Differentiation between Aspergillus flavus and Aspergillus parasiticus from pure culture and aflatoxin-contaminated grapes using PCR-RFLP analysis of aflR-aflJ intergenic spacer

Aflatoxins (AFs) represent the most important single mycotoxin-related food safety problem in developed and developing countries as they have adverse effects on human and animal health. They are produced mainly by Aspergillus flavus and A. parasiticus. Both species have different aflatoxinogenic profile. In order to distinguish between A. flavus and A. parasiticus, gene-specific primers were designed to target the intergenic spacer (IGS) for the AF biosynthesis genes, aflJ and aflR. Polymerase chain reaction (PCR) products were subjected to restriction endonuclease analysis using BglII to look for restriction fragment length polymorphisms (RFLPs). Our result showed that both species displayed different PCR-based RFLP (PCR-RFLP) profile. PCR products from A. flavus cleaved into 3 fragments of 362, 210, and 102 bp. However, there is only one restriction site for this enzyme in the sequence of A. parasiticus that produced only 2 fragments of 363 and 311 bp. The method was successfully applied to contaminated grapes samples. This approach of differentiating these 2 species would be simpler, less costly, and quicker than conventional sequencing of PCR products and/or morphological identification.     

PROTEIN PRECIPITATION DURING WORT BOILING: QUALITY ASPECTS OF DIMINISHED WORT BOILING TIMES OF BREWS PREPARED FROM PROANTHOCYANIDIN-FREE OR REGULAR RAW MATERIALS

When using proanthocyanidin-free materials for the production of beer, a reduction of the wort boiling time can be considered. In worts prepared with regular malt and tannin-free hop extract there is a continuous precipitation of the malt flavanoids while in brews prepared from proanthocyanidin-free malt and regular hops there is a simultaneous extraction and removal of the hop flavanoids leading to constant levels of these hop flavanoids. The results also show that the level of Kjeldahl nitrogen in worts boiled with hops will be the same as that in worts boiled with n-hexane tannin-free hop extracts. These results and the fact that more protein precipitates in brews containing no malt or hop proanthocyanidins suggest that, unlike what is the case during the development of beer haze, polyphenols are not necessary for an effective protein precipitation during wort boiling.     

Calibration and evaluation of nitric acid and ammonia permeation tubes by UV optical absorption

An ultraviolet (UV) optical absorption system has been developed for absolute calibrations of nitric acid (HNO3) and ammonia (NH3) permeation tube emission rates. Using this technique, dilute mixtures containing NH3 or HNO3, both of which interact strongly with many surfaces, are accurately measured at levels below a part per million by volume. This compact and portable instrument operates continuously and autonomously to rapidly (<1 h) quantify the emission of trace gases from permeation devices that are commonly used to calibrate air-monitoring instruments. The output from several HNO3 and NH3 permeation tubes, with emission rates that ranged between 13 and 150 ng/min, was examined as a function of temperature, pressure, and carrier gas flow. Absorptions of 0.015% can be detected which allows a precision (3sigma) of +/-1 ng/min for the HNO3 and NH3 permeation tubes studied here. The accuracy of the measurements, which relies on published UV absorption cross sections, is estimated to be +/-10%. Measurements of permeation tube emission rates using ion chromatography analysis are made to further assess measurement accuracy. The output from the HNO3 and NH3 permeation tubes examined here was stable over the study period, which ranged between 3 months and 1 year for each permeation tube.     

The effect of acid dextrinisation on enzyme-resistant starch content in extruded maize starch

The enzyme-resistant starch (ERS) content in processed high amylose and regular maize starches has been studied, with and without acid dextrinisation. The physicochemical and structural characteristics of the starches were analysed using a variety of techniques. The increase in ERS in high amylose maize starch with dextrinisation was related to the formation of a critical molecular weight fraction (MW ∼ 20,000) that could rearrange structurally. Further dextrinisation reduced the processed starch MW to below where it could still form ERS. Regular maize starch containing less than 30% amylose did not increase its resistance to amylase digestibility with acid dextrinisation, probably due to impairment of amylose rearrangement by the numerous branched amylopectin chains. The ERS, which is likely to form during the enzyme-digestion process, is a linear molecule with a maximum degree of polymerisation (DP) of 30, irrespective of the starch source, processing conditions applied or type and amount of acid used.     

Bromamine decomposition kinetics in aqueous solutions

The objectives of this study are to investigate the kinetics of bromamine decomposition and to identify the corresponding relevant reactions. Experiments were performed with a stopped-flow spectrophotometer system. Experimental variables investigated included pH (6.5-9.5), bromamines concentration (0.15-0.50 mM), ammonia to bromine ratio (5-100), and phosphate and carbonate buffers concentration (5-40 mM). The experimental results were consistent with a reaction scheme that involved the reversible disproportionation of monobromamine into dibromamine and ammonia (2NH2Br (k1)<=>(k(-1)) NHBr2 + NH3), followed by irreversible decomposition of monobromamine and dibromamine into products (2NHBr2 (k2) --> products and NH2Br + NHBr2 (k3) --> products). The monobromamine disproportionation reaction was found to undergo general acid catalysis, and the two subsequent decomposition reactions were found to experience base catalysis. Experimental results were analyzed for the determination of catalysis terms corresponding to H+, NH4+, H2PO4-, HCO3-, and H2O for rate constants k1 and k(-1); HPO4(2-) and H2O for k2; and OH-, CO3(2-), and H2O for k3. These constants were fitted with the Br?nsted relationship, and the resulting fitting expressions were used to calculate any relevant catalysis rate constants that could not be determined at the range of experimental conditions used.     

