A fast, sensitive, and specific reverse-transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay was developed that involved a single tube and a 1-step reaction for detecting infectious bursal disease virus (IBDV). Four specific primers were used for amplification of the VP2 gene of IBDV. The amplified LAMP products were detected by DNA electrophoresis and by direct observation with the naked eye in the presence of SYBR Green I. The sensitivity of RT-LAMP was determined to be 0.01 fg of IBDV viral RNA. This assay for IBDV is more sensitive than the conventional RT-polymerase chain reaction assay, which has a detection limit of 1 ng. The LAMP assay was also assessed for specificity and was found to precisely discriminate between positive and negative test samples. This newly established LAMP assay, combined with RT, is a practical diagnostic tool because IBDV-infected and uninfected clinical samples collected from an experimental farm could be discriminated. Full verification of a sample’s IBDV status was obtained within 40 min of extraction of the viral RNA, which could then be directly added to the RT-LAMP reaction mixture.

Objective—To determine values for total body water (TBW), extracellular fluid volume (ECFV), intracellular fluid volume (ICFV), and plasma volume (PV) in healthy neonatal (< 24 hours old) foals and to create a multifrequency bioelectrical impedance analysis (MF-BIA) model for use in neonatal foals. Animals—7 healthy neonatal foals. Procedures—Deuterium oxide (0.4 g/kg, IV), sodium bromide (30 mg/kg, IV), and Evans blue dye (1 mg/kg, IV) were administered to each foal. Plasma samples were obtained following an equilibration period, and the TBW, ECFV, ICFV, and PV were calculated for each foal. An MF-BIA model was created by use of morphometric measurements from each foal. Results—Mean ± SD values were obtained for TBW (0.744 ± 0.024 L/kg), ICFV (0.381 ± 0.018 L/kg), ECFV (0.363 ± 0.014 L/kg), and PV (0.096 ± 0.015 L/kg). The 95% limits of agreement between the MF-BIA and indicator dilution techniques were within ± 2 L for TBW and ECFV. Conclusions and Clinical Relevance—Fluid volumes in neonatal foals were found to be substantially larger than fluid volumes in adult horses. Multifrequency bioelectrical impedance analysis may be a useful technique for predicting TBW, ICFV, and ECFV in neonatal foals.

Objective—To determine effects of various concentrations of retinoic acid (RA) or a synthetic RA receptor antagonist (LE135) on equine chondrocytes or bone marrow—derived equine mesenchymal stem cells (BMDMSCs) in monolayer cultures. Sample—Articular cartilage and BMDMSCs from 5 clinically normal horses. Procedures—Monolayers of chondrocytes cultured in standard media and of BMDMSCs cultured in chondrogenic media were treated with RA at concentrations of 0, 0.1, 1, or 10μM or LE135 at concentrations of 0, 0.1, 1, or 10μM on day 0. On days 7 and 14, samples were analyzed for DNA concentration, chondrocyte morphology or features consistent with chondrogenesis (ie, chondral morphology [scored from 0 to 4]), and gene expression of collagen type Ia (CI), collagen type II (CII), and aggrecan. Results—Chondrocytes treated with RA had more mature chondral morphology (range of median scores, 3.0 to 4.0) than did untreated controls (range of median scores, 0.5 to 0.5). Chondrocytes treated with LE135 did not sustain chondrocyte morphology. All BMDMSCs had evidence of chondral morphology or high CII:CI ratio. Retinoic acid (1 or 10μM) or LE135 (10μM) treatment decreased DNA content of BMDMSC cultures. At 0.1 and 1μM concentrations, LE135 weakly but significantly increased chondral morphology scores, compared with untreated controls, but lack of aggrecan expression and lack of increased CII:CI ratio, compared with that of controls, did not affect chondrogenesis. Conclusions and Clinical Relevance—RA promoted maturation and hypertrophy in chondrocytes but not BMDMSCs in monolayer cultures. Deficiency or blockade of RA may prevent hypertrophy and maturation of differentiated chondrocytes.

