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. 2020 Dec;13(12):2649-2657.
doi: 10.14202/vetworld.2020.2649-2657. Epub 2020 Dec 14.

Relationship of frozen-thawed semen quality with the fertility rate after being distributed in the Brahman Cross Breeding Program

Affiliations

Relationship of frozen-thawed semen quality with the fertility rate after being distributed in the Brahman Cross Breeding Program

Berlin Pandapotan Pardede et al. Vet World. 2020 Dec.

Abstract

Background and aim: Various factors can reduce the quality of semen used for artificial insemination and have an impact on fertility decline, such as poor handling during frozen semen distribution. This study was aimed at assessing the quality of frozen-thawed semen after distribution in the field and its importance in maintaining fertility.

Materials and methods: The Brahman Cross (BX) breeding program of PT Lembu Jantan Perkasa, Indonesia, was used. This program was preferred due to its adherence to guidelines that limit the effects of extraneous factors that may affect semen quality. Frozen-thawed semen samples from eight bulls with the same production code were analyzed and compared between the production site (artificial insemination [AI] center) and the field (BX breeding program). Total and progressive motility (PM) of sperm were determined using computer-assisted semen analysis. Plasma membrane integrity (PMI) was assessed using hypoosmotic swelling test, sperm viability using Eosin-Nigrosin staining, acrosome integrity using trypan blue-Giemsa staining, morphological abnormalities using William staining, and DNA fragmentation using toluidine blue staining. The fertility rate was determined using the conception rate (%) derived from AI data based on 502 AI services and 478 cows in the BX breeding program. A t-test was used to compare the quality of frozen-thawed semen before and after distribution. The relationship between the qualities of frozen semen after distribution in the field with fertility was analyzed using Pearson correlation.

Results: There was no significant difference (p>0.05) in the quality of frozen-thawed semen (sperm motility, PMI, viability, acrosome integrity, abnormalities, and DNA fragmentation) between the production site (AI center) and after distribution in the field (BX breeding program). The semen met the minimum standards for AI programs. Total motility (r=0.986), PM (r=0.961), sperm viability (r=0.971), PMI (r=0.986), and acrosome integrity (r=0.992) were all positively correlated (p<0.05) with fertility rate; while sperm abnormalities (r=-0.996) and sperm DNA fragmentation (r=0.975) were negatively correlated (p<0.05) with fertility rate.

Conclusion: The study showed that to achieve the maximal and optimal fertility rate in bulls in an AI program, the overall quality of frozen-thawed semen in all aspects is critical. This can be achieved if the handling during distribution and storage, as well as the various factors that may affect the quality of semen in the field, can be controlled properly.

Keywords: Brahman cross; artificial insemination; distribution; fertility rate; frozen semen quality.

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Figures

Figure-1
Figure-1
(a) Sperm viability assessment by Eosin-Nigrosin staining. The living sperm showed a colorless-head; the dead sperm will be colored by eosin and showed a redhead (arrow). (b) Sperm acrosome integrity assessment by trypan blue-Giemsa staining. Sperm with an intact acrosome showed a purple head, while a non-intact acrosome showed a pale or faded lavender head (arrow). (c) Sperm morphology assessment by carbol fuchsin eosin staining to distinguish between normal and abnormal sperm (arrow). (d) Sperm DNA fragmentation assessment by toluidine blue staining. Non-fragmented sperm showed a bright blue head, while fragmented sperm showed a dark blue head (arrow).
Figure-2
Figure-2
The results of the assessment of sperm viability (a), acrosome integrity (b), sperm abnormality (c), and sperm DNA fragmentation (d) in frozen semen before and after distribution in the Brahman Cross breeding program.
Figure-3
Figure-3
The relationship between total and progressive motility, sperm viability, plasma membrane integrity, and acrosome integrity with fertility rate (p<0.05) in frozen-thawed semen after distribution in the BX breeding program.
Figure-4
Figure-4
The relationship between sperm abnormalities and sperm DNA fragmentation with fertility rate (p<0.05) in frozen semen after distribution in the BX breeding program.

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