Tracy L. Drazenovich

Molecular Detection of Microbes in Nasal Tissue of Dogs with Idiopathic Lymphoplasmacytic Rhinitis

Lymphoplasmacytic rhinitis (LPR) is a common histologic finding in dogs with chronic nasal disease; however, potential etiologies of this disorder have not been examined. We investigated the hypothesis that specific microbes contribute to clinical disease in dogs with LPR. Paraffin-embedded nasal biopsies were obtained from 19 dogs with LPR, 10 dogs with nasal neoplasia, and 10 dogs with nasal aspergillosis. Nucleic acids were extracted from paraffin blocks, and real-time quantitative polymerase chain reaction (PCR) was employed for detection of target genes for bacterial and fungal DNA, canine adenovirus 2 (CAV-2), parainfluenza virus 3 (PI-3), Chlamydial Chlamydophila spp., and Bartonella spp. Conventional PCR was used for detection of Mycoplasma spp. Statistical analysis was performed using the Mann-Whitney U-test for nonparametric data, and significance was set at P < 0.05. DNA or RNA for CAV-2, PI-3, Bartonella, Mycoplasma, and Chlamydophila was not detected in any nasal biopsy. DNA loads for bacterial DNA did not differ among disease groups. Detection of fungal DNA in nasal biopsies was highest in dogs with aspergillosis (P < 0.0001); however, nasal biopsies of LPR dogs also displayed higher fungal DNA levels than samples from dogs with nasal neoplasia (P = 0.016). Detection of high levels of fungal DNA in nasal biopsies of dogs with LPR suggests that fungal organisms may be causally associated with the inflammation observed, although the possibility of entrapment or accumulation of fungi in the nasal cavity due to chronic inflammation cannot be excluded. Further investigations are required to elucidate the underlying etiopathogenesis of LPR.

Effects of dietary lysine supplementation on upper respiratory and ocular disease and detection of infectious organisms in cats within an animal shelter.

OBJECTIVE To determine within a cat shelter effects of dietary lysine supplementation on nasal and ocular disease and detection of nucleic acids of Chlamydophila felis, feline calicivirus (FCV), and feline herpesvirus (FHV-1). ANIMALS 261 adult cats. PROCEDURES Cats were fed a diet containing 1.7% (basal diet; control cats) or 5.7% (supplemented diet; treated cats) lysine for 4 weeks. Plasma concentrations of lysine and arginine were assessed at the beginning (baseline) and end of the study. Three times a week, cats were assigned a clinical score based on evidence of nasal and ocular disease. Conjunctival and oropharyngeal swab specimens were tested for FHV-1, FCV, and C felis nucleic acids once a week. RESULTS Data were collected from 123, 74, 59, and 47 cats during study weeks 1, 2, 3, and 4, respectively. By study end, plasma lysine concentration in treated cats was greater than that in control cats and had increased from baseline. There was no difference between dietary groups in the proportion of cats developing mild disease. However, more treated cats than control cats developed moderate to severe disease during week 4. During week 2, FHV-1 DNA was detected more commonly in swab specimens from treated versus control cats. CONCLUSIONS AND CLINICAL RELEVANCE Dietary lysine supplementation in the amount used in our study was not a successful means of controlling infectious upper respiratory disease within a cat shelter. Rather, it led to increases in disease severity and the incidence of detection of FHV-1 DNA in oropharyngeal or conjunctival mucosal swab specimens at certain time points.

Evaluation of thermal antinociceptive effects after intramuscular administration of hydromorphone hydrochloride to American kestrels (Falco sparverius)

