Proposed criteria for classification of acute myeloid leukemia in dogs and cats

Blood and bone marrow smears from 49 dogs and cats, believed to have myeloproliferative disorders (MPD), were examined by a panel of 10 clinical pathologists to develop proposals for classification of acute myeloid leukemia (AML) in these species. French-American-British (FAB) group and National Cancer Institute (NCI) workshop definitions and criteria developed for classification of AML in humans were adapted. Major modifications entailed revision of definitions of blast cells as applied to the dog and cat, broadening the scope of leukemia classification, and making provisions for differentiating erythremic myelosis and undifferentiated MPD. A consensus cytomorphologic diagnosis was reached in 39 (79.6%) cases comprising 26 of AML, 10 of myelodysplastic syndrome (MDS), and 3 of acute lymphoblastic leukemia (ALL). Diagnostic concordance for these diseases varied from 60 to 81% (mean 73.3 +/- 7.1%) and interobserver agreement ranged from 51.3 to 84.6% (mean 73.1 +/- 9.3%). Various subtypes of AML identified included Ml, M2, M4, M5a, M5b, and M6. Acute undifferentiated leukemia (AUL) was recognized as a specific entity. M3 was not encountered, but this subclass was retained as a diagnostic possibility. The designations M6Er and MDS-Er were introduced where the suffix "Er" indicated preponderance of erythroid component. Chief hematologic abnormalities included circulating blast cells in 98% of the cases, with 36.7% cases having >30% blast cells, and thrombocytopenia and anemia in approximately 86 to 88% of the cases. Bone marrow examination revealed panmyeloid dysplastic changes, particularly variable numbers of megaloblastoid rubriblasts and rubricytes in all AML subtypes and increased numbers of eosinophils in MDS. Cytochemical patterns of neutrophilic markers were evident in most cases of Ml and M2, while monocytic markers were primarily seen in M5a and M5b cases. It is proposed that well-prepared, Romanowsky-stained blood and bone marrow smears should be examined to determine blast cell types and percentages for cytomorphologic diagnosis of AML. Carefully selected areas of stained films presenting adequate cellular details should be used to count a minimum of 200 cells. In cases with borderline diagnosis, at least 500 cells should be counted. The identity of blast cells should be ascertained using appropriate cytochemical markers of neutrophilic, monocytic, and megakaryocytic differentiation. A blast cell count of > 30% in blood and/or bone marrow indicates AML or AUL, while a count of < 30% blasts in bone marrow suggests MDS, chronic myeloid leukemias, or even a leukemoid reaction. Myeloblasts, monoblasts, and megakaryoblasts comprise the blast cell count. The FAB approach with additional criteria should be used to distinguish AUL and various subtypes of AML (Ml to M7 and M6Er) and to differentiate MDS, MDS-ER, chronic myeloid leukemias, and leukemoid reaction. Bone marrow core biopsy and electron microscopy may be required to confirm the specific diagnosis. Immunophenotyping with lineage specific antibodies is in its infancy in veterinary medicine. Development of this technique is encouraged to establish an undisputed identity of blast cells. Validity of the proposed criteria needs to be substantiated in large prospective and retrospective studies. Similarly, clinical relevance of cytomorphologic, cytochemical, and immunophenotypic characterizations of AML in dogs and cats remains to be determined.     

A histomorphologic and immunohistochemical study of chordoma in twenty ferrets (Mustela putorius furo).

The histomorphologic and immunohistochemical features of chordoma in 20 ferrets were evaluated. The mean age was 3.4 years, and, in the cases for which sex was known, females (n = 10) outnumbered males (n = 5) two to one. All 20 tumors occurred on the tip of the tail. Nineteen of 20 tumors (95%) were composed of three tissue components, often arranged concentrically with lobules of physaliferous cells at the periphery, trabecular bone in the center, and cartilage in between. The bone often contained marrow and hematopoietic cells. One tumor lacked chondromatous or osseous tissue. Immunohistochemical results were consistent with previous studies of chordoma. All 20 tumors (100%) were positive for keratin and vimentin intermediate filaments; 15 (75%) were positive for S-100 protein; and 17 (85%) were positive for neuron specific enolase. This neoplasm shares morphologic and immunohistochemical features with "classic," as well as chondroid chordoma, of human beings, making it a potential animal model.     

The anti-proteolytic activity of blood and ovarian follicular fluid in sheep after stimulation with serum gonadotropin (PMSG) and administration of Antisergon]

The synthesis and secretion of trypsin (trypsin model serine protease) inhibitors are regulated in ovarian follicles by gonadotropins. The superovulation stimulations with 400 IU FSH, 1000 IU PMSG, 1000 IU HCG, 750 IIU PMSG + 750 IU HCG influence in a different way the trypsin inhibiting activities (TIA) of blood plasma (BP) (Figs 1 and 2) and follicular fluid (fig. 3); this points to a possibility of local effects. An increase in the average values of TIA in BP was statistically significant during the whole experiment: P less than 0.05 to P less than 0.001 (following the administration of PMSG+HCG, or PMSG, and HCG); Antisergon administered in 68 hours after PMSG reduced this increase. The changes in the fraction of low-molecular TIA in BP (after BP treatment with perchloric acid) were of converse nature; a decrease in the average values ranged from P less than 0.02 to P less than 0.001 (following PMSG or other stimulations). Antisergon did not influence this decrease. The changes observed on particular days of the trial (Figs. 1 and 2) also indicate different effects of the preparations, mainly of the component LH, which resulted in the occurrence of large nonovulating follicles (greater than 10 mm--"cystic" ones). No such follicles were observed in nonstimulated ewes and after FSH stimulation. The administration of antisergon (goat's antiserum against PMSG) 68 hours after PMSG administration did not prevent their creation. The TIA of follicular fluid (FF) of antral follicles was on average tenfold in comparison with that of blood plasma; and the TIA FF of follicles greater than 10 mm was higher (up to P less than 0.001) than the TIA FF of follicles less than 10 mm. The administration of Antisergon in shorter intervals following PMSG administration (12, 24, 48 and 58 hours) influenced the average values of TIA BP in 120 hours (since PMSG administration) in dependence on time (Tab. I). The effects of Antisergon administered in 12 and 24 hours after PMSG administration on the TIA BP were insignificant if it was administered in 48 and 58 hours the TIA BP increased (P less than 0.02; P less than 0.001) in comparison with the interval of 12 hours. The TIA FF of follicles less than 5 mm, 5-10 mm and greater than 10 mm varied in dependence on the time intervals of Antisergon administration (Fig. 4). The statistical significance of these changes in shown in Tab. II.(ABSTRACT TRUNCATED AT 400 WORDS)     

