Nouws J F, Broex N J, den Hartog J M
Tijdschr Diergeneeskd. 1988 Mar 1;113(5):243-6.
The New Netherlands Kidney test is designed for the detection of antibacterial residues in slaughtered animals. It is a single-plate system in which the culture medium used is composed of Standard II nutrient agar, supplemented with 1 per cent of NaCl, 1 per cent of dextrose, 0.2 per cent of KH2PO4 and 0.28 per cent of Na2HPO4.12H2O. After autoclaving, this medium is cooled to 55 degrees C, which is followed by the addition of trimethoprim (0.12 microgram/ml of agar) and spores of Bacillus subtilis BGA (10(4) ml of agar). The pH should be adjusted to 7.00 +/- 0.05, the prepared medium then being poured out into plates specially designed for this purpose, in such a manner that the thickness of the layer of agar is 2.2 mm throughout. The following method is used in examining slaughtered animals for the presence of residues showing antibacterial activity. Four paper discs are placed in the renal pelvis for a minimum period of thirty minutes; two discs impregnated with fluid from the renal pelvis are frozen at -20 degrees C for possible re-inspection and the other two are laid diagonally on a test plate. 25 microliter of a trimethoprim solution (containing 2 micrograms of TMP per ml of a 10 per cent NaCl solution) are dropped on each of the two last-named discs. The test plates are incubated at 37 degrees C for a period ranging from thirteen to eighteen hours.(ABSTRACT TRUNCATED AT 250 WORDS)
新荷兰肾脏检测法旨在检测屠宰动物体内的抗菌残留。它是一种单平板系统,所用培养基由标准II营养琼脂组成,并添加1%的氯化钠、1%的葡萄糖、0.2%的磷酸二氢钾和0.28%的十二水磷酸氢二钠。高压灭菌后,将该培养基冷却至55摄氏度,随后加入甲氧苄啶(0.12微克/毫升琼脂)和枯草芽孢杆菌BGA孢子(10⁴个/毫升琼脂)。pH值应调至7.00±0.05,然后将制备好的培养基倒入专门为此设计的平板中,使琼脂层厚度始终为2.2毫米。采用以下方法检查屠宰动物体内是否存在具有抗菌活性的残留。将四张纸圆盘置于肾盂中至少30分钟;两张浸有肾盂液体的圆盘在-20摄氏度下冷冻以备重新检测,另外两张对角放置在试验平板上。在最后提到的两张圆盘上各滴加25微升甲氧苄啶溶液(每毫升10%氯化钠溶液含2微克甲氧苄啶)。将试验平板在37摄氏度下孵育13至18小时。(摘要截选至250字)