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.

Pregnancy in many mammals, including mares, is associated with physiological changes that are reflected in hematological and biochemical profiles. Understanding those physiological changes and differentiating them from pathological changes is imperative for providing care and medical therapy in pregnant mares. Our objective was to compare normal hematological and biochemical profiles in healthy non-pregnant as well as healthy pregnant mares during the 1st and 2nd trimesters of pregnancy. Blood was collected by jugular venipuncture into ethylenediaminetetraacetic acid and serum tubes. Whole blood was analyzed using an ADVIA 120 hematologic analyzer and serum was analyzed using a Beckman Coulter AU5400. Statistical differences were detected using analysis of variance (ANOVA) and independent Student's t-test; P < 0.05 was considered significant. Results demonstrated higher red blood cell counts and hemoglobin concentrations and lower eosinophil counts (P < 0.001) in non-pregnant mares (n = 32) compared with pregnant mares at both 1st (n = 25) and 2nd (n = 17) trimesters. Biochemical analysis showed a significant decrease (P < 0.001) in albumin and blood urea nitrogen (P < 0.001) in the 2nd trimester and higher potassium levels (P = 0.03) in pregnant mares. Understanding such physiological changes is imperative to providing optimum care and medical treatment in mares. These data will assist clinicians to better evaluate and treat pregnant mares.

OBJECTIVE To evaluate the effect of an immunotherapeutic product on concentrations of anti–Pythium insidiosum antibodies in dogs. ANIMALS 7 healthy hound-crossbreds. PROCEDURES Antibody concentrations were evaluated before (day 0) and after administration of the immunotherapeutic product. The immunotherapeutic product was administered on days 0, 7, and 21. Serum was obtained on days 0, 7, 14, 21, 28, 35, 42, 49, and 56. Anti–P insidiosum antibody concentrations were measured and reported as the percentage positivity relative to results for a strongly positive control serum. RESULTS Mean ± SD percentage positivity before administration of the immunotherapeutic product was 7.45 ± 3.02%. There was no significant change in anti–P insidiosum antibody concentrations after administration of the product, with percentage positivity values in all dogs remaining within the range expected for healthy dogs (3% to 15%). CONCLUSIONS AND CLINICAL RELEVANCE Administration of the immunotherapeutic product to healthy dogs in accordance with the manufacturer's suggested protocol did not induce a significant change in anti–P insidiosum antibody concentrations. These results suggested that administration of the immunotherapeutic product may not interfere with postadministration serologic monitoring. However, further investigations will be required to determine whether there is a similar effect in naturally infected dogs.

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 optimal protocol for acquisition of CT images of the dentition in alpacas. ANIMALS 3 healthy adult male alpacas. PROCEDURES Each alpaca was anesthetized with an IM injection of a combination of ketamine, xylazine, and butorphanol and positioned in sternal recumbency on the CT couch with its legs folded in a natural cush position and its head positioned within the isocenter of the gantry of a 64-slice CT scanner. Images were acquired by means of 6 protocols (sequential and helical modes at slice thicknesses of 1.25, 2.5, and 5 mm). Five images (2 molar, 2 premolar, and mandibular incisor teeth) were selected from each protocol for evaluation by 3 veterinary radiologists. For each image, tooth root visibility and sharpness and image noise artifact were subjectively evaluated on a 3-point scoring system. RESULTS Slice thickness significantly affected tooth root visibility and tooth root sharpness but did not affect image noise artifact. Acquisition mode significantly affected tooth root visibility and tooth root sharpness as well as image noise artifact. Tooth root visibility and sharpness did not differ significantly between the helical and sequential images when the slice thickness was 1.25 mm. Image noise artifact was greater for helical images than sequential images but did not differ by slice thickness within either acquisition mode. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that for a 64-slice CT scanner, the optimal protocol for the acquisition of CT images of the dentition in alpacas was a sequential scan with a slice thickness of 1.25 mm.

OBJECTIVE To use high-field and low-field MRI to describe the anatomy of the proximal portion of the tarsal region (proximal tarsal region) of nonlame horses. SAMPLE 25 cadaveric equine tarsi. PROCEDURES The proximal portion of 1 tarsus from each of 25 nonlame horses with no history of tarsal lameness underwent high-field (1.5-T) and low-field (0.27-T) MRI. Resulting images were used to subjectively describe the anatomy of that region and obtain measurements of the collateral ligaments of the tarsocrural joint. RESULTS Long and short components of the lateral and medial collateral ligaments of the tarsocrural joint were identified. Various bundles of the short collateral ligaments were difficult to delineate on low-field images. Ligaments typically had low signal intensity in all sequences; however, multiple areas of increased signal intensity were identified at specific locations in most tarsi. This signal intensity was attributed to focal magic angle effect associated with orientation of collagen fibers within the ligaments at those locations. Subchondral bone of the distal aspect of the tibia was uniform in thickness, whereas that of the medial trochlear ridge of the talus was generally thicker than that of the lateral trochlear ridge. In most tarsi, subchondral bone of the talocalcaneal joint decreased in thickness from proximal to distal. CONCLUSIONS AND CLINICAL RELEVANCE Results generated in this study can be used as a reference for interpretation of MRI images of the proximal tarsal region in horses.

