OBJECTIVE To quantify fatigue-induced electromyographic changes in hind limb muscles in horses. ANIMALS 8 Thoroughbreds. PROCEDURES The left and right hind limb longissimus dorsi, tensor fasciae latae, gluteus medius, and biceps femoris muscles were instrumented for surface electromyography. Hoof strain gauges were attached to confirm stride cycle. Each horse was galloped on a treadmill (grade, 3%) at a constant speed (12.6 to 14.7 m/s) to achieve fatigue after approximately 360 seconds. Before and after this exercise, the horses were trotted at 3.5 m/s. At 30-second intervals during galloping an integrated electromyography (iEMG) value for a stride and the median frequency of muscle discharge (MF) in each limb were measured. The mean of stride frequency (SF), iEMG value, and MF of 5 consecutive strides at the start and end of galloping for the lead and trailing limbs were compared. For trotting, these variables were compared at 60 seconds before and after galloping. RESULTS The mean ± SD value for SF decreased over time (2.14 ± 0.06 to 2.05 ± 0.07 stride/s). In both the lead and trailing limbs, fatigue decreased the iEMG values of the gluteus medius and biceps femoris muscles but not those of the longissimus dorsi and tensor fasciae latae muscles. The MF did not change for any muscle during galloping with fatigue. The SF, iEMG value, and MF did not change during trotting with fatigue. CONCLUSIONS AND CLINICAL RELEVANCE Fatigue induced by high-speed galloping decreased the gluteus medius and biceps femoris muscles' iEMG values in Thoroughbreds. Fatigue of these less fatigue-resistant hind limb muscles would affect a horse's speed.

OBJECTIVE To determine the in vitro effect of calcitriol on indicators of immune system function in endotoxin-primed blood samples from healthy dogs. SAMPLE Blood samples from 6 healthy adult dogs. PROCEDURES Leukocytes were primed by incubation of blood samples with lipopolysaccharide (LPS; endotoxin) or PBS solution (unprimed control group) for 1 hour. Following priming, blood samples were incubated with calcitriol (2 × 10(−7)M) or ethanol (control substance) for 24 hours. After sample incubation, LPS-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) was measured with a canine-specific multiplex assay, and apoptosis and toll-like receptor 4 (TLR4) expression were evaluated via flow cytometry. RESULTS LPS stimulation of unprimed leukocytes but not endotoxin-primed leukocytes resulted in a significant increase in TNF and IL10 production, confirming the presence of endotoxin tolerance in dogs in vitro. Endotoxin priming significantly increased neutrophil viability with no effect on lymphocyte viability or TLR4 expression by neutrophils and monocytes. Calcitriol exposure significantly decreased LPS-stimulated production of TNF by unprimed and endotoxin-primed leukocytes. Conversely, calcitriol exposure had no effect on IL10 production by unprimed leukocytes but did significantly increase IL10 production by endotoxin-primed leukocytes. Calcitriol had no significant effect on the degree of neutrophil or lymphocyte apoptosis, nor was neutrophil and monocyte TLR4 expression affected in unprimed or endotoxin-primed leukocytes. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in unprimed and endotoxin-primed canine leukocytes in vitro, without compromising neutrophil and monocyte TLR4 expression or altering the viability of neutrophils and lymphocytes in canine blood samples.

OBJECTIVE To compare bronchoalveolar lavage (BAL) accomplished by use of a bronchoscopic (B-BAL) and a nonbronchoscopic (NB-BAL) technique in healthy cats. ANIMALS 12 healthy cats. PROCEDURES Two BALs were performed in a randomized order 2 weeks apart in each cat. Cats were anesthetized, and a 2.9-mm fiberoptic bronchoscope (B-BAL) or 8F red rubber catheter (NB-BAL) was wedged in a bronchus. Two 5-mL aliquots of saline (0.9% NaCl) solution were infused into the left and right caudal lung fields and aspirated manually with a 20-mL syringe. Proportion of BAL fluid (BALF) retrieved, depth of wedging, and anesthetic complications were recorded. Total nucleated cell count, differential cell count, and semiquantitative scores of cytologic slide quality were determined for all BALF samples. Results were compared with ANOVAs and Wilcoxon signed rank tests. RESULTS Proportion of retrieved BALF and depth of wedging were significantly greater for B-BAL than NB-BAL. Differential cell counts and cytologic slide quality did not differ significantly between techniques. Complications included transient hemoglobin desaturation (24/24 [100%] BALs) and prolonged anesthetic recovery time (4/24 [17%] BALs). Anesthetic recovery scores did not differ significantly between techniques. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that NB-BAL was noninferior to B-BAL with regard to ease of performance, anesthetic variables, and cytologic slide quality for cats without clinical respiratory tract disease.

