Resolution of aflatoxins B1, B2, G1, and G2 by high-pressure liquid chromatography.

Aflatoxins were completely resolved as sharp peaks in the order BU-B2-G1-G2 by high-pressure liquid chromatography on a small particle (10 mum) porous silica gel column in 7-13 min (B1 through G2) by a water-saturated chloroform-cyclohexane-acetonitrile elution solvent (25+7.5+1.0), with detection by ultraviolet absorbance at 360 nm. The relationship between peak height and amount injected was linear over a 5-400 ng range for each aflatoxin. Both retention times and peak heights were highly reproducible, multiple injections of mixed standards giving coefficients of variation of 1.0-1.4% (retention time) and 1.6-2.8% (peak height) for the 4 aflatoxins. Detection was highly sensitive, with mean peak height, mm/ng, of 7.1 (B1), 6.4 (B2), 4.5 (G1), and 4.1(G2), allowing detection of 1-2 ng of each aflatoxin.     

Multiple forms of acetyl-naphthyl-esterase activity in soil organic matter

Acetyl-naphthyl-esterase activity has been identified and characterized in organic matter extracted from an A₁ horizon of an Alpine podzol. The temperature optimum of the esterase is about 75° C and its activity rises with increasing pH, without reaching a maximum value in the tested range. The Michaelis constant has been determined as Km = 2.950 mM. Pronase does not disrupt esterase activity. Electrofocusing in acrylamide gel shows several peaks of enzyme activity, which correspond with humic isoelectric bands. The location of acetyl-esterase activity in organic matter is discussed.     

Studies on soil fumigation—III: Effects on enzyme activities,bacterial numbers and extractable ninhydrin reactive compounds

Fumigation of a field soil with chloropicrin and methyl bromide, either singly or in combination, differentially decreased soil enzyme activities and viable bacterial numbers and increased the amounts of ninhydrin reactive compounds extractable with acidified Tris buffer. Chloropicrin treatment was more effective than methyl bromide.The rates of hydrolysis of both an amide and a peptide derivative were decreased by chloropicrin treatment and remained relatively low despite changes in activities over 325 days. By contrast, caseinase activity initially was decreased by both chloropicrin and methyl bromide fumigation, but activities of the fumigated soils recovered to exceed those of untreated soils. Thereafter, caseinase activities of fumigated and untreated soils exhibited relatively large fluctuations, which were partly associated with seasonal drying of the soils in the field.Chloropicrin but not methyl bromide fumigation markedly depressed the viable bacterial populations, which subsequently increased to be above those of the untreated soils. There was no consistent relationship between the release of ninhydrin reactive compounds following fumigation and changes in bacterial numbers or changes in enzyme activity. Autolytic reactions are probably important in the early stages of amino-nitrogen release in fumigated soils. Net gains in caseinase activity may be partly due to the production de novo of extracellular proteases by microorganisms or to the release of intracellular proteases from killed cells.     

Studies on soil fumigation—I: Effects on ammonium,nitrate and phosphate in soil and on the growth,nutrition and yield of wheat

Fumigation of field soil with chloropicrin alone or followed by methyl bromide, each at 220Kg·ha^?1, released 20–30 parts/10⁶ NH⁺₄-N which persisted for 75 days; such fumigation also doubled the amount of bicarbonate-extractable phosphate 28 days after fumigation. Soil fumigation increased both the vegetative and grain yields as well as increasing the content of N in the grain and the content of K and Cl in the tops at ear emergence. Root growth and the phosphate uptake activity of the roots were increased by soil fumigation.     

The effect on young beef cattle of supplementing hay with dried grass or concentrates

Forty-eight Friesian steers with an average live-weight of 168 kg were given hay ad libitum supplemented with 0, 0.6, 1.2, 1.8, 2.4 and 30 kg artifically dried grass or concentrates. In vitro dry matter digestibility of hay was 55%. Liveweight gain increased with each successive increase in the level of supplementation. The intake of hay decreased but the total DM was greatest at the higher level of supplementation. The results suggest that dried grass pellets combine well with hay and may be used to replace concentrates. It was shown that 1 kg of either dried grass or concentrates given as a supplement to hay fed ad libitum, will increase the live-weight gain by 0.16 kg d^?1 and decrease hay intake by 0.25 kg d^?1.     

Detection of twelve mycotoxins in mixed animal feedstuffs, using a novel membrane cleanup procedure.