Pseudomonas aeruginosa is an important animal pathogen and contributes to hemorrhagic pneumonia in mink. Between April 2011 and December 2016, samples of lung, liver, and spleen were collected from mink with this disease on 11 mink farms in 5 Chinese provinces. From these samples, we obtained 98 isolates of P. aeruginosa that belonged to 5 serotypes: G (n = 58), I (n = 15), C (n = 8), M (n = 5), and B (n = 2); 10 isolates were not typeable (10/98). More than 90% of the isolates formed biofilms, and 85% produced slime. All 98 isolates were resistant to 10 antibiotics (oxacillin, ampicillin, penicillin G, amoxicillin, ceftriaxone, cefazolin, cefaclor, tilmicosin, tildipirosin, and sulfonamide). However, almost all were susceptible to gentamicin, polymyxin B, and amikacin. We identified 56 unique genotypes by pulsed-field gel electrophoresis. These findings have revealed genetic diversity and high antimicrobial resistance in P. aeruginosa isolated from mink with hemorrhagic pneumonia and will facilitate the prevention and control of this disease.

Genus Artemisia occurs as a hardy plant and has a wide range of culinary and medicinal features. In this study, we aimed to describe the melanin inhibitory activity of one Artemisia species, i.e., Artemisia capillaris Thunb. Ethanol extracts of fermented Artemisia capillaris (Art.EtOH.FT) and non-fermented Artemisia capillaris (Art.EtOH.CT) were tested for their ability to inhibit tyrosinase activity and melanin pigmentation. Both extracts showed dose-dependent inhibition against alpha-melanocyte stimulating hormone−stimulated melanin formation and tyrosinase activity, without cytotoxicity. At 100 ㎍/mL, both extracts showed greater inhibition than kojic acid, the positive control. Protein expressions of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) at the transcriptional level were determined by using real-time and semi-quantitative polymerase chain reaction. To complete the mechanistic study, presences of upstream elements of MITF, the phosphorylated-extracellular signal-regulated kinase (p-ERK), and phosphorylated−mitogen-activated protein kinase kinase (p-MEK) were confirmed by using western blot analysis. Expressions of p-TYR, p-TRP-1 and p-TRP2, downstream factors for p-ERK and p-MITF, were translationally inhibited by both extracts. Art.EtOH.FT induced more potent effects than Art.EtOH.CT, especially signal transduction effects. In summary, Artemisia capillaris extracts appear to act as potent hypopigmentation agents.

A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, CD3+ CD4+ , CD3+ CD8+ , CD3+ CD4+ CD8+ T-lymphocytes, and PRRSVspecific interferon (IFN)-γ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages (CD172α+ PRRSV-N+ PAM) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of CD172α+ PRRSV-N+ PAM (p 0.05) as well as with macroscopic lung lesion (p 0.01). Plasma and tissue viral loads had significant (p 0.05) negative correlations with CD3+ CD4+ CD8+ T-lymphocyte percentage in PBMC. Frequencies of CD3+ CD8+ and CD3+ CD4+ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p

Canine coronavirus is a single-stranded RNA virus that causes enteritis in dogs of any age. Coronaviral enteritis is seldom definitively diagnosed, since it is usually much less severe than many other types of enteritis and is self-limiting. Conventional diagnostics for the canine coronaviral enteritis such as polymerase chain reaction (PCR), virus isolation, and electron microscopic examination are inappropriate for small animal clinics due to the complicated experimental processes involved. Therefore, a commercially available lateral flow test kit based on chromatographic immunoassay techniques was tested to evaluate its performance as a first-line diagnostic test kit that could be used in clinics. The coronavirus antigen test kit detected canine coronavirus-infected dogs with 93.1% sensitivity and 97.5% specificity. The detection limit of the test kit was between 1.97 × 104 /mL and 9.85 × 103 /mL for samples with a 2- fold serial dilution from 1.25 × 106 TCID50 (TCID50, 50% tissue culture infectious dose). Additionally, the test kit had no cross-reactivity with canine parvovirus, distemper virus, or Escherichia coli. Overall, the commercially available test kit showed good diagnostic performance in a clinical setting, with results similar to those from PCR, confirming their potential for convenient and accurate use in small animal clinics.