| 1. |
- Deori, Sourabh, et al.
(författare)
-
Comparison of single layer centrifugation and magnetic activated cell sorting for selecting viable boar spermatozoa after thawing
- 2022
-
Ingår i: Livestock Science. - : Elsevier BV. - 1871-1413 .- 1878-0490. ; 257
-
Tidskriftsartikel (refereegranskat)abstract
- Sperm selection techniques, such as magnetic activated cell sorting (MACS) and colloid centrifugation, are reported to select good quality spermatozoa from semen samples of various species. Although the sperm quality of fresh boar semen is usually good, cryopreservation has a negative effect on parameters such as plasma membrane integrity and mitochondrial activity. Therefore, the objective of the present study was to determine whether MACS or centrifugation through a single layer of colloid (Single Layer Centrifugation, SLC) would be beneficial in enriching thawed boar sperm samples for viable spermatozoa with active mitochondria and good chromatin integrity. Frozen samples from three boars, three ejaculates per boar, were thawed and split. One part was selected by MACS, one was prepared by SLC, and the remainder served as the control. Controls and the selected sperm samples were evaluated for sperm quality (plasma membrane integrity, chromatin integrity, mitochondrial membrane potential and production of reactive oxygen species). Although several aspects of sperm quality were improved in the SLC-selected sperm samples compared to control, the flow-through MACS samples were only improved in having a lower proportion of spermatozoa with immature chromatin (Hi green fluorescence) compared to the labeled control. Sperm quality in the SLC samples was better than in the flow-through samples from MACS. Therefore, despite promising reports of the use of MACS for selecting good quality spermatozoa from semen in other species, the method was not useful for improving sperm quality in the thawed boar sperm samples in this experiment.
|
|
| 2. |
- Deori, Sourabh, et al.
(författare)
-
Fast protein liquid chromatography profiles of seminal plasma proteins in young bulls: A biomarker of sperm maturity?
- 2021
-
Ingår i: Livestock Science. - : Elsevier BV. - 1871-1413 .- 1878-0490. ; 250
-
Tidskriftsartikel (refereegranskat)abstract
- Breeding companies want to use semen from bulls as soon as possible to take advantage of their desirable genetics. It takes several weeks for the sperm quality of young bulls to stabilize and for post-thaw sperm quality to become acceptable for artificial insemination. Seminal plasma proteins protect spermatozoa during cryopreservation; it may take some time for the seminal plasma protein profile to stabilize. The purpose of this study was to determine if the seminal plasma protein profile can be used as a marker of likely seminal maturity in young bulls. A comparison was made of the seminal plasma protein profile in the ejaculates of 10 bulls of 9-10 months old (Sample I), with the profiles from ejaculates taken from the same bulls at 13-16 months old (Sample II) using fast protein liquid chromatography. This is a method for separating classes of proteins according to their binding ability. The peak area and peak height of different classes of proteins did not differ significantly between the two samples for each bull, except for peak 5 (heparin-binding proteins) and total peak area (p<0.05). The heparinbinding protein peak height and area were significantly higher (p<0.05) in Sample II than in Sample I. In conclusion, levels of fertility associated heparin-binding proteins increase with age in young bulls and might serve as a biomarker of sperm maturity. .
|
|
| 3. |
|
|
| 4. |
- Deori, Sourabh, et al.
(författare)
-
Single Layer Centrifugation with 20% or 30% Porcicoll separates the majority of spermatozoa from a sample without adversely affecting sperm quality
- 2020
-
Ingår i: Reproduction in Domestic Animals. - : Wiley. - 0936-6768 .- 1439-0531. ; 55, s. 1337-1342
-
Tidskriftsartikel (refereegranskat)abstract
- Centrifugation of boar semen through one layer of 40% colloid (Porcicoll) was previously shown to separate spermatozoa from bacteria without having a detrimental effect on sperm quality. However, some spermatozoa were lost. The purpose of the present study was to determine whether 20% or 30% Porcicoll could be used to recover most of the spermatozoa without impacting on sperm quality. Insemination doses (n = 10) from a commercial boar station were sent to the laboratory at the Swedish University of Agricultural Sciences and processed by Single Layer Centrifugation with 20% and 30% Porcicoll approximately 7 hr after semen collection. The resulting sperm samples and controls were evaluated for sperm quality immediately and again after storage at 16-18 degrees C for 4 and 7 days. Sperm recovery was 94 +/- 18% and 87 +/- 15% for 20% and 30% Porcicoll, respectively (p > .05). Sperm mitochondrial membrane potential and chromatin integrity were unaffected (p > .05). The proportion of live spermatozoa producing superoxide (9 +/- 8%, 7 +/- 6% and 3 +/- 1%;p < .05), and the proportion of spermatozoa with high stainability DNA (0.68 +/- 19%, 0.61 +/- 0.22% and 0.96 +/- 0.23%;p < .05- <0.01), were marginally increased whereas membrane integrity, although high, was lower in the centrifuged samples than in the controls (82 +/- 8%, 83 +/- 5% versus 92 +/- 4%;p < .05). In conclusion, centrifugation through 20% or 30% Porcicoll enables most spermatozoa to be recovered, without having a major effect on sperm quality. These results are encouraging for further studies involving microbiological investigation of the processed samples, and scaling-up to process larger volumes of boar ejaculates.
|
|
