Timmons E H, Olmsted G M, Kaplan H M
Lab Anim Sci. 1977 Aug;27(4):518-21.
Eggs and sperm were aseptically collected from leopard frogs (Rana pipiens) and passed into sterilized flexible film isolators. Egg fertilization was accomplished, and germfree tadpoles were produced. The germfree tadpoles were active and free-swimming for 3 weeks, but only 6.2% survived to 4 weeks of age. None survived after the 4th week.
从豹蛙（北美豹蛙）无菌采集卵子和精子，并将其转移到经过消毒的柔性薄膜隔离器中。完成卵子受精后，培育出无菌蝌蚪。这些无菌蝌蚪在3周内活动自如且能自由游动，但只有6.2%存活到4周龄。第4周后无一存活。