Objective—To determine the mode of inheritance for cerebellar abiotrophy (CA), a neurologic disease in Arabians. Animals—804 Arabians, including 29 horses (15 males and 14 females) with CA. Procedures—Most horses (n = 755) belonged to 1 of 4 paternal families. Among the 29 CA-affected horses, all had clinical signs consistent with the disease; the disease was confirmed histologically following euthanasia in 8 horses. From the pedigree information, inbreeding coefficients were calculated for 16 affected horses and compared with coefficients for a subgroup of 16 unaffected horses. Complex segregation analysis was used to determine the effect of a putative Mendelian locus on the development of the disease and the probable mode of inheritance of CA. Results—The mean inbreeding coefficient was 0.0871 for CA-affected and unaffected horses, suggesting that all of the Arabians were inbred to the same degree and that affected horses were not more inbred than were unaffected horses. Results of the complex segregation analysis were consistent with a single Mendelian autosomal recessive mode of inheritance. Conclusions and Clinical Relevance—Knowledge of the mode of inheritance of CA should help breeders to make informed decisions regarding the selection of animals for mating when closely related horses have developed CA or produced CA-affected foals.

Objective: To noninvasively evaluate physiologic postprandial adaptations of the heart in snakes. Animals: 6 juvenile Paraguay anacondas (Eunectes notaeus). Procedures: The heart of each anaconda was echocardiographically evaluated after food was withheld for 28 days as well as 3 and 10 days after feeding. Physical measurements included body length, weight, and circumference at the level of the heart. Echocardiographic measurements included heart rate and 2-D total and internal ventricular area. From these measurements, total ventricular volume as well as the myocardial area as a surrogate of myocardial mass was calculated. Results: No significant changes in body length, weight, and circumference were found. Significant increases in heart rate (from 45 to 58 beats/min), total ventricular volume (from 4.63 to 5.54 mL), and myocardial area (from 0.7 to 0.81 cm2) were detected 10 days after feeding, compared with results obtained prior to feeding after food had been withheld for 28 days. No pericardial effusion was detected at any time point. Conclusions and Clinical Relevance: Echocardiographic evaluation of the heart of anacondas was performed, and feeding resulted in concentric cardiac hypertrophy. Physiologic fluctuation of cardiac dimensions should be considered when cardiac imaging is performed in snakes.

Objective—To determine effects of duration and type of anesthetic on tear production in dogs. Animals—8 female Beagles. Procedures—Each dog was randomly allocated into 1 of 4 groups according to a Latin square design to receive anesthesia as follows: 1 hour with isoflurane, 1 hour with desflurane, 4 hours with isoflurane, and 4 hours with desflurane. Each dog was anesthetized with the selected inhalant 4 times during a 4-week period, with at least 5 days separating anesthetic episodes. Aqueous tear production was measured via the Schirmer I tear test at baseline and 10 minutes, 30 minutes, and 1 hour after induction of anesthesia as well as 2, 3, and 4 hours after induction for the 4-hour groups. Tear production was also measured after the dogs were standing after recovery from anesthesia and 2, 10, and 22 hours after recovery from anesthesia. Results—Aqueous tear production was significantly reduced in dogs during anesthesia and returned to baseline values immediately after recovery and until 10 hours after anesthesia in all treatment groups. Inhalant type and duration had no significant effect. Neither lateral recumbency nor left versus right eyes had a significant effect. Conclusions and Clinical Relevance—Results suggested that inhalant anesthetics did not reduce tear production after anesthesia and that longer-duration anesthesia did not cause decreased tear production, compared with shorter-duration anesthesia.

This study evaluated the analgesia effects of the epidural administration of 0.1 mg/kg bodyweight (BW) of morphine or 5 micrograms/kg BW of buprenorphine in ponies with radiocarpal joint synovitis. Six ponies were submitted to 3 epidural treatments: the control group (C) received 0.15 mL/kg BW of a 0.9% sodium chloride (NaCl) solution; group M was administered 0.1 mg/kg BW of morphine; and group B was administered 5 micrograms/kg BW of buprenorphine, both diluted in 0.9% NaCl to a total volume of 0.15 mL/kg BW administered epidurally at 10 s/mL. The synovitis model was induced by injecting 0.5 ng of lipopolysaccharide (LPS) in the left or right radiocarpal joint. An epidural catheter was later introduced in the lumbosacral space and advanced up to the thoracolumbar level. The treatment started 6 h after synovitis induction. Lameness, maximum angle of carpal flexion, heart rate, systolic arterial pressure, respiratory rate, temperature, and intestinal motility were evaluated before LPS injection (baseline), 6 h after LPS injection (time 0), and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after treatments. Although the model of synovitis produced clear clinical signs of inflammation, the lameness scores in group C were different from the baseline for only up to 12 h. Both morphine and buprenorphine showed a reduction in the degree of lameness starting at 0.5 and 6 h, respectively. Reduced intestinal motility was observed at 0.5 h in group M and at 0.5 to 1 h in group B. Epidural morphine was a more effective analgesic that lasted for more than 12 h and without side effects. It was concluded that morphine would be a valuable analgesic option to alleviate joint pain in the thoracic limbs in ponies.