A multimycotoxin thin layer chromatographic screening method is described which is applicable to most animal feedstuffs. Interference from nonspecific lipid, pigment, and other components of simple and mixed feeds is reduced to a minimum by using a membrane cleanup step. Aflatoxins B1, B2, G1, and G2, citrinin, diacetoxyscirpenol, ochratoxin A, patulin, penitrem A, sterigmatocystin, T-2 toxin, and zearalenone may be reliably detected. The sensitivity of the method is generally low for mixed feeds but even so aflatoxin B1 can be detected at a level of 3 ppb and ochratoxin A at 80 ppb. While the basic method is less sensitive for sterigmatocystin (330 ppb), patulin (600 ppb), zearalenone (1000 ppb), and the trichothecenes (1000-4000 ppb), it may be adapted so as to reduce the above detection limits when the presence of these toxins is suspected. Lower levels may be detected in extracts of simple feeds.     

Laboratory study of phosphorus forms and test procedures on soils under aerobic and anaerobic conditions

The inorganic forms of phosphorus in nine samples of surface soils from the Nainital Tarai of India were determined by a series of extractions with different reagents after incubation for 45 days under aerobic or anaerobic conditions. The forms of phosphorus in samples incubated under aerobic conditions and the pH range of 7.2–8.5 indicated that the soils had been subject to slight chemical weathering. Anaerobic incubation to simulate conditions in lowland rice paddies converted part of the inorganic phosphorus into more labile forms, based on changes in amounts of isotopically exchangeable phosphorus. Amounts of the element converted to more labile forms differed among the nine soil samples.A better correlation was found between amounts of phosphorus extracted by 0.5M NaHCO³ (Olsen's method) than by 0.03N NH₄F in 0.25N HCl (Bray's method) and the isotopically exchangeable phosphorus. The better correlation suggests that the bicarbonate extraction would be the better method for estimating “availablelrd phosphorus in soils of lowland rice paddies. Recalibration of the test prior to such use seems desirable to allow for increases in amounts of phosphorus in labile forms after soils have been shifted from aerobic to anaerobic conditions.     

Automated method for the determination of niacin and niacinamide in cereal products: collaborative study.

In a collaborative study, an automated method for the determination of niacin and niacinamide in cereal products was compared with the official final action microbiological (43.121-43.125) and chemical (43.044-43.046) methods. Ten samples of cereal products, including enriched flour, yeast-leavened baked products, fortified breakfast cereals, and baked pet food products, were submitted to 14 laboratories. Nine laboratories reported values by the automated method, 6 reported values by the microbiological method, and 7 reported values by the chemical method. The results from the microbiological method were not subjected to analysis of variance because of the unusually large between-laboratory variation. The between-laboratory coefficients of variation for the automated and chemical methods were 10.90 and 10.18%, on the basis of results from 7 and 4 laboratories, respectively. There was no significant (p greater than 0.05) difference between methods when results from the 4 laboratories who used both methods were compared. The automated chemical method has been adopted as official first action.     

Cherry leaf roll virus causing a disease of Betula spp. in the United Kingdom

A virus, serologically related to isolates of cherry leaf rollvirus (CLRV) from Prunus avium (L.) L. and Sambucus nigra L.was transmitted to a range of herbaceous plants from leavesor roots of thirty-one of forty-seven trees of Betula spp. growingat two heathland sites in Berkshire. Ageing leaves of virus-infectedindividuals which had mild chiorotic blotches or ring and linepatterns in July showed bright yellow ring and line patternsin October. In the season after B. verrucosa seedlings wereinoculated with CLRV isolates, leaves developed chlorotic ringand blotch patterns; the symptoms being similar whether theinocula were originally isolated from S. nigra, P. avium orB. verrucosa. Palisade and mesophyll cells of naturally infectedleaves of B. verrucosa contained chloroplasts in which granalorganization was appreciably less distinct than that of chloroplastsfrom virus-free leaves. In many instances, virus-infected leafcells had spherical virus-like particles about 30 mn in diametercontained within tubular structures in cell wall projections;these structures were not seen in virus-free material. An isolate of CLRV from B. verrucosa, purified from systemicallyinfected leaves of Nicotiana clevelandii Gray by ammonium sulphateprecipitation and differential centrifugation, contained twomajor components having sedimentation coefficients of 118s and130s. Each component was composed of spherical particles withdiameters in the range 26–30 nm and having buoyant densitiesin caesium chloride of 1·460 and 1·493 gm/cm³.Polyacrylamide gel electrophoresis of preparations from bothbuoyant components indicated that they each contained a singlespecies of protein whose molecular weight was in the range 53,000–54,000daltons.