OBJECTIVE To characterize expression profiles of circulating microRNAs via genome-wide sequencing for dogs with congestive heart failure (CHF) secondary to myxomatous mitral valve degeneration (MMVD). ANIMALS 9 healthy client-owned dogs and 8 age-matched client-owned dogs with CHF secondary to MMVD. PROCEDURES Blood samples were collected before administering cardiac medications for the management of CHF. Isolated microRNAs from plasma were classified into microRNA libraries and subjected to next-generation sequencing (NGS) for genome-wide sequencing analysis and quantification of circulating microRNAs. Quantitative reverse transcription PCR (qRT-PCR) assays were used to validate expression profiles of differentially expressed circulating microRNAs identified from NGS analysis of dogs with CHF. RESULTS 326 microRNAs were identified with NGS analysis. Hierarchical analysis revealed distinct expression patterns of circulating microRNAs between healthy dogs and dogs with CHF. Results of qRT-PCR assays confirmed upregulation of 4 microRNAs (miR-133, miR-1, miR-let-7e, and miR-125) and downregulation of 4 selected microRNAs (miR-30c, miR-128, miR-142, and miR-423). Results of qRT-PCR assays were highly correlated with NGS data and supported the specificity of circulating microRNA expression profiles in dogs with CHF secondary to MMVD. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested that circulating microRNA expression patterns were unique and could serve as molecular biomarkers of CHF in dogs with MMVD.

OBJECTIVE To evaluate the coding regions of ADAMTS17 for potential mutations in Chinese Shar-Pei with a diagnosis of primary open-angle glaucoma (POAG), primary lens luxation (PLL), or both. ANIMALS 63 Shar-Pei and 96 dogs of other breeds. PROCEDURES ADAMTS17 exon resequencing was performed on buccal mucosal DNA from 10 Shar-Pei with a diagnosis of POAG, PLL, or both (affected dogs). A candidate causal variant sequence was identified, and additional dogs (53 Shar-Pei [11 affected and 42 unaffected] and 95 dogs of other breeds) were genotyped for the variant sequence by amplified fragment length polymorphism analysis. Total RNA was extracted from ocular tissues of 1 affected Shar-Pei and 1 ophthalmologically normal Golden Retriever; ADAMTS17 cDNA was reverse transcribed and sequenced, and ADAMTS17 expression was evaluated by quantitative reverse-transcription PCR assay. RESULTS All affected Shar-Pei were homozygous for a 6-bp deletion in exon 22 of ADAMTS17 predicted to affect the resultant protein. All unaffected Shar-Pei were heterozygous or homozygous for the wild-type allele. The variant sequence was significantly associated with affected status (diagnosis of POAG, PLL, or both). All dogs of other breeds were homozygous for the wild-type allele. The cDNA sequencing confirmed presence of the expected variant mRNA sequence in ocular tissue from the affected dog only. Gene expression analysis revealed a 4.24-fold decrease in the expression of ADAMTS17 in ocular tissue from the affected dog. CONCLUSIONS AND CLINICAL RELEVANCE Results supported that the phenotype (diagnosis of POAG, PLL, or both) is an autosomal recessive trait in Shar-Pei significantly associated with the identified mutation in ADAMTS17.

OBJECTIVE To determine the optimal anatomic site and directional aim of a penetrating captive bolt (PCB) for euthanasia of goats. SAMPLE 8 skulls from horned and polled goat cadavers and 10 anesthetized horned and polled goats scheduled to be euthanized at the end of a teaching laboratory. PROCEDURES Sagittal sections of cadaver skulls from 8 horned and polled goats were used to determine the ideal anatomic site and aiming of a PCB to maximize damage to the midbrain region of the brainstem for euthanasia. Anatomic sites for ideal placement and directional aiming were confirmed by use of 10 anesthetized horned and polled goats. RESULTS Clinical observation and postmortem examination of the sagittal sections of skulls from the 10 anesthetized goats that were euthanized confirmed that perpendicular placement and firing of a PCB at the intersection of 2 lines, each drawn from the lateral canthus of 1 eye to the middle of the base of the opposite ear, resulted in consistent disruption of the midbrain and thalamus in all goats. Immediate cessation of breathing, followed by a loss of heartbeat in all 10 of the anesthetized goats, confirmed that use of this site consistently resulted in effective euthanasia. CONCLUSIONS AND CLINICAL RELEVANCE Damage to the brainstem and key adjacent structures may be accomplished by firing a PCB perpendicular to the skull over the anatomic site identified at the intersection of 2 lines, each drawn from the lateral canthus of 1 eye to the middle of the base of the opposite ear.

