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.     

Comparison of bronchoalveolar lavage findings and measurements of gas exchange during exercise in horses with poor racing performance

Twenty-four Thoroughbred and twelve Standardbred racehorses aged between 2 and 6 years, presented for reported poor racing performance, underwent clinical exercise testing. During the last 10 s of exercise at each speed throughout an incremental speed exercise test on a treadmill inclined at a 10% slope, samples of arterial blood and expired gases were collected. Maximum oxygen uptake and the partial pressures of oxygen and carbon dioxide in arterial blood were determined. These values were compared between the two breeds of horses and also with reference to cytological findings of bronchoalveolar lavage samples, including neutrophil, erythrocyte and haemosiderophage percentage and the total nucleated cell concentration. The results revealed an inverse relationship (Spearman R = -0.45, p < 0.05) between the total nucleated cell count in bronchoalveolar lavage samples and arterial oxygen partial pressure during exercise at 11 m.s(-1). This result suggests that subclinical pulmonary disease may be a more important cause of poor racing performance than previously thought. Also of note was a positive correlation (Spearman R = 0.50, p < 0.05) between maximum oxygen uptake and the percentage of erythrocytes.     

Cytochemical staining characteristics of lymph nodes from normal and lymphoma-affected dogs

Frozen sections and imprint smears were used to evaluate the presence and pattern of cytochemical staining reactions in the B- and T-cell regions of lymph nodes from normal dogs and dogs with lymphoma. Staining procedures evaluated included peroxidase (PER), Sudan black B (SBB), naphthol AS-D chloroacetate esterase (CAE), alpha-naphthyl butyrate esterase (NBE), acid phosphatase (ACP), and leukocyte alkaline phosphatase (LAP). In normal lymph nodes, macrophages and some lymphocytes within the interfollicular (T-cell) region and medulla stained positive with ACP and NBE. Smaller numbers of macrophages also occurred sporadically within the germinal follicles. Cells positive for PER, SBB, and CAE were scattered infrequently throughout all regions of the normal lymph node, consistent with granulocytes and mast cells. The LAP stained cells were predominantly and prominently located within the mantle zone of secondary follicles and to a much lesser extent within the germinal centers, compatible with B-cell lymphocytes derived from follicular center cells. Of the 12 dogs with lymphoma, 7 cases (4 immunoblastic, 2 large noncleaved, 1 small noncleaved) stained diffusely positive with LAP, 4 cases (all lymphoblastic) had numerous focally positive lymphocytes using ACP and NBE, and 1 case (immunoblastic) did not stain positive with any of the cytochemical reactions. Cytochemical staining of canine lymph nodes with NBE, ACP, and LAP proved useful in distinguishing between B- or T-cell regions and detecting different cell types of canine lymphoma.     

Soil CO2 emission in sugarcane management systems

Sugarcane management systems affect soil attributes such as the carbon cycle. This fact has stimulated the sugar and alcohol industry to refine the sugarcane production systems by replacing the pre-harvest burning (PB) and manual harvest with mechanized harvesting followed by residue deposition. The aim of this study was to evaluate different management systems with respect to C cycling carbon dioxide and soil parameters (chemical, physical and biological) which were determined over the season. Three sugarcane cultivation systems were evaluated at the following periods: (a) PB, (b) 5-year green harvest and (c) 10-year green harvest. The results indicated that CO₂ emission was 36% greater in the 10-year sugarcane green harvest system than in the PB system. The bulk density and macroporosity were the factors that were most affected by the different sugarcane management systems and that significantly influenced soil CO₂ emissions. The principal component analysis showed that soil CO₂ emission was 18% influenced by base saturation (V%) and 14% by pH, especially in the PB area. Additionally, 19% was affected by carbon and macroporosity in the 5- and 10-year green harvest areas, respectively. From our results, it can be concluded that the most CO₂ emissions are in the areas of sugarcane green, this is due to the higher carbon concentration when compared with the area of burning sugarcane. The parameters that most influenced the CO₂ emissions were bulk density, porosity, macroporosity, pH and V%.     

Identification of infrared absorption spectral characteristics of synovial fluid of horses with osteochondrosis of the tarsocrural joint

OBJECTIVE: To determine the feasibility of the use of Fourier-transform infrared (FTIR) spectroscopy within the midinfrared range to differentiate synovial fluid samples of joints with osteochondrosis from those of control samples. ANIMALS: 33 horses with osteochondrosis of the tarsocrural joint and 31 horses free of tarsocrural joint disease. PROCEDURES: FTIR spectroscopy of synovial fluid was used. Sixty-four synovial fluid samples from the tarsocrural joint were collected. Of these, 33 samples were from horses with radiographic evidence of osteochondrosis of the tarsocrural joint and 31 from control joints. Disease-associated features within infrared spectra of synovial fluid were statistically selected for spectral classification, and the variables identified were used in a classification model. Linear discriminant analysis and leave-one-out cross-validation were used to develop a classifier to identify joints with osteochondrosis. RESULTS: 12 significant subregions were identified that met the selection criteria. The stepwise discriminant procedure resulted in the final selection of 6 optimal regions that most contributed to the discriminatory power of the classification algorithm. Infrared spectra derived from synovial fluid of joints with osteochondrosis were differentiated from the control samples with accuracy of 77% (81% specificity and 73% sensitivity). CONCLUSIONS AND CLINICAL RELEVANCE: The disease-associated characteristics of infrared spectra of synovial fluid from joints with osteochondrosis may be exploited via appropriate feature selection and classification algorithms to differentiate joints with osteochondrosis from those of control joints. Further study with larger sample size including age-, breed-, and sex-matched control horses would further validate the clinical value of infrared spectroscopy for the diagnosis of osteochondrosis in horses.     

