Journal - Association of Official Analytical Chemists最新文献

The use of the lipid globule stain to aid in differentiating the Bacillus cereus group (i.e., B. cereus, B. cereus var. mycoides, and B. thuringiensis) from other Bacillus species was investigated. Smears from colonies grown on suitable agar were made on precleaned slides, stained, and examined microscopically for characteristic deep blue lipid globules. The study included a total of 649 cultures of Bacillus species plus 143 incompletely characterized Bacillus isolates from food. Only B. cereus, B. cereus var. mycoides, B. thuringiensis, B. megaterium, and B. sphaericus were consistently positive for lipid globules, although at times, a few cells of B. aneurinolyticus and B. thiaminolyticus were also positive. The lipid globule stain procedure is of value in differentiating Bacillus species, especially when performed by an experienced analyst and used in conjunction with tests for cell and spore morphology.

A simple and precise method of detecting brominated vegetable oil (BVO) in soft drinks is described. After extraction of BVO using diethyl ether, the concentrated ethereal solution was treated with a small quantity of zinc dust to convert the organic bromide to inorganic form; the solution was subsequently treated with lead dioxide to liberate bromine. The bromine evolved was detected by means of fluorescein-impregnated filter paper strip that turns pink because eosin is formed. The test can detect as low as 10 ppm (2 mg/200 ml) of BVO under experimental conditions. Gas chromatography was carried out on sodium methoxide derivatives prepared from ether extract for quantitation.

The analytical methods for the detection of the staphylococcal enterotoxins can be divided into 2 categories: (1) methods for detection of enterotoxin-producing staphylococcal strains; (2) methods for detection of enterotoxin in foods. Gel diffusion methods (Ouchterlony, microslide), in which the enterotoxin produced by any given strain is compared to one of the identified enterotoxins, are used most frequently for strain testing. The sensitivity of these methods is from 0.1 to 0.5 micrograms enterotoxin/mL, which is normally adequate to determine the enterotoxigenicity of strains. The methods for the detection of enterotoxin in foods need to be much more sensitive to detect less than 1 ng of enterotoxin/g of food that may be present. The radioimmunoassay (RIA), the enzyme-linked immunosorbent assay (ELISA), and the reversed passive latex agglutination (RPLA) method have the necessary sensitivity to detect 1 ng/g of enterotoxin in foods without the use of complicated extraction-concentration procedures. Kits based on the ELISA and RPLA methods are now available commercially for the detection of enterotoxins in foods. Tests have shown that the ELISA methods are somewhat more sensitive than the RPLA method.

In the United States and Canada, Clostridium perfringens remains a leading cause of bacterial food poisoning in humans. It has been primarily associated with meat and poultry products prepared in food service establishments. Fecal spore levels of 10(6) or more per g are considered indicative of a food poisoning outbreak. However, elevated spore levels of this organism are frequently seen in healthy elderly individuals, an observation that complicates investigations of suspected outbreaks. Recent studies with this population indicate that fecal enterotoxin levels are a valuable and effective assay for confirming outbreaks due to this organism. With regard to the toxin itself, a membrane protein of 50 000-70,000 molecular weight has been isolated as a possible enterotoxin-receptor site. It is the subsequent action of the toxin on membrane structure that results in the loss of ions and fluid associated with illness. In addition, the enterotoxin gene has been cloned in E. coli and sequenced. Using toxin-specific DNA probes, only 6% of non-symptomatic farm animals were found to possess the enterotoxin gene, disproving the hypothesis that all strains of this organism can produce the toxin.