Collaborative study of the colorimetric determination of nitrate and nitrite in cheese.

A quantitative colorimetric method for the determination of nitrate and nitrite in cheese has been subjected to collaborative study. The method includes clarification of an aqueous extract of cheese with zinc hydroxide, reduction of nitrate to nitrite via a spongy cadmium collumn (the nitrite originally present is unaltered), diazotization of sulfanilic acid with the nitrite, and coupling with 1-naphthylamine hydrochloride to form a pink azo dye whose absorbance is measured at 522 nm. The spectrophotometric responses are compared to a standard curve. In samples containing both nitrate and nitrite, nitrate is determined by difference. A standard deviation of 5.5 was obtained (5 of 6 collaborators) when a cheese sample spiked with 276 ppm sodium nitrate was analyzed by the method. The method has been adopted as official first action.     

Substances interfering with the gas-liquid chromatographic determination of T-2 mycotoxin.

Two substances interfering with the gas-liquid chromatographic (GLC) detection of T-2 mycotoxin were identified as 1-glyceryl-monooleate and 1-glycerylmonolinoleate. These monoglycerides are natural products formed by species of Fusarium growing on cereal grains and are also additives contained in liquid vegetable and animal fats added to the feed mixture. The monoglycerides can be removed from the analytical sample by resolution by thin layer chromatography prior to separation by GLC. Trimethylsilyl ether derivatives of the monoglycerides and T-2 toxin have almost identical retention times on 3% OV-1 columns, whereas the trifluoroacetyl and pentafluoropropionyl derivatives give baseline separation on the same column. The monoglycerides can be misidentified as the T-2 toxin in analyses involbing GLC.     

Production and persistence of urease activity in soils

Addition of urea to Iowa soils did not induce urease activity, but production of urease activity was observed on addition of glucose and other organic materials that promote microbial activity. The persistence of the urease activity produced on addition of these materials varied with the soil, but, with each soil studied, the urease activity after addition of organic materials eventually was identical to that of the unamended soil. No increase or decrease in urease activity was observed when unamended or urea-treated soils were incubated under aerobic conditions for several months. It is concluded that soil constituents protect urease against microbial degradation and other processes leading to inactivation of enzymes and that every soil has a stable level of urease activity determined by the ability of its constituents to provide this protection.     

Sodium bicarbonate as an extractant for soil phosphate III. Effects of the buffering capacity of a soil for phosphate

An index of the buffering capacity for phosphate of a group of soils was obtained by measuring adsorption of phosphate from dilute solutions of calcium chloride. The effect of buffering capacity on the amount of phosphate initially displaced by solutions of sodium bicarbonate and on the amount of secondary adsorption from bicarbonate was then studied. These two effects were separated using a regression procedure in which the soil: solution ratio was the independent variable.As buffering capacity increased the amount of phosphorus initially displaced decreased and the amount of secondary adsorption increased. Both these changes resulted in a decrease in the amount of phosphate in the extract. The effect of buffering capacity was greater with the Olsen method (soil : solution ratio 1 : 20; 30 min) than with the Colwell method (soil : solution ratio 1 : 100; 16 h). The relation between phosphorus extracted and buffering capacity was of a similar shape to that between effectiveness of fertilizer and buffering capacity. However, the first relation depends on the conditions of extraction and the second on the kind of plant grown and on the conditions of growth. Hence the two relations do not necessarily coincide.The effect of buffering capacity on the proportion of added phosphate initially displaced from the soil became more marked as the period of incubation prior to extraction was increased. Bicarbonate soil tests would therefore indicate that, on soils of low buffering capacity, the decrease with time in availability of applied phosphate would be smaller than on soils of high buffering capacity. This effect differs from that observed with plants.     

Comparison of dieldrin, lindane, and DDT extractions from serum, and gas-liquid chromatography using glass capillary columns.

Rats were given an oral dose of 14C-labeled chlorinated pesticides to obtain serum containing p,p'-DDT, dieldrin, or lindane. Simple hexane and formic acid-hexane extraction methods, involving pretreatment of the serum with formic acid, were compared by radiometric and by paper chromatographic and gas chromatographic analysis. In vivo binding of chlorinated pesticides to constituents of the serum does not necessarily prohibit their isolation by simple hexane extractions, provided that the extraction is very vigorous and at least 5 min long. Stable emulsions were broken by cooling in liquid nitrogen or Dry Ice-acetone. The hexane extraction method described yields quantitative recovery of the pesticides studied, whereas the formic acid-hexane method is quantitative for p,p'-DDT, 93% for dieldrin, and 89% for lindane. Gas chromatographic comparison of both methods, using human serum, shows that the hexane method extracts 16% more beta-BHC, 7% more dieldrin and HCB, and 4% more p,p'-DDE from serum than does the formic acid-hexane method. The difference for p,p'-DDT is not significant. Gas chromatography with glass capillary columns and an all-glass solids injection system yielded detection limits as low as 15 fg. Data show that the use of an internal standard considerably improves the precision of quantitation.     

