interpretation 1: the twice-born become twice-born when the undergo upanayana.
interpretation 2: the twice-born were human in their previous incarnation also.
Are both interpretations supported by scripture?
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The first interpretation is of course supported by scriptures. A twice-born is so called because he is considered to have been born two times. I do not know about the 2nd one.
There are four classes: Brahmin, Kshatriya, Vaishya and the Shudra. 2 Three classes are twice-born: Brahmin, Ksatriya and Vaisya. Of these:
The first birth is from the mother and the second at the tying of the Muñja-grass girdle. At the latter birth, the Savitrı verse is his mother, while the teacher is said to be his father.
Vashishta Smriti 18.104.22.168-3
I did not see the second interpretation anywhere. However, a second type of interpretation is implied by the Kularnavatantra regarding different kinds of higher forms of Tantriki diksha. For example, its mentioned that Sparsha Diksha by Guru is like the mother-bird nurturing the baby-birds (14/35) ,Vedhadiksha like tortoise-mother nurturing the babies by mind(14/37).
By the way the concept of tortoise-mother's nurturing the tortoise-babies by mind has been used by many of our saints including Sri Ramakrishna (Kathamrita,Udbodhan, page 20, March, 1882).There is no scientific basis of this view so far as I know. Babies of all reptiles are self sufficient from the time of birth and sustain even if their mother die before they are born.