Uses giemsa's dye and microscope observe bursa copulatrix sperm of silkworm directly, artificial insemination using the sperm, the studies shows: The seminal fluid is similar in the male moth seminal vesicle and in the female bursa copulatrix after mating 20 to 30 minute, and the seminal fluid is contains large number of eupyrene sperm bundles and not be activated. The result of microscope observation and artificial insemination using sperm treated with diffirent trypsin concentration shows that 0.2μg/ml trypsin in 50% semen solution, the fertilization rate is highest. The sperm treated with trypsin, plasmin, pepsin and succinic acid sodium respectively, only trypsin can dissociate the eupyrene sperm bundles and improve the motility of apyrene sperm synchronously , and olso only the trypsin-treated sperm have the ability of fertilization. The optimum concentration of sperm for artificial insemination is 2-fold diluted still be found in the research.
Chen Tianfei, Wu Dayang
. Activation and artificial insemination using bursa copulatrix sperm of silkworm, Bombyx mori[J]. Chinese Agricultural Science Bulletin, 2005
, 21(1)
: 20
-20
.
DOI: 10.11924/j.issn.1000-6850.050120
1Watson L R. Demonstration of insemination of the queenbee. Journal of Economic Entomology,1927,20:3
2Abraham R. Das Verhalten der Spermien in der weiblichen bettwanze (Cimex lectularius L.), und Verbleib der uberschussigen Spermamasse. Zeitschrift Parasitenknunde, 1934,(6):559~591
3Davis N T. Studies on the reproductive physiology of Cimicidae(Hemiptera). II. Artificial insemination and the function of the seminal fluid. Journal of Insect Physiology, 1965,(11):355~366
4Burcham E. Artificial insemination of Aedes aegypti (L.). Annals Entomological Society America,1957,49: 467~493
5Gottschewski G. Kunstliche Befruchtung bei Drosophila. American Naturalist,1937,70:650
6Omura S. Artificial insemination of Bombyx mori. Japan Journal of Applied Zoology, 1933,(6):281~285
7Suda K, Omura S. Artificial insemination of Bombyx mori. Journal of Sericultural Science of Japan,1933,4:2
8Omura S. Spermatozoa of Lepidoptera. I. Specific differentiation in the spermato-prostatic reaction. Japan Journal of Genetics,1936b,15:148~152
9Omura S. Studies of the reproductive system of the male of Bombyx mori II. Post-testicular organs and post-testicular behaviour of the spermatozoa. Journal of Faculty of Agriculture. Hokkaido Imperial University, 1938,40:129~170
10Kasuga H, Aigaki T, Osanai M. System for supply of free arginine in the spermatophore of Bombyx mori. Arginine-liberating activities of contents of male reproductive glands. Insect Biochemistry, 1987,17:317~322
11Osanai M. Arginine degradation cascade, a novel energy-yielding system for spermatozoa of the silkworm, Bombyx mori (Bombycidae). Frontiers and New Horizons in Amino Acid Research,1992,15:295~302
12Osanai M, Isono M. Dissociation of eupyrene sperm bundle by acids, especially succinate accumulated in the spermatophore of the silkmoth, Bombyx mori. Invertebrate Reproduction and Development,1997,31:99~108
13Takemura Y, Kanda T, Tamura T, Shinbo H, Horie Y. Development of new method for artificial insemination of the silkworm, Bombyx mori. Journal of Sericultural Science of Japan,1996,65:456~463
14Takemura Y, Kanda T, Horie Y. Artificial insemination using trypsin-treated sperm in the silkworm. Bombyx mori. J Insect Physiol,1999,45:471~477
15Meves F. Ueber oligopyrene und apyrene Spermien undu¨ber ihre Entsehung, nach Beobachtungen an Paludina und Pygaera. Archv Mikrosk Anat Entwiclungsmech,1903,61:1~82
16Iriki S. On the function of apyrene spermatozoa in the silkworm. Zoological Magazine of Japan,1941,53:123~124
17He L, Nakagaki M, Kajiura Z, Takei R. Influence of low temperature during egg incubation on spermatogenesis of the silkworm, Bombyx mori. J Seric Sci Jpn,1996, 65:467~471
18Kawamura N, Sahara K. In vitro cultivation of spermatocysts to mature sperm in the silkworm Bombyx mori. Dev Growth Differ,2002,44: 273~280
19冯家新.蚕种学.北京:中国农业出版社,1993.76
20Takemura Y, Kanda T, Horie Y. Artificial insemination using cryopreserved sperm in the silkworm, Bombyx mori. J Insect Physiol, 2000,46:491~497
21文才艺,葛小荆.家蚕精液漂洗技术及人工授精研究.湖北农业科学, 1996,(5):38~40
22Sahara K, Takemura Y. Application of Artificial Insemination Technique to Eupyrene and/or Apyrene Sperm in Bombyx mori. Journal of experimental zoology,2003, 297A:196~200