ADJUNCTIVE THERAPY FOR FETLOCK FLEXURAL DEFORMITIES ACCOMPANIED BY A DEEP TISSUE INJURY IN A CALF: A CASE REPORT

A five-day-old Holstein Friesian male calf was presented for the evaluation of severe fetlock flexural deformities with a large pressure ulcer over the right metacarpal bone. Superficial and deep digital flexor tenotomy was performed. The surgical wound recovered well, but 15 days after surgery, the skin over the pressure ulcer was starting to fall away and exposing tendon and muscle. A hoof toe-extension shoe was applied to decrease pressure on the wound. The wound had a moderate bacterial infection and slowly responded to cleansing twice a day with 0.9% normal saline, 10% povidone iodine and a topical antibiotic. Topical autologous platelet-rich plasma (PRP) was then applied to the lesion ([Formula: see text][Formula: see text]cm) twice. The wound showed rapid tissue regeneration, good quality wound healing and no complications. After PRP treatment, the lesion size decreased ([Formula: see text][Formula: see text]cm). At this point, 0.9% normal saline cleansing and a bandage were applied until the wound had completely healed. This case report suggests that the hoof toe-extension shoe and topical dressing with autologous PRP, as a combination treatment, could be effective for fetlock flexural deformities with skin defects or delayed wound healing in large animals.

GRANGER CAUSALITY ANALYSIS BETWEEN INTRA-OSSEOUS PRESSURE AND ARTERIAL BLOOD PRESSURE IN AFRICAN GREY PARROTS (PSITTACUS ERITHACUS)

Monitoring blood pressure helps a clinical veterinarian assess various conditions in birds. Blood pressure is not only a bio-indicator of renal or cardiovascular disease but is also a vital indicator for anesthesia. Anesthetic- and sedation-related mortality is higher in birds than dogs or cats. The traditional method of blood pressure measurement in mammals mainly relies on indirect methods. However, indirect blood pressure measurement is not reliable in birds, making the direct method the only gold standard. Although an arterial catheter can provide continuous real-time arterial pressure in birds, the method requires technical skill and is limited by bird size, and is thus not practical in birds with circulatory collapse. Intra-osseous (IO) blood pressure is potentially related to arterial pressure and may be a much easier and safer technique that is less limited by animal size. However, the relationship between IO pressure and arterial blood pressure has not been established. This study used mathematical methods to determine the relationship between IO pressure and arterial blood pressure. The Granger causality (G.C.) theory was applied in the study and used to analyze which pressure signal was leading the other. Our findings suggest that IO pressure is G.C. by arterial blood pressure; thus, the use of IO pressure measurements as an alternative to arterial blood pressure measurement is a rational approach.

THE EFFECTS OF OXIDATIVE STRESS ON THE EYES OF SPRAGUE–DAWLEY RATS

An imbalance between oxidants and antioxidants in favor of the oxidants, potentially leading to damage, is termed as ‘oxidative stress’. Some studies indicate that reactive oxygen species (ROS) may increase in resistance of the aqueous humor drainage or induce the rearrangement of human trabecular meshwork and result in glaucoma finally. An eight-week d-galactose injection was used to mimic high oxidative stress rats. Behavior tests, fundus images, anterior chamber depth, anterior chamber diameter (ACDia) and pupil diameter (PD) were investigated. Intraocular pressure (IOP) measurement and serum were taken for antioxidant capability of superoxide dismutase (SOD) and the amount of nitric oxide (NO). After perfusion, Müller’s cells and astrocyte, microglial and inducible Nitric Oxide Synthase (iNOS) were labeled. The activities of SOD were decreased after induction. The spatial memory ability and muscular endurance were impaired through Morris water maze (MWM) and weight-loaded forced swimming test. These results demonstrated that oxidative stress was exacerbated by d-galactose. The IOP was gradually increased in the third week after d-galactose induction. Immunohistochemical stains showed that the density of Müller’s cells, astrocytes and microglia increased in d-galactose group. Microglia mostly became active. These data indicate that oxidative stress may result in the impairment of behavior, increase of Müller’s cells, astrocyte and microglia which is induced by d-galactose. The correlation between oxidative stress and elevation of IOP should be further studied. These may involve in the pathologies of aging or other neurodegenerative disease.

