Objective-To compare in vitro expansion, explant colonization, and matrix synthesis of equine tendon- and bone marrow-derived cells in response to insulin-like growth factor-I (IGF-I) supplementation. Sample-Cells isolated from 7 young adult horses. Procedures-Tendon- and bone marrow-derived progenitor cells were isolated, evaluated for yield, and cultured on autogenous cell-free tendon matrix for 7 days. Samples were analyzed for cell viability and expression of collagen type I, collagen type III, and cartilage oligomeric matrix protein mRNAs. Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period. Results-Tendon- and bone marrow–derived cells required 17 to 19 days of monolayer culture to reach 2 passages. Mean +/- SE number of monolayer cells isolated was higher for tendon-derived cells (7.9 +/- 0.9 × 10(6)) than for bone marrow–derived cells (1.2 +/- 0.1 × 10(6)). Cell numbers after culture for 7 days on acellular tendon matrix were 1.6- to 2.8-fold higher for tendon-derived cells than for bone marrow-derived cells and 0.8- to 1.7-fold higher for IGF-I supplementation than for untreated cells. New collagen and glycosaminoglycan syntheses were significantly greater in tendon-derived cell groups and in IGF-I-supplemented groups. The mRNA concentrations of collagen type I, collagen type III, and cartilage oligomeric matrix protein were not significantly different between tendon- and bone marrow-derived groups. Conclusions and Clinical Relevance-In vitro results of this study suggested that tendon-derived cells supplemented with IGF-I may offer a useful resource for cell-based strategies in tendon healing.

Objective-To evaluate the osseous structures of the external acoustic meatus, tympanic cavity, and tympanic bulla of llamas (Lama glama) by use of computed tomography (CT) and establish measurement values for use in detection of abnormalities associated with the external or middle ear in llamas. Animals-10 adult llama heads without any evidence of ear disease. Procedures-Heads of 10 healthy llamas euthanized by use of a captive bolt striking the dorsal aspect of the skull were collected. Transverse images of the heads were acquired with 1-mm slice thickness, and images were reconstructed in sagittal and dorsal planes. Measurements of the bony structures of the external and middle ear of each head were obtained. Results-The osseous external acoustic meatus curved ventrally as it tracked medially. Its narrowest portion was located at the level of the tympanic annulus. The tympanic bulla conformation differed widely from the bubble-shaped tympanic bulla in dogs and cats. The bulla was divided by the stylohyoid fossa into a larger caudolateral and a smaller caudomedial process; its interior had a honeycombed structure with pneumatized cells similar to the honeycombed appearance of the human mastoid process. Conclusions and Clinical Relevance-Results provided new information regarding the shape and dimensions of the osseous external and middle ear structures in adult llamas without ear disease. Specific landmarks for location of the external acoustic meatus, tympanic cavity, and tympanic bulla in relation to each other were identified. Knowledge of the CT appearance of normal structures will help clinicians to identify changes attributable to middle ear otitis, external ear canal stenosis, or congenital malformations of the ear in this species.

Objective: To compare leakage and maximum intraluminal pressures for a novel suture material with pressures for comparable suture material when used in closure of intestinal anastomoses in canine cadavers. Sample: Healthy intestines from cadavers of dogs euthanized for reasons unrelated to the study. Procedures: 18 anastomoses were performed on intestinal sections within 72 hours after dogs were euthanized and intestinal samples collected. Anastomoses were performed with a simple continuous suture pattern. Leakage and maximum intraluminal pressures were measured and recorded for 6 control segments and 18 anastomosed sections. A barbed glycomer 631 suture (size 4–0 United States Pharmacopeia [USP]) was compared with glycomer 631 sutures (sizes 3–0 and 4–0 USP). Results for leakage and maximum intraluminal pressures were compared via an ANOVA. Results: The barbed glycomer 631 suture material leaked at a significantly higher pressure than did the comparable glycomer 631 suture materials. Maximum intraluminal pressures were not significantly different among the suture materials. Conclusions and Clinical Relevance: Barbed glycomer 631 4–0 USP suture material was as effective as glycomer 631 suture materials and may be a safe alternative for use in closure of enterectomies in dogs.

Objective: To determine the effect of various UVB radiation sources on plasma 25-hydroxyvitamin D3 concentrations in Hermann's tortoises (Testudo hermanni). Animals: 18 healthy Hermann's tortoises. Procedures: Tortoises were exposed to sunlight in an outdoor enclosure located in the natural geographic range of Hermann's tortoises (n = 6 tortoises) or a self-ballasted mercury-vapor lamp (6) or fluorescent UVB-emitting lamp (6) in an indoor enclosure for 35 days. Plasma samples were obtained from each tortoise on the first (day 0) and last (day 35) days of the study, and concentrations of 25-hydroxyvitamin D3 were determined. Amount of UVB radiation in enclosures was measured. Results: Mean ± SD plasma 25-hydroxyvitamin D3 concentrations for tortoises exposed to the mercury-vapor and fluorescent lamps were significantly lower on day 35 (155.69 ± 80.71 nmol/L and 134.42 ± 51.42 nmol/L, respectively) than they were on day 0 (368.02 ± 119.34 nmol/L and 313.69 ± 109.54 nmol/L, respectively). Mean ± SD plasma 25-hydroxyvitamin D3 concentration for tortoises exposed to sunlight did not differ significantly between days 0 (387.74 ± 114.56 nmol/L) and 35 (411.51 ± 189.75 nmol/L). Mean day 35 plasma 25-hydroxyvitamin D3 concentration was significantly higher for tortoises exposed to sunlight versus those exposed to mercury-vapor or fluorescent lamps. Sunlight provided significantly more UVB radiation than did the mercury-vapor or fluorescent lamps. Conclusions and Clinical Relevance: Plasma 25-hydroxyvitamin D3 concentrations differed between tortoises exposed to sunlight and those exposed to artificial UVB sources. Exposure to sunlight at a latitude similar to that of the natural geographic range is recommended for healthy and calcium-deficient tortoises.

