Development of Similar Broth Microdilution Methods to Determine the Antimicrobial Susceptibility of Flavobacterium columnare and F. psychrophilum

Flavobacterium columnare and F. psychrophilum are major fish pathogens that cause diseases that may require antimicrobial therapy. Choice of appropriate treatment is dependent upon determining the antimicrobial susceptibility of isolates. Therefore we optimized methods for broth microdilution testing of F. columnare and F. psychrophilum to facilitate standardizing an antimicrobial susceptibility test. We developed adaptations to make reproducible broth inoculums and confirmed the proper incubation time and media composition. We tested the stability of potential quality-control bacteria and compared test results between different operators. Log phase occurred at 48 h for F. columnare and 72–96 h for F. psychrophilum, confirming the test should be incubated at 28°C for approximately 48 h and at 18°C for approximately 96 h, respectively. The most consistent susceptibility results were achieved with plain, 4-g/L, dilute Mueller–Hinton broth supplemented with dilute calcium and magnesium. Supplementing the broth with horse serum did not improve growth. The quality-control strains, Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658, yielded stable minimal inhibitory concentrations (MIC) against all seven antimicrobials tested after 30 passes at 28°C and 15 passes at 18°C. In comparison tests, most MICs of the isolates agreed 100% within one drug dilution for ampicillin, florfenicol, and oxytetracycline. The agreement was lower with the ormetoprim–sulfdimethoxine combination, but there was at least 75% agreement for all but one isolate. These experiments have provided methods to help standardize antimicrobial susceptibility testing of these nutritionally fastidious aquatic bacteria.

Received June 24, 2015; accepted October 2, 2015.     

A technique for central venous pressure measurement in normal horses

Objective – To investigate a technique of central venous pressure (CVP) measurement using a newly developed catheter in healthy adult horses. Design – Prospective experimental study. Setting – University research facility. Animals – Twenty healthy adult horses. Interventions – An equine central venous catheter was inserted into the jugular vein to a length of approximately 80 cm from the mid‐cervical region in an attempt to catheterize the pulmonary artery. Pulmonary arterial catheterization was confirmed by echocardiography. Insertion distance and pressure were measured at this location with a disposable manometer. The catheter was then withdrawn until presence in the right atrium was confirmed by echocardiography. Insertion distance and pressure were also measured at this location. The catheter was then withdrawn in 5 cm increments until exiting the jugular insertion site with pressure measured at each location. All pressure measurements were taken with the manometer zero position at the point of the shoulder. Measurements and Main Results – Pulmonary artery catheterization was successful in 16 of 20 horses. Mean pulmonary arterial pressure was 23.8 cm H₂O (17.5 mm Hg) (95% confidence interval [CI] 20.9–26.7 cm H₂O [15.4–19.6 mm Hg]). Mean right atrial pressure was 8.3 cm H₂O (6.1 mm Hg) (95% CI 7.1–9.4 cm H₂O [5.2–6.9 mm Hg]). Right atrial pressure was compared with pressures recorded at sequential insertion distances and resulted in a recommendation for catheter insertion of at least 40 cm for CVP measurement in adult horses. Jugular venous pressure measurement was statistically different from CVP measurement. Conclusions – This catheter measurement technique is well tolerated in normal horses. Routine clinical use of this equine central venous catheter may improve our ability to monitor patients and improve patient care and outcomes of ill horses in hospital.     

Hybrid technique for ventricular septal defect closure in a dog using an Amplatzer® Duct Occluder II

A left-to-right shunting muscular ventricular septal defect (VSD) was diagnosed in a 4-month-old, female, 1.8 kg Bichon Frise – poodle mix dog. Echocardiographic evidence of cardiac remodeling, calculated pulmonary blood flow (Qp) to systemic blood flow (Qs) ratio of 2.8, and radiographic evidence of pulmonary edema supported the diagnosis of a hemodynamically important VSD. Using a combination of surgery and interventional catheter-based techniques to approach the VSD through the right ventricle, the VSD was occluded with an Amplatzer^® Duct Occluder (ADO) II device. The ADO II is a low profile, flexible device originally developed for patent ductus arteriosus closure in humans that has been used to close muscular and perimembranous VSD in children. This report describes the hybrid procedure and imaging that was essential for successful occlusion of the VSD in this dog.     

