Cryoprotectant is the crucial factor in the cryopreservation process. In general, there are two types of cryoprotectant, permeating and non‐permeating cryoprotectants. Dimethyl sulfoxide (DMSO) and egg yolk are common permeating and non‐permeating cryoprotectants respectively. Hence, the objective of the present study was to determine the best proportion of DMSO and egg yolk for the cryopreservation of Rasbora tawarensis sperm. A completely randomized experimental design was used in this study which involves two types of cryoprotectant and their combination at different concentrations, namely 5% DMSO, 5% egg yolk, 5% DMSO + 5% egg yolk and 2.5% DMSO + 2.5% egg yolk. Every treatment was conducted in three replicates. Combination of 5% DMSO + 5% egg yolk gave the best results cryoprotectant treatment had significant effects on sperm motility, fertilization and hatching rate of the R. tawarensis eggs (p < .05). It is concluded that the best proportion of cryoprotectants for sperm cryopreservation in this species is 5% DMSO + 5% egg yolk.
Sperms are vulnerable to oxidative stress due to the high proportion of polyunsaturated fatty acids within their membranes. This condition could decrease sperms quality during preservation. β-carotene is among antioxidants able to neutralize reactive oxygen species, natural by-products of oxygen metabolism in sperms. This study was done to investigate the capability and protective effect of this antioxidant on motility, viability, acrosomal integrity, and plasma membrane integrity of frozen-thawed sperms of Aceh swamp buffalo. The study was conducted using completely randomized design with four treatments and five replications. Fresh semen was diluted in egg-yolk tris-based extenders with the addition of antioxidant of 0% (w/v) (control, P0), 0.625% (w/v) (P1), 1.25% (w/v) (P2), and 2.5% (w/v) (P3) prior to freezing. Variables of sperms quality examined were motility, viability, acrosomal integrity, and plasma membrane integrity of semen after dilution, equilibration, and thawing. Semen having the best post-thawing quality after β-carotene treatment was used for artificial insemination (AI) for accessing its pregnancy rate. The results showed that the addition of 0.625% (w/v) β-carotene (P1) resulted in the highest motility, viability, acrosomal integrity, and plasma membrane integrity. Pregnancy rate of buffalo inseminated with semen treated with 0.625% β-carotene (P1) was 50%. In conclusion, administration of 0.625% β-carotene was able to maintain and could protect motility, viability, acrosomal integrity, and plasma membrane integrity of frozen sperms of Aceh swamp buffalo that are good for artificial insemination.
Eriani K, Rahmi R, Jamil I, Rosnizar R, Azhar A. 2019. Body size characteristics and polymorphism in GH and GHRH genes of Simeulue Buffalo of Aceh, Indonesia. Biodiversitas 20: 236-242. This study was done to determine body size characteristics, to identify the presence of polymorphism in the GH/MspI and GHRH/HaeIII genes, and to analyze the relationship between GH and GHRH gene polymorphisms to body size characteristics in Simeulue buffalo. Experimental animals used were 45 Simeulue buffalo randomly sampled from three subdistricts, i.e., Salang, Alafan and Simeulue Cut, fifteen from each subdistrict. Body size characteristics measured were withered height, body length, chest circumference, chest depth, and hip height. A 5 mL whole blood was collected from fifteen out of forty five by using EDTA as an anticoagulant. Extraction of genomic DNA and analysis of GH/MspI and GHRH/HaeIII polymorphisms was done using GeneAid DNA extraction kit and polymerase chain reaction-restriction length fragment polymorphism (PCR-RFLP) method, respectively. The data of body size characteristics which includes mean value, standard deviation, and coefficient of variation were analyzed descriptively. The molecular data represented by DNA banding pattern was used to calculate allele and genotype frequencies, expected (He) and observed (Ho) heterozygosities, and Hardy-Weinberg equilibrium. The relationship between genotype and body size characteristic was analyzed using t-test. The results showed that coefficient of variation in body size characteristics was low, ranging from 5.39-16.06% in the age category of 4-6 years old and ranging from 5.25-11.27% in the age category of 2- 4 years old. The results of the molecular analysis indicated that GH/MspI locus was monomorphic by the presence of one (+/+) genotype. GHRH/HaeIII locus was polymorphic as shown by the presence of 3 genotypes namely AA (0.13), AB (0.80) and BB (0.07). Expected heterozygosity (0.80) was higher than observed heterozygosity (0.49). The frequency of GHRH/HaeIII agreed with Hardy Weinberg equilibrium. GHRH/HaeIII polymorphism did not significantly influence (P>0.05) body size characteristics of Simeulue buffalo.
Abstract. Eriani K, Sari N, Ihdina M, Rosnizar. 2017. The effect of equilibration time on semen freezing of local swamp buffalo (Bubalus bubalis) with combination extender of lactose and glycerol. Nusantara Bioscience 9: 77-82. This study was to determine the different time of equilibration on semen of swamp buffalo (Bubalus bubalis) which was diluted by adding Tris-Egg Yolk extender of lactose and glycerol cryoprotectant combination. Fresh sperm of the swamp buffalo (B. bubalis) was diluted by using Tris-Egg Yolk extender of lactose cryoprotectants combination 0 mM (L 0 ), 60 mM (L 60 ), 120 mM (L 120 ) and glycerol 3% (G 3 ), 5% (G 5 ), 7% (G 7 ) with the equilibration of 2,5, 3, and 4 hours. The parameter used in this study was the percentage of sperm motility and live treatment. The results of the 2.5-hour equilibration with a combination of cryoprotectants L 60 G 7 showed the best quality in all parameters. The results of the 3-hour equilibration with combination of cryoprotectants L 60 G 7 showed the best quality in all parameters whereas the best quality in all parameters in 4-hour equilibration was the combination of cryoprotectants L 120 G 7 .
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