Objective: To determine morphological and mechanical properties of trabecular bone of horses with a bone fragility syndrome (BFS; including silicate-associated osteoporosis). Sample: Cylindrical trabecular bone samples from the distal aspects of cadaveric third metacarpal bones of 39 horses (19 horses with a BFS [BFS bone samples] and 20 horses without a BFS [control bone samples]). Procedures: Bone samples were imaged via micro-CT for determination of bone volume fraction; apparent and mean mineralized bone densities; and trabecular number, thickness, and separation. Bone samples were compressed to failure for determination of apparent elastic modulus and stresses, strains, and strain energy densities for yield, ultimate, and failure loads. Effects of BFS and age of horses on variables were determined. Results: BFS bone samples had 25% lower bone volume fraction, 28% lower apparent density, 18% lower trabecular number and thickness, and 16% greater trabecular separation versus control bone samples. The BFS bone samples had 22% lower apparent modulus and 32% to 33% lower stresses, 10% to 18% lower strains, and 41 % to 52% lower strain energy densities at yield, ultimate, and failure loads, compared with control bone samples. Differences between groups of bone samples were not detected for mean mineral density and trabecular anisotropy. Conclusions and Clinical Relevance: Results suggested that horses with a BFS had osteopenia and compromised trabecular bone function, consistent with bone deformation and pathological fractures that develop in affected horses. Effects of this BFS may be systemic, and bones other than those that are clinically affected had changes in morphological and mechanical properties.

Objective: To optimize the use of CT-guided modeling for the calculation of body surface area (BSA) in domestic rabbits (Oryctolagus cuniculus). Animals: 12 domestic rabbits. Procedures: Adult rabbits (body weight, 1 to > 4 kg) that were client-owned animals undergoing CT for disease diagnosis or deceased laboratory animals donated from other research projects were scanned with a CT scanner. Images were transferred to a radiation therapy planning software program. Image slices were captured as contiguous slices at 100 kVp and 100 mA and processed to 0.1-cm-thick sections. The length of each contoured slice was summed to calculate a final BSA measurement. Nonlinear regression analysis was then used to derive an equation for the calculation of BSA in rabbits. Results: The constant calculated by use of this method was 9.9 (range, 9.59 to 10). The R2 for the goodness of fit was 0.9332. The equation that best described BSA as a function of body weight for domestic rabbits with this method was as follows: BSA = (9.9 × [body weight {in grams}]2/3)/10,000. Conclusions and Clinical Relevance: The BSA calculated via the CT-guided method yielded results similar to those obtained with equations for other similarly sized mammals and verified the use of such equations for rabbits. Additionally, this technique can be used for species that lack equations for the accurate calculation of BSA.

Objective: To determine the tissue distribution of enrofloxacin after intramammary or simulated systemic administration in isolated perfused sheep udders by measuring its concentration at various sample collection sites. Sample: 26 udders (obtained following euthanasia) from 26 healthy lactating sheep. Procedures: For each isolated udder, 1 mammary gland was perfused with warmed, gassed Tyrode solution. Enrofloxacin (1 g of enrofloxacin/5 g of ointment) was administered into the perfused gland via the intramammary route or systemically via the perfusion fluid (equivalent to a dose of 5 mg/kg). Samples of the perfusate were obtained every 30 minutes for 180 minutes; glandular tissue samples were obtained at 2, 4, 6, and 8 cm from the teat base after 180 minutes. The enrofloxacin content of the perfusate and tissue samples was analyzed via high-performance liquid chromatography with UV detection. Results: After intramammary administration, maximun perfusate enrofloxacin concentration was detected at 180 minutes and, at this time, mean tissue enrofloxacin concentration was detected and mean tissue enrofloxacin concentration was 123.80, 54.48, 36.72, and 26.42 μg/g of tissue at 2, 4, 6, and 8 cm from the teat base, respectively. Following systemic administration, perfusate enrofloxacin concentration decreased with time and, at 180 minutes, tissue enrofloxacin concentrations ranged from 40.38 to 35.58 μg/g of tissue. Conclusions and Clinical Relevance: By 180 minutes after administration via the intramammary or systemic route in isolated perfused sheep mammary glands, mean tissue concentration of enrofloxacin was greater than the minimum inhibitory concentration required to inhibit growth of 90% of many common mastitis pathogens in sheep. Use of either route of administration (or in combination) appears suitable for the treatment of acute mastitis in sheep.

