Objective- To assess kinetic 2-([(18)F]fluoro)-2-deoxy-d-glucose ((18)FDG) uptake in the brain of anesthetized healthy adult dogs by use of positron emission tomography (PET) and to determine whether (18)FDG uptake differs among anatomic regions of the brain. Animals- 5 healthy Beagles. Procedures- Each isoflurane-anesthetized dog was administered (18)FDG IV (dose range, 3.0 to 5.2 mCi), and PET data were acquired for 2 hours. A CT scan (without contrast agent administration) was performed to allow more precise neuroanatomic localization. Defined regions of interest within the brain were drawn on reconstructed image data. Standard uptake values (SUVs) for (18)FDG were calculated to generate time-activity curves and determine time to peak uptake. Results- Time-activity curve analysis identified 4 regional uptake patterns: olfactory, gray matter, white matter, and other (brainstem, cerebellum, and occipital and frontal regions). The highest maximum SUVs were identified in the olfactory bulbs and cerebral gray matter, and the lowest maximum SUV was identified in cerebral white matter. Mean time to peak uptake ranged from 37.8 minutes in white matter to 82.7 minutes in the olfactory bulbs. Conclusions and Clinical Relevance- Kinetic analysis of (18)FDG uptake revealed differences in uptake values among anatomic areas of the brain in dogs. These data provide a baseline for further investigation of (18)FDG uptake in dogs with immune-mediated inflammatory brain disease and suggest that (18)FDG-PET scanning has potential use for antemortem diagnosis without histologic analysis and for monitoring response to treatment. In clinical cases, a 1-hour period of PET scanning should provide sufficient pertinent data.

Objective-To determine associations of blood analysis variables and orbit and nasal planum surface temperatures with the onset and severity of Mycoplasma bovis pneumonia in calves. Animals-28 healthy calves. Procedures-Calves were challenged with M bovis (n = 24) on day 0 or not challenged (4). Blood samples were obtained for cardiac troponin I, CBC, and serum biochemical analyses on various days. Orbit and nasal planum surface temperatures were determined with infrared thermography on various days. Calves were euthanized, gross necropsies were performed, heart and lung samples were collected for histologic evaluation, and microbial cultures of lung samples were performed on day 14. Pneumonia severity was categorized as mild (< 10% lung consolidation) or moderate (≥ 10% lung consolidation). Associations between measured variables and severity of pneumonia or sample collection day were determined. Results-Plasma cardiac troponin I concentration for the 28 calves was significantly higher on day 14 than it was on day 0 or 7 (least squares mean, 0.02, 0, and 0 ng/mL, respectively). No other variables changed significantly during the study. No substantial gross or histologic abnormalities were identified in cardiac muscle samples. Day 14 plasma fibrinogen concentration was significantly different between calves with mild pneumonia and those with moderate pneumonia (mean, 0.44 and 0.74 g/dL, respectively). Calves with moderate pneumonia had significantly lower least squares mean surface temperature of the dorsal aspect of the nasal planum (18.7°C) versus calves with mild pneumonia (22.9°C). Conclusions and Clinical Relevance-Results indicated the evaluated variables had low value for assessment of bovine respiratory disease complex in calves.

Objective-To determine the accuracy of neurologic data, survey radiographic results, or both for localization of the site of thoracolumbar intervertebral disk herniation in dogs. Sample-338 dogs with surgically confirmed intervertebral disk herniation from disk spaces T10-11 to L6-7. Procedures-Medical records and archived survey radiographs were reviewed for each case. Data were analyzed with multivariable logistic regression models. Three models were fit to develop subsets of the data consisting of survey radiographic data, neurologic examination data, and a combination of survey radiographic and neurologic examination data. The resulting models were validated by evaluating predictive performance against a validation subset of the data. Results-Models incorporating survey radiographic data and a combination of survey radiographic and neurologic data had similar predictive ability and performed better than the model based solely on neurologic data but resulted in substantial errors in predictions. Conclusions and Clinical Relevance-A combination of neurologic examination data as recorded in the medical records and radiographic data did not enhance predictive performance of multivariable logistic regression models over models limited to radiographic data. Neurologic and radiographic findings should not be used to completely exclude areas in an abnormal spinal cord region from further evaluation with advanced imaging.