Delayed embolization of an Amplatz® canine duct occluder in a dog

A 5-year old, 5.8 kg, castrated male Pomeranian was diagnosed with a type IIa patent ductus arteriosus (PDA) with a minimal ductal diameter of 3.5 mm and ampulla width of 7.1 mm based on angiographic assessment. A 6 mm Amplatz^® Canine Duct Occluder (ACDO) was deployed within the PDA. Once deployed, the device assumed it's native shape and back-and-forth maneuvering was performed with the delivery cable to assess device stability. Device position and complete occlusion were confirmed with both angiography and transesophageal echocardiography prior to and after release of the device. The device location was confirmed within the ductus arteriosus by echocardiography prior to discharge. The dog was discharged with instructions for strict activity restriction. Two days after discharge, the dog was left unsupervised in the backyard and shortly afterwards was found coughing with severe respiratory distress. The dog was evaluated at an emergency hospital and thoracic radiographs documented embolization of the ACDO to the main pulmonary artery along with a severe alveolar pattern throughout the right lung fields. Shortly after obtaining thoracic radiographs, the dog experienced cardiopulmonary arrest with unsuccessful resuscitation. This case describes a possible complication of transcatheter PDA occlusion with an ACDO, which has not been previously reported. An incident report, or catalog of adverse events with these devices, may prove useful in identifying additional fatal complications that others may have encountered, but are not reported in the literature. The report of this complication emphasizes the importance of strict activity restriction after device placement in dogs.     

Inflammatory aural polyp in a horse

An inflammatory aural polyp was identified in a 1-year-old standardbred filly, which presented with otorrhea and head rubbing. The polyp was removed by traction-avulsion, and the filly showed no subsequent signs of otorrhea. Aural polyps have not been reported in horses, but they are commonly seen in companion animals and humans.     

Pharmacodynamics of carvedilol in conscious, healthy dogs

The purpose of the study reported here was to determine the magnitude and duration of beta-blocking efficacy, determine an effective dose and dosing interval, and document safety and tolerability of carvedilol given orally in clinically normal dogs. Pharmacodynamic data were evaluated in conscious, unrestrained, healthy hound dogs at baseline and after long-term oral administration of carvedilol (1.5 mg/kg of body weight PO q12h for >5 days). At baseline, heart rate (HR) and blood pressure (BP) data were collected continuously for 24 hours, and complete echocardiography was performed. This protocol was repeated after long-term oral carvedilol administration. Additionally, isoproterenol was administered to evaluate the magnitude and duration of the nonselective beta-blocking efficacy of carvedilol. An isoproterenol challenge was performed 0.75, 1.5, 2.25, 4, 6, 12, and 24 hours after carvedilol administration, with echocardiography being performed once at 2 hours. Plasma samples were obtained prior to each challenge time point for determination of plasma carvedilol concentration. Time series regression analysis indicated no difference between baseline and carvedilol-induced HR or BP trend lines in 6 of 8 dogs. In 2 of 8 dogs, HR, after long-term carvedilol administration, was reduced. Carvedilol attenuated isoproterenol-induced changes in HR by 54-76% through 12 hours and by 30% at 24 hours. The BP changes were attenuated by 80-100% through 12 hours. These data suggest that carvedilol (1.5 mg/kg PO q12h) in healthy, conscious dogs confers nonselective beta blockade for 12 hours, with minimal effects on resting HR, BP, and echocardiographic variables. Additionally, the magnitude of beta blockade correlated strongly to peak plasma carvedilol concentration, suggesting that therapeutic drug monitoring may be clinically useful.     

Restoring important hair properties with wool keratin proteins and peptides

In this work the effect on hair of two types of keratin samples obtained from wool, was investigated. Fiber surface changes were evaluated by contact angle measurements. The effectiveness of these keratin ingredients to restore the mechanical properties and the moisture content of the fibers was also determined. Modifications of hair properties due to some conventional chemical treatments were demonstrated with lower values of contact angle and detrimental effects on the mechanical properties. Application of keratin peptides and proteins to pretreated hair improved the fibers’ moisture content and their mechanical properties.