Vibriosis caused by Vibrio anguillarum and edwardsiellosis caused by Edwardsiella tarda are septicemic diseases of many commercially important freshwater and marine fishes, and threaten the aquaculture industry in Korea. Early diagnosis and accurate identification of these two bacterial species could help to prevent these diseases and minimize the damage to cultured marine species. This study designed a duplex polymerase chain reaction (PCR) method for the simultaneous detection of two major fish pathogens: V. anguillarum and E. tarda. Each pair of oligonucleotide primers exclusively amplified the target groEL gene of the specific microorganism. Twenty-two Vibrio and ten non-Vibrio enteric species were used to check the specificity of the primers, which were found to be highly specific for the target species, even among closely related species. The detection limit was 400 pg for V. anguillarum and 4 ng for E. tarda when mixed purified DNA was used as the template. This assay showed high specificity and sensitivity in the simultaneous detection of V. anguillarum and E. tarda from artificially inoculated seawater and fish.
Yogurt powder is fermented milk processed in the form of dry yogurt, and has advantages such as stability, storability, convenience, and portability. China and Vietnam are important export target countries because of the increased demand for dairy products. Therefore, we surveyed dairy product standardization in order to establish an export strategy. Lactic acid bacteria counts are unregulated in Korea and Vietnam. In China, lactic acid bacteria counts are regulated at 1×106 colony-forming units (CFU)/mL and detected at 6.24±0.33 Log CFU/mL. All three countries have regulated standards for total bacterial counts. In China, total bacterial counts of milk powder are regulated to n=5, c=2, m=50,000, M=200,000 and detected at 6.02±0.12 Log CFU/mL, exceeding the acceptable level. Lactic acid bacterial counts appeared to exceed total bacterial counts. Coliform group counts, Staphylococcus aureus, Listeria monocytogenes, and Salmonella species were not detected. Acidity is not regulated in Korea and Vietnam. In China, acidity was regulated to over 70°T and detected 352.38±10.24°T. pH is unregulated in all three countries. pH was compared to that of general fermented milk, which is 4.2, and that of the sample was 4.28±0.01. Aflatoxin levels are not regulated in Korea and China. In Vietnam, aflatoxin level is regulated at 0.05 ppb. Therefore, all ingredients of the yogurt powder met the safety standards. This data obtained in this study can be used as the basic data in assessing the export quality of yogurt powder.
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