The use of cytochemistry,immunophenotyping, flow cytometry,and in vitro differentiation to determine the ontogeny of a canine monoblastic leukemia

We evaluated the utility of cytochemistry, immunophenotyping, flow cytometry, and in vitro culture with forced differentiation of leukemic cells as diagnostic aids to identify the malignant cell ontogeny in a dog with leukemia. A tentative diagnosis of monoblastic leukemia was established by microscopic examination of Romanowsky-stained blood smears and bone marrow aspirate smears. This diagnosis also was supported by the light scatter signature that identified the blast cells as large, non-granular monocytic cells using a CellDyn 3500 automated hematology analyzer; as well as by the detection of N-butyrate esterase and the lack of choloroacetate esterase or leukocyte peroxidase by cytochemical staining. Subsequently, leukemic cells were isolated from the dog's peripheral blood and placed into tissue culture or cryopreserved. The leukemic cells grew in suspension cultures and proliferated spontaneously for up to 4 days. By day 7, proliferation was negligible. Upon culture with conditioned supernatant using mitogen-stimulated human T cells as a source of cytokines, an increased proportion of cells entered S phase by day 2 of culture; however, proliferation declined markedly by day 4, at which time the cells had apparently differentiated to adherent, vacuolated macrophages. The cytokine-stimulated leukemic cells were positive for the monocyte/macrophage specific markers alpha-1-antitrypsin, alpha-1-antichymotrypsin, lysozyme, CD14, MHC class II, and calprotectin, an antigen found in differentiated macrophages and granulocytes. Despite the strong tendency of the leukemic cells towards monocytic differentiation, our results suggested that they retained some features of a myelomonocytic precursor. These data show that cytochemistry, immunophenotyping, flow cytometry, and in vitro differentiation of canine leukemia cells are useful tools for confirming the lineage of malignant hematopoietic cells.     

Effects of Postharvest Processing on Texture Profile Analysis of Cooked Rice

The effects of drying conditions, final moisture content, and degree of milling on the texture of cooked rice varieties, as measured by texture profile analysis, were investigated. Instrumentally measured textural properties were not significantly (α = 0.05) affected by drying conditions, with the exception of cohesiveness. Cohesiveness was lower in rice dried at lower temperatures (18°C or ambient) than in that dried at the higher commercial temperatures. Final moisture content and degree of milling significantly (α = 0.05) affected textural property values for adhesiveness, cohesiveness, hardness, and springiness; their effects were interdependent. The effects of deep milling were more pronounced in the rice dried to 15% moisture than that dried to 12%. In general, textural property values for hardness were higher and those for cohesiveness, adhesiveness, and springiness were lower in regular-milled rice dried to 15% moisture than in that dried to 12%. In contrast, hardness values were lower and cohesiveness, adhesiveness, and springiness values were higher in deep-milled rice dried to 15% moisture than in that dried to 12% moisture. Deep milling resulted in rice with lower hardness values and higher cohesiveness, adhesiveness, and springiness values.     

Capture efficiency of a multi-species survey trawl for Snow Crab (Chionoecetes opilio) in the Newfoundland region

An experiment was carried out during September of 2001 to estimate the relative catching efficiency of the NAFC's (Northwest Atlantic Fisheries Centre) standard survey bottom trawl, the Campelen 1800 shrimp trawl, for Snow Crab (Chionoecetes opilio). Secondary trawls were mounted underneath the main trawl directly behind the footgear to estimate escapement of crab passing underneath the main trawl. Overall, the capture efficiency of the survey trawl was found to be much lower than 1.0 and highly variable, with variability being strongly related to substrate type and Snow Crab size. The probability of capture monotonically increased with crab size, and was higher on soft than on hard substrates. Three substrate-specific capture efficiency functions were defined; (1) efficiency was highest (about 0.73–0.95), and least dependent on crab size, on the softest mud substrate; (2) efficiency was lower (about 0.39–0.70) and linearly related to crab size throughout a soft mud–sand substrate; and (3) efficiency was lowest (about 0.05–0.33) on a variety of hard substrates within relatively shallow water. Our models over-estimated true efficiency because secondary trawls were not fully efficient, i.e., crab escaped underneath their footgear. Nevertheless, our experiment demonstrated that substrate type can affect capture efficiency and annual changes in trawl configuration or in Snow Crab distribution in relation to substrate type will affect survey biomass estimates. Additional studies using various approaches could provide a basis for standardizing survey catches for varying capture efficiency effects and refining survey-based estimates of biomass and exploitation rates of Snow Crab.     

