© International Digital Organization for Scientific Information ISSN: 1817-3098 Online Open Access W J Zool ogy Corresponding Author: Dr. Rossen Stefanov, Institute of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences, 73 Tzarigradsko shosse, 1113 Sofia, Bulgaria 36 Volume 1 Number(1) : 36-39, Jan-Jun, 2006 Artificial Insemination of Sheep and Cow with Semen Treated by Cu/ Zn-superoxide Dismutase from the Fungal Strain Humicola lutea 103 R. Stefanov, E. Krumova, M. Angelova, P. Dolashka, W. Voelter and Z. Zachariev 1 2 2 3 4 1 Institute of Biology and Immunology of Reproduction Acad. Kiril Bratanov, 1 Bulgarian Academy of Sciences, 73 Tzarigradsko Shosse, 1113 Sofia, Bulgaria The Stefan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences Acad. G. Bonchev str., bl.26, 1113 Sofia, Bulgaria 2 Institute of Organic Chemistry Bulgarian Academy of Sciences Acad. G. Bonchev str., bl.9, 1113 Sofia, Bulgaria 3 Abteilung für Physikalische Biochemie des Physiologisch-chemischen Institut der Universität Tübingen, 4 Hoppe-Seyler - Straße 4, D-72076 Tübingen, Germany Abstract: The aim of present study was to examine the potential protective role of a novel antioxidant enzyme preparation on the viability and fertility of bull and ramen spermatozoa. Cu/ Zn-SOD, isolated from the fungal strain Humicola lutea 103 (HLSOD), was used. Data analysis showed that HLSOD significantly influenced spermatozoa viability during incubation at 39°C for 300 min. A total number of 103 ewes and 168 cows were inseminated. The conception rates of frozen bull semen treated with HLSOD were by 8.53 and 8.01% better compared to the control after 1-st and 1+2-nd insemination, respectively. Similar results were obtained in the experiments on artificial insemination of ewes. Addition of the antioxidant enzyme to both chilled and frozen-thawed ramen semen resulted in 6-8% increase of fertility after cervical insemination compared to nontreated semen. Using frozen/ thawed ram semen with HLSOD for intrauterine insemination 10% higher conception rates were established. Key words: Spermatozoa %superoxide dismutase %artificial insemination %fertility INTRODUCTION An important reason for the decrease in fertility during Sperm cells of domestic animals have been the object of radicals [5]. The sperm plasma membrane contains a high various investigations all over the world. The capacity of amount of unsaturated fatty acids and is therefore spermatozoa to perform independent movement is a particularly susceptible to peroxydative damage with characteristic physiological capacity of theirs related to their subsequent loss of membrane integrity, impaired cell biological function - to fertilize the ovocyte. The successful function and decreased motility of spermatozoa [6, 7]. In freezing of bull s semen was achieved after discovering the addition to these damaging effects, reactive oxygen species ’ protective effect of glycerine and of the yolk of hen s egg [1]. are physiologically involved in hyperactivation of ’ This was followed by a period of intensive empirical spermatozoa and their continuous production is a study with numerous manuscript published demonstrating prerequisite for capacitation [8]. Semen contains antioxidants the effect of different storage diluents, protective agents and including ascorbic acid, tocopherol, B-carotine, catalasa, other additives [2] cooling rate, storage temperature, warming glutation peroxidase and superoxide dismutase, that rates and extending semen preservation methods across many balance lipid peroxidation and prevent excessive peroxide livestock species. The researchers on the influence of low formation [9, 10]. temperatures on the physiological properties and the fertilizing In the present study, we compared the effect of ability of ram spermatozoa are contradictory. Some authors Cu/ Zn-SOD from the fungal strain Humicola lutea 103 report that these characteristics do not change greatly for 24 h (HLSOD) on viability and fertility rates of frozen-thawed if stored at 4°C [3]. Other authors find that the ram bull spermatozoa, chilled and frozen-thawed ram spermatozoa considerably worsen their fertilizing ability in spermatozoa. these conditions. The researchers show that the physiological properties of MATERIAL AND METHODS ram spermatozoa are greatly influenced by the composition of the diluents [4]. Production of the HLSOD: The fungal strain H. lutea 103 The problem however, of ram spermatozoa preservation from the Mycological Collection of Institute of Microbiology, in vitro at low temperatures is still-unclear. Sofia was used for production of Cu/ Zn SOD [11]. storage of semen is formation of lipid peroxides of oxygen