Analytical methods for mycotoxins

Biological, physico-chemical and immunochemical methods are available today for solving the multiple problems which turn up in the course of mycotoxin diagnosis and mycotoxin research. In this review, selected analysis techniques are presented and their limits and possibilities are discussed. The biological methods (chicken-embryo-test, skin irritation test, cell cultures) indicate only toxicities, mostly without further specification for mycotoxins. In contrast, modern physico-chemical analysis (DC, HPLC, CLC, MS) allows the detection of defined mycotoxins in the picogram range; these methods should be employed or official and judicial purposes. The recently developed immunochemical methods (RIA, EIA) are characterized by a simple sample preparation, a short test duration and a high sensitivity; they could be used as a screening method in the course of mycotoxicological control of food and feed.     

Anti-erythrocyte membrane antibodies detected in sera of dogs naturally infected with Babesia gibsoni.

Due to the potential for anti-erythrocyte membrane antibodies as possible enhancers of erythrocyte destruction, the presence of serum anti-erythrocyte membrane antibodies in 31 dogs with Babesia gibsoni infection admitted to a veterinary hospital was investigated by an enzyme linked immunosorbent assay (ELISA) and immunoblotting analyses. This infection resulted in an increase of anti-erythrocyte membrane antibodies in 84% (IgG) and 74% (IgM) of 31 infected dogs, respectively. This was confirmed by the similarity in the protein profiles of the erythrocyte membrane antigens immunoblotted with rabbit antiserum to dog erythrocyte membrane antigens and infected dog serum. These results suggest the production of anti-erythrocyte membrane antibodies was induced by B. gibsoni infection.     

Stoffbildung, CO2-Gaswechsel, Kohlenhydrat- und Stickstoffgehalt von Winterweizen bei erhöhter CO2-Konzentration und Trocken-streß

Dry Matter Production, CO₂ Exchange, Carbohydrate and Nitrogen Content of Winter Wheat at Elevated CO₂ Concentration and Drought Stress
Methods of mathematical modelling and simulation are being used to an increasing degree in estimating the effects of rising atmospheric CO₂ concentration and changing climatic conditions on agricultural ecosystems. In this context, detailed knowledge is required about the possible effects on crop growth and physiological processes. To this aim, the influence of an elevated CO₂ concentration and of drought stress on dry matter production, CO₂ exchange, and on carbohydrate and nitrogen content was studied in two winter wheat varieties from shooting to milk ripeness. Elevated CO₂ concentration leads to a compensation of drought stress and at optimal water supply to an increase of vegetative dry matter and of yield to the fourfold value. This effects were caused by enhanced growth of secondary tillers which were reduced in plants cultivated at atmospheric CO₂ concentration. Analogous effects in the development of ear organs were influenced additionally by competitive interactions between the developing organs. The content and the mass of ethanol soluble carbohydrates in leaves and stems were increased after the CO₂ treatment and exhausted more completely during the grain filling period after drought stress. Plants cultivated from shooting to milk ripeness at elevated CO₂ concentration showed a reduced response of net photosynthesis rate to increasing CO₂ concentration by comparison with untreated plants. The rate of dark respiration was increased in this plants.     

Plasma Cell Myeloma in the Horse

Plasma cell myelomas in horses have been reported infrequently. Data from 10 cases, 9 from the literature and 1 new case, are used to characterize the disease in the horse. Hot-blooded horses (7/10), specifically Quarter Horses (4/10), were most often affected. Median age at diagnosis was 11 years (range, 3 mo-22 yr) and both male (5) and female horses (5) were represented equally. Clinical findings included weight loss (6/8), anorexia (4/8), fever (4/8), limb edema (4/8), pneumonia (3/8), rear leg paresis/ataxia (3/8), epistaxis (3/8), palpable lymphadenopathy (2/8), and bone pain (2/8). Anemia (8/8) was present routinely, and in three horses, RBCs were macrocytic. Leukopenia (2/8), thrombocy-topenia (2/8), and circulating plasma cells (3/8) were variable findings. Except for abnormal protein concentrations and hyponatremia (3), abnormal results from serum biochemical analysis including hypo-cholesterolemia (1), hypercalcemia (1), and azotemia (1) were reported infrequently. Hyperproteinemia (8/9), hypoalbuminemia (7/9), and hyperglobulinemia (8/9) were characteristic but not invariable findings. Monoclonal proteins (7/7) were detected in the α₂, β, or γ region by serum electrophoresis. The paraprotein's heavy chain, determined in four horses, was a subclass of IgG. Three horses had decreased concentrations of normal immunoglobulins. Variable proteinuria (trace to 4+) was detected by routine urinalysis in four of six horses. Bence Jones proteinuria was detected in one of five horses (heat precipitation) and monoclonal proteins were detected in two of three electrophoresed urine samples. Three of the horses had lytic bone lesions detected radiographically. Bone marrow aspirates were diagnostic in two of five horses. Atypical plasma cells or increased numbers of plasma cells or both were present in histologic sections of bone marrow in six of eight horses. Common extraosseous sites of plasma cell infiltration included lymph nodes (8/8), kidney (5/8), spleen (5/8), liver (3/8), lung (3/8), brain (2/8), and orbit (2/8). Two horses had intracellular and extracellular crystalline deposits.     

Thrombocyte aggregation in the cat

Platelet aggregation in healthy and sick cats after adding various aggregating agents is described. Feline platelets aggregate irreversibly in response to 0.15-1.0 micrograms/ml collagen, 1 microM ADP, 0.3 IU/ml test-thrombin and 0.71 NIH/ml Topostasin. Epinephrine, ristocetin and kaolin failed to cause aggregation. The aggregation function was decreased in a cat with liver damage and icterus; in 2 cats with uremia platelet aggregation was normal. Acetylsalicylic acid (ASA) (10 mg/kg iv) inhibits platelet aggregation in the presence of collagen in low concentrations; high concentrations of collagen succeeded in inducing platelet aggregation.     

Peritoneo-pericardial diaphragmatic hernia in a Persian cat--a congenital defect]

In this clinical study, a diaphragmatic peritoneo-pericardial hernia in an eight month-old Persian cat is reported. The diagnostic procedure is described, and the clinical symptoms and diagnostic possibilities are discussed.     

Measurement of nitrate leaching losses from arable plots under different nitrogen input regimes

Abstract. Leaching losses of nitrate-nitrogen were measured from a set of eight hydrologically isolated plots on a clay loam soil over the period from September 1987 to February 1990. Variable drainflow recovery from the plots hampered accurate estimation of nitrate loading, but results suggest that, when inorganic nitrogen fertilizer is applied up to the recommended amount, there is little influence of the amount applied on the amount leached. We did, however, observe the following effects on nitrate leaching: leguminous green manure incorporated in autumn increased leaching of nitrate-nitrogen by 10–15 kg per hectare during the winter; autumn cultivation caused some increase in leaching compared with no cultivation in one year; some systematic variations in nitrate leaching occurred between years and between plots, but were unrelated to treatments.
From the results we conclude that green manuring does not provide sufficient nitrogen for organically grown crops on this soil but contributes significantly to nitrate leaching, and that growing spring cereals, with the land remaining in stubble as long as possible in autumn, may be the best strategy to minimize nitrate leaching.     

Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Cystocentesis in dogs and cats]

The technique of cystocentesis in dogs and cats was studied. The indications and contraindications of this method are discussed, using illustrations.