Trehalose and a Calcium Chelator for Ram Semen Cryopreservation

Bittencourt, Rodrigo Freitas; Oba, Eunice; Filho, Antônio de Lisboa Ribeiro; Biscarde, Carmo Emanuel Almeida; Chalhoub, Marcos; Bicudo, Sony Dimas

doi:10.5380/avs.v19i2.31064

Cited by 5 publications

(3 citation statements)

References 19 publications

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“…The total amount of spermatozoa per treatment group was sufficient for at least two insemination doses of 100 × 10 6 spermatozoa with motility and packed in French straws (0.25 mL). Subsequently, the samples were cooled to 5 °C (0.46 °C/min) (Model 518C, Minitub do Brasil Ltda, Porto Alegre, Brazil), left to equilibrate for 2 h ( Bittencourt et al, 2014 ), and were then frozen in liquid nitrogen vapor.…”

Section: Methodsmentioning

confidence: 99%

Dimethylacetamide alone or in combination with glycerol can be used for cryopreservation of ovine semen

et al. 2020

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Dimethylacetamide has been included in different extenders for the cryopreservation of semen from species with promising results. The objective of this study was to evaluate the use of dimethylacetamide (DMA) in different concentrations, associated or not with glycerol (GLY), for the cryopreservation of ovine semen, and its effects on in vitro sperm parameters and post-thaw in vivo fertility. Five semen samples of five adult Santa Ines sheep (n=25) were used. The collected ejaculates were divided among the seven treatments for subsequent cryopreservation. The treatments presented different concentrations of DMA and GLY, being divided as G1: GLY 6%; G2: DMA 3%; G3: GLY 5% + DMA 1%; G4: GLY 4% + DMA 2%; G5: GLY 3% + DMA 3%; G6: GLY 2% + DMA 4%; G7: GLY 1% + DMA 5%. %. Post-thawing of the straws, aliquots were evaluated for computerized sperm kinetics (CASA) and plasma membrane integrity, using fluorescent probes and flow cytometry. After the in vitro evaluation of the sperm parameters, in vivo testing was performed by laparoscopic artificial insemination of 72 females. The post-thaw total motility (%) evaluated by CASA were 51.4, 51.4, 50.1, 53.6, 52.3, 52.8 and 46.9, respectively, for the seven groups. And the plasma membrane integrity (%) were 19.7, 28.4, 22.3, 29.4, 24.3, 17.9 and 16.9, respectively. There were no differences (P> 0.05) between the treatments for the parameters of spermatic kinetics and membrane integrity. For females inseminated with semen from the control group (G1, GLY6%), the percentage of pregnant females was 36.1%, a result similar to that obtained with G3 treatment (GLY5% + DMA1%). In conclusion, dimethylacetamide, either alone or in combination with glycerol, can be used for cryopreservation of ovine semen.

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Section: Methodsmentioning

confidence: 99%

Dimethylacetamide alone or in combination with glycerol can be used for cryopreservation of ovine semen

et al. 2020

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“…Compuseram o estudo o tratamento 1 (T1-6% de LDL), o tratamento 2 (T2-8% de LDL), o tratamento 3 (T3-12% de LDL), o tratamento 4 (T4-16% de LDL) e o tratamento 5 (T5-20% de LDL). Para a composição do meio base, foi utilizada a solução TRIS-gema de ovo e glicerol (Bittencourt et al, 2014), substituindo-se totalmente a gema de ovo por LDL para os diferentes tratamentos.…”

Section: Methodsunclassified

Utilização da lipoproteína de baixa densidade, em diferentes concentrações, em meio diluidor para criopreservação do sêmen ovino

Lima

Bittencourt

Filho

et al. 2019

Arq. Bras. Med. Vet. Zootec.

