Isolation of Cokeromyces recurvatus, initially misidentified as Coccidioides immitis, from peritoneal fluid in a cat with jejunal perforation.

Cokeromyces recurvatus, a zygomycete, was isolated by fungal culture from the peritoneal fluid of a cat with jejunal perforation secondary to intestinal lymphosarcoma. This organism has not been recovered previously from a veterinary patient. The tissue form of C. recurvatus is morphologically similar to those of Coccidioides immitis and Paracoccidioides brasiliensis and may be misdiagnosed as 1 of these organisms on the basis of cytologic or histopathologic specimens, particularly in geographic regions where these organisms are not endemic.     

Optimization and validation of a flow cytometric method for immunophenotyping peripheral blood lymphocytes from cynomolgus monkeys (Macaca fascicularis)

BACKGROUND: Guidelines published by the Food and Drug Administration and Center for Human Medicinal Products describe the need to assess immunotoxic effects in nonclinical studies that evaluate drug toxicity, including the use of immunophenotyping to measure immunotoxicity. We are not aware of previous studies, however, that have validated methods for immunophenotyping peripheral blood lymphocyte subsets in whole blood samples from cynomolgus monkeys. OBJECTIVE: The purpose of this study was to optimize and validate a flow cytometric assay for immunophenotyping lymphocytes in the peripheral blood of cynomolgus monkeys. METHODS: A series of prevalidation experiments were done to determine optimal reagents, volumes, timing, and other procedural details of the flow cytometric assay. Using the optimized method, we then determined precision, interindividual variation, laboratory-to-laboratory variability, and sample stability. Stabilized human blood was used as a positive control for staining, processing, and analysis. The percentage and number of pan-T cells (CD3+), T-helper cells (CD3+4+), T cytotoxic/suppressor cells (CD3+8+), natural killer cells (CD3-16+), and B-cells (CD3-20+) were determined in 146 male and 140 female, clinically healthy monkeys and reference intervals were calculated. RESULTS: By doing 4-color staining with a lyse-wash method, intra- and interassay precision were <5% for all lymphocyte subsets. Variability between technicians and laboratories was minimal (CVs<3%). Samples were stable for up to 24 hours after staining and fixing. CONCLUSIONS: The validated method is extremely robust and can be performed under good laboratory practice conditions to support nonclinical studies. Reference intervals for lymphocyte subsets were similar to those previously reported.     

Surgical management of gallbladder mucoceles in dogs: 22 cases (1999-2003)

OBJECTIVES: To describe preoperative, surgical, and postoperative findings and determine prognostic indicators and treatment recommendations in dogs treated surgically for gallbladder mucocele. DESIGN: Retrospective study. ANIMALS: 22 client-owned dogs. PROCEDURES: Medical records of dogs with gallbladder mucoceles that were treated surgically were reviewed. History, clinical signs, results of selected clinicopathologic analyses and abdominal ultrasonography, surgical procedure performed, results of histologic examination of a liver biopsy specimen, and survival time were recorded. Follow-up information was obtained via telephone interview with owners and referring veterinarians. RESULTS: Dogs were 7 to 15 years of age and had non-specific clinical signs (vomiting, anorexia, and lethargy). Physical examination findings included icterus, signs of depression, and signs of discomfort on palpation of the abdomen. Sixteen dogs had a definitive diagnosis and 6 dogs were strongly suspected of having a gallbladder mucocele on the basis of results of abdominal ultrasonography. Fifteen dogs survived after surgery; 3 of these dogs had bile-induced peritonitis, and 4 had pancreatitis. One dog was euthanatized as a result of severe pancreatitis, and 1 was euthanatized because of acute renal failure; 5 dogs died as a result of pancreatitis, cholecystitis, or bile-induced peritonitis. Hepatic abnormalities were detected histologically in all dogs. CONCLUSIONS AND CLINICAL RELEVANCE: No predictors of survival were identified. No associations between outcome of surgical treatment (survival vs nonsurvival) and preoperative findings, biliary rupture, surgical procedure performed, results of histologic examination of the liver, or development of pancreatitis were found. Cholecystoduodenostomy and cholecystectomy appear to be acceptable treatments for gallbladder mucocele.     

Measurement of serum IgG in foals by radial immunodiffusion and automated turbidimetric immunoassay