Objective-To ultrasonographically quantify experimentally induced effusion of the distal interphalangeal (DIP) joint of horses and compare results with those obtained with palpation. Sample-8 forelimbs from equine cadavers and forelimbs of 5 mares. Procedures-Preliminary ex vivo experiments were performed to validate the methods. Then, the DIP joints of the forelimbs of standing horses were serially distended with saline (0.9% NaCl) solution (1, 4, and 10 mL) by injection through an intra-articular catheter. Two ultrasonographers measured distension of the dorsal recess of the DIP joint, and 2 surgeons, who were not aware of the volume injected, graded joint effusion by palpation. Results-Intraobserver and interobserver repeatability was excellent for ultrasonographic measurements. Interobserver agreement for use of palpation to detect joint distension was moderate (κ = 0.45). There was an overall increase in the palpation distension grade with an increase in injected volume. Sensitivity for detection with palpation of larger volumes (4 and 10 mL) was high (92% and 100%, respectively). However, sensitivity was lower (57%) for detection with palpation of minimal distension (1 mL). Conclusions and Clinical Relevance-Although palpation provided a reliable clinical assessment of DIP joint effusion for volumes of 4 to 10 mL, ultrasonographic measurements were easy to obtain, more accurate, and able to detect smaller amounts of distension. This may be clinically relevant for the assessment of effusion of the DIP joint that can arise in horses with early osteoarthritis or infectious arthritis with concomitant soft tissue swelling that precludes accurate assessment with palpation.

Objective—To determine the skin temperature of the metacarpus in horses associated with the use of bandages and tendon boots, compared with the bare limb, at rest and after 20 minutes of lunging. Animals—10 adult horses. Procedures—Skin temperature on the bare metacarpus of both forelimbs was measured at rest and after lunging. Subsequently, a bandage was applied to the left metacarpus and a tendon boot to the right metacarpus and skin temperature was measured at rest and after lunging. Skin temperature was measured with fixed sensors and thermographically. Results—Mean ± SD skin temperatures of the bare metacarpi were 14.1 ± 2.4°C (left) and 14.1 ± 3.4°C (right) at rest, and 14.4 ± 1.8°C (left) and 13.6 ± 2.6°C (right) after exercise. Skin temperatures under the bandage were 15.3 ± 1.6°C at rest and 24.8 ± 3.6°C after exercise. Skin temperatures under the tendon boot were 15.3 ± 2.6°C at rest and 20.6 ± 2.9°C after exercise. Skin temperatures under the bandage and tendon boot were significantly higher after exercise than at rest. Skin temperatures at rest were not significantly different with a bare limb, bandage, or tendon boot. Conclusions and Clinical Relevance—Skin temperature of the metacarpus in horses increased significantly during exercise but not at rest when a bandage or tendon boot was used. The authors speculate that both a bandage and a tendon boot accelerate the warmup phase of exercise. Further research should focus on the effects of warmup and maximum exercise on the temperature of other anatomic structures such as tendons.

Objective—To establish a nonterminal semen collection method for use in captive Chilean rose tarantulas (Grammostola rosea) and to evaluate tools for investigating morphology and viability of spermatozoa. Animals—7 mature male Chilean rose tarantulas. Procedures—Each tarantula was anesthetized in a 500-mL induction chamber containing a cotton ball infused with 2 mL of isoflurane. Semen collection was performed by applying direct pressure to the palpal bulbs (sperm storage organs) located on the distal segment of the palpal limbs. Morphology of spermatozoa was examined by light microscopy and transmission and scanning electron microscopy. Propidium iodide and a fluorescent membrane-permeant nucleic acid dye were used to evaluate cell viability. Results—Semen was collected successfully from all 7 tarantulas. Microscopic examination of semen samples revealed coenospermia (spherical capsules [mean ± SD diameter, 10.3 ± 1.6 μm] containing many nonmotile sperm cells [mean number of sperm cells/capsule, 18.5 ± 3.8]). Individual spermatozoa were characterized by a spiral-shaped cell body (mean length, 16.7 ± 1.4 μm; mean anterior diameter, 1.5 ± 0.14 μm). Each spermatozoon had no apparent flagellar structure. The fluorescent stains identified some viable sperm cells in the semen samples. Conclusions and Clinical Relevance—The described technique allowed simple and repeatable collection of semen from Chilean rose tarantulas. Semen from this species was characterized by numerous spherical capsules containing many nonmotile spermatozoa in an apparently quiescent state. Fluorescent staining to distinguish live from dead spermatozoa appeared to be a useful tool for semen evaluation in this species.