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Utilização da lipoproteína de baixa densidade, em diferentes concentrações, em meio diluidor para criopreservação do sêmen ovino [Use of low density lipoprotein in different concentrations in diluent medium for cryopreservation of ovine semen]A. RESUMOA utilização da gema de ovo dificulta a padronização de meios diluidores e apresenta riscos biológicos. Assim, este estudo avaliou diferentes concentrações da lipoproteína de baixa densidade (LDL), em substituição à gema de ovo, para a confecção de diluentes para criopreservação espermática em ovinos. Foram utilizados um diluidor controle (CTR= 20% de gema de ovo) e cinco tratamentos, substituindo-se a gema pelas diferentes proporções de LDL (T1=6%; T2=8%; T3=12%; T4=16%; T5=20%), todos à base de TRIS-glicerol. Para o estudo, utilizaram-se dois ejaculados, de seis reprodutores da raça Santa Inês. Sessenta dias após a criopreservação, as amostras foram descongeladas e avaliadas subjetivamente quanto à motilidade total (MT, %) e progressiva (MP, %), ao vigor (1-5) e à integridade funcional (choque hisposmótico com água destilada, %) e estrutural (corante supravital eosina, %) das membranas espermáticas. As avaliações de vigor e funcionalidade de membrana não diferiram (P>0,05) entre os grupos. Entretanto, os grupos T4 (P<0,01) e T5 (P<0,05) foram superiores ao CTR para os parâmetros MT, MP e integridade estrutural de membrana, o que confirma que as LDLs podem ser alternativas eficientes para substituição da gema de ovo em diluidores para criopreservação de sêmen ovino. ABSTRACTThe use of egg yolk makes it difficult to standardize extenders and presents biological hazards. Thus, this study evaluated different concentrations of low-density lipoprotein (LDL) to replace yolk extenders for production of sperm for cryopreservation in ovine. A control extender was used (CTR= 20% yolk) and five treatments, replacing the yolk by different ratios of LDL (T1= 6%; T2= 8%, T3= 12%; T4= 16%; T5= 20%) all based on TRIS-glycerol. For the study, two ejaculates from six Santa Ines breeding were used. Sixty days after cryopreservation, the samples were thawed and evaluated for total motility (MT, %) and progressive motility (MP, %), vigor (1-5) and the functional integrity (hyposmotic shock with distilled water, %) and structural (supravital dye eosin, %) of the sperm membranes. The evaluations of strength and membrane functionality didn't differ (P> 0.05) between groups. However, T4 (P< 0.01) and T5 (P< 0.05) groups were superior to the CTR for the MT, MP, and membrane structural integrity parameters, which confirms that LDLs can be efficient alternatives for yolk replacement in extenders for cryopreservation of ovine semen.

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“…Egg yolk and/or milk represent a risk of contamination if microorganisms, such as bacteria and fungi, are present in the fresh product. Such contamination can release endo-toxins that reduce the fertilization capacity of sperm (Bittencourt et al, 2008). Furthermore, high density lipoproteins (HDLs) in egg yolk are one factor that decreases the quality of semen by causing efflux of cholesterol from the sperm plasma membrane and resulted in change in fluidity that increase the sensitivity to cold shock (Amirat et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Impact of Replacing Egg Yolk With Lecithin on Sperm Characteristics, Bacterial Count and Fertilizing Ability of Cryopreserved Buffalo Semen.

El-Sherbieny

2014

Journal of Animal and Poultry Production

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Soy bean lecithin has been used to replace egg yolk in the semen extender. The aim of this study was to evaluate the effect of replacing egg yolk by soy-lecithin in tris extender on freezability and fertilizing capacity of buffalo semen. Semen from five buffalo bulls was frozen in tris extender containing 0.5, 1.0 and 2% soy-lecithin (treatments) or 15% egg yolk (control). Percentage of motility, livability and intact acrosome spermatozoa and also bacterial count was assessed in diluted, postequilibrated and post-thawed semen. Results revealed insignificant differences in percentage of sperm motility in diluted semen. However, percentages of sperm motility, livability and intact acrosome spermatozoa increased (P<0.05) and bacterial count decreased (P<0.05) in post-equilibrated and post-thawed semen extended with 0.5% soy-lecithin. These results reflected in the highest conception rate (66.7%) of buffalo cows inseminated with semen containing 05% soy-lecithin compared with 53.3% in semen containing egg yolk (control). In conclusion, egg yolk could be replaced by soy-lecithin in tris-based extender at a level of 0.5% to increase fertilizing capacity of cryopreserved buffalo semen.

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Trehalose and a Calcium Chelator for Ram Semen Cryopreservation

Cited by 5 publications

References 19 publications

Dimethylacetamide alone or in combination with glycerol can be used for cryopreservation of ovine semen

Dimethylacetamide alone or in combination with glycerol can be used for cryopreservation of ovine semen

Utilização da lipoproteína de baixa densidade, em diferentes concentrações, em meio diluidor para criopreservação do sêmen ovino

Impact of Replacing Egg Yolk With Lecithin on Sperm Characteristics, Bacterial Count and Fertilizing Ability of Cryopreserved Buffalo Semen.

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