Abstract
Abolition of the salt monopoly has lead to increases in the distribution and the import of natural salts and foods containing these salts. There is a possibility of the contaminations of these natural salt products with halophiles such as extremely halophilic archaea under the circumstances. The contaminants can putrefy the salt products, and damage the food manufacturers and importers dealing in salts. Therefore, we were convinced of the necessity of detection procedures for contaminated halophiles, and examined the conditions for detecting halophiles specifically using a polymerase chain reaction (PCR) in the present study. Three pairs of oligodeoxyribonucleotides (ODN) reported previously and novel twelve pairs of ODN were constructed for PCR primers. The latter were derived from twelve insertion sequences of halophile (ISH) in the whole genome of Halobacterium sp. NRC-1. The chromosomal DNA of two extremely halophilic archaea, Halobacterium sp. NRC-1and H. salinarum, and four other food contaminating eubacteria such as Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Vibrio parahaemolyticus were extracted and purified for PCR templates. The amplified products were analyzed on agarose gel electrophoreses.The results indicated that the four pairs of PCR primers showed halophile-specific amplification. It is suggested that these four pairs of primers are useful for the specific detection of the contaminating halophiles in natural salt products.