Stocktake Sale on now: wide range of books at up to 70% off!
Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Impact of bull age, sperm processing, and microclimatic conditions on the viability and DNA integrity of cryopreserved bovine sperm

Burcu Cinar A , Heinrich Bollwein A , Mathias Siuda A , Matthias Lautner B , Claus Leiding B and Eleni Malama https://orcid.org/0000-0003-1433-7336 A *
+ Author Affiliations
- Author Affiliations

A Clinic of Reproductive Medicine, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, Zurich 8057, Switzerland.

B Besamungsverein Neustadt a.d. Aisch e.V., Karl-Eibl-Straße 17-27, Neustadt a.d. Aisch 91413, Germany.

* Correspondence to: emalama@vetclinics.uzh.ch

Handling Editor: Marc Yeste

Reproduction, Fertility and Development 36, RD23219 https://doi.org/10.1071/RD23219
Submitted: 7 December 2023  Accepted: 11 April 2024  Published online: 7 May 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Context

Seasonal microclimatic fluctuations can cause changes in sperm quality even in dairy bulls bred under temperate climate. These changes can vary between sires of different age and affect sperm freezability.

Aims

We aimed to evaluate the modulating effect of bull age and equilibration time before freezing on the seasonal pattern of sperm viability and DNA integrity post-thaw.

Methods

In the frame of systematic sperm quality control, we assessed the integrity of sperm plasma membrane and acrosome (PMAI) in 15,496 cryopreserved bovine batches, and the percentage of sperm with high DNA fragmentation index (%DFI) after 0 h and 3 h incubation at 38°C post-thaw (3 h) in 3422 batches. Semen was equilibrated for 24 h before freezing if collected on Monday or Wednesday and 72 h if produced on Friday. We investigated the effect of season, bull age, equilibration, and temperature-humidity index (THI) on the day of semen collection on sperm traits using mixed-effects linear models.

Key results

PMAI and %DFI (0 h and 3 h) deteriorated with increasing THI. The effect of THI on %DFI was detected with a 30-day time lag. Seasonal fluctuations of sperm quality were similar between young, mature, and older sires. Prolonged equilibration did not affect PMAI but was linked to elevated %DFI (3 h) in summer.

Conclusions

Extending equilibration from 24 to 72 h is compatible with commercial standards of bovine sperm quality post-thaw; however, it could interfere with the seasonal pattern of the latter.

Implications

Systematic monitoring of bovine sperm quality enables the prompt detection of stress factors related to microclimate and semen processing.

Keywords: bull, bull age, cryopreservation, DNA integrity, microclimate, season, sperm quality control, sperm viability.

References

Aguilar-Mahecha A, Hales BF, Robaire B (2001) Expression of stress response genes in germ cells during spermatogenesis. Biology of Reproduction 65, 119-127.
| Crossref | Google Scholar | PubMed |

Ahmadi A, Ng S-C (1999) Fertilizing ability of DNA-damaged spermatozoa. Journal of Experimental Zoology 284, 696-704.
| Crossref | Google Scholar | PubMed |

Aitken RJ (2017) Reactive oxygen species as mediators of sperm capacitation and pathological damage. Molecular Reproduction and Development 84, 1039-1052.
| Crossref | Google Scholar | PubMed |

Amann RP, DeJarnette JM (2012) Impact of genomic selection of AI dairy sires on their likely utilization and methods to estimate fertility: a paradigm shift. Theriogenology 77, 795-817.
| Crossref | Google Scholar | PubMed |

Argov N, Sklan D, Zeron Y, Roth Z (2007) Association between seasonal changes in fatty-acid composition, expression of VLDL receptor and bovine sperm quality. Theriogenology 67, 878-885.
| Crossref | Google Scholar | PubMed |

Argov-Argaman N, Mahgrefthe K, Zeron Y, Roth Z (2013) Season-induced variation in lipid composition is associated with semen quality in Holstein bulls. Reproduction 145, 479-489.
| Crossref | Google Scholar | PubMed |

Arslan HO, Herrera C, Malama E, Siuda M, Leiding C, Bollwein H (2019) Effect of the addition of different catalase concentrations to a TRIS-egg yolk extender on quality and in vitro fertilization rate of frozen-thawed bull sperm. Cryobiology 91, 40-52.
| Crossref | Google Scholar | PubMed |