OBJECTIVE To evaluate the antinociceptive and sedative effects and duration of action of hydromorphone hydrochloride after IM administration to American kestrels (Falco sparverius). ANIMALS 11 healthy 2-year-old American kestrels. PROCEDURES Hydromorphone (0.1, 0.3, and 0.6 mg/kg) and an equivalent volume of saline (0.9% NaCl) solution (control treatment) were administered IM to kestrels in a masked randomized complete crossover study design. Foot withdrawal response to a thermal stimulus was determined 30 to 60 minutes before (baseline) and 0.5, 1.5, 3, and 6 hours after treatment administration. Agitation-sedation scores were determined 3 to 5 minutes before each thermal test. RESULTS Hydromorphone at 0.6 mg/kg, IM, significantly increased the thermal foot withdrawal threshold, compared with the response after administration of saline solution, for up to 3 hours, and hydromorphone at 0.1, 0.3, and 0.6 mg/kg, IM, significantly increased withdrawal responses for up to 6 hours, compared with baseline values. No significant differences in mean sedation-agitation scores were detected between hydromorphone and saline solution treatments; however, appreciable sedation was detected in 4 birds when administered 0.6 mg of hydromorphone/kg. CONCLUSIONS AND CLINICAL RELEVANCE Hydromorphone at the doses evaluated significantly increased the thermal nociception threshold for American kestrels for 3 to 6 hours. Additional studies with other types of stimulation, formulations, dosages, routes of administration, and testing times are needed to fully evaluate the analgesic and adverse effects of hydromorphone in kestrels and other avian species and the use of hydromorphone in clinical settings.

Evaluation of thermal antinociceptive effects and pharmacokinetics after intramuscular administration of butorphanol tartrate to American kestrels (Falco sparverius)

OBJECTIVE To evaluate antinociceptive effects and pharmacokinetics of butorphanol tartrate after IM administration to American kestrels (Falco sparverius). ANIMALS Fifteen 2- to 3-year-old American kestrels (6 males and 9 females). PROCEDURES Butorphanol (1, 3, and 6 mg/kg) and saline (0.9% NaCl) solution were administered IM to birds in a crossover experimental design. Agitation-sedation scores and foot withdrawal response to a thermal stimulus were determined 30 to 60 minutes before (baseline) and 0.5, 1.5, 3, and 6 hours after treatment. For the pharmacokinetic analysis, butorphanol (6 mg/kg, IM) was administered in the pectoral muscles of each of 12 birds. RESULTS In male kestrels, butorphanol did not significantly increase thermal thresholds for foot withdrawal, compared with results for saline solution administration. However, at 1.5 hours after administration of 6 mg of butorphanol/kg, the thermal threshold was significantly decreased, compared with the baseline value. Foot withdrawal threshold for female kestrels after butorphanol administration did not differ significantly from that after saline solution administration. However, compared with the baseline value, withdrawal threshold was significantly increased for 1 mg/kg at 0.5 and 6 hours, 3 mg/kg at 6 hours, and 6 mg/kg at 3 hours. There were no significant differences in mean sedation-agitation scores, except for males at 1.5 hours after administration of 6 mg/kg. CONCLUSION AND CLINICAL RELEVANCE Butorphanol did not cause thermal antinociception suggestive of analgesia in American kestrels. Sex-dependent responses were identified. Further studies are needed to evaluate the analgesic effects of butorphanol in raptors.

Composition and characteristics of urinary calculi from guinea pigs

OBJECTIVE To determine the mineral composition of calculi, anatomic locations of the calculi, and findings of urinalysis and bacteriologic culture of urine and calculi in guinea pigs with urolithiasis. DESIGN Cross-sectional study. ANIMALS 127 guinea pigs. PROCEDURES Records of urinary calculi that had been submitted to the University of California Stone Laboratory from 1985 through 2003 were reviewed. In addition, submissions of urinary calculi for evaluation by the laboratory were prospectively solicited from 2004 through 2007. Prospectively obtained calculi were accompanied by a urine sample for urinalysis and bacteriologic culture and a completed questionnaire. All calculi were analyzed by use of polarized light microscopy and infrared spectroscopy. A subset of calculi was examined by means of x-ray diffractometry (XRD). RESULTS 83% (43/52) of calculi from the laboratory database and 93% (70/75) of calculi that were prospectively solicited were composed of 100% calcium carbonate. Analysis via XRD confirmed that 5 of 6 calculi from a subset that had the greatest gross morphologic variation were composed of 100% calcite. Although many guinea pigs had received anti-microbials before bacteriologic cultures of urine were performed, Corynebacterium renale was isolated from 5 urine samples. CONCLUSIONS AND CLINICAL RELEVANCE Contrary to findings of other studies, urinary calculi analyzed for the present study were most commonly composed of 100% calcium carbonate, and infrared spectroscopy or XRD was necessary to differentiate this mineral from others. Treatments, including diet and husbandry practices, should be developed to help prevent development of calcium carbonate calculi in guinea pigs.

