International Guidelines for Veterinary Tumor Pathology: A Call to Action.

Standardization of tumor assessment lays the foundation for validation of grading systems, permits reproducibility of oncologic studies among investigators, and increases confidence in the significance of study results. Currently, there is minimal methodological standardization for assessing tumors in veterinary medicine, with few attempts to validate published protocols and grading schemes. The current article attempts to address these shortcomings by providing standard guidelines for tumor assessment parameters and protocols for evaluating specific tumor types. More detailed information is available in the Supplemental Files, the intention of which is 2-fold: publication as part of this commentary, but more importantly, these will be available as "living documents" on a website (www.vetcancerprotocols.org), which will be updated as new information is presented in the peer-reviewed literature. Our hope is that veterinary pathologists will agree that this initiative is needed, and will contribute to and utilize this information for routine diagnostic work and oncologic studies. Journal editors and reviewers can utilize checklists to ensure publications include sufficient detail and standardized methods of tumor assessment. To maintain the relevance of the guidelines and protocols, it is critical that the information is periodically updated and revised as new studies are published and validated with the intent of providing a repository of this information. Our hope is that this initiative (a continuation of efforts published in this journal in 2011) will facilitate collaboration and reproducibility between pathologists and institutions, increase case numbers, and strengthen clinical research findings, thus ensuring continued progress in veterinary oncologic pathology and improving patient care.

Comparison of bovine hematology reference intervals from 1957 to 2006.

BACKGROUND: The most commonly used bovine hematology reference intervals were published in 1965. We found the results from healthy cattle in 2001 differed from those in many ways. Discovery of the original laboratory book used to calculate the 1965 values gave us the opportunity to evaluate whether hematology values of healthy cattle have changed over time. OBJECTIVE: The purpose of this study was to establish hematology reference intervals for Holstein cows, compare selected hematologic results with similar population data from 1957, and compare these reference intervals with those of other North American veterinary schools and published values. METHODS: Reference intervals were developed in 2001 using clinically healthy, bovine leukemia virus-negative, mid-lactation Holstein cows. Selected parts of the hemograms and neutrophil:lymphocyte (N:L) ratio were compared with those from healthy, age-matched Holstein cows evaluated in 1957. Bovine reference intervals were solicited from clinical pathology laboratories in North American veterinary colleges and analyzed for population characteristics and method of analysis. RESULTS: Between 1957 and 2001, mean neutrophil counts increased significantly, whereas lymphocyte, monocyte, and eosinophil counts and hemoglobin concentration decreased significantly. Mean N:L ratio increased significantly to 1.17. Most surveyed laboratories were using the 1965 reference intervals. Two other institutions that had developed reference intervals after 2000 had results similar to ours. CONCLUSIONS: Continued use of older bovine hematology reference intervals could lead to misinterpretation of within-reference neutrophil counts as neutrophilia and under-recognition of neutropenia, eosinophilia, monocytosis, or lymphocytosis. Use of N:L>1 as evidence of inflammation should be discontinued or used with great caution.

Circulating siderophagocytes and erythrophagocytes in a corn snake (Elaphe guttata) after coelomic surgery.

Leukocytes containing nonheme iron and phagocytosed fragments of erythrocytes were found in blood smears from a corn snake (Elaphe guttata) collected 20 and 79 days after coelomic surgery (ovariosalpingectomy). Numerous immature and mitotic erythrocytes also were seen in the sample taken 20 days postsurgically. Siderophagocytes and erythrophagocytes had not been observed before surgery and were not found in multiple subsequent blood samples collected 112-602 days after surgery. Other than these hematologic abnormalities, laboratory findings were unremarkable and the snake recovered uneventfully. Based on examination of sequential blood smears, the circulating siderophagocytes were interpreted as recirculating macrophages involved in the removal of blood from the coelomic cavity after mild postsurgical hemorrhage.

Serum biochemical abnormalities in goats with uroliths: 107 cases (1992-2003).

OBJECTIVE: To characterize serum biochemical abnormalities in goats with uroliths. DESIGN: Retrospective case-control series. ANIMALS: 107 male goats with uroliths and 94 male goats with various nonrenal diseases (controls). PROCEDURES: For male goats, results of serum biochemical analyses collected from 1992 through 2003 were retrieved from computerized records, as were signalment, clinical diagnoses, and discharge status. Results of analyses for BUN, creatinine, phosphorus, calcium, Na, K, Cl, total CO2, anion gap, and glucose were compared between goats with uroliths and control goats. RESULTS: Goats with uroliths had higher mean BUN, creatinine, total CO2, K, and glucose concentrations and lower mean phosphorus, Na, and Cl concentrations than control goats, with no difference in mean calcium concentration and anion gap. Goats with uroliths had higher frequency of azotemia, hypophosphatemia, hypochloridemia, and increased total CO2 and lower frequency of decreased total CO2 than control goats. Urolithiasis occurred more frequently in castrated males than in sexually intact males and in dwarf African breeds than in other breeds. CONCLUSIONS AND CLINICAL RELEVANCE: Goats with uroliths often had hypophosphatemia at admission. Hypochloridemic metabolic alkalosis was the most common acid-base disorder. Rupture in the urinary tract system was associated with increased prevalence of hyponatremia and hyperkalemia. Clinicians should be aware of these abnormalities when determining fluid therapy.

Identification of a novel hemotropic mycoplasma in a splenectomized dog with hemic neoplasia.

