A Method for Cytogenetic and Cytological Examination of Small Shelled Larvae of Bivalves and Other Zooplankton

Studies on chromosomes and nuclei of very small bivalve larvae have been impeded by the veliger shell. It has been determined that the alcohokacetic acid fixative commonly used in cytogenetic techniques can be made to act as a decalcifying agent upon repeated heating. In addition, transfer of formalin fixed shelled specimens, routinely used as marine bioassay organisms, into ethyl alcohohacetic acid (3:1) fixative also yields clear cells for cytological examination of decalcified but otherwise intact oyster larvae and other zoo-plankton. Identification of cell type, such as germ-line primordia, in, for example, reproductive and ploidy level studies, and observations on the presence of bacteria can be made from the preparations. Material can be examined up to at least a year after preservation. The method is evaluated and its modifications are discussed.     

Initiation and characterization of five embryonic cell lines from the cotton boll weevil anthonomus grandis in a commercial serum- free medium

Summary Five continuous cell lines were initiated from embryonic tissue of the cotton boll weevilAnthonomus grandis Boheman in a commercially available, serum-free medium (Excell 401) and have undergone in excess of 60 passages. Isoenzyme analysis confirmed that the lines originated from boll weevil tissue. Four of the lines grew as single attached cells of either epithelioid or fibroblastoid morphology. The fifth line, BRL-AG-2, grew primarily as cell aggregates and was found to release ecdysteroids (primarily ecdysone) into the culture medium. Evidence was also obtained suggesting that line BRL-AG-2 synthesizes chitin. Three lines, BRL-AG-1, BRL-AG-3A, and BRL-AG-3C, could be induced to produce an antibacterial factor(s) which was released into the culture medium.