The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure
One of the critical points in the cryopreservation process is the use of a proper diluent while lowering the temperature following the resuspension and thawing processes. Here, we tested an alternative diluent for the process of freezing boar semen. We used skimmed dried milk (SDM) during the coolin...
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| Format: | Article |
| Language: | English |
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Universidade Estadual de Londrina
2015-07-01
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| Series: | Semina: Ciências Agrárias |
| Subjects: | |
| Online Access: | http://www.uel.br/revistas/uel/index.php/semagrarias/editor/submission/18766 |
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| author | Tatyane Bandeira Barros Daianny Barboza Guimarães Ludymila Furtado Cantanhêde Aline Viana Dias Leonardo Peres de Souza Jean Magloire Feugang Ricardo Toniolli |
| author_facet | Tatyane Bandeira Barros Daianny Barboza Guimarães Ludymila Furtado Cantanhêde Aline Viana Dias Leonardo Peres de Souza Jean Magloire Feugang Ricardo Toniolli |
| author_sort | Tatyane Bandeira Barros |
| collection | DOAJ |
| description | One of the critical points in the cryopreservation process is the use of a proper diluent while lowering the temperature following the resuspension and thawing processes. Here, we tested an alternative diluent for the process of freezing boar semen. We used skimmed dried milk (SDM) during the cooling and post-thawed resuspension steps. To do so, we collected semen from 15 Dalland boars using the glovedhand technique, and incubated each ejaculate sample at 30 °C. We then removed two semen aliquots from a pre-dilution. We diluted one of the aliquots in Beltsville thawing solution (BTS - control), and the remaining sample was diluted in SDM. Both aliquots were subsequently held at 30º C for 45 min (1st period of stabilization). At the end of this period, we analysed vigor and motility to determine sperm metabolic activity. We then held the diluted semen at 25° C for 30 min (2nd period of stabilization) and at 17° C for 2 h (3rd period stabilization). We centrifuged the semen at 800 × g and 1600 × g at 5º C for 15 min, discarded the supernatant, and resuspended the sperm pellet in 2 mL of the cooling diluent at 5° C for 1h. We again diluted the samples in 2 mL of the freezing diluent, poured them into straws, and cooled and plunged them into liquid N2. The sêmen samples were thawed in a 39º C water bath, and were resuspended in their respective diluents at the same temperature. We determined the following sperm features: vigor, motility, vitality, acrosomal integrity and membrane functionality. During the first phase of temperature cooling (30º C), semen diluted in SDM exhibited a higher vigor (3.4 ± 0.6) and motility (78.6 ± 13.0) than those diluted BTS (vigor: 3.1 ± 0.7; motility: 69.4±14.3). However, after the thawing procedure, the inverse was observed in that: BTS samples exhibited a higher vigor (2.1 ± 0.6) and motility (35.5 ± 21.0) than SDM samples (vigor: 1.7 ± 0.9; motility: 22.8 ± 18.1). Regarding membrane functionality and acrosomal integrity, we did not find a significant difference between BTS and SDM, although SDM provided a higher percentage of living cells at the end of the freeze/thaw procedures (72.3 ± 16.4). In summary, we suggest that BTS should be considered a better option than SDM for the cooling and post-thaw resuspension steps of boar semen freezing process. |
| first_indexed | 2024-12-11T12:20:33Z |
| format | Article |
| id | doaj.art-bbe27ccd1ae440baa2c53ef97c43507e |
| institution | Directory Open Access Journal |
| issn | 1676-546X 1679-0359 |
| language | English |
| last_indexed | 2024-12-11T12:20:33Z |
| publishDate | 2015-07-01 |
| publisher | Universidade Estadual de Londrina |
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| series | Semina: Ciências Agrárias |
| spelling | doaj.art-bbe27ccd1ae440baa2c53ef97c43507e2022-12-22T01:07:33ZengUniversidade Estadual de LondrinaSemina: Ciências Agrárias1676-546X1679-03592015-07-01363Supl12023203010.5433/1679-0359.2015v36n3Supl1p2023The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedureTatyane Bandeira Barros 0Daianny Barboza Guimarães 1Ludymila Furtado Cantanhêde 2Aline Viana Dias 3Leonardo Peres de Souza 4Jean Magloire Feugang 5Ricardo Toniolli 6Universidade Estadual do CearáUniversidade Estadual do CearáUniversidade Estadual do CearáUniversidade Estadual do CearáUniversidade Federal do PiauíUniversity State MississipiUniversidade Estadual do CearáOne of the critical points in the cryopreservation process is the use of a proper diluent while lowering the temperature following the resuspension and thawing processes. Here, we tested an alternative diluent for the process of freezing boar semen. We used skimmed dried milk (SDM) during the cooling and post-thawed resuspension steps. To do so, we collected semen from 15 Dalland boars using the glovedhand technique, and incubated each ejaculate sample at 30 °C. We then removed two semen aliquots from a pre-dilution. We diluted one of the aliquots in Beltsville thawing solution (BTS - control), and the remaining sample was diluted in SDM. Both aliquots were subsequently held at 30º C for 45 min (1st period of stabilization). At the end of this period, we analysed vigor and motility to determine sperm metabolic activity. We then held the diluted semen at 25° C for 30 min (2nd period of stabilization) and at 17° C for 2 h (3rd period stabilization). We centrifuged the semen at 800 × g and 1600 × g at 5º C for 15 min, discarded the supernatant, and resuspended the sperm pellet in 2 mL of the cooling diluent at 5° C for 1h. We again diluted the samples in 2 mL of the freezing diluent, poured them into straws, and cooled and plunged them into liquid N2. The sêmen samples were thawed in a 39º C water bath, and were resuspended in their respective diluents at the same temperature. We determined the following sperm features: vigor, motility, vitality, acrosomal integrity and membrane functionality. During the first phase of temperature cooling (30º C), semen diluted in SDM exhibited a higher vigor (3.4 ± 0.6) and motility (78.6 ± 13.0) than those diluted BTS (vigor: 3.1 ± 0.7; motility: 69.4±14.3). However, after the thawing procedure, the inverse was observed in that: BTS samples exhibited a higher vigor (2.1 ± 0.6) and motility (35.5 ± 21.0) than SDM samples (vigor: 1.7 ± 0.9; motility: 22.8 ± 18.1). Regarding membrane functionality and acrosomal integrity, we did not find a significant difference between BTS and SDM, although SDM provided a higher percentage of living cells at the end of the freeze/thaw procedures (72.3 ± 16.4). In summary, we suggest that BTS should be considered a better option than SDM for the cooling and post-thaw resuspension steps of boar semen freezing process.http://www.uel.br/revistas/uel/index.php/semagrarias/editor/submission/18766Seminal freezing; Skimmed dried Milk; Sperm viability |
| spellingShingle | Tatyane Bandeira Barros Daianny Barboza Guimarães Ludymila Furtado Cantanhêde Aline Viana Dias Leonardo Peres de Souza Jean Magloire Feugang Ricardo Toniolli The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure Semina: Ciências Agrárias Seminal freezing; Skimmed dried Milk; Sperm viability |
| title | The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure |
| title_full | The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure |
| title_fullStr | The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure |
| title_full_unstemmed | The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure |
| title_short | The use of skimmed dried milk as an alternative diluent for the cooling step during the boar sêmen freezing procedure |
| title_sort | use of skimmed dried milk as an alternative diluent for the cooling step during the boar semen freezing procedure |
| topic | Seminal freezing; Skimmed dried Milk; Sperm viability |
| url | http://www.uel.br/revistas/uel/index.php/semagrarias/editor/submission/18766 |
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