Arunkumar R, Kumaresan A, Sinha MK, Elango K, Ebenezer Samuel King JP, Nag P, Karuthadurai T, Baithalu RK, Mohanty TK, Kumar R, Datta TK (2022) The cryopreservation process induces alterations in proteins associated with bull sperm quality: the equilibration process could be a probable critical control point. Frontiers in Endocrinology 13, 1064956.
| Crossref | Google Scholar |

Barnett AG, Dobson AJ (2010) ‘Analysing seasonal health data.’ (Springer: Berlin, London) doi:10.1007/978-3-642-10748-1

Barth AD, Bowman PA (1994) The sequential appearance of sperm abnormalities after scrotal insulation or dexamethasone treatment in bulls. The Canadian Veterinary Journal 35, 93-102.
| Google Scholar | PubMed |

Bhakat M, Mohanty TK, Raina VS, Gupta AK, Khan HM, Mahapatra RK, Sarkar M (2011) Effect of age and season on semen quality parameters in Sahiwal bulls. Tropical Animal Health and Production 43, 1161-1168.
| Crossref | Google Scholar | PubMed |

Brito LFC, Silva AEDF, Rodrigues LH, Vieira FV, Deragon LAG, Kastelic JP (2002) Effects of environmental factors, age and genotype on sperm production and semen quality in Bos indicus and Bos taurus AI bulls in Brazil. Animal Reproduction Science 70, 181-190.
| Crossref | Google Scholar | PubMed |

Bucher K, Malama E, Siuda M, Janett F, Bollwein H (2019) Multicolor flow cytometric analysis of cryopreserved bovine sperm: a tool for the evaluation of bull fertility. Journal of Dairy Science 102, 11652-11669.
| Crossref | Google Scholar |

Carreira JT, Trevizan JT, Carvalho IR, Kipper B, Rodrigues LH, Silva C, Perri SHV, Drevet JR, Koivisto MB (2017) Does sperm quality and DNA integrity differ in cryopreserved semen samples from young, adult, and aged Nellore bulls? Basic and Clinical Andrology 27, 12.
| Crossref | Google Scholar |

Cormier N, Bailey JL (2003) A differential mechanism is involved during heparin- and cryopreservation-induced capacitation of bovine spermatozoa. Biology of Reproduction 69, 177-185.
| Crossref | Google Scholar | PubMed |

Dancho M, Vaughan D (2023) anomalize: tidy anomaly detection. R package version 0.3.0. Available at https://cran.r-project.org/package=anomalize

Evenson D, Jost L (2000) Sperm chromatin structure assay for fertility assessment. Current Protocols in Cytometry 13, 7.13.1-7.13.27.
| Crossref | Google Scholar |

Fatehi AN, Bevers MM, Schoevers E, Roelen BAJ, Colenbrander B, Gadella BM (2006) DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after the first cleavages. Journal of Andrology 27, 176-188.
| Crossref | Google Scholar | PubMed |

Finocchiaro R, Van Kaam JBCHM, Portolano B, Misztal I (2005) Effect of heat stress on production of mediterranean dairy sheep. Journal of Dairy Science 88, 1855-1864.
| Crossref | Google Scholar | PubMed |

Fleisch A, Malama E, Witschi U, Leiding C, Siuda M, Janett F, Bollwein H (2017) Effects of an extension of the equilibration period up to 96 hours on the characteristics of cryopreserved bull semen. Theriogenology 89, 255-262.
| Crossref | Google Scholar | PubMed |

Fuerst-Waltl B, Schwarzenbacher H, Perner C, Sölkner J (2006) Effects of age and environmental factors on semen production and semen quality of Austrian Simmental bulls. Animal Reproduction Science 95, 27-37.
| Crossref | Google Scholar | PubMed |

Garcia-Oliveros LN, de Arruda RP, Batissaco L, Gonzaga VHG, Nogueira VJM, Florez-Rodriguez SA, Almeida FdS, Alves MBR, Pinto SCC, Nichi M, Losano JDdA, Kawai GKV, Celeghini ECC (2020) Heat stress effects on bovine sperm cells: a chronological approach to early findings. International Journal of Biometeorology 64, 1367-1378.
| Crossref | Google Scholar | PubMed |

Garner DL, Thomas CA, Gravance CG (1999) The effect of glycerol on the viability, mitochondrial function and acrosomal integrity of bovine spermatozoa. Reproduction in Domestic Animals 34, 399-404.
| Crossref | Google Scholar |

Gasparrini A (2011) Distributed lag linear and non-linear models in R: the package dlnm. Journal of Statistical Software 43, 1-20.
| Crossref | Google Scholar | PubMed |