INVESTIGATION OF EFFECTS OF PORCINE CIRCOVIRUS TYPE 2 (PCV2) VACCINATION ON CELLULAR IMMUNITY IN PIGS IN A TAIWAN PIG FARM

Porcine circovirus type 2 (PCV2) is the major cause of serious porcine circovirus-associated diseases (PCVADs) causing significant economic losses in the pig industry. The PCV2 vaccines have proved effective by lowering serum and tissue viral loads. However, little is known about its effects on the cellular functions of viral target cells in pigs. For this, blood samples from 38 vaccinated and 34 non-vaccinated, 12–14 week-old, clinically healthy and PCV2-infected pigs in a commercial farm were sampled and anticoagulated. The viral load in plasma and the cellular proliferation and mRNA expression levels of IL-1 [Formula: see text], IL-2, IL-4, IL-8, IL-10 and IFN-[Formula: see text] of peripheral blood mononuclear cells (PBMCs) with or without concanavalin A (Con A) and PCV2 stimulation in vitro were then investigated by real-time polymerase chain reaction and flow cytometry. Compared with the non-vaccinated pigs, the average viral loads in the plasma and cultivated PBMCs were reduced in the PCV2-vaccinated pigs. Significant differences in the aspects of increased cell proliferation, decreased average viral loads in PBMCs three days after Con A stimulation, increased mRNA expression levels of IL-2 and IFN-[Formula: see text] after re-exposure to PCV2, and decreased levels of IL-1 [Formula: see text], IL-4 and IL-8 after Con A or PCV2 stimulation were also noted in the PCV2-vaccinated pigs. The results suggested that the PCV2-vaccinated pigs were likely to develop better cell-mediated immunity than the non-vaccinated pigs.

DEVELOPMENT OF A UPL PROBE-BASED REAL-TIME PCR ASSAY OF PORCINE DELTACORONAVIRUS IN TAIWAN

Porcine deltacoronavirus (PDCoV) causes clinical symptoms characterized by severe diarrhea and vomiting in neonatal piglets and pregnant sows, which is similar to those resulted from transmissible gastroenteritis coronavirus (TGEV) and porcine epidemic diarrhea virus (PEDV). Since PEDV was considered as the dominant enteric virus all round the world, PDCoV has been unwittingly overlooked due to its indistinguishable clinical signs with other porcine coronaviruses and relatively low death rates in the pig farm. Specimens which have been previously performed for the detection of PEDV in Animal Disease and Diagnostic Center, National Pingtung University of Science and Technology (NPUST) from January 5, 2015 to January 11, 2016 were examined by a novel universal probe library (UPL) probe-based real-time polymerase chain reaction (PCR). A total of 527 clinical specimens from pigs with diarrhea suspected were examined for PDCoV. Positive rates of PDCoV in small intestine and rectal swab were 4.3% (13/305) and 1.8% (4/222), respectively. Collectively, as to the total specimens, the detection rate is 3.2% (17/527). Our results provide development of a UPL probe-based real-time PCR assay and retrospective investigation of potentially circulating PDCoVs in the field in the whole 2015 and early 2016.

CLINICAL AND HISTOPATHOLOGICAL FEATURES OF SCOPOLAMINE HYDROBROMIDE INDUCED DRY EYE SYNDROME IN SPRAGUE-DAWLEY RATS

Dry eye syndrome (DES), is one of the most common and irritating ocular diseases in humans and animals due to deficits in quantities or/and quality of tear film. In this study, a rat model of experimental DES has been developed using the cholinergic inhibitor, scopolamine hydrobromide (SCOP), at the dose of 25[Formula: see text]mg/rat/day via subcutaneous injection, for a consecutive 21 days without low humidity environment. Clinical ophthalmic evaluations were performed by tear volume assessment using endodontic paper point, slit-lamp biomicroscope, and fluorescein staining at day 0, 7, 14, and 21 post-inductions. The results of ophthalmic examination showed that rats with SCOP treatment reduced about 40% of tear secretion. Half of the SCOP-treated rats exhibited diffuse corneal fluorescein staining involving 80% of the corneal surface, minimal keratoconjunctivitis, roughened corneal surface and thin corneal epithelium under histopathological examination. About 30% of the rats showed variable infiltration of lymphocytes in between the tubular acinar glands. This animal model with significant reduction of tear production and diffuse corneal fluorescein staining in rats could be used for the preclinical assessment of therapeutic interventions.

