잔류 항균물질에 대한 미생물학적 간이검사법의 검출감도 비교

Comparison of Detectable Levels for Screening Residual Antibacterial Agents by Bioassay

본 연구는 EEC 4-plate법과 그 변법 그리고 식품공전상 간이검사법으로써 어류질병의 예방 및 치료에 사용되는 28종류의 항균물질에 대한 최저 검출한계를 서로 비교하여 어체내 잔류 항균물질의 최적 간이검사법 (bioassay)으로서의 유효성을 확인하고자 하였다. 식품공전상 간이검사법은 PCs에 뛰어난 검출감도를 나타내었으며 AGs에 대하여 좋은 감도를 보였으나 TCs, MLs, NFs, QNs에 대해서는 낮은 검출감도를 나타내었다. 한편, SAs에 대하여는 대단히 저조한 감도를 보였다. EEC 4-plate변법은 TCs에 대하여 우수한 검출감도를 나타내었다. EEC 4-plate법 및 그 변법은 PCs, MLs, NFs, QNs, SAs에 대하여 상대적으로 높은 검출감도를 나타내었다. 시험법들은 모두 CMs에 대하여는 검출감도가 낮았다. 결국 EEC 4-plate 변법이 여러계열의 항균물질에 대해 검출감도가 뛰어나고 항균활성의 범위가 넓어 어체내 잔류 항균성물질을 가장 유효하게 스크리닝할 수 있는 간이검사법으로 확인되었다.

Minimun-detectable levels to 28 antibacterial agents used for the prevention and the treatment of fish diseases were determined to establish optimal detective method of bioassay in fish by the EEC 4-plate method, the modified method of EEC 4-plate and the standard method of analysis in food safety regulation. The test organisms used in the methods of bioassay were as follows: Bacillus subtilis BGA (B. subtilis) and Micrococcus luteus ATCC 9341 (M. luteus) in the EEC 4-plate method, B. subtilis, M. luteus and Bacillus cereus var. mycoides ATCC 11778 (B. cereus) in the modified of EEC 4-plate, and B. subtilis, M. luteus, B. cereus and Bacillus stearothermophilis var. calidolactis C-953 (B. stearothermophilis) in the standard method. The standard method showed predominant sensitivity in the detection of penicillins (PCs), and was also highly sensitive to aminoglycosides (AGs). The sensitivity of standard method in the detection of tetracyclines (TCs), marrolides (MLs), nitrofuran derivatives(NFs) and quinolones (QNs) was very low, and against sulfonamides (SAs), however, was extremely low. The modified method of EEC 4-plate showed very high sensitivity to TCs. Both the EEC 4-plate and the modified method of EEC 4-plate showed competitively high sensitivity in the detection of PCs, MLs, NFs, QNs and SAs. All the methods studied in the experiment showed very low sensitivity against chloramphenicol (CMs). Consequently, the modified method of EEC 4-Plate was the best bioassay method with a wide range of sensitivity for the optimal detection of the residual antibacterial agents in fish.