| 1. |
- Goodla, Lavanya, et al.
(författare)
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Quality of bull spermatozoa after preparation by single-layer centrifugation
- 2014
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Ingår i: Journal of Dairy Science. - : American Dairy Science Association. - 0022-0302 .- 1525-3198. ; 97, s. 2204-2212
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Tidskriftsartikel (refereegranskat)abstract
- The present study aimed to evaluate the effect of single-layer centrifugation (SLC) through a species-specific colloid (Androcoll-B; patent pending, J. M. Morrell) on bull sperm quality. Computer-assisted sperm analysis of motility and flow cytometric analysis of sperm viability (SYBR-14/propidium iodide staining), chromatin integrity (acridine orange staining), reactive oxygen species production [Hoechst 33258-hydroethidine-2',7'-dichlorodihydrofluorescein diacetate (HO-HE-DCFDA) staining], mitochondrial membrane potential (staining with JC-1 probe), and protein tyrosine phosphorylation (specific antibody staining) were performed on unselected and SLC-selected sperm samples. Single-layer centrifugation of bull spermatozoa resulted in the selection of a sperm population that had high mitochondrial membrane potential, a higher content of phosphorylated protein, and more reactive oxygen species than control samples. Sperm chromatin damage was lower in the SLC samples although sperm viability and motility did not differ between SLC samples and controls. These observations suggest that SLC of bull semen in a soybean-containing extender improved some, but not all, parameters of sperm quality.
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| 2. |
- Yusnizar, Yulnawati
(författare)
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Frozen-Thawed Epididymal Sperm Quality and the Success Rate of Artificial Insemination in Spotted Buffaloes (Bubalus bubalis carabanensis)
- 2013
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Ingår i: Buffalo Bulletin. - 0125-6726. ; 32, s. 494-497
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Tidskriftsartikel (refereegranskat)abstract
- Spotted buffalo is an exotic species in Indonesia, especially in Torajaland, South Sulawesi. Population of this species is close to become extinct due to a high slaughtering rate of this animal in the funeral ceremonies. The extinction risks are become worse since the farmer kept the bull with special treatment and prevent them from mating activity. Therefore, we tried to conserve gamete by collecting the sperm from caudal epididymides of bulls soon after slaughtering. Soya bean-based (A) and two egg yolk-based (B and C) extenders were used in order to find out the optimal freezing extender that could maintain the quality and fertility of post-thawed epididymal sperm. The post-thawed progressive motility of epididymal sperm was 45, 40 and 39.2%, while livability was 64.8, 66 and 63.3% and functional membrane integrity was 65.8, 65.4 and 63% for A, B and C extenders, respectively. In general, there were no significantly different (P>0.05) of all observed parameters in different phospholipid-based extenders. The pregnancy rate obtained from soya bean-based extender (37.5%) was also similar (P>0.05) to that of egg yolk-based (40%) extender. In conclusion, soya bean-based and egg yolk-based extenders could maintain the post-thawed quality of spotted buffalo epididymal sperm and yielded similar pregnancy rate following artificial insemination program.
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| 3. |
- Yusnizar, Yulnawati, et al.
(författare)
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Microphthalmia-associated transcription factor mutations are associated with white-spotted coat color in swamp buffalo
- 2015
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Ingår i: Animal Genetics. - : Wiley. - 0268-9146 .- 1365-2052. ; 46:6, s. 676-682
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Tidskriftsartikel (refereegranskat)abstract
- A candidate gene analysis of the microphthalmia-associated transcription factor (MITF) gene was used in an attempt to identify the genetic basis for a white-spotted coat color phenotype in the Asian swamp buffalo (Bubalus bubalis carabanensis). Ninety-three buffaloes32 solid, 38 spotted and 23 white individualswere Sanger-sequenced for all MITF exons as well as highly conserved intronic and flanking regions. MITFcDNA representing skin and iris tissue from six spotted, nine solid and one white buffaloes was also Sanger-sequenced to confirm detected mutations. Two independent loss-of-function mutations, a premature stop codon (c.328C>T, p.Arg110*) and a donor splice-site mutation (c.840+2T>A, p.Glu281_Leu282Ins8), both of which cause white-spotted coat color in swamp buffaloes, were identified. The nonsense mutation leads to a premature stop codon in exon 3, and likely removal of the resulting mRNA via nonsense-mediated decay pathway, whereas the donor splice-site mutation leads to aberrant splicing of exon 8 that encodes part of a highly conserved region of MITF. The resulting insertion of eight amino acid residues is expected to perturb the leucine zipper part in the basic helix-loop-helix leucine zipper (bHLH-Zip) domain and will most likely influence dimerization and DNA binding capacity. Electrophoretic mobility shift assay was performed using mutant and wild-type MITF proteins and showed that the mutant MITF protein resulting from the splice-site mutation decreased invitro DNA binding capacity compared to wild-type MITF. White-spotted buffalo bulls are sacrificed in funeral ceremonies in Tana Toraja, Indonesia, because they are considered holy, and our results show that genetic variation causes a tie to the cultural use of these buffaloes.
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