A CASE OF BILATERAL RENAL B-CELL LYMPHOMA WITH UVEITIS AND SECONDARY POLYCYTHEMIA IN A JACK RUSSELL TERRIER

An 8-year-old, castrated male, Jack Russell Terrier was presented with clinical signs of uveitis and glaucoma in the right eye (oculus dexter (OD)) for 2–3 months. Ophthalmic examination revealed OD episcleral congestion, corneal vascularization, aqueous flare, iris bombe, and vision loss. OD glaucoma and uveitis were under control by medical treatment. After 8 months, the dog had acute uveitis in the left eye (oculus sinister (OS)). General physical examination, blood examinations, radiography, and urinalysis were unremarkable. No improvement was observed after for 1 month of uveitis treatment, and more examinations were strongly suggested. Abdominal ultrasonography revealed several hypoechoic masses or cysts on both kidneys. Computed tomography (CT) showed diffuse nonenhancing hypodense patches in both kidneys. Renal tumor was highly suspected, but biopsy was declined by the owner. Ten days after CT examination, the dog suddenly developed anorexia, weakness, uremia, hyperkalemia, and polycythemia. The dog was euthanatized because of the worsening clinical condition, poor prognosis, and financial constraints, and both kidneys were biopsied for histopathology. Immunohistochemically, the neoplastic cells were positive for B-cell marker (CD20) and negative for T-cell marker (CD3). Bilateral renal B-cell origin lymphoma was diagnosed by histologic and immunophenotyping examinations. In conclusion, bilateral renal lymphoma may be diagnosed with no systemic signs except bilateral uveitis. Renal function impairment was acutely developed with severe clinical signs at the end stage of the disease. Early, aggressive, complete examinations, and treatments are strongly recommended.

EFFECT OF EFFLUX PUMP INHIBITORS ON ANTIMICROBIAL ACTIVITY OF FLORFENICOL AGAINST RESISTANT STRAINS OF PORCINE ACTINOBACILLUS PLEUROPNEUMONIAE AND PASTEURELLA MULTOCIDA

The aim of this study is to investigate whether combinational use of efflux pump inhibitors (EPIs) could improve florfenicol (FF) antimicrobial activity. Five EPIs including Carbonyl Cyanide Chlorophenylhydrazone (CCCP), omeprazole, Phenylalanine-arginine [Formula: see text]-naphthylamide (PA[Formula: see text]N), reserpine and verapamil were individually combined with FF and the minimum inhibitory concentration (MIC) against porcine Actinobacillus pleuropneumoniae and Pasteurella multocida were evaluated by the broth microdilution assay. The results indicated that CCCP demonstrated substantial improvement on the antimicrobial activity (FF MIC reduction [Formula: see text] folds) while PAßN showed minimal effect at high concentrations (80–120[Formula: see text][Formula: see text]g/mL). The MICs of FF were further examined with CCCP at 5[Formula: see text][Formula: see text]g/mL against resistant A. pleuropneumoniae and at 2[Formula: see text][Formula: see text]g/mL against resistant P. multocida. With this combination, a total of 75% (6/8) of A. pleuropneumoniae and 100% (8/8) of P. multocida resistant isolates showed significantly reduced ([Formula: see text] folds) FF MICs. In addition, 100% of the 16 resistant bacterial strains carried the floR gene that regulates the phenicol-specific efflux pump (FloR pump). The time kill experiments were used to verify the effect of CCCP demonstrating a bactericidal effect in resistant A. pleuropneumoniae and synergistic effect in resistant P. multocida. In contrast, the FF MICs were not significantly affected in the FF susceptible strains of these two bacteria. These findings suggested that despite small sample sizes, consistent beneficial effects of CCCP were evident such that the combinational use of selective EPI with FF might be an alternative antibacterial strategy against FF resistant A. pleuropneumoniae and P. multocida. The anti-efflux mechanism was indicated, but more researches to further decipher the mechanisms of CCCP’s effects are warranted.

MULTIPLEX POLYMERASE CHAIN REACTION ASSAY FOR THE DETECTION OF FIVE COMMON ENDOPARASITE INFESTATIONS IN LABORATORY RODENTS

Rodent pinworms, Spironucleus muris and Tritrichomonas muris are the endoparasites that should be monitored and excluded from laboratory animal colonies. Nevertheless, traditional diagnostic methods may not efficiently detect and accurately demonstrate the endoparasite infestation status. In this study, we developed a multiplex PCR assay targeting the rRNA genes to simultaneously detect and differentiate five endoparasites, including Syphacia obvelata, Syphacia muris, Aspiculuris tetraptera, Spironucleus muris, and T. muris, as well as a housekeeping gene in feces. The multiplex PCR could identify an equivalent infection of pinworm, Spironucleus muris and T. muris, with a detection limit of as few as 10 copies. Furthermore, dual infections with up to 100-fold differences and triple infections with 10-fold differences in parasite loads can also be detected. In comparison of traditional methods with the multiplex PCR assay, 76 rodents from 11 research colonies and 3 pet shops and additional 27 fecal samples from laboratory rodents were screened for the infestation status of the five endoparasites. The multiplex PCR had higher sensitivity (97.2–100%) and accuracy (99–100%) than those of the traditional antemortem (sensitivity: 83–100%; accuracy: 94–100%) and postmortem methods (sensitivity: 75–100%; accuracy: 92.1–100%). In addition, an early stage of S. obvelata contamination in a SPF laboratory animal colony was also successfully detected by this multiplex PCR assay. This Pinworm/Spironucleus/Tritrichomonas/Actin Multiplex PCR assay should be a powerful tool to screen endoparasite infestations in laboratory colonies without animal sacrifice.