Objective: To provide values for gastrointestinal permeability and absorptive function tests (GIPFTs) with chromium 51 (51Cr)-labeled EDTA, lactulose, rhamnose, d-xylose, 3-O-methyl-d-glucose, and sucrose in Beagles and to evaluate potential correlations between markers. Animals: 19 healthy adult male Beagles. Procedures: A test solution containing 3.7 MBq of 51Cr-labeled EDTA, 2 g of lactulose, 2 g of rhamnose, 2 g of d-xylose, 1 g of 3-O-methyl-d-glucose, and 8 g of sucrose was administered intragastrically to each dog. Urinary recovery of each probe was determined 6 hours after administration. Results: Mean ± SD (range) percentage urinary recovery was 6.3 ± 1.6% (4.3% to 9.7%) for 51Cr-labeled EDTA, 3.3 ± 1.1% (1.7% to 5.3%) for lactulose, 25.5 ± 5.0% (16.7% to 36.9%) for rhamnose, and 58.8% ± 11.0% (40.1% to 87.8%) for 3-O-methyl-d-glucose. Mean (range) recovery ratio was 0.25 ± 0.06 (0.17 to 0.37) for 51Cr-labeled EDTA to rhamnose, 0.13 ± 0.04 (0.08 to 0.23) for lactulose to rhamnose, and 0.73 ± 0.09 (0.60 to 0.90) for d-xylose to 3-O-methyl-d-glucose. Median (range) percentage urinary recovery was 40.3% (31.6% to 62.7%) for d-xylose and 0% (0% to 0.8%) for sucrose. Conclusions and Clinical Relevance: Reference values in healthy adult male Beagles for 6 of the most commonly used GIPFT markers were determined. The correlation between results for 51Cr-labeled EDTA and lactulose was not as prominent as that reported for humans and cats; thus, investigators should be cautious in the use and interpretation of GIPFTs performed with sugar probes in dogs with suspected intestinal dysbiosis.

Objective: To compare effects of 3 methods of topically applied cold treatment (cryotherapy) on digital laminar and venous temperatures in horses. Animals: 9 healthy adult Thoroughbreds. Procedures: Thermocouples were placed in palmar digital veins and digital laminae of both forelimbs of horses. Three methods of cryotherapy were applied to the distal aspects of the limbs: wader boot (63-cm-tall vinyl boot filled with ice and water [ice slurry]), ice bag (5-L fluid bag filled with ice slurry), and a gel pack boot (boot containing frozen gel packs). Gel packs and ice slurries were replenished every hour during cryotherapy. The forelimb that received the first treatment was randomly assigned; thereafter, control and treated forelimbs were alternated for each treatment. For each treatment, temperatures were recorded every minute during 15-minute pretreatment, 2-hour treatment, and ≥ 30 minute rewarming periods. Once temperatures had returned to within 3°C below pretreatment values, the experiment was repeated in a similar manner for other cryotherapy methods. Results: Digital venous temperatures were similar to laminar temperatures during each treatment. Ice bag and wader boot treatments caused similar cooling of digits. Gel boot treatment did not cause substantial cooling of digits. Conclusions and Clinical Relevance: Ice bag treatment caused laminar and digital venous cooling equivalent to that of wader boot treatment. Cryotherapy by use of 5-L fluid bags with an ice slurry may be a readily available, practical, and efficient method for prevention of laminitis in horses. Digital laminar and venous temperatures were similar in forelimbs of horses before and during cryotherapy.

The objective of this study was to compare bone marrow (BM) aspirates from the sternum and the tuber coxae of middle-aged horses. Bone marrow was obtained from the sternum and both tubera coxae of 12 healthy, 13-year-old geldings. Two different puncture techniques were used for the tuber coxae. The 2 syringes used for sternal sampling were evaluated separately. The mononuclear cell (MNC) fraction of the BM was isolated and the mesenchymal stem cells (MSCs) were culture-expanded. At the sternum, BM aspiration was always possible. Bone marrow aspiration at the tuber coxae required straight and deep needle penetration combined with high negative pressure. With this technique a median sample amount of 11.0 mL with large individual variation was obtained. A median of 3.06 × 10(6) MNC/mL BM (1st syringe) and 2.46 × 10(6) MNC/mL BM (2nd syringe) was isolated from sternal samples. In contrast, the tuber coxae yielded a median of 0.27 × 10(6) MNC/mL BM. The first passage yielded a median of 2.19 × 10(6) MSC (1st syringe) and 1.13 × 10(6) MSC (2nd syringe) from sternal samples, compared to a significantly lower median number of MSC from tuber coxae BM (0.06 × 10(6) MSC). The number of MNC and MSC obtainable from the BM aspirates taken from the tuber coxae is significantly lower than that obtained from the sternal BM aspirates. Autologous BM for the equine athlete is particularly clinically relevant at an advanced age. Based on our findings, the tuber coxae cannot be recommended for BM aspiration in middle-aged horses.