Objective—To determine immunomodulatory effects of synbiotics administered in ovo on immune-related gene expression in adult chickens. Animals—30 Green-legged Partridgelike chickens. Procedures—On incubation day 12, eggs were injected with 3 synbiotics (Lactococcus lactis subsp lactis IBB SL1 with raffinose family oligosaccharides [RFOs; S1], Lactococcus lactis subsp cremoris IBB SC1 with RFOs [S2], and Lactobacillus acidophilus and Streptococcus faecium with lactose [S3]). Control eggs were injected with RFOs prebiotic or saline (0.9% NaCl) solution. Gene expression of 6 cytokines (interleukin [IL]-4, IL-6, IL-12p40, IL-18, interferon [IFN]-β, and IFN-γ) and 1 chemokine (IL-8) was analyzed in the cecal tonsils and spleen of 6-week-old chickens by means of reverse transcription quantitative PCR assays. Results—Gene expression for IL-4, IL-6, IFN-β, and IL-18 was significantly upregulated in the spleen of chickens in groups S2 and S3. In contrast, IL-12 expression was downregulated in group S2 and IFN-γ expression was downregulated in group S3. Expression of IL-8 did not change in chickens treated with synbiotics in ovo. Gene expression of all cytokines, except for IL-18, was downregulated in cecal tonsils. Conclusions and Clinical Relevance—In ovo administration of synbiotics activated the immune system in adult chickens. The intestinal immune system (cecal tonsils) had downregulation of expression for the cytokines evaluated, which indicated an increase in oral tolerance, whereas in the peripheral part of the immune system (spleen), expression of IL-4 and IL-6 was upregulated. Evaluation of immune-related gene expression patterns may be useful when monitoring the effectiveness of synbiotic selection with respect to immunobiotic properties.

Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered.

The objective of this study was to validate non-equilibrium gravitational field-flow fractionation (GrFFF), an immunotag-less method of sorting mesenchymal stem cells (MSCs) into subpopulations, for use with MSCs derived from equine muscle tissue, periosteal tissue, bone marrow, and adipose tissue. Cells were collected from 6 young, adult horses, postmortem. Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and left supragluteal subcutaneous adipose tissue. Aliquots of 800 3103 MSCs from each tissue source were separated and injected into a ribbon-like capillary device by continuous flow (GrFFF proprietary system). Cells were sorted into 6 fractions and absorbencies [optical density (OD)] were read. Six fractions from each of the 6 aliquots were then combined to provide pooled fractions that had adequate cell numbers to seed at equal concentrations into assays. Equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells were consistently sorted into 6 fractions that remained viable for use in further assays. Fraction 1 had more cuboidal morphology in culture when compared to the other fractions. Statistical analysis of the fraction absorbencies (OD) revealed a P-value of ,0.05 when fractions 2 and 3 were compared to fractions 1, 4, 5, and 6. It was concluded that non-equilibrium GrFFF is a valid method for sorting equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells into subpopulations that remain viable, thus securing its potential for use in equine stem cell applications and Veterinary medicine.

Mice were intranasally inoculated at various times to optimize the vaccination strategy with a new live candidate vaccine expressing the antigens CP39, FimA, PtfA, and ToxA of Pasteurella multocida and F1P2 of Bordetella bronchiseptica in an attenuated live Salmonella system to protect against progressive atrophic rhinitis (PAR). Sixty BALB/c mice were divided equally into 4 groups. The group A mice were vaccinated only at 12 wk of age, the group B mice received a primary vaccination at 9 wk of age and a booster at 12 wk of age, the group C mice received a primary vaccination at 6 wk of age and boosters at 9 and 12 wk of age, and the group D mice were inoculated intranasally with sterile phosphate-buffered saline as a control. The humoral and mucosal immune responses of groups A, B, and C increased significantly compared with those of the control group. Expression of the cytokines interleukin-4 and interferon-g in splenocytes also increased significantly. In addition, the group B mice exhibited significantly fewer gross lesions in lung tissue compared with the other vaccinated groups after challenge with a virulent P. multocida strain. These results indicate that a strategy of double intranasal vaccination can optimize protection against PAR.