The use of antibiotics in livestock production in North America and possible association with elevated abundance of detectable antimicrobial resistance genes (ARG) is a growing concern. Real-time, quantitative PCR (RT-qPCR) was used to determine the relative abundance and diversity of ARG in fecal composite and catch basin samples from 4 beef feedlots in Alberta. Samples from a surrounding waterway and municipal wastewater treatment plants were also included to compare the ARG profile of urban environments and fresh water with that of feedlots. The relative abundance of 18 resistance genes across 5 antibiotic families including sulfonamides, tetracyclines, macrolides, fluoroquinolones, and β-lactams was examined. Sulfonamide, fluoroquinolone, and β-lactam resistance genes predominated in wastewater treatment samples, while tetracycline resistance genes predominated in cattle fecal composite samples. These results reflect the types of antibiotic that are used in cattle versus humans, but other factors such as co-selection of ARG and variation in the composition of bacterial communities associated with these samples may also play a role.

To obtain immunogenic conjugate antigens, adipic acid dihydrazide (ADH), as a bridge, and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimidehydrochloride (EDAC), as a coupling agent, were used to conjugate the purified fusion protein, clumping factor A-fibronectin binding protein ClfA A-FnBPA, and type 5 capsular polysaccharide (CP5). The conjugates were mixed with an adjuvant, and mice were immunized 3 times and challenged with Staphylococcus aureus 1 week later. Antibody titers were determined by indirect enzyme-linked immunosorbent assay (ELISA). At 14 days after the first immunization, antibodies against the purified protein and conjugate were detected; after 28 days, antibody levels increased; and a week after the third immunization, antibody levels continued to increase. However, the conjugate antibody titers were higher than those of the purified protein during the study, and no IgG antibodies against purified CP5 were detected during the entire experiment. The protection rate increased to 90% in the conjugate group, indicating that the conjugate imparts a relatively higher protective efficacy than the purified protein and purified CP5.

OBJECTIVE To validate that dogs become hypocoagulable following rattlesnake envenomation and to determine whether thromboelastographic abnormalities are correlated with envenomation severity for dogs bitten by rattlesnakes native to southern California. ANIMALS 14 dogs with observed or suspected rattlesnake envenomation (envenomated dogs) and 10 healthy control dogs. PROCEDURES For each dog, a citrate-anticoagulated blood sample underwent kaolin-activated thromboelastography. For each envenomated dog, a snakebite severity score was assigned on the basis of clinical findings, and prothrombin time, activated partial thromboplastin time, and platelet count were determined when the attending clinician deemed it necessary and owner finances allowed. RESULTS For 12 of 14 envenomated dogs, the thromboelastographically determined clot strength was below the 25th percentile for the clot strength of control dogs, which was indicative of a hypocoagulable state. No envenomated dog had thromboelastographic results indicative of a hypercoagulable state. One envenomated dog had a prolonged prothrombin time, but the activated partial thromboplastin time and all thromboelastographic variables were within the respective reference ranges for that dog. Seven of 13 envenomated dogs were thrombocytopenic (platelet count, ≤ 170,000 platelets/μL). Snakebite severity score was negatively correlated with platelet count but was not correlated with any thromboelastographic variable. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that dogs generally become hypocoagulable following rattlesnake envenomation. Thromboelastography might provide an objective measure of the coagulation status of envenomated dogs and aid in the identification of dogs that are in a hypocoagulable state and in need of antivenin treatment prior to the onset of progressive clinical signs.

OBJECTIVE To quantify the effect of time and recumbency on CT measurements of lung volume and attenuation in healthy cats under general anesthesia. ANIMALS 8 healthy research cats. PROCEDURES Anesthetized cats were positioned in sternal recumbency for 20 minutes and then in left, right, and left lateral recumbency (40 minutes/position). Expiratory helical CT scan of the thorax was performed at 0 and 20 minutes in sternal recumbency and at 0, 5, 10, 20, 30, and 40 minutes in each lateral recumbent position. For each lung, CT measurements of lung volume and attenuation and the extent of lung areas that were hyperaerated (−1,000 to −901 Hounsfield units [HU]), normoaerated (−900 to −501 HU), poorly aerated (−500 to −101 HU), or nonaerated (−100 to +100 HU [indicative of atelectasis]) were determined with a semiautomatic threshold-based technique. A restricted maximum likelihood analysis was performed. RESULTS In lateral recumbency, the dependent lung had significantly greater attenuation and a lower volume than the nondependent lung. Within the dependent lung, there was a significantly higher percentage of poorly aerated lung tissue, compared with that in the nondependent lung. These changes were detected immediately after positioning the cats in lateral recumbency and remained static with no further significant time-related change. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that once anesthetized healthy cats were positioned in lateral recumbency, the dependent lung lobes underwent a rapid reduction in lung volume and increase in lung attenuation that did not progress over time, predominantly attributable to an increase in poorly aerated lung tissue.