MD1068Z - Bull sperm thawing medium - Google Patents
Bull sperm thawing medium Download PDFInfo
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- MD1068Z MD1068Z MDS20160037A MDS20160037A MD1068Z MD 1068 Z MD1068 Z MD 1068Z MD S20160037 A MDS20160037 A MD S20160037A MD S20160037 A MDS20160037 A MD S20160037A MD 1068 Z MD1068 Z MD 1068Z
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- Moldova
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- thawing
- medium
- bull sperm
- sperm
- thyroxine
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- 238000010257 thawing Methods 0.000 title claims abstract description 24
- 244000309464 bull Species 0.000 title claims abstract description 15
- XUIIKFGFIJCVMT-LBPRGKRZSA-N L-thyroxine Chemical compound IC1=CC(C[C@H]([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-LBPRGKRZSA-N 0.000 claims abstract description 13
- 229950008325 levothyroxine Drugs 0.000 claims abstract description 13
- 239000012153 distilled water Substances 0.000 claims abstract description 12
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 12
- 239000001509 sodium citrate Substances 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000000877 morphologic effect Effects 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 230000003187 abdominal effect Effects 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 2
- 230000037149 energy metabolism Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 102000001390 Fructose-Bisphosphate Aldolase Human genes 0.000 description 1
- 108010068561 Fructose-Bisphosphate Aldolase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 108010019077 beta-Amylase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000009027 insemination Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 230000002366 lipolytic effect Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 210000005132 reproductive cell Anatomy 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000019100 sperm motility Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- DVBIMNUZCMCPRL-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;pentahydrate Chemical class O.O.O.O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O DVBIMNUZCMCPRL-UHFFFAOYSA-K 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Description
Invenţia se referă la criobiologie şi zootehnie, şi anume la un mediu pentru decongelarea spermei de taur. The invention relates to cryobiology and zootechnics, namely to a medium for thawing bull sperm.
Este cunoscut un mediu pentru decongelarea spermei de taur, care conţine citrat de sodiu şi apă distilată [1]. A medium for thawing bull sperm is known, which contains sodium citrate and distilled water [1].
Dezavantajul acestui mediu constă în aceea că el conţine doar electrolit, ceea ce asigură o protejare mai slabă la acţiunea factorilor de decongelare, în consecinţă indicii fiziologici şi morfologici ai spermei după decongelare rămân la un nivel scăzut comparativ cu sperma nativă. The disadvantage of this medium is that it contains only electrolyte, which provides weaker protection against the action of thawing factors, consequently the physiological and morphological indices of sperm after thawing remain at a low level compared to native sperm.
De asemenea este cunoscut un procedeu de sporire a calităţii spermei decongelate de taur, care constă în adăugarea lichidului peritoneal (abdominal) în procesul de decongelare [2]. A process for increasing the quality of thawed bull sperm is also known, which consists of adding peritoneal (abdominal) fluid to the thawing process [2].
Dezavantajul acestei soluţii constă în aceea că lichidul abdominal al embrionilor după conţinutul chimic şi biochimic diferă de indicii analogici ai plasmei seminale de taur. Pe de o parte, lichidul abdominal este o sursă suplimentară de dialdehidă malonică, ce se formează în procesul peroxidării lipidelor şi are un efect toxic, pe de altă parte reprezintă un solvent, care poate influenţa negativ asupra structurilor membranare ale celulelor reproductive. The disadvantage of this solution is that the abdominal fluid of embryos differs in chemical and biochemical content from the analogous indices of bull seminal plasma. On the one hand, the abdominal fluid is an additional source of malonic dialdehyde, which is formed in the process of lipid peroxidation and has a toxic effect, on the other hand, it is a solvent, which can negatively affect the membrane structures of reproductive cells.
Cea mai apropiată soluţie după esenţa invenţiei propuse este mediul pentru decongelarea spermei de taur, care conţine citrat de sodiu, glucoză, trilon B, β-amilază, catalază, aldolază şi apă distilată [3]. The closest solution according to the essence of the proposed invention is the medium for thawing bull sperm, which contains sodium citrate, glucose, trilon B, β-amylase, catalase, aldolase and distilled water [3].
Dezavantajul acestui mediu constă în aceea că el conţine multe componente, trei dintre care sunt enzime, ceea ce complică tehnologia preparării mediului, deoarece necesită o respectare strictă a condiţiilor de pregătire şi aplicare, totodată enzimele conţinute în mediu sunt costisitoare, ceea ce reduce accesibilitatea lui. The disadvantage of this medium is that it contains many components, three of which are enzymes, which complicates the technology of preparing the medium, as it requires strict adherence to the conditions of preparation and application, while the enzymes contained in the medium are expensive, which reduces its accessibility.