Hypogammaglobulinemia as a result of failure of transfer of passive immunity (FTPI) is an important risk factor for infectious disease in neonatal foals. The current gold standard for determining serum immunoglobulin concentrations is radial immunodiffusion (RID). The purpose of this study was to compare immunoglobulin concentrations measured by RID with those determined by an automated turbidimetric immunoassay (TIA), which has a much shorter turnaround time. Immunoglobulin concentrations were measured by both RID and TIA in serum collected from 84 neonatal foals. Sixty-seven foals had results within the linear range for both assays. Sensitivity and specificity of TIA for diagnosis of FTPI with IgG < or = 800 mg/dL were 0.81 (95% CI 0.70-0.88) and 0.86 (95% CI 0.76-0.93) and with IgG < or = 400 mg/dL were 0.63 (95% CI 0.35-0.86) and 0.92 (95% CI 0.87-0.95), respectively. A significant linear relationship was found between IgG concentrations determined by TIA and RID (TIA = 0.9511RID + 8.4354; R2 = .59, P < .0001). The coefficients of variation for between-run and within-run precision for the TIA were 2.5 and 3%, respectively. Storage of samples from 10 foals at -20 degrees C for 10-12 months resulted in a reduction in TIA-measured serum IgG concentration of -17.6% (SD = 3.7%), indicating that long-term storage of samples at -20 degrees C should be avoided. The results of this study indicate that measurement of serum IgG by TIA can be used to evaluate foals for FTPI.     

Clinical outcome of congenital extrahepatic portosystemic shunt attenuation in dogs aged five years and older: 17 cases (1992-2005)

OBJECTIVE: To assess the outcome of extrahepatic portosystemic shunt (EHPSS) treatment in dogs aged 5 years and older. DESIGN: Retrospective case series. ANIMALS: 17 client-owned dogs. PROCEDURES: Medical records for dogs (> or = 5 years old) that underwent surgical attenuation of an EHPSS (1992 through 2005) were evaluated; data, including clinical signs, clinicopathologic findings, surgical procedure, and outcome, were recorded. Follow-up information was obtained via patient examination or telephone interview with veterinarians and owners. RESULTS: Dogs (5 to 9 years old [median age, 6.6 years]) had neurologic (n = 12), urinary tract (8), and gastrointestinal tract (6) EHPSS-associated clinical signs. Serum bile acids and ammonia concentrations were abnormal in all evaluated dogs. Treatment of EHPSSs included complete (n = 6 dogs) or partial (2) suture attenuation or ameroid constrictor placement (9). Two dogs died following surgery. Follow-up information (6 to 120 months) was available for 13 dogs. Deaths were attributable to heart failure (n = 1), bacterial hepatitis (2; with pyelonephritis in 1 dog), and unknown causes (3). At a median of 23 and 25 months, serum bile acids concentrations had almost normalized in 5 of 8 dogs and ammonia concentrations were within reference limits in 3 of 5 dogs, respectively; dogs with abnormal liver function test results had no associated clinical signs. Median long-term survival time was 72 months. CONCLUSIONS AND CLINICAL RELEVANCE: Attenuation of EHPSS in > or = 5-year-old dogs ameliorated signs of liver dysfunction in surviving dogs, although return of normal liver function occurred less frequently than expected.     

Biodiversity management approaches for stream–riparian areas: Perspectives for Pacific Northwest headwater forests,microclimates, and amphibians

Stream–riparian areas represent a nexus of biodiversity, with disproportionate numbers of species tied to and interacting within this key habitat. New research in Pacific Northwest headwater forests, especially the characterization of microclimates and amphibian distributions, is expanding our perspective of riparian zones, and suggests the need for alternative designs to manage stream–riparian zones and their adjacent uplands. High biodiversity in riparian areas can be attributed to cool moist conditions, high productivity and complex habitat. All 47 northwestern amphibian species have stream–riparian associations, with a third being obligate forms to general stream–riparian areas, and a quarter with life histories reliant on headwater landscapes in particular. Recent recognition that stream-breeding amphibians can disperse hundreds of meters into uplands implies that connectivity among neighboring drainages may be important to their population structures and dynamics. Microclimate studies substantiate a “stream effect” of cool moist conditions permeating upslope into warmer, drier forests. We review forest management approaches relative to headwater riparian areas in the U.S. Pacific Northwest, and we propose scenarios designed to retain all habitats used by amphibians with complex life histories. These include a mix of riparian and upslope management approaches to address the breeding, foraging, overwintering, and dispersal functions of these animals. We speculate that the stream microclimate effect can partly counterbalance edge effects imposed by upslope forest disturbances, hence appropriately sized and managed riparian buffers can protect suitable microclimates at streams and within riparian forests. We propose one approach that focuses habitat conservation in headwater areas – where present management allows extensive logging – on sensitive target species, such as tailed frogs and torrent salamanders that often occur patchily. Assuming both high patchiness and some concordance among the distribution of sensitive species, protecting areas with higher abundances of these animals could justify less protection of currently unoccupied or low-density habitats, where more intensive forest management for timber production could occur. Also, we outline an approach that protects juxtaposed headwater patches, retaining connectivity among sub-drainages using a 6th-field watershed spatial scale for assuring well-distributed protected areas across forested landscapes. However, research is needed to test this approach and to determine whether it is sufficient to buffer downstream water quality and habitat from impacts of headwater management. Offering too-sparse protection everywhere is likely insufficient to conserve headwater habitats and biodiversity, while our alternative targeted protection of selected headwaters does not bind the entire forest landscape into a biodiversity reserve.