author(s)
G Lokesh, Geetha N Murthy, Gargi, M Maheswari, Jayaprakash Pandey and Alok Sahay

abstract
The techniques of cryopreservation of semen and artificial insemination of the female can be helpful in solving the reproductive problems and is also desirable for the selective breeding based on desired traits in wild silkworms. Moreover the genetic resources can be conserved ex situ for a longer period. In this context, a collection protocol for cryopreservation of semen from the wild silkworm Antheraea mylitta a tropical tasar silkworm at -196ºC in liquid nitrogen was standardized. Further, artificial insemination of virgin female moth was also pursued to get viable embryos. Under standardization, the semen was collected from the seminal vesicle of freshly emerged male moth and from the bursa copulatrix (BC) and spermatheca of the female moth after 1 hour mating. The morphology and behaviour of semen was recorded through microscopic examination. The semen consisting of sperms entangled in a membrane in the form of bundles. Hence, the sperm bundles were treated with trypsin to dissociate the sperm bundles for activation of sperms before introducing into the female reproductive system for further fertilization. Microscopic observation of semen sample recovered from the bursa copulatrix showed rigorous motility of the spermatozoa. Artificial insemination using freshly collected semen sample treated with trypsin, preserved semen sample (-80ºC) and semen collected from bursa copulatrix was carried out through the vaginal opening of the female virgin moth. Fecundity was recorded in each group which did not reveal significant difference. However, hatching of larvae recorded significant difference with 82% hatching under natural mating on the day of hatching (82%) compared to under artificial insemination of freshly collected semen (26%), preserved semen (25%) and semen from bursa copulatrix (39%). The study enumerates that, standardization of cryopreservation techniques followed by artificial insemination lead to the development of hybrids in tropical tasar through crossing different ecoraces.