Objective—To assess relationships of acetabular volume (AV), femoral head volume (FV), and portion of the femoral head within in the acetabulum (FVIA) with each other and with degrees of hip joint laxity and degenerative joint disease from youth to maturity in dogs predisposed to developing hip joint osteoarthritis (OA). Animals—46 mixed-breed half- or full-sibling hound-type dogs. Procedures—The distraction index (DI), AV, FV, FVIA, and degree of osteoarthritis (OA score) were quantified in 1 hip joint at 16, 32, and 104 weeks of age. Relationships among variables were evaluated within and between ages. Ratios corresponding to OA scores were compared within ages. Differences among 16-week ratios corresponding to 32-week OA scores and among 16- and 32-week ratios corresponding to 104-week OA scores were evaluated. Results—Significant positive relationships existed between FV and AV across ages as well as between FVIA/FV and FVIA/AV and between DI and OA score across and within most ages. Such relationships also existed within these variables across most ages. Negative relationships of DI and OA scores with FVIA/FV and FVIA/AV within and among all ages were significant. Sixteen-week AVs, FVs, and FVIAs were greater and FV/AVs and OA scores were less than 32- and 104-week values. The 32-week FVIA/FV was less than 16- and 104-week values, and the 32-week FVIA/AV was less than the 104-week value. The FVIA/FV and FVIA/AV were lower and the DI was higher with higher OA scores within and among most ages. Conclusions and Clinical Relevance—Structural volumes in lax canine hip joints changed predictably relative to each other during growth, despite degenerative changes. Measures developed in this study may augment current diagnosis and treatment strategies for hip dysplasia in dogs.

This study established a disease model and protocol for bacterial challenge with Escherichia coli serogroup O2 strain EC317 in Japanese quail. Five groups of 10 birds each were injected subcutaneously in the breast with 200 μL of a brain–heart infusion (BHI) culture containing 1 × 10(8), 1 × 10(7), 1 × 10(6), 1 × 10(5), or 1 × 10(4) colony-forming units/mL of the test organism, which had been isolated from a turkey with cellulitis and septicemia. Birds in a 6th group were controls that received sterile BHI alone. Localized lesions of cellulitis developed in all of the birds that received E. coli. The morbidity and mortality rates were highest (100%) in the birds receiving the highest dose of E. coli and decreased linearly with decreasing dose (P < 0.05). Severity of disease, including lesions of pericarditis and perihepatitis, was also directly proportional to the dose of E. coli. These findings indicate that this disease challenge protocol can be used to study disease resistance and immunologic consequences of contaminant exposure or other stressors in birds.

In this study the tonB2 gene was cloned from Actinobacillus pleuropneumoniae JL01 (serovar 1) and expressed as a glutathione-S-transferase (GST) fusion protein in Escherichia coli BL21(DE3). The GST fusion protein was recognized by antibodies in serum positive for A. pleuropneumoniae by Western blot analysis. Purified soluble GST-TonB2 was assessed for its ability to protect BALB/c mice against A. pleuropneumoniae infection. Mice were vaccinated with GST-TonB2 subcutaneously and challenged intraperitoneally with either approximately 4.0 × 10(5) colony-forming units (CFU) or approximately 1.0 × 10(6) CFU of A. pleuropneumoniae 4074. They were examined daily for 7 d after challenge. The survival rate of the TonB2-vaccinated mice was significant higher than that of the mice given recombinant GST or adjuvant alone. These results demonstrate that A. pleuropneumoniae TonB2 is immunogenic in mice and should be further assessed as a potential candidate for a vaccine against A. pleuropneumoniae infection. In addition, an indirect enzyme-linked immunosorbent assay (ELISA) based on the GST-TonB2 recombinant protein was developed. Compared with the ApxIVA ELISA, the TonB2 ELISA provided earlier detection of antibodies in pigs at various times after vaccination with A. pleuropneumoniae live attenuated vaccine. When compared with an indirect hemagglutination test, the sensitivity and specificity of the TonB2 ELISA were 95% and 88%, respectively. The TonB2 ELISA provides an alternative method for rapid serologic diagnosis of A. pleuropneumoniae infection through antibody screening, which would be especially useful when the infection status or serovar is unknown.