Induction of lactogenic immunity to transmissible gastroenteritis virus of swine using an attenuated coronavirus mutant able to survive in the physicochemical environment of the digestive tract.

A transmissible gastroenteritis (TGE) coronavirus mutant (188-SG), selected as attenuated and resistant to acidity and proteases of the digestive tract of adult pigs, was used as vaccine ("Nouzilly strain") in sows to protect suckling piglets against a challenge exposure carried out with a highly virulent TGEV strain. The pregnant sows were immunized once (42-49 days before farrowing) or twice (42-49 and 7-15 days before farrowing) by the oral, intramuscular or conjunctival route with the 188-SG strain. Sows exposed to virulent TGEV in the field and experimentally infected sows (two oral inoculations during pregnancy) were used as positive controls leading to high protection. The neutralizing antibody response to vaccination and/or infection was studied in serum and milk. No protection against mortality was observed in the litters of (1) the nine seronegative, susceptible sows, with piglet mortality of 65/70, (2) the seven once orally vaccinated sows, with mortality of 44/54, (3) the seven sows vaccinated twice by the conjunctival route, with mortality of 55/76. Moderate protection was observed in (1) the eight sows vaccinated intramuscularly twice with piglet mortality of 36/90, (2) the seven orally and intramuscularly vaccinated sows with piglet mortality of 31/51. In of 3 contrast, improved protection was observed in (1) the 10 sows vaccinated twice orally, with piglet mortality of 23/95, (2) the four naturally infected sows with piglet mortality of 6/41, (3) the six sows experimentally infected with virulent TGEV with piglet mortality of 1/59. No correlation was found between neutralizing antibodies titers in serum and milk and protection rate of the piglets. The results indicate that relative protective lactogenic immunity against TGEV is induced only by repeated ingestion of the attenuated 188-SG strain of TGEV.     

Comparative studies of BHV-1 variants by in vivo--in vitro tests.

The new encephalitogenic BHV-1.3 and previously characterized BHV-1 strains were studied with reference to their immunogenic and protective potency and their antigenic relationships using "in vitro" and "in vivo" tests. The "in vitro" results obtained by neutralization kinetics showed that the Los Angeles (LA) strain (BHV-1.1) and a vaginal isolate L-114 strain (BHV-1.2) had antigenic similarities. Conversely, the behavior of the encephalitogenic strain A-663 (BHV-1.3), was significantly distinct. The "in vivo" protection test was carried out in calves using LA and A-663 strains. Post-vaccination antibodies and challenge with A-663 strain showed that the immunogenic behavior and protective capacities of both strains were similar. Neutralization kinetics differences between BHV-1.1 and BHV-1.3 did not alter the "in vivo" protection against BHV-1.3 challenge.     

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 10⁸ colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×10³ and 2.3×10⁴ cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R ²=0.74).     

Transmission of the agent causing a melon yellowing disease by the greenhouse whitefly Trialeurodes vaporariorum in southeast Spain

The agent causing a yellowing disease of melon (Cucumis melo), which results in severe losses in crops under plastic on the coastal plains of southeast Spain, was shown to be transmitted in a semipersistent manner by the greenhouse whitefly (Trialeurodes vaporariorum Westwood). The agent was transmitted by grafting, but not by mechanical inoculation or through seeds. The agent was acquired in the minimum period tested (2 h) and could infect plants in an infection feeding interval of 6 h. Capsella bursa-pastoris, Cucumis melo, C. sativus, Cucurbita moschata, Cichorium endivia, Lactuca sativa andTaraxacum officinale were found susceptible.Results suggest that the yellowing disease affecting melon crops in the southeast of Spain is due to a pathogen similar to beet pseudo yellows virus, but this has to be confirmed by serology.     

Landscape-ecological mapping of the Netherlands

The Landscape-ecological Mapping of the Netherlands project (LMN project) started in 1983 with the aim of establishing a landscape-ecological database for use in developing and evaluating national land-use plans. The project, working with grid cells of 1 km², has four working objectives: a) development of mapping potential for basic landscape-ecological data, b) assessment of susceptibility to interventions, c) evaluation of significance for nature conservation and d) production of vulnerability maps, as a combination of susceptibility and significance. In addition to information on soil, groundwater, ecotopes, flora and fauna, the database also incorporates information on physiographical features and entire landscapes. The resulting database is a geographic information system (GIS). This article describes the second phase of the project (1985–1989), covering the Randstad area, and focusses on the methods and the applications potential of the database.