval following tulathromycin administration in goats was 34 days.

Objective: To determine the minimum anesthetic concentration (MAC) of sevoflurane in thick-billed parrots (Rhynchopsitta pachyrhyncha) and compare MAC obtained via mechanical and electrical stimulation. Animals: 15 healthy thick-billed parrots. Procedures: Anesthesia was induced in each parrot by administration of sevoflurane in oxygen. An end-tidal sevoflurane concentration of 2.5% was established in the first bird. Fifteen minutes was allowed for equilibration. Then, 2 types of noxious stimulation (mechanical and electrical) were applied; stimuli were separated by 15 minutes. Responses to stimuli were graded as positive or negative. For a positive or negative response to a stimulus, the target end-tidal sevoflurane concentration of the subsequent bird was increased or decreased by 10%, respectively. The MAC was calculated as the mean end-tidal sevoflurane concentration during crossover events, defined as instances in which independent pairs of birds evaluated in succession had opposite responses. A quantal method was used to determine sevoflurane MAC. Physiologic variables and arterial blood gas values were also measured. Results: Via quantal analysis, mean sevoflurane MAC in thick-billed parrots determined with mechanical stimulation was 2.35% (90% fiducial interval, 1.32% to 2.66%), which differed significantly from the mean sevoflurane MAC determined with electrical stimulation, which was 4.24% (90% fiducial interval, 3.61% to 8.71%). Conclusions and Clinical Relevance: Sevoflurane MAC in thick-billed parrots determined by mechanical stimulation was similar to values determined in chickens and mammals. Sevoflurane MAC determined by electrical stimulation was significantly higher, which suggested that the 2 types of stimulation did not induce similar results in thick-billed parrots.

Objective: To evaluate use of a diode laser to induce tendinopathy in the superficial digital flexor tendon (SDFT) of horses. Animals: 4 equine cadavers and 5 adult horses. Procedures: Cadaveric SDFT samples were exposed to a diode laser at various energy settings to determine an appropriate energy for use in in vivo experiments; lesion size was assessed histologically. In vivo experiments involved laser energy induction of lesions in the SDFT (2 preliminary horses [0, 25, 75, and 87.5 J] and 3 study horses [0 and 125 J]) and assessment of lesions. Study duration was 21 days, and lesions were assessed clinically and via ultrasonography, MRI, and histologic evaluation. Results: Lesion induction in cadaveric tissues resulted in a spherical cavitated core with surrounding tissue coagulation. Lesion size had a linear relationship (R2 = 0.9) with the energy administered. Size of in vivo lesions in preliminary horses indicated that larger lesions were required. In study horses, lesions induced with 125 J were ultrasonographically and histologically larger than were control lesions. At proximal and distal locations, pooled (preliminary and study horses) ultrasonographically assessed lesions were discrete and variable in size (mean ± SEM lesion percentage for control lesions, 8.5 ± 3%; for laser lesions, 12.2 ± 1.7%). Ultrasonography and MRI measurements were associated (R2 > 0.84) with cross-sectional area measurements. Conclusions and Clinical Relevance: In vivo diode laser–induced lesions did not reflect cadaveric lesions in repeatable size. Further research is required before diode lasers can reliably be used for inducing tendinopathy.

Objective: To evaluate 2 plate designs for pancarpal arthrodesis and their effects on load transfer to the respective bones as well as to develop a computational model with directed input from the biomechanical testing of the 2 constructs. Sample: Both forelimbs from the cadaver of an adult castrated male Golden Retriever. Procedures: CT imaging was performed on the forelimb pair. Each forelimb was subsequently instrumented with a hybrid dynamic compression plate or a castless pancarpal arthrodesis plate. Biomechanical testing was performed. The forelimbs were statically loaded in the elastic range and then cyclically loaded to failure. Finite element (FE) modeling was used to compare the 2 plate designs with respect to bone and implant stress distribution and magnitude when loaded. Results: Cyclic loading to failure elicited failure patterns similar to those observed clinically. The mean ± SD error between computational and experimental strain was < 15% ± 13% at the maximum loads applied during static elastic loading. The highest bone stresses were at the distal extent of the metacarpal bones at the level of the screw holes with both plates; however, the compression plate resulted in slightly greater stresses than did the arthrodesis plate. Both models also revealed an increase in bone stress at the proximal screw position in the radius. The highest plate stress was identified at the level of the radiocarpal bone, and an increased screw stress (junction of screw head with shaft) was identified at both the most proximal and distal ends of the plates. Conclusions and Clinical Relevance: The FE model successfully approximated the biomechanical characteristics of an ex vivo pancarpal plate construct for comparison of the effects of application of different plate designs.