The way to obtain serum for inducing superovulation in cows
(57) Abstract:Serum for inducing superovulation in cows was obtained as follows. Mature bull intramuscularly injected under the scheme progesterone and 2.5% dose of 0.10-0.15 mg/kg of body weight 2 times a day for 3 days, and then estradiol 2% dose of 0.10 mg/kg wet weight, after this bull castrated and take his blood, from which the serum. The method provides the maximum number of embryos from donor cows. The invention relates to the field of biotechnology. Some of its elements can be used in the practice of veterinary obstetrics and gynecology.Currently, the team there are 3 types of natural hormones, can cause superovulation: hormonal serum of pregnant mares, pituitary gonadotropin and gonadotropin menopause man (centuries Madison, B. L. Madison. Embryo transfer in the practice of breeding dairy cattle. M: VO "Agropromizdat", 1988, page 48).A method of obtaining a blood serum for inducing superovulation in cows, including obtaining serum from blood collected from the jugular vein (see L. K. Ernst, N. And.Sergeev. Transplantation S="ptx2">But, as the authors state, when hormonal treatment according to this scheme, respond with superovulate 72% of the animals, and the embryos receive 47% of donor cows from the number of processed (see ibid, page 120).Technical solution to the problem is to create the least time-consuming, more cheap, do not result in a disruption of pregnancy in producers of blood and ultimately a more effective method, which allows you to get the maximum number of embryos from donor cows.The task is achieved in that in the method of obtaining blood serum for inducing superovulation in cows, including obtaining serum from blood collected from the jugular vein, Mature bull intramuscularly injected under the scheme progesterone 2.5%, a dose of 0.10 - 0.15 mg/kg of body weight 2 times a day for 3 days, and then estradiol 2% within 2 days dose of 0.10 mg/kg wet weight, after this bull castrated, take from him the blood, and receive from her the serum. The introduction of this serum increases the output of a full-fledged embryos.According to patent scientific and technical literature not found a similar set of features that allows to judge about the level of invention proposals.An example of a specific implementation of sleduushie 2 day estradiol 2% - a 3.0 milThe next day (day 7) is produced by castration of the animal surgically and the blood sample is 12-14 hours. The activity in this way the resulting serum is determined by the reaction of the uterus in immature mice (similar to the determination of the activity of serum geropsych mares). The way to obtain serum for inducing superovulation in cows, including obtaining serum from blood collected from the jugular vein, characterized in that the Mature bull intramuscularly injected under the scheme progesterone and 2.5% dose of 0.10 - 0.15 mg/kg of body weight 2 times a day for 3 days, and then estradiol 2% dose of 0.10 mg/kg wet weight, after this bull castrated and take his blood.
SUBSTANCE: method involves using product containing spermatozoa treated by drying with freezing to humidity level of 1% and having injured membrane or spermatozoon head so that spermatozoon nucleus retains its genetic validity enough for fertilization. The spermatozoon heads fertilize an isolated oocyte after rehydration and microinjection being done. The retained genetic integrity is enough for fertilizing an oocyte and producing living descendants. Method involves collecting living mature spermatozoa, making spermatozoa suspension in special purpose physiological medium, freezing the spermatozoa suspension for producing frozen spermatozoa, drying the frozen spermatozoa or spermatozoa heads in vacuum to humidity level of 1%, making rehydration of spermatozoa or spermatozoa heads with injured membrane and selecting those retaining nucleus of genetic integrity. The selected spermatozoa or spermatozoa heads are used for fertilizing isolated oocytes with living descendants being produced.
EFFECT: enhanced effectiveness in producing living descendants.
39 cl, 5 dwg, 1 tbl
SUBSTANCE: the present innovation deals with individual matching donor sheep to recipient sheep at similar antigenic composition of blood types being correspondent to the value of antigenic similarity index being ra=0.51-1.00, where ra - antigenic similarity index. Moreover, the mentioned antigenic similarity index should be calculated by the following formula: where S - the number of similar antigens in a donor sheep and in a recipient sheep, n1 - the number of detected antigens in a donor sheep, n2 - the number of detected antigens in a recipient sheep. The present innovation enables to increase the level of adaptability of transferred ovine embryos by 25%.
EFFECT: higher efficiency of embryo transfer.
3 ex, 4 tbl
SUBSTANCE: method includes ovary removal after animal slaughtering, their transportation, ovocytes extraction from ovary, their cultivation to metaphase II stage, preparation of boar sperm, combined cultivation of ovocytes and spermatozoids and cultivation of ovulums. Besides, ovary is separated from animals no later than 30 minutes after the moment of animal immobilisation. Ovary is transplanted in free of antibiotics salt solution at 35-37°C during 1 hour. Ovocytes are taken from ovary by dissectioning of visible antrum-containing follicles. Received ovocytes are split into groups by quantity of surrounding cumuluce cell layers, namely: ovocytes surrounded by 4-5 layers of cumuluce cells, 2-3 layers and one layer of cumuluce layer. Each group of ovocytes is cultivated in medium containing 10 IU of human chorionic gonadotropin. To fertilise ovocytes in vitro, ejaculated sperm of pigs is used, which is dissolved with glucose-chelate-citrate-sulphate diluter and cleaned from semen plasma dissolved with mTBM (mouse thymic virus) medium and incubated during 25 minutes under incubator conditions at 5% CO2 and 38.5°C. Produced zygotes are cultivated in NCSU-23 medium free of phenol red and enriched with 0.1% of amino acids during 6-7 days; in addition, 0.17 mM Na-pyruvate and 2.75 mM Na-lactate are introduced into NCSU-23 medium during the first three days of cultivation.
EFFECT: increase of mature ovocytes yield and their fertilisation ratio in vitro.
1 tbl, 1 ex
FIELD: package industry.
SUBSTANCE: container 10 includes vessel (21) with biological medium, germinal cells and/or one or more embryos. Vessel (21) has CO2 permeable wall, CO2 permeable gasket (28) and closure medium (30) for selective access to internal vessel cavity. Buffer chamber (60) for air saturated with CO2 surrounds vessel at least partially and is formed by shell (61). Buffer chamber is connected to CO2 permeable wall. Such container construction is intended specifically for intravaginal cultivation, and permeable gasket (28) prevents vaginal secretion from entering buffer chamber. Buffer chamber (60) normalises water pH in vessel after the vessel leaves environment saturated with CO2.
EFFECT: prevented negative effect for long-term embryo(s) cultivation.
53 cl, 19 dwg
SUBSTANCE: method includes oophorectomy in animals after slaughter, transportation of oophorons, extraction of oocyte-cumulus complexes by aspiration of follicles content, irrigation of the complexes and their cultivation in a medium not covered with oil, in the presence of serum gonadotropin of pregnant mares, human chorionic gonadotropin and 5% estrous serum of bovine cattle. At that, follicles of diametre from 2 to 8 mm are used to obtain oocytes. After extraction of the oocyte-cumulus complexes are immediately placed in a washing medium containing 0.5 mM of isobutylmethylxanthine to synchronise reinitiation of meiosis. Before the cultivation the selection of oocytes based on their morphological evaluation is carried out. All the procedures of isolation, irrigation and selection are carried out in a medium containing 0.5 mM of isobutylmethylxanthine. Oocyte-cumulus complexes were cultured in a medium modified by insertion of 50 ng/ml of bovine prolactin hormone.
EFFECT: increase of proportion of mature oocytes in vitro, and their ability for further development.
1 ex, 1 tbl
SUBSTANCE: method comprises extracting the oocytes-cumulus complexes from ovaries, cultivating them to the stage of metaphase II, co-cultivating of oocytes and sperm cells, cultivating the impregnated ootids. And additionally the follicle-stimulating and luteinising hormone is administered in the form of the combined preparation Menopur, estradiol, heparin, caffeine and antibiotic. The cultivating is carried out in the base medium. Extraction of the oocytes-cumulus complexes from ovaries is carried out from live sheep under anaesthesia by puncturing ovarian follicles during laparotomy. Superovulation stimulation is preliminary carried out. In the maturation medium the preparation Menopur is added, the concentration of both components that are part of the preparation, is 0.075 IU/ml, and estradiol is added to the maturation medium in the concentration of 10 mcg/ml. For impregnation the newly obtained sperm of tups is used. For tup sperm capacitation and to the maturation medium the combination of heparin is added in an amount of 5 units/ml and caffeine in an amount of 0.2 mg/ml. The cultivation and impregnation is carried out in four well Petri dishes in 500 mcl nutrient medium under 200 mcl paraffin oil.
EFFECT: use of the method enables to increase the proportion of output of oocytes.
3 cl, 11 dwg, 3 ex