Gasparrini A, Armstrong B, Kenward MG (2010) Distributed lag non-linear models. Statistics in Medicine 29, 2224-2234.
| Crossref | Google Scholar | PubMed |

Gliozzi TM, Turri F, Manes S, Cassinelli C, Pizzi F (2017) The combination of kinetic and flow cytometric semen parameters as a tool to predict fertility in cryopreserved bull semen. Animal 11, 1975-1982.
| Crossref | Google Scholar | PubMed |

Green CE, Watson PF (2001) Comparison of the capacitation-like state of cooled boar spermatozoa with true capacitation. Reproduction 122, 889-898.
| Crossref | Google Scholar | PubMed |

Gürler H, Malama E, Heppelmann M, Calisici O, Leiding C, Kastelic JP, Bollwein H (2016) Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology 86, 562-571.
| Crossref | Google Scholar | PubMed |

Hallap T, Nagy S, Håård M, Jaakma Ü, Johannisson A, Rodriguez-Martinez H (2005) Sperm chromatin stability in frozen-thawed semen is maintained over age in AI bulls. Theriogenology 63, 1752-1763.
| Crossref | Google Scholar | PubMed |

Haugan T, Reksen O, Gröhn YT, Kommisrud E, Ropstad E, Sehested E (2005) Seasonal effects of semen collection and artificial insemination on dairy cow conception. Animal Reproduction Science 90, 57-71.
| Crossref | Google Scholar | PubMed |

Karabinus DS, Vogler CJ, Saacke RG, Evenson DP (1997) Chromatin structural changes in sperm after scrotal insulation of Holstein bulls. Journal of Andrology 18, 549-555.
| Crossref | Google Scholar | PubMed |

Karoui S, Díaz C, González-Marín C, Amenabar ME, Serrano M, Ugarte E, Gosálvez J, Roy R, López-Fernández C, Carabaño MJ (2012) Is sperm DNA fragmentation a good marker for field AI bull fertility? Journal of Animal Science 90, 2437-2449.
| Crossref | Google Scholar | PubMed |

Koivisto MB, Costa MTA, Perri SHV, Vicente WRR (2009) The effect of season on semen characteristics and freezability in Bos indicus and Bos taurus bulls in the Southeastern region of Brazil. Reproduction in Domestic Animals 44, 587-592.
| Crossref | Google Scholar | PubMed |

Kumaresan A, Johannisson A, Al-Essawe EM, Morrell JM (2017) Sperm viability, reactive oxygen species, and DNA fragmentation index combined can discriminate between above- and below-average fertility bulls. Journal of Dairy Science 100, 5824-5836.
| Crossref | Google Scholar | PubMed |

Leite TG, do Vale Filho VR, de Arruda RP, de Andrade AFC, Emerick LL, Zaffalon FG, Martins JAM, Andrade VJd (2010) Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry. Animal Reproduction Science 120, 31-38.
| Crossref | Google Scholar | PubMed |

Llamas Luceño N, de Souza Ramos Angrimani D, de Cássia Bicudo L, Szymańska KJ, Van Poucke M, Demeyere K, Meyer E, Peelman L, Mullaart E, Broekhuijse MLWJ, Van Soom A (2020) Exposing dairy bulls to high temperature-humidity index during spermatogenesis compromises subsequent embryo development in vitro. Theriogenology 141, 16-25.
| Crossref | Google Scholar | PubMed |

Malama E, Kiossis E, Theodosiou T, Boscos C, Bollwein H (2012) Lag effect of microclimatic conditions on DNA integrity of frozen-thawed bovine sperm. Animal Reproduction Science 136, 33-41.
| Crossref | Google Scholar | PubMed |

Malama E, Zeron Y, Janett F, Siuda M, Roth Z, Bollwein H (2017) Use of computer-assisted sperm analysis and flow cytometry to detect seasonal variations of bovine semen quality. Theriogenology 87, 79-90.
| Crossref | Google Scholar | PubMed |

Malama E, Siuda M, Wyck S, Witschi U, Bollwein H (2020) Multicolor flow cytometry for quality control of cryopreserved bovine sperm. Experience from a Swiss semen production center. Animal Reproduction Science 220, 106454.
| Crossref | Google Scholar |

Mathevon M, Buhr MM, Dekkers JCM (1998) Environmental, management, and genetic factors affecting semen production in Holstein bulls. Journal of