Interpretation of solute profile dynamics in irrigated soils

Summary Knowledge of the flux of water flowing through macropores in soils is required to devise management strategies for efficient fertiliser use and to prevent fast movement of solutes and pollutants to groundwaters. Water and solute balances in soil profiles were used to develop a simple model for assessing the magnitude of macropore flow. Fluxes of water bypassing the soil matrix were calculated at 35 sites to be between 0 and 415 mm y^–1, with the flux being < 200 mm y^–1 at most sites. The maximum flux was three times the flux flowing through the soil matrix but only one third of that infiltrating the soil. The flux of macropore flow was not simply related to soil types or soil properties, although the highest fluxes did occur in cracking soils. A qualitative method of using soil chloride profiles to indicate the occurrence (but not magnitude) of bypass flux was also demonstrated. Both these quantitative and qualitative assessments of bypass flow should assist in interpreting root-zone hydrology in soils.     

Determining resistance to <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> and fumonisin accumulation in African maize inbred lines resistant to <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and aflatoxins

Fusarium verticillioides and Aspergillus flavus cause Fusarium ear rot (FER) and Aspergillus ear rot (AER) of maize, respectively. Both pathogens are of concern to producers as they reduce grain yield and affect quality. F. verticillioides and A. flavus also contaminate maize grain with the mycotoxins fumonisins and aflatoxins, respectively, which has been associated with mycotoxicosis in humans and animals. The occurrence of common resistance mechanisms to FER and AER has been reported. Hence, ten Kenyan inbred lines resistant to AER and aflatoxin accumulation were evaluated for resistance to FER, F. verticillioides colonisation and fumonisin accumulation; and compared to nine South African lines resistant to FER and fumonisin accumulation. Field trials were conducted at three localities in South Africa and two localities in Kenya. FER severity was determined by visual assessment, while F. verticillioides colonisation and fumonisin content were quantified by real-time PCR and liquid chromatography tandem mass spectrometry, respectively. Significant genotype x environment interactions was determined at each location (P ≤ 0.05). Kenyan inbred CML495 was most resistant to FER and F. verticillioides colonisation, and accumulated the lowest concentration of fumonisins across localities. It was, however, not significantly more resistant than Kenyan lines CML264 and CKL05015, and the South African line RO549 W, which also exhibited low FER severity (≤5%), fungal target DNA (≤0.025 ng μL^?1) and fumonisin levels (≤2.5 mg kg^?1). Inbred lines resistant to AER and aflatoxin accumulation appear to be promising sources of resistance to F. verticillioides and fumonisin contamination.     

Landscape dynamics of floral resources affect the supply of a biodiversity-dependent cultural ecosystem service

Context

Cultural ecosystem services, many of which depend on biodiversity, are recognized as important but seldom quantified biophysically across landscapes. Furthermore, many ecosystem service models are static, and the supply of cultural ecosystem services may be misrepresented if seasonal shifts in biotic communities are ignored.

Objectives

We modeled landscape dynamics of wildflower blooms in a temperate montane landscape to determine (1) how floral resources (wildflower species richness, abundance, timing, and presence of charismatic species) changed over the growing season, (2) how projected wildflower viewing hotspots varied over space and time, and (3) how spatial shifts in floral resources affected potential public access to wildflower viewing.

Methods

Data were collected at 63 sites across a rural-to-urban gradient in the Southern Appalachian Mountains (USA). Generalized linear models were used to identify factors affecting floral resources at two temporal scales. Floral resources were projected across the landscape and hotspots of wildflower viewing were quantified using overlay analysis.

Results

Floral resources were affected by topoedaphic conditions, climate, and surrounding building density and changed seasonally. Seasonal models revealed locational shifts in ecosystem service hotspots, which changed the proportion of hotspots accessible to the public and identified wildflower-viewing opportunities unnoticed by static models.

Conclusion

Relationships between landscape gradients, biodiversity, and ecosystem service supply varied seasonally, and our models identified cultural ecosystem service hotspots otherwise obscured by simple proxies. Landscape models of biodiversity-based cultural ecosystem services should include seasonal dynamics of biotic communities to avoid under- or over-emphasizing the importance of particular locations in ecosystem service assessments.

     

Environmental heterogeneity and biotic interactions as potential drivers of spatial patterning of shorebird nests

Context

Species distributions are driven by a wide variety of abiotic and biotic factors, including nest placement for breeding individuals. As such, the spatial distribution of nests within a landscape can reflect environmental heterogeneity, habitat preferences, or even interactions with predators and other species.

Objectives

We determined the extent to which environmental heterogeneity and predation risk accounted for the observed spatial distribution of nests.

Methods

We assessed the spatial distribution of 112 nests of a migratory shorebird, the Hudsonian Godwit (Limosa haemastica), at Beluga River, Alaska, from 2009 to 2012, and explicitly tested for the relative influence of habitat characteristics and predation risk on nest locations. We also evaluated the effect of nest location, distance to conspecific nests, and proximity to roads on nest fate using 64 nests that were monitored through completion.

Results

Hudsonian Godwit nests were clustered across the landscape despite a lack of significant spatial autocorrelation (i.e., patchiness) in vegetation characteristics at either the micro- or landscape scale. Nest fate also was not predicted by either the distance to the nearest conspecific neighbor or proximity to roads. Thus, neither habitat characteristics nor predation risk explained the clustering of godwit nests.

Conclusions

These results suggest that godwits may select nest locations based more on social cues than underlying heterogeneity in vegetation or predation risk. As such, intra- and inter-specific interactions should be considered when developing management plans for species of conservation concern.