Widespread Recombination, Reassortment, and Transmission of Unbalanced Compound Viral Genotypes in Natural Arenavirus Infections

Arenaviruses are one of the largest families of human hemorrhagic fever viruses and are known to infect both mammals and snakes. Arenaviruses package a large (L) and small (S) genome segment in their virions. For segmented RNA viruses like these, novel genotypes can be generated through mutation, recombination, and reassortment. Although it is believed that an ancient recombination event led to the emergence of a new lineage of mammalian arenaviruses, neither recombination nor reassortment has been definitively documented in natural arenavirus infections. Here, we used metagenomic sequencing to survey the viral diversity present in captive arenavirus-infected snakes. From 48 infected animals, we determined the complete or near complete sequence of 210 genome segments that grouped into 23 L and 11 S genotypes. The majority of snakes were multiply infected, with up to 4 distinct S and 11 distinct L segment genotypes in individual animals. This S/L imbalance was typical: in all cases intrahost L segment genotypes outnumbered S genotypes, and a particular S segment genotype dominated in individual animals and at a population level. We corroborated sequencing results by qRT-PCR and virus isolation, and isolates replicated as ensembles in culture. Numerous instances of recombination and reassortment were detected, including recombinant segments with unusual organizations featuring 2 intergenic regions and superfluous content, which were capable of stable replication and transmission despite their atypical structures. Overall, this represents intrahost diversity of an extent and form that goes well beyond what has been observed for arenaviruses or for viruses in general. This diversity can be plausibly attributed to the captive intermingling of sub-clinically infected wild-caught snakes. Thus, beyond providing a unique opportunity to study arenavirus evolution and adaptation, these findings allow the investigation of unintended anthropogenic impacts on viral ecology, diversity, and disease potential.

Flexible bronchoscopy and bronchoalveolar lavage in 68 cats (2001-2006)

BACKGROUND Bronchoscopy is an important tool for identifying an underlying etiology for respiratory disease in cats. However, the procedure is challenging, because feline airways are small and prone to bronchoconstriction. HYPOTHESIS Bronchoscopy and bronchoalveolar lavage (BAL) are appropriate and safe diagnostic procedures in the cat. ANIMALS Sixty-eight cats. METHODS Flexible bronchoscopy was performed in all cats with the cats under propofol infusion with jet ventilation. The procedures were reviewed for BAL volumes instilled and recovered and for the number and type of complications with the use of 3 flexible endoscopes < 5.0-mm outer diameter. The BAL procedure was compared among scopes by using a one-way analysis of variance. Complication rates were compared by using chi-square analysis. Significance was set at P < .05. RESULTS Clinical diagnoses included inflammatory airway disease in 46 of 68 cats, pneumonia in 10 of 68, neoplastic disease in 8 of 68, and other conditions in 4 of 68 cats. Mean lavage volumes instilled for the 3 scopes were 2.62-5.05 mL/kg (range, 0.77-9.38 mL/kg). Mean percent fluid recovered for the 3 scopes was 51-73%, (range, 0-140%). BAL cell counts were adequate for cytologic assessment (> 300 cells/microL) in 61 of 64 cats (97%), and in 107 of 120 samples (89%) collected. Complications occurred in 38% of procedures; however, these were mild in 24% of cats; 6% of cats died or were euthanized after the procedure. Complications were not associated with fluid volume instilled or recovered, and could not be related to the underlying disease process. CONCLUSIONS AND CLINICAL IMPORTANCE Flexible bronchoscopy with BAL was well tolerated in most cats examined.

Evaluation of thermal antinociceptive effects after oral administration of tramadol hydrochloride to American kestrels (Falco sparverius)