Salmonella in uncooked retail meats in New Zealand

A national quantitative survey of Salmonella in five types of uncooked retail meats in New Zealand was undertaken from August 2003 to May 2005 to establish baseline proportionality data. The overall prevalence of Salmonella in 1,108 meat samples was 1.1% (95% confidence interval, 0.6 to 1.9). Low prevalences of Salmonella in each meat type were observed, with 3% (1.2 to 6.1) in chicken, 1.3% (0.3 to 3.8) in lamb and mutton, 0.5% (0 to 3.0) in unweaned veal, 0.4% (0 to 2.4) in beef, and 0% (0 to 1.6) in pork. The Salmonella serotypes isolated were Salmonella Infantis from beef; Salmonella Typhimurium PT1 from unweaned veal and chicken; Salmonella sp. 6,7:k:-, Salmonella Enteritidis PT9a, Salmonella sp. 4,5,12:-:-, Salmonella sp. 4,12:-:-, and Salmonella Typhimurium PT160 from chicken; and Salmonella sp. 4:-:2 and Salmonella Brandenburg from lamb. Four of the isolates from chicken, Salmonella sp. 4,5,12:-:- (two isolates), Salmonella sp. 4,12:-:-, and Salmonella Typhimurium PT1, were very similar phenotypically and serologically to the attenuated Salmonella vaccine strain used in MeganVacl for poultry. One lamb sample yielded a count of Salmonella Brandenburg of 4.24 most probable number (MPN)/g, while all other positive samples were <1.0 MPN/g. The results provide baseline proportionality data for Salmonella in retail uncooked meats that will contribute invaluably toward future risk assessment in light of other information, such as consumption data that can be used for risk characterization.     

Copper accumulation in soils from two different‐aged apricot orchards in Central Otago,New Zealand

In recent years there has been a move in New Zealand to convert small, uneconomic orchards to other agricultural uses. Overseas research has demonstrated that copper from fungicide sprays often accumulates in the soils of orchards and can cause certain problems for plant growth and livestock. To date, no assessments appear to have been made of the degree of copper accumulation in such older orchards in New Zealand. This study presents the results of an assessment of two well‐established apricot orchards in the Central Otago region of South Island, New Zealand.

Two aspects of the results are surprising. The first is the relatively low levels of copper in the soil when compared with results from another orchard in the region. That orchard is 18 years old and mean soil copper has already reached 100.0 μg/g, indicating a much faster rate of accumulation than in the two orchards in the current study. The second unexpected result is that the younger orchard has a significantly higher level of copper than the older orchard. Reasons for these findings are discussed in relation to apparent differences in soil properties.     

Water treatment with silicofluorides and lead toxicity

Toxic metals like lead, manganese, copper and cadmium damage neurons and deregulate neurotransmitters like serotonin and dopamine (which are essential to normal impulse control and learning). Earlier studies show that — controlling for socio‐economic and demographic factors — environmental pollution with lead is a highly significant risk factor in predicting higher rates of crime, attention deficit disorder or hyperactivity, and learning disabilities. Exposure and uptake of lead has been associated with industrial pollution, leaded paint and plumbing systems in old housing, lead residues in soil, dietary habits (such as shortages of calcium and iron), and demographic factors (such as poverty, stress, and minority ethnicity). We report here on an additional “risk co‐factor” making lead and other toxic metals in the environment more dangerous to local residents: the use of silicofluorides as agents in water treatment. The two chemicals in question — fluosilicic acid and sodium silicofluoride — are toxins that, despite claims to the contrary, do not dissociate completely and change water chemistry when used under normal water treatment practices. As a result, water treatment with siliconfluorides apparently functions to increase the cellular uptake of lead. Data from lead screening of over 280,000 children in Massachusetts indicates that silicofluoride usage is associated with significant increases in average lead in children's blood as well as percentage of children with blood lead in excess of 10μg/dL. Consistent with the hypothesized role of silicofluorides as enhancing uptake of lead whatever the source of exposure, children are especially at risk for higher blood lead in those communities with more old housing or lead in excess of 15 ppb in first draw water samples where silicofluorides are also in use. Preliminary findings from county‐level data in Georgia confirm that silicofluoride usage is associated with higher levels of lead in children's blood. In both Massachusetts and Georgia, moreover, behaviors associated with lead nurotoxicity are more frequent in communities using silicofluorides than in comparable localities that do not use these chemicals. Because there has been insufficient animal or human testing of silicofluoride treated water, further study of the effect of silicofluorides is needed to clarify the extent to which these chemicals are risk co‐factors for lead uptake and the hazardous effects it produces.