Infection of wheat seminal roots by varieties of Phialophora radicicola and Gaeumannomyces graminis

The growth of isolates of Phialophora radicicola var. radicicola, P. radicicola var. graminicola, Gaeumannomyces graminis var. graminis, G. graminis var. tritici and Leptosphaeria narmari was compared on the coleoptiles and roots of wheat seedlings. Fungal growth was measured as the extent and density of dark runner hyphae. All except P. radicicola var. graminicola grew on coleoptiles and all grew on roots although only G. graminis var. tritici extensively colonized the root stele. Growth rate on roots was positively correlated with that on agar, P. radicicola var. graminicola and L. narmari growing at about half the rate of the other fungi; hyphal density was high for P. radicicola var. graminicola but relatively low for the other fungi. For P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici growing from buried inocula, the extent and density of hyphae up roots towards the seed was similar to that down, but G. graminis var. tritici caused chocolate-brown stelar discoloration up roots only.Root invasion by P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici was described from sections. Each gave a different pattern of hyphae and host response within an inoculum layer, and progressive changes occurred away from the inoculum. Studies of the rate of penetration by each fungus and the rate and pattern of death of cortical cells explained the differences between fungi. G. graminis var. tritici penetrated living cells in advance of other soil micro-organisms, and hence by hyaline hyphae inducing much lignituber formation as a host resistance reaction. P. radicicola var. graminicola penetrated only senescent or dead cells in association with other soil microorganisms, and hence by dark hyphae, inducing little lignituber formation. P. radicicola var. radicicola was intermediate in all these respects. The high hyphal density of P. radicicola var. graminicola was due to the colonization of cortical cells and spaces by dark, clearly visible, rather than hyaline hyphae, which are invisible in unstained roots. Cell death in the outer cortex explained the observed progressive restriction of growth by all fungi to the inner cortex with increasing distance from the inoculum. Spread by G. graminis var. tritici up roots was ectotrophic relative to the stele but down roots hyphae spread rapidly within the stele. Stelar reactions suggested as resistance mechanisms occurred up roots only. Their absence down roots is attributed to infection disrupting stelar transport.     

Decomposition of algal cells and components and their stabilization through complexing with model humic acid-type phenolic polymers

Axenic cultures of Anacystis, Microcoleus, Plectonema and Synechococcus isolated from Greenfield sandy loam and of Anabaena flos-aquae, Nostoc muscorum and Chlorella pyrenoidosa from other sources were cultured under light and constant aeration and with [U-¹⁴C]-glucose in the nutrient medium. Whole cells, cell walls, cytoplasm and extracellular polysaccharides of selected species readily decomposed in the soil and after 22 weeks between 61 and 81% of the added C had evolved as CO₂. Complexing of cell wall and cytoplasmic preparations from A. flos-aquae and N. muscorum with model humic acid-type phenolic polymers reduced decomposition of the cell walls by 40% and of the cytoplasm by 70%. Over 50% of the residual 14C activity in the soil amended with whole algal cells remained in the 0.5% NaOH-extracted soil. With exception of Microcoleus sp. more of the residual ¹⁴C from cell walls, cytoplasm and polysaccharide fractions was present in the humic acid or fulvic acid fractions.     

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.     

Pathogens occurring on ten varieties of ryegrass at Rothamsted in 1973–74

Six varieties oi Loliumperenne, three of L. multiflorum and one hybrid (L. multiflorum × L. perenne) were assessed for diseases over two harvest years. Crown rust (Puccinia coronata) and mildew (Erysiphe graminis) occurred on some varieties in 1973. Leaf spotting (Drechslera spp.) was common, but slight on most varieties, in both years. Ryegrass mosaic virus (RMV) and its mite vector, Abacarus hystrix were present on all varieties in the second year. The relative susceptibility of varieties to RMV was similar in the field and in glasshouse tests.