. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. Vol. 8, p. 76 Asiatic Herpeiological Research 1999 Material and Methods Animals and samples collection The samples used in our experiments were provided by Chang Zhou Breeding Center of Soft-Shelled Tur- tle (near Nangjing in Jiangsu Province in east China). The breeding environment was similar to their natural living condition. Except December and January every month we obtained blood plasma, testes and epid- idymides that those were from six adult male soft- shelled turtles, Pelodiscus sinensis (
. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. Vol. 8, p. 76 Asiatic Herpeiological Research 1999 Material and Methods Animals and samples collection The samples used in our experiments were provided by Chang Zhou Breeding Center of Soft-Shelled Tur- tle (near Nangjing in Jiangsu Province in east China). The breeding environment was similar to their natural living condition. Except December and January every month we obtained blood plasma, testes and epid- idymides that those were from six adult male soft- shelled turtles, Pelodiscus sinensis (there body weights were about 650-1000g) in one year. The blood plasma samples were stored at -24° until the testosterone analysis was performed. The tissue sam- ples of testes and epididymides were weighted and fixed in Bouin fluid. Histological procedures For the histological examination, a piece of testis and a piece of epididymis (about 25mm") were fixed in Bouin fluid for 24 hours. After discolored, the sam- ples were embedded in paraffin, cut in 5um sections, and then stained with haematoxylin and eosin. Testosterone assay We determined the plasma concentration of test- osterone by radioimmunoassay (RIA) using a tech- nique provided by WHO. The kits used in our experiments were from Hua Mei Biological Engineer- ing Co. (a Sino-US joint venture). Briefly, the hor- mone was extracted from plasma samples (250-7501) twice with 5 ml of anhydrous ether, with an average recovery of 96%. The extract was dried at 40° by con- stant temperature bath, and dissolved in 2 ml phos- phate-buffered saline containing gelatin (GPBS). we placed ml of this solution in duplicate assay tubes and incubated with radio labeled testosterone (H-T cpm) and anti-testosterone antiserum ( ml) for 18-24 h at 4°C. After removing the unbound frac- tion with dextran-coated chorale, the samples were placed in scintillation fluid (TP-POPOP-toluene) and the radioactivity (cpm) was detecte
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