A 3-year-old sexually intact male Bull Mastiff underwent splenectomy for splenic thrombosis; prior to and after splenectomy, multiple blood transfusions were administered. Two weeks after the procedure, T-cell lymphoproliferative disease was diagnosed. Treatment with prednisone and chlorambucil was initiated, and 2 weeks later, cytologic examination of a blood smear revealed small (0.3 microm), coccoid basophilic bodies on the surface of approximately 70% of the RBCs. Morphologically, these resembled "Candidatus Mycoplasma haemominutum." A polymerase chain reaction assay was used to amplify a partial 16S rRNA sequence in blood obtained from the dog; the product was sequenced and compared with 16S rRNA gene sequences of other hemotropic mycoplasmas. The sequence was 98% homologous to that of "Candidatus M haemominutum", but only 77% homologous to that of M haemocanis and M haemofelis.

Effect of preexisting FeLV infection or FeLV and feline immunodeficiency virus coinfection on pathogenicity of the small variant of Haemobartonella felis in cats.

OBJECTIVE: To investigate the effects of preexisting FeLV infection or FeLV and feline immunodeficiency (FIV) coinfection on the pathogenicity of the small variant of Haemobartonella felis (Hfsm, California variant) in cats. ANIMALS: 20 FeLV infected, 5 FeLV-FIV coinfected, and 19 retrovirus-free cats. PROCEDURES: A client-owned cat, coinfected with FeLV and Hfsm, was the source for Hfsm. Inoculum 1 (FeLV free) was obtained by passage of source Hfsm through 4 FeLV-resistant cats. Inoculum 2 was obtained by further passage of Hfsm (inoculum 1) through 2 specific pathogen-free cats. RESULTS: A mild-to-moderate anemia started 21 days after inoculation, with its nadir occurring at 35 to 42 days after inoculation. Infection with Hfsm induced greater decrease in hemoglobin concentration in FeLV infected cats, compared with retrovirus free cats. Reticulocytosis, macrocytosis, and polychromasia of erythrocytes developed in anemic cats regardless of retrovirus infection status. Mean neutrophil counts decreased during the hemolytic episode. For most cats, the anemia was transient. Four FeLV infected cats, 1 of which was also FIV infected, developed fatal FeLV-associated myeloproliferative diseases. Of the surviving cats, 8 died over the next 24 months from other FeLV-related diseases. Hemolysis did not recur after the initial episode. Inoculum 1 induced more severe anemia than inoculum 2. CONCLUSIONS AND CLINICAL RELEVANCE: Our results support the clinical observation that cats coinfected with FeLV and H felis develop more severe anemia than cats infected with H felis alone. Infection with Hfsm may induce myeloproliferative disease in FeLV infected cats. The small variant of H felis may lose pathogenicity by passage through FeLV-free cats.

The usefulness and limitations of hand-held refractometers in veterinary laboratory medicine: an historical and technical review.

Medical hand-held refractometers have been used in veterinary practice since their development in the 1960s. They have become ubiquitous for the measurement of protein and urine solute concentrations because of their rapidity of analysis, ease of use, and relatively low cost. Refraction of light offers advantages for the determination of solute concentrations because the measurement requires no chemical alteration of the specimen. Numerous authors have reported that the results of protein estimation by refractometry for domestic mammals correlate well with those obtained by the biuret method, although others have reported both higher and lower refractometric results compared with biuret results. Major discrepancies between biuret and refractometric results have been reported for avian samples. Some of the variation in reported results may be due to differences in design by refractometer manufacturers. Another possible source may be variation in the biuret reagent mixture and assay conditions. Refractometers also can be used to calculate serum water concentration. A table that converts index of refraction to serum water concentration can be used to convert electrolyte concentration from mmol/L of serum to mmol/L of serum water, a more accurate indicator of effective electrolyte concentration. Refractometers are especially useful for determining urine specific gravity on veterinary samples because they require relatively small sample volumes. Specific gravity continues to be the most common unit for reporting total solids concentration. Some solutes, such as acetone, may cause false increases in specific gravity by refractometry, as they increase refraction but are less dense than water.

Comparison of refractometer and biuret methods for total protein measurement in body cavity fluids.

Most hand-held medical refractometers have internal scales that limit protein measurement to results >/=2.5 g/dL. Tables for conversion of refraction (r) to protein concentration for values as low as 0.1 g/dL were published in the 1960s, but their accuracy for use on body fluids has not been established. The purpose of this study was to assess the reliability of body cavity fluid protein determination by refractometry. We compared the protein concentration of 25 body cavity fluids as determined by 2 Goldberg type hand-held refractometers with results obtained by the biuret method. Published charts converting refraction (r) to protein concentration were used to determine protein concentration in samples with protein <2.5 g/dL. Higher protein values were read directly from the instruments. The range of comparison was limited to >/=0.6 g/dL, the lowest concentration of the biuret method's standard curve. Twenty-one peritoneal fluid, 2 pleural fluid and 2 pericardial fluid samples from 16 horses, 5 cattle, 3 dogs, 2 llamas and 1 cat were tested. The results obtained by the two refractometers were closely and linearly related to biuret results (P<.001), with slopes by linear regression analysis close to 1, and correlation coefficients >0.977. Based on this study, the range for quantification of body cavity fluid protein concentration by refractometry can be extended below 2.5 g/dL, allowing for quantitative assessment of most clinical samples.