OBJECTIVE To evaluate effects of various concentrations of collagenase and dimethyl sulfoxide (DMSO) on yield of equine adipose-derived multipotent stromal cells (ASCs) before and after cryopreservation. SAMPLE Supragluteal subcutaneous adipose tissue from 7 Thoroughbreds. PROCEDURES Tissues were incubated with digests containing 0.1%, 0.05%, or 0.025% type I collagenase. Part of each resulting stromal vascular fraction was cryopreserved in 80% fetal bovine serum (FBS), 10% DMSO, and 10% Dulbecco modified Eagle medium F-12 and in 95% FBS and 5% DMSO. Half of each fresh and cryopreserved heterogeneous cell population was not immunophenotyped (unsorted) or was immunophenotyped for CD44+, CD105+, and major histocompatability complex class II (MHCII; CD44+-CD105+-MHCII+ cells and CD44+-CD105+-MHCII− cells). Cell proliferation (cell viability assay), plasticity (CFU frequency), and lineage-specific target gene and oncogene expression (reverse transcriptase PCR assays) were determined in passage 1 cells before and after culture in induction media. RESULTS Digestion with 0.1% collagenase yielded the highest number of nucleated cells. Cell surface marker expression and proliferation rate were not affected by collagenase concentration. Cryopreservation reduced cell expansion rate and CD44+-CD105+-MHCII− CFUs; it also reduced osteogenic plasticity of unsorted cells. However, effects appeared to be unrelated to DMSO concentrations. There were also variable effects on primordial gene expression among cell isolates. CONCLUSIONS AND CLINICAL RELEVANCE Results supported the use of 0.1% collagenase in an adipose tissue digest and 5% DMSO in cryopreservation medium for isolation and cryopreservation, respectively, of equine ASCs. These results may be used as guidelines for standardization of isolation and cryopreservation procedures for equine ASCs.

The aim of this study was to determine the efficacy and pharmacokinetics of bupivacaine in combination with epinephrine or dexmedetomidine after intraperitoneal administration in cats undergoing ovariohysterectomy. Sixteen healthy adult cats (3.3 ± 0.6 kg) were included in a prospective, randomized, masked clinical trial after obtaining owners' consent. Anesthetic protocol included buprenorphine-propofol-isoflurane. Meloxicam [0.2 mg/kg body weight (BW)] was administered subcutaneously before surgery. Cats were randomly divided into 2 groups to receive 1 of 2 treatments. Intraperitoneal bupivacaine 0.25% (2 mg/kg BW) was administered with epinephrine (BE group; 2 μg/kg BW) or dexmedetomidine (BD group; 1 μg/kg BW) before ovariohysterectomy (n = 8/group). A catheter was placed in the jugular vein for blood sampling. Blood samples were collected for up to 8 h after bupivacaine was administered. Plasma concentrations and pharmacokinetics of bupivacaine were determined using liquid chromatography tandem mass spectrometry (LC-MS/MS) and non-compartmental model, respectively. Pain was evaluated using the UNESP-Botucatu multidimensional composite pain scale (MCPS), the Glasgow composite feline pain scale (GPS), and a dynamic visual analog scale up to 8 h after extubation. Rescue analgesia was provided with buprenorphine if MCPS was ≥ 6. Repeated measures linear models were used for analysis of pain and sedation scores (P < 0.05). Maximum bupivacaine plasma concentrations (Cmax) for BE and BD were 1155 ± 168 ng/mL and 1678 ± 364 ng/mL (P = 0.29) at 67 ± 13 min (Tmax) and 123 ± 59 min (P = 0.17), respectively. Pharmacokinetic parameters and pain scores were not different between treatments (P > 0.05). One cat in the BE group received rescue analgesia (P = 0.30). Intraperitoneal bupivacaine with epinephrine or dexmedetomidine produced concentrations below toxic levels and similar analgesic effects. It is therefore safe to administer these drug combinations in cats undergoing ovariohysterectomy.