Problema pe care o soluţionează invenţia constă în menţinerea la un nivel înalt a mobilităţii gameţilor şi a integrităţii acrozomului după decongelare, crearea unui mediu pentru decongelarea spermei de taur accesibil, necostisitor, care să asigure stabilizarea structurilor morfologice ale spermatozoizilor după decongelare, stimularea metabolismului energetic şi sporirea indicilor morfologici şi fiziologici ai materialului seminal după decongelare. The problem solved by the invention consists in maintaining a high level of gamete mobility and acrosome integrity after thawing, creating an accessible, inexpensive environment for thawing bull sperm, which ensures the stabilization of the morphological structures of sperm after thawing, stimulating energy metabolism and increasing the morphological and physiological indices of semen after thawing.
Problema tehnică este soluţionată în invenţia propusă prin simplificarea componenţei mediului cunoscut, în care se include L-tiroxină. The technical problem is solved in the proposed invention by simplifying the composition of the known medium, which includes L-thyroxine.
Mediul pentru decongelarea spermei de taur conţine citrat de sodiu, L-tiroxină şi apă distilată, în următorul raport al componentelor: The medium for thawing bull sperm contains sodium citrate, L-thyroxine and distilled water, in the following ratio of components:
citrat de sodiu, g 1,5…2,0 L-tiroxină, µg 30…50 apă distilată, ml până la 100.sodium citrate, g 1.5…2.0 L-thyroxine, µg 30…50 distilled water, ml up to 100.
Rezultatul constă în menţinerea la un nivel înalt a mobilităţii gameţilor şi a integrităţii acrozomului după decongelare. The result is the maintenance of a high level of gamete mobility and acrosome integrity after thawing.
Efectul mediului este asigurat de proprietatea L-tiroxinei de a stimula activitatea metabolică şi regenerarea fiziologică a celulelor, de a regla permeabilitatea membranelor şi spori formarea energiei celulare. Prezenţa legăturilor conjugate în structura L-tiroxinei, responsabile de stabilitatea energetică şi termodinamică a sistemelor, poate determina şi rezistenţa celulelor la influenţa factorilor nefavorabili ai congelării. Deoarece acrosoma conţine setul de enzime proteolitice şi lipolitice ce determină procesul de fecundare, menţinerea integrităţii acrosomei şi, deci, a conţinutului acestor enzime după decongelare la un nivel înalt sporeşte capacitatea de fecundare a spermei decongelate. The effect of the environment is provided by the property of L-thyroxine to stimulate metabolic activity and physiological regeneration of cells, to regulate membrane permeability and enhance the formation of cellular energy. The presence of conjugated bonds in the structure of L-thyroxine, responsible for the energetic and thermodynamic stability of systems, can also determine the resistance of cells to the influence of unfavorable factors of freezing. Since the acrosome contains the set of proteolytic and lipolytic enzymes that determine the fertilization process, maintaining the integrity of the acrosome and, therefore, the content of these enzymes after thawing at a high level increases the fertilizing capacity of thawed sperm.
Conţinutul optimal al L-tiroxinei în componenţa mediului elaborat a fost determinat în mod experimental. The optimal content of L-thyroxine in the composition of the developed medium was determined experimentally.
Pregătirea mediului se realizează în modul următor. Într-un vas de sticlă, cotat de 100 ml, în condiţii sterile se adaugă 50 ml apă distilată, în care se dizolvă prin agitare citrat de sodiu triplu substituit pentahidratat (C6H5O7Na3x5H2O). După dizolvarea totală a citratului de sodiu se adaugă L-tiroxina conform exemplelor de realizare a invenţiei, soluţia se amestecă energic, după care se aduce cu apa distilată până la cotă. Mediul obţinut este omogen, transparent, fără precipitat sau fulgi. Mediul se pregăteşte nemijlocit înainte de întrebuinţare, având următorul raport al componentelor: The preparation of the medium is carried out as follows. In a glass vessel, with a capacity of 100 ml, under sterile conditions, 50 ml of distilled water is added, in which triple-substituted sodium citrate pentahydrate (C6H5O7Na3x5H2O) is dissolved by stirring. After the complete dissolution of the sodium citrate, L-thyroxine is added according to the embodiments of the invention, the solution is mixed vigorously, then it is brought to the mark with distilled water. The medium obtained is homogeneous, transparent, without precipitate or flakes. The medium is prepared immediately before use, having the following ratio of components:
Exemplul 1 Example 1
citrat de sodiu, g 1,5 L-tiroxină, µg 50 apă distilată, ml până la 100.sodium citrate, g 1.5 L-thyroxine, µg 50 distilled water, ml to 100.