Cinnamamide,an avian repellent,reduces woodpigeon damage to oilseed rape

Woodpigeons Columba palumbus cause significant damage to oilseed rape Brassica napus. (Cruciferae) in Britain, especially between January and March. The potential of cinnamamide, a non-lethal chemical repellent, to protect growing oilseed rape from woodpigeon damage was assessed in 20×20 m plots laid out in a 9-ha field frequented by >500 woodpigeons. Despite a low initial loading of cinnamamide and low persistence of the compound after spraying, the inner leaves of treated plants received less damage than the inner leaves of untreated plants (P<0·05). This was reflected later in the trial in a decline in damage to the outer leaves of treated plants (P<0·05). No signs of phytotoxicity were seen on treated leaves during the trial. Cinnamamide did not completely protect the rape, and some plants in treated plots received considerable damage. This was probably due to the compound's poor persistence on the leaf and because it was applied late in the growing season (late February) when birds had already made inroads into the crop. © 1998 SCI.     

Serological Survey of Leptospiral Antibodies in Swine in Quebec

The prevalence of reactors to different serovars of Leptospira interrogans was determined in 117 swine premises in Quebec. A total of 926 sera were tested, using the microscopic agglutination method, against six leptospiral serovars: pomona, icterohaemorrhagiae, grippotyphosa, hardjo, canicola and ballum. Of the sera tested, 30 (3.2%) reacted to one of the three following serovars: pomona (2%), icterohaemorrhagiae (1.1%) and ballum (0.1%).     

Latitudinal variation in growth and otolith‐inferred field metabolic rates of Canadian young‐of‐the‐year Arctic charr

Countergradient variation (CGV) is defined as genetic variation that counteracts the negative influences of the physical environment, minimising phenotypic variability along an environmental gradient. CGV is thought to have relevance in predicting the response of organisms to climate variability and change. To test the hypothesis that growth rate increased with latitude, consistent with CGV, young‐of‐the‐year (YOY) Arctic charr, Salvelinus alpinus, were examined along a ~27° latitudinal gradient in central and eastern Canada. Growth rates were estimated from fork lengths standardised by the thermal opportunity for growth based on experienced water temperatures derived using otolith oxygen stable isotopes. Results demonstrated patterns consistent with CGV, where northern populations demonstrated faster growth rates. A secondary aim was to test for similar geographical patterns in otolith‐inferred metabolic rates, which reflect the energetic costs of standard metabolic rate (SMR) and other processes such as feeding, locomotion, thermoregulation, reproduction and growth. Results demonstrated a significant, positive relationship between otolith‐inferred metabolic rate and latitude, which may reflect an increase in one, or a combination, of the above‐noted physiological processes. The similar latitudinal pattern in growth and otolith‐inferred metabolic rates suggests greater intake of food per unit of time by northern fish. The phenotypic variation in physiological traits observed here demonstrates the significant adaptability of Arctic charr to different thermal regimes with different growing season lengths. Determining the relative contributions of phenotypic plasticity and genetic variation to the observed latitudinal variation will be critical to predicting the responses of Arctic charr to climate change more accurately.     

A dose-finding,long-term study on the use of calcium chloride in saline solution as a method of nonsurgical sterilization in dogs: evaluation of the most effective concentration with the lowest risk

Background

Canine overpopulation is a global issue with serious health and welfare implications. Nonsurgical methods of sterilization could yield positive impacts on this problem, but no long-term data on such methods are available. The objective of the current investigation was to determine the effects of intratesticular injections of calcium chloride dihydrate (CaCl₂) in saline in dogs over a one year period. Five concentrations (0%, 10%, 20%, 30%, 60%) of CaCl₂ in saline were administered via intratesticular injection to groups of 10 dogs each. Total sperm count and motility, blood levels of testosterone, and side effects were examined at 0, 2, 6, and 12 months post-injection (PI). Testicular size and semen volume were examined at 0 and 12 months PI.

Results

Total sperm count, semen volume and testosterone showed significant dose-dependent decreases upon treatment with 10%-60% CaCl₂ compared with either the control group (0% CaCl₂) or baseline for each treatment group. Azoospermia was achieved for at least 12 months PI in 60% and 80% of treated dogs after administration of a 10% and 20% CaCl₂, respectively. Treatment with 30% or 60% CaCl₂ resulted in azoospermia in 100% of dogs, but more side effects were observed, while no side effects were noticed at lower doses. For each treatment group, testosterone levels had decreased an average of 35%-70% at 6 months following treatment. However, testosterone levels rebounded by the 12-month time point in all groups except the highest dosage group (60% CaCl₂), which remained at the low end of physiological range throughout the study. Sperm motility dropped to zero or near zero in all dogs treated with CaCl₂. Testicular size was significantly smaller at 12 months PI for all groups when compared to baseline.

Conclusions

This first long-term study confirms reports of the efficacy of CaCl₂ sterilization. However, at dosages free of adverse events, calcium chloride in saline may not provide permanent sterilization as previously believed. Future work should explore optimized solvents to increase the permanence of the well-tolerated 20% formulation.