OBJECTIVE To identify whether age, sex, or breed is associated with crown height of the left and right maxillary first molar tooth (M1) measured on CT images, to develop a mathematical model to determine age of horses by use of M1 crown height, and to determine the correlation between M1 crown height measured on radiographic and CT images. SAMPLE CT (n = 735) and radiographic images (35) of the heads of horses. PROCEDURES Crown height of left and right M1 was digitally measured on axial CT views. Height was measured on a lateral radiographic image when available. Linear regression analysis was used to identify factors associated with crown height. Half the data set was subsequently used to generate a regression model to predict age on the basis of M1 crown height, and the other half was used to validate accuracy of the predictions. RESULTS M1 crown height decreased with increasing age, but the rate of decrease slowed with increasing age. Height also differed by sex and breed. The model most accurately reflected age of horses < 10 years old, although age was overestimated by a mean of 0.1 years. The correlation between radiographic and CT crown height of M1 was 0.91; the mean for radiographic measurements was 2.5 mm greater than for CT measurements. CONCLUSIONS AND CLINICAL RELEVANCE M1 crown height can be used to predict age of horses. Results for CT images correlated well with those for radiographic images. Studies are needed to develop a comparable model with results for radiographic images.

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.

In this study, it was aimed to investigate the presence and the prevalence of Enterococcus faecalis and Enterococcus faecium strains isolated from the urine and stool samples. A total of 500 routine urine and feces samples were used for testing as the study materials, and a total of 349 Enterococcus spp. were collected for investigation. For the isolation, blood agar and bile esculin agar were used. DNA isolations of the 24-hour growth cultures of possible enterococci were carried out using a DNA isolation kit.Out of 350 routine urine and 150 stool samples taken with the approval of the patients, 235 (67.1%) and 114 (76%) Enterococcus spp. were isolated respectively. Using the multiplex PCR method with species specific primers, 136 (57.8%) of urine and 22 (19.2%) of stool originated enterococcal strains were identified as Enterococcus faecalis; on the other hand, 17 (7.2%) of urine and 61 (53.5%) of stool originated enterococci were identified as Enterococcus faecium.As a result of the study in Van, Turkey, the isolation rate of Enterococcus faecalis and Enterococcus faecium strains were found to be lower than other regions.

Squamous cell carcinomas are malign tumors composed of squamous epithelium of skin and mucous membranes and showing squamous differentiation. A male, 11-year-old Chinchilla race cat was brought to Ankara University, Veterinary Faculty, Veterinary Hospital, Department of Internal Diseases with complaints of anorexia, weakness, dysphagia, fatigue and difficulty in swallowing solid food. Metronidazole (15 mg/kg, iv), amoxicillin-clavulanic acid (25 mg/kg, sc) and fluid treatment were administered to the patient, respectively. As a predisposing factor in the development of oral squamous cell carcinoma in cats, exposure to cigarette smoke, the use of flea collar, and especially nutrition with tuna fish- containing ingredients are mentioned.

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

Bovine mastitis (BM) has resulted in enormous economic loss in the dairy industry and coagulase-negative staphylococci (CNS) have caused subclinical BM. Although VITEK 2 GP ID card (VITEK 2) has been used for CNS identification, the probability of identification varies. The rpoB sequence typing (RSTing) method has been used for molecular diagnosis and epidemiology of bacterial infections. In this study, we undertook RSTing of CNS and compared the results with those of VITEK2 and 16S rRNA gene sequencing. As compared VITEK2, the molecular-based methods were more reliable for species identification; moreover, RSTing provided more molecular epidemiological information than that from 16S rRNA gene sequencing.

Three aged dogs, all over 15 years old, with multiple seizure events and various neurologic signs such as head turn and ataxia were brought to the referring hospital. Two cases were tentatively diagnosed with intracranial meningioma based on magnetic resonance imaging, and one case was histopathologically confirmed as meningioma at necropsy. All dogs improved after initiation of pulsed high-dose hydroxyurea (HU) treatment. Clinical sign relapses were managed by repeating high-dose HU without obvious side effects. Pulsed high-dose HU can be applied when clinical signs are refractory to the traditionally recommended dosage for intracranial meningioma in dogs.