OBJECTIVE To evaluate the thermal antinociceptive and sedative effects and duration of action of tramadol hydrochloride after oral administration to American kestrels (Falco sparverius). ANIMALS 12 healthy 3-year-old American kestrels. PROCEDURES Tramadol (5, 15, and 30 mg/kg) and a control suspension were administered orally in a masked randomized crossover experimental design. Foot withdrawal response to a thermal stimulus was determined 1 hour before (baseline) and 0.5, 1.5, 3, 6, and 9 hours after treatment. Agitation-sedation scores were determined 3 to 5 minutes before each thermal stimulus test. RESULTS The lowest dose of tramadol evaluated (5 mg/kg) significantly increased the thermal foot withdrawal thresholds for up to 1.5 hours after administration, compared with control treatment values, and for up to 9 hours after administration, compared with baseline values. Tramadol at doses of 15 and 30 mg/kg significantly increased thermal thresholds at 0.5 hours after administration, compared with control treatment values, and up to 3 hours after administration, compared with baseline values. No significant differences in agitation-sedation scores were detected between tramadol and control treatments. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated oral administration of 5 mg of tramadol/kg significantly increased thermal nociception thresholds for kestrels for 1.5 hours, compared with a control treatment, and 9 hours, compared with baseline values; higher doses resulted in less pronounced antinociceptive effects. Additional studies with other types of stimulation, formulations, dosages, routes of administration, and testing times would be needed to fully evaluate the analgesic and adverse effects of tramadol in kestrels and other avian species.

Interpretation of Multisegment Bronchoalveolar Lavage in Cats (1/2001–1/2011)

BACKGROUND Cytologic results from bronchoscopic BAL in cats with naturally occurring respiratory disease have not been reported, and the clinical utility of multisegment lavage has not been evaluated. HYPOTHESIS BAL cytology from 2 separate lung segments in cats will have similar cell counts, cytologic interpretation, or both. ANIMALS Eighty-seven cases in 85 cats (2 examined twice) with naturally occurring lower respiratory disease. METHODS A combined prospective/retrospective evaluation of all cats with multisegment BAL was performed. BAL fluid was evaluated for total nucleated cell counts, differential cell counts, and cytologic characteristics at each lavage site. BAL fluid was categorized as eosinophilic, neutrophilic, lymphocytic, hypercellular, or mixed. Radiographs were assessed for diffuse or focal disease. RESULTS Clinical diagnoses included inflammatory airway disease (n = 63), pneumonia (n = 15), neoplasia (n = 6), and undetermined (n = 3). Total nucleated cell counts varied between sites regardless of radiographic evidence of focal or diffuse radiographic disease. In 28/87 cases (32%), cell counts differed between lavage sites by 2.2-40 fold. BAL yielded similar cytologic interpretation of inflammation in 45/87 (52%) cases. In 8/14 cases that had BAL performed at the site of a focal radiographic infiltrate, as well as at a site of diffuse infiltrates, the same inflammatory interpretation was made at each site. CONCLUSIONS AND CLINICAL IMPORTANCE Total and differential cell counts in BAL fluid often differ between lung segments in cats with lower respiratory disease, and caution is warranted when using a single BAL cytology to define the inflammatory response in cats with spontaneously occurring lower respiratory tract disease.

Pharmacokinetics of hydromorphone hydrochloride after intravenous and intramuscular administration of a single dose to American kestrels (Falco sparverius).

OBJECTIVE To determine the pharmacokinetics of hydromorphone hydrochloride after IV and IM administration in American kestrels (Falco sparverius). ANIMALS 12 healthy adult American kestrels. PROCEDURES A single dose of hydromorphone (0.6 mg/kg) was administered IM (pectoral muscles) and IV (right jugular vein); the time between IM and IV administration experiments was 1 month. Blood samples were collected at 5 minutes, 1 hour, and 3 hours (n = 4 birds); 0.25, 1.5, and 9 hours (4); and 0.5, 2, and 6 hours (4) after drug administration. Plasma hydromorphone concentrations were determined by means of liquid chromatography with mass spectrometry, and pharmacokinetic parameters were calculated with a noncompartmental model. Mean plasma hydromorphone concentration for each time was determined with naïve averaged pharmacokinetic analysis. RESULTS Plasma hydromorphone concentrations were detectable in 2 and 3 birds at 6 hours after IM and IV administration, respectively, but not at 9 hours after administration. The fraction of the hydromorphone dose absorbed after IM administration was 0.75. The maximum observed plasma concentration was 112.1 ng/mL (5 minutes after administration). The terminal half-life was 1.25 and 1.26 hours after IV and IM administration, respectively. CONCLUSION AND CLINICAL RELEVANCE Results indicated hydromorphone hydrochloride had high bioavailability and rapid elimination after IM administration, with a short terminal half-life, rapid plasma clearance, and large volume of distribution in American kestrels. Further studies regarding the effects of other doses, other administration routes, constantrate infusions, and slow release formulations on the pharmacokinetics of hydromorphone hydrochloride and its metabolites in American kestrels may be indicated.