Exemplul 2 Example 2
citrat de sodiu, g 1,7 L-tiroxină, µg 40 apă distilată, ml până la 100.sodium citrate, g 1.7 L-thyroxine, µg 40 distilled water, ml to 100.
Exemplul 3 Example 3
citrat de sodiu, g 2,0 L-tiroxină, µg 30 apă distilată, ml până la 100.sodium citrate, g 2.0 L-thyroxine, µg 30 distilled water, ml to 100.
Mediul a fost experimentat în condiţii de laborator. În prima serie de experienţe pentru decongelare au fost selectate variantele de mediu conform exemplelor 1-3. Rezultatele decongelării spermei de taur în mediul din variantele propuse în invenţie sunt incluse în tabelul 1. The medium was tested under laboratory conditions. In the first series of experiments for thawing, the medium variants according to examples 1-3 were selected. The results of thawing bull sperm in the medium from the variants proposed in the invention are included in table 1.
Tabelul 1 Table 1
Indicii morfologici şi fiziologici studiaţi Variantele experimentale Exemplul 1 Exemplul 2 Exemplul 3 Mobilitatea, bal 4,20 ± 0,14 5,00 ± 0,14 4,50 ± 0,15 Integritatea acrosomei, % 61,20 ± 1,30 61,40 ± 2,07 61,40 ± 1,52Morphological and physiological indices studied Experimental variants Example 1 Example 2 Example 3 Mobility, bal 4.20 ± 0.14 5.00 ± 0.14 4.50 ± 0.15 Acrosome integrity, % 61.20 ± 1.30 61.40 ± 2.07 61.40 ± 1.52
Varianta optimală a compoziţiei mediului în care s-au manifestat cele mai pronunţate proprietăţi stabilizatoare este cea de a doua. The optimal variant of the medium composition in which the most pronounced stabilizing properties were manifested is the second one.
În a doua serie de experienţe decongelarea spermei de taur a fost realizată în trei variante: martor, cea mai apropiată soluţie, invenţia propusă. Sperma de taur a fost decongelată în conformitate cu varianta martor în soluţie izotonică de citrat de sodiu, în mediul conform celei mai apropiate soluţii şi în mediul propus în invenţie. In the second series of experiments, the thawing of bull sperm was carried out in three variants: control, the closest solution, the proposed invention. Bull sperm was thawed according to the control variant in isotonic sodium citrate solution, in the medium according to the closest solution and in the medium proposed in the invention.
În tabelul 2 sunt prezentaţi indicii morfologici şi fiziologici ai spermei decongelate conform celei mai apropiate soluţii şi invenţiei propuse. Table 2 presents the morphological and physiological indices of thawed sperm according to the closest solution and the proposed invention.
Tabelul 2 Table 2
Indicii morfologici şi fiziologici studiaţi Sperma nativă Martor Cea mai apropiată soluţie Invenţia propusă Mobilitatea, bal 7,20 ± 0,11 3,70 ± 0,23 4,80 ± 0,10 5,20 ± 0,07 Integritatea acrosomei, % 70,40 ± 1,34 50,20 ± 1,48 55,00 ± 1,00 61,20 ± 0,84Morphological and physiological indices studied Native sperm Control Closest solution Proposed invention Mobility, bal 7.20 ± 0.11 3.70 ± 0.23 4.80 ± 0.10 5.20 ± 0.07 Acrosome integrity, % 70.40 ± 1.34 50.20 ± 1.48 55.00 ± 1.00 61.20 ± 0.84
Analiza rezultatelor obţinute demonstrează că după decongelare indicii studiaţi conform invenţiei sunt superiori în comparaţie cu cea mai apropiată soluţie. Astfel, mobilitatea spermatozoizilor conform invenţiei, în comparaţie cu varianta martor, constituie 40,5%, pe când mobilitatea conform celei mai apropiate soluţii, comparativ cu martorul, constituie 29,7%. Integritatea acrosomei conform invenţiei constituie 21,9%, conform celei mai apropiate soluţii - 9,56%. The analysis of the obtained results demonstrates that after thawing the studied indices according to the invention are superior in comparison with the closest solution. Thus, the mobility of spermatozoa according to the invention, in comparison with the control variant, constitutes 40.5%, while the mobility according to the closest solution, in comparison with the control, constitutes 29.7%. The integrity of the acrosome according to the invention constitutes 21.9%, according to the closest solution - 9.56%.
