Objective: To determine whether withholding of food affects autonomic nervous system balance by analysis of heart rate (HR), HR variability (HRV), and frequency of second-degree atrioventricular block in horses. Animals: 5 healthy Thoroughbreds. Procedures: For two 24-hour periods in a crossover study, food was withheld from horses or horses were maintained on their regular feeding schedule (control conditions) in their stalls and Holter monitor ECG recordings were obtained. The ECGs were analyzed by use of fast-Fourier transformation, and power spectrum densities were calculated for low-frequency (0.01 to 0.07 Hz) and high-frequency (0.07 to 0.6 Hz) variations in HR. Serum cortisol and plasma ACTH, norepinephrine, and glucose concentrations were measured at predetermined time points. Results: Withholding of food resulted in significantly lower HR and more frequent second-degree atrioventricular block (the frequency of which was inversely related to the HR), compared with findings for control conditions. Circadian rhythms were similar during food-withholding and control conditions; peak HR was detected from 7:00 pm to 8:00 pm, and the lowest HR was detected in the early morning. During food-withholding conditions, the low-frequency and high-frequency components of HRV were significantly higher, and the low-frequency-to-high-frequency ratio was lower than during control conditions. Serum cortisol concentration was higher and plasma glucose concentration was lower at 6:00 pm in horses when food was withheld, compared with findings during control conditions. Conclusions and Clinical Relevance: Indices of HRV seemed to be sensitive to changes in autonomic nervous activity and may be useful as clinical indices of the neuroendocrine response to stressors in horses.

Objective: To validate the use of radiography to determine the length of sagittal otoliths in intact bogue (Boops boops; a sparid fish [commonly called sea bream]). Sample: 52 bogue cadavers. Procedures: Weight and standard lengths (from the tip of the snout to the caudal end of the last vertebra) of fish were measured. The radiographic beam was centered over the postorbital area, and images were obtained via ventrodorsal, 30° right dorsal–left ventral oblique, and 30° left dorsal–right ventral oblique projections. Otoliths were removed from the fish; sagittae were measured directly and on radiographic images by use of a vernier caliper. The relationship between direct and radiographic measurements of sagittal otoliths was calculated. Results: Paired sagittal, lapillus, and asteriscus otoliths were identified. Sagittal otoliths were comma-shaped on ventrodorsal projections; in oblique projections, they appeared ovoid and pairs of otoliths were located in the same dorsal plane. Radiographic length of sagittal otoliths was significantly correlated with directly measured length, and radiographic measurements were not significantly different between ventrodorsal and oblique radiographic projections. Conclusions and Clinical Relevance: Radiographic examination of sagittae was useful to determine the length of sagittal otoliths and, consequently, the fish length. In ecological applications, the radiographic measurement of sagittal otolith length may be useful for studies on the aquatic diet of organisms (fish, pinnipeds, and marine birds) because it allows for quick back-calculation to the size of prey.

Objective: To establish a safe anesthetic protocol with little effect on blood biochemical values and IV glucose tolerance test (IVGTT) results in Japanese black bears (Ursus thibetanus japonicus). Animals: 16 captive female Japanese black bears (5 to 17 years of age). Procedures: Bears were randomly assigned to 4 treatment groups (4 bears/group) in which various treatment combinations were administered via blow dart: tiletamine HCl and zolazepam HCl (9 mg/kg) alone (TZ), TZ (6 mg/kg) and acepromazine maleate (0.1 mg/kg), TZ (6 mg/kg) and butorphanol tartrate (0.3 mg/kg), or TZ (3 mg/kg) and medetomidine HCl (40 μg/kg). Glucose injection for the IVGTT was started 130 minutes after TZ administration. Blood samples were obtained before, at, and intermittently after glucose injection for measurement of biochemical variables as well as plasma glucose and serum insulin concentrations during the IVGTT. Rectal temperature, pulse rate, and respiratory rate were assessed every 15 minutes during the experiment. Results: Induction and maintenance of anesthesia were safely achieved with little adverse effect on cardiopulmonary function when each of the 4 anesthetic regimens was used, although mild hypothermia was induced. No difference was evident between treatment groups in blood biochemical values. Blood glucose and insulin concentration profiles during the IVGTT were similar among the bears given TZ, with or without acepromazine or butorphanol, but hyperglycemia and hypoinsulinemia developed in bears given TZ with medetomidine. Conclusions and Clinical Relevance: All 4 anesthetic regimens yielded chemical restraint without affecting clinical and biochemical values in bears, but medetomidine appeared to affect IVGTT results. For this reason, medetomidine should not be used when anesthetizing bears for IVGTTs.

Objective: To determine associations between serum concentrations of omega-3 polyunsaturated fatty acids or body condition and serum concentrations of adiponectin, leptin, insulin, glucose, or triglyceride in healthy dogs. Animals: 62 healthy adult client-owned dogs. Procedures: Body condition score and percentage of body fat were determined. Blood samples were collected after food was withheld for 12 hours. Serum was harvested for total lipid determination, fatty acid analysis, and measurement of serum concentrations of adiponectin, leptin, insulin, glucose, and triglyceride. Associations between the outcome variables (adiponectin, leptin, insulin, glucose, and triglyceride concentrations) and each of several variables (age, sex, percentage of body fat, and concentrations of total lipid, α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid) were determined. Results: Serum concentrations of docosapentaenoic acid were significantly positively associated with concentrations of adiponectin and leptin and negatively associated with concentrations of triglyceride. Serum concentrations of α-linolenic acid were significantly positively associated with concentrations of triglyceride. No significant associations were detected between serum concentrations of eicosapentaenoic acid or docosahexaenoic acid and any of the outcome variables. Percentage of body fat was significantly positively associated with concentrations of leptin, insulin, and triglyceride but was not significantly associated with adiponectin concentration. Age was positively associated with concentrations of leptin, insulin, and triglyceride and negatively associated with concentrations of adiponectin. Sex did not significantly affect serum concentrations for any of the outcome variables. Conclusions and Clinical Relevance: Docosapentaenoic acid may increase serum concentrations of adiponectin and leptin and decrease serum triglyceride concentration in healthy dogs.

Objective: To assess the prevalence of Mycobacterium bovis infection in cattle and wild ruminants (WRs) in a wildlife-livestock interface area (WLIA) of the Mexican highland plateau. Animals: 24,400 cattle from 793 herds (including 17,351 commercially slaughtered cattle) and 142 WRs (110 white-tailed deer [Odocoileus virginianus], 20 red deer [Cervus elaphus], and 12 North American elk [Cervus canadensis]) harvested via controlled hunting. Procedures: Cattle were serially tested for M bovis infection via caudal fold tuberculin and comparative cervical tuberculin tests during field surveillance. Carcasses of cattle and WRs were inspected for gross lesions; samples suggestive of tuberculosis were analyzed via histologic evaluation and mycobacterial culture (HMC). A PCR assay to detect Mycobacterium tuberculosis complex organisms was performed to confirm positive results of HMC. Results: WRs had inflammatory lesions in lungs and lymph nodes, although HMC results did not indicate M bovis infection. Eight cattle had positive results for both tuberculin tests, and 31 had positive results for HMC of grossly detected lesions; all were from 7 herds, and ≥ 1 cow in each herd had positive PCR assay results. These 7 herds were depopulated; adjacent herds and herds related via commerce were quarantined. Calculated true prevalence of M bovis infection was 0.86% (95% confidence interval, 0.24% to 1.49%) in cattle; M bovis was not detected in any WRs. Conclusions and Clinical Relevance: M bovis infection was present in cattle. Although transmission to WRs in this WLIA was not detected, diagnosis and prevention activities should be implemented and consolidated to prevent potential M bovis transmission between cattle and WRs.

Objective: To determine mechanical properties of various prosthetic materials secured to cadaveric canine femurs via various methods and to compare results with those for isolated loops of prosthetic material. Sample: 80 femurs obtained from cadavers of skeletally mature large-breed dogs. Procedures: 10 femoral constructs in each of 8 groups (single circumfabellar loop of polyethylene cord, double loop of polyethylene tape secured via a bone anchor [BAPT], single or double circumfabellar loops of nylon leader material [CNL] or polyethylene tape [CPT], or single or double loops of polyethylene tape secured via a toggle placed through a bone tunnel [BTPT]) and 10 isolated loops of prosthetic material in each corresponding configuration were tested. Stress relaxation, creep, elongation, load at 3 mm of displacement, stiffness, and peak load at failure were determined. Results: 5 single CNL constructs failed before completion of testing. Double CNL and single circumfabellar polyethylene cord constructs had the lowest loads at 3 mm of displacement. Single and double CPT constructs had the highest stiffness. Double BTPT and CPT constructs had the highest peak loads at failure. Double BTPT, double CPT, and BAPT constructs were mechanically superior on the basis of lower creep and stress relaxation and higher stiffness and load at 3 mm of displacement versus other constructs. Stiffness of femoral constructs was 28% to 69% that of corresponding isolated prosthetic loops. Conclusions and Clinical Relevance: Double BTPT, double CPT, and BAPT constructs were mechanically superior to other constucts. Mechanical properties and methods of anchorage and securing of free ends of prostheses contributed to mechanical properties of constructs.

Objective: To determine concentration-dependent effects of tiludronate on cartilage explants incubated with or without recombinant equine interleukin-1β (rEq IL-1). Sample: Articular cartilage explants from the femorotibial joints of 3 young adult horses. Procedures: Cartilage explants were incubated with 1 of 6 concentrations (0, 0.19, 1.9, 19, 190, or 1,900 mg/L) of tiludronate and with or without rEq IL-1 (0.01 ng/mL) for 96 hours. Prostaglandin E2 (PGE2) concentrations in culture medium and explant digests were analyzed via PGE2 enzyme immunoassay. Sulfated glycosaminoglycan (sGAG) concentrations in culture medium were quantified via 1,9-dimethylmethylene blue assay. Chondrocyte apoptosis in paraffin embedded explant sections was measured via terminal deoxynucleotidyl transferase-mediated dUTP nick end–labeling assay. Relative gene expression of matrix metalloproteinases (MMPs), interleukin (IL)-6, and IL-8 was determined via the comparative cycle threshold method. Results: rEq IL-1 increased PGE2 concentration, sGAG release from explants, chondrocyte apoptosis, and MMP gene expression. Lower tiludronate concentrations reduced rEq IL-1–induced sGAG release and chondrocyte apoptosis, whereas the higher tiludronate concentrations increased sGAG release and chondrocyte apoptosis. At the highest tiludronate concentration evaluated, IL-8 gene expression was increased independent of whether rEq IL-1 was present. Conclusions and Clinical Relevance: Tiludronate had biphasic concentration-dependent effects on cartilage explants that were independent of PGE2 secretion or MMP gene expression. Low tiludronate concentrations had some chondroprotective effects, whereas high tiludronate concentrations were detrimental to equine articular cartilage. Administration of tiludronate intra-articularly to horses may be detrimental, dependent on the dose used. In vivo studies are needed before intra-articular tiludronate administration to horses can be recommended.

Objective: To determine plasma pharmacokinetics of penciclovir following oral and rectal administration of famciclovir to young Asian elephants (Elephas maximus). Animals: 6 healthy Asian elephants (5 females and 1 male), 4.5 to 9 years old and weighing 1,646 to 2,438 kg. Procedures: Famciclovir was administered orally or rectally in accordance with an incomplete crossover design. Three treatment groups, each comprising 4 elephants, received single doses of famciclovir (5 mg/kg, PO, or 5 or 15 mg/kg, rectally); there was a minimum 12-week washout period between subsequent famciclovir administrations. Serial blood samples were collected after each administration. Samples were analyzed for famciclovir and penciclovir with a validated liquid chromatography–mass spectroscopy assay. Results: Famciclovir was tolerated well for both routes of administration and underwent complete biotransformation to the active metabolite, penciclovir. Mean maximum plasma concentration of penciclovir was 1.3 μg/mL at 1.1 hours after oral administration of 5 mg/kg. Similar results were detected after rectal administration of 5 mg/kg. Mean maximum plasma concentration was 3.6 μg/mL at 0.66 hours after rectal administration of 15 mg/kg; this concentration was similar to results reported for humans receiving 7 mg/kg orally. Conclusions and Clinical Relevance: Juvenile Asian elephants are susceptible to elephant endotheliotropic herpesvirus. Although most infections are fatal, case reports indicate administration of famciclovir has been associated with survival of 3 elephants. In Asian elephants, a dose of 8 to 15 mg of famciclovir/kg given orally or rectally at least every 8 hours may result in penciclovir concentrations that are considered therapeutic in humans.

Objective: To examine the distribution of water in hoof wall specimens of horses via nuclear magnetic resonance (NMR) microscopy and determine changes in water distribution during hydration. Sample: 4 hoof wall specimens (2 obtained from the dorsum and 1 each obtained from the lateral quarter and lateral heel regions) of the stratum medium of healthy hooves of 1 horse. Procedures: Equine hoof wall specimens were examined via NMR microscopy. Proton density–weighted 3-D images were acquired. Changes during water absorption were assessed on sequential images. Results: The inner zone of the stratum medium had higher signals than did the outer zone. Areas of high signal intensity were evident in transverse images; these corresponded to the distribution of horn tubules. During water absorption, the increase in signal intensity started at the bottom of a specimen and extended to the upper region; it maintained the localization pattern observed before hydration. The relationship between the local maximal signals in areas corresponding to the horn tubules and minimal signal intensities in areas corresponding to the intertubular horn was similar and maintained approximately a linear distribution. Conclusions and Clinical Relevance: Based on the premise that signal intensity reflects water content, hydration in the equine hoof wall during water absorption occurred concurrently in the tubules and intertubular horn, and there was maintenance of the original water gradients. This technique can be applied for the assessment of pathophysiologic changes in the hoof wall on the basis of its hydration properties.

Objective: To evaluate the precision and accuracy of assessing bone mineral density (BMD) by use of mean gray value (MGV) on digitalized and digital images of conventional and digital radiographs, respectively, of ex vivo bovine and equine bone specimens in relation to the gold-standard technique of dual-energy x-ray absorptiometry (DEXA). Sample: Left and right metatarsal bones from 11 beef cattle and right femurs from 2 horses. Procedures: Bovine specimens were imaged by use of conventional radiography, whereas equine specimens were imaged by use of computed radiography (digital radiography). Each specimen was subsequently scanned by use of the same DEXA equipment. The BMD values resulting from each DEXA scan were paired with the MGVs obtained by use of software on the corresponding digitalized or digital radiographic image. Results: The MGV analysis of digitalized and digital x-ray images was a precise (coefficient of variation, 0.1 and 0.09, respectively) and highly accurate method for assessing BMD, compared with DEXA (correlation coefficient, 0.910 and 0.937 for conventional and digital radiography, respectively). Conclusions and Clinical Relevance: The high correlation between MGV and BMD indicated that MGV analysis may be a reliable alternative to DEXA in assessing radiographic bone density. This may provide a new, inexpensive, and readily available estimate of BMD.