Comparativ cu sperma nativă indicii studiaţi după decongelare conform invenţiei sunt majoraţi faţă de cea mai apropiată soluţie - mobilitatea spermatozoizilor constituie 72,2%, iar integritatea acrosomei atinge 86,9%, contra 66,6% şi 78,1% respectiv. Compared to native sperm, the indices studied after thawing according to the invention are increased compared to the closest solution - sperm motility is 72.2%, and acrosome integrity reaches 86.9%, compared to 66.6% and 78.1% respectively.
Mediul propus stabilizează structurile morfologice ale spermatozoizilor, stimulează metabolismul energetic şi restabilirea fiziologică a gameţilor, ceea ce duce la creşterea mobilităţii spermatozoizilor după decongelare. Menţinerea integrităţii acrosomei la un nivel înalt asigură o capacitate sporită de fecundare a spermei de taur. Mediul este necostisitor, accesibil, aplicarea lui asigură o calitate eficientă a materialului reproducător decongelat. The proposed medium stabilizes the morphological structures of spermatozoa, stimulates energy metabolism and physiological restoration of gametes, which leads to increased sperm mobility after thawing. Maintaining acrosome integrity at a high level ensures an increased fertilization capacity of bull sperm. The medium is inexpensive, accessible, its application ensures an efficient quality of thawed reproductive material.
1. Инструкция по организации и технологии работы станций и предприятий по исскуственному осеменению сельскохозяйственных животных. Москва, Колос, 1981 1. Instruction on organization and technology of work stations and enterprises on artificial insemination of agricultural animals. Moscow, Kolos, 1981
2. RU 2358689 C1 2009.06.20 2. RU 2358689 C1 2009.06.20
3. SU 880420 A1 1981.11.15 3. SU 880420 A1 1981.11.15
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Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| MDS20160037A MD1068Z (en) | 2016-03-18 | 2016-03-18 | Bull sperm thawing medium |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| MDS20160037A MD1068Z (en) | 2016-03-18 | 2016-03-18 | Bull sperm thawing medium |
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| Publication Number | Publication Date |
|---|---|
| MD1068Y MD1068Y (en) | 2016-09-30 |
| MD1068Z true MD1068Z (en) | 2017-04-30 |
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| Application Number | Title | Priority Date | Filing Date |
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| MDS20160037A MD1068Z (en) | 2016-03-18 | 2016-03-18 | Bull sperm thawing medium |
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| MD (1) | MD1068Z (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU880420A1 (en) * | 1980-02-27 | 1981-11-15 | За витель 880420 (И) | Medium for defrosting bull sperm |
| MD638F1 (en) * | 1996-05-03 | 1996-12-31 | Inst De Fiziol Al Academiei De | Medium for bull's sperm decongelation |
| RU2358689C1 (en) * | 2007-10-02 | 2009-06-20 | Волгоградская государственная сельскохозяйственная академия | Method for increasing activity, survivability and breeding efficiency of frozen bulls' semen |
| UA47713U (en) * | 2009-07-09 | 2010-02-25 | Национальный Университет Биоресурсов И Природопользования Украины | Medium for defrosting deep-frozen sperm of bulls |
| UA50908U (en) * | 2009-12-29 | 2010-06-25 | Национальный Университет Биоресурсов И Природопользования Украины | Method for preserving sperm of hogs |
| UA51245U (en) * | 2009-12-29 | 2010-07-12 | Национальный Университет Биоресурсов И Природопользования Украины | method for decryoconservation of bull sperm |
-
2016
- 2016-03-18 MD MDS20160037A patent/MD1068Z/en not_active IP Right Cessation
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU880420A1 (en) * | 1980-02-27 | 1981-11-15 | За витель 880420 (И) | Medium for defrosting bull sperm |
| MD638F1 (en) * | 1996-05-03 | 1996-12-31 | Inst De Fiziol Al Academiei De | Medium for bull's sperm decongelation |
| RU2358689C1 (en) * | 2007-10-02 | 2009-06-20 | Волгоградская государственная сельскохозяйственная академия | Method for increasing activity, survivability and breeding efficiency of frozen bulls' semen |
| UA47713U (en) * | 2009-07-09 | 2010-02-25 | Национальный Университет Биоресурсов И Природопользования Украины | Medium for defrosting deep-frozen sperm of bulls |
| UA50908U (en) * | 2009-12-29 | 2010-06-25 | Национальный Университет Биоресурсов И Природопользования Украины | Method for preserving sperm of hogs |
| UA51245U (en) * | 2009-12-29 | 2010-07-12 | Национальный Университет Биоресурсов И Природопользования Украины | method for decryoconservation of bull sperm |
Non-Patent Citations (1)
| Title |
|---|
| Инструкция по организации и технологии работы станций и предприятий по исскуственному осеменению сельскохозяйственных животных. Москва, Колос, 1981 * |
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| MD1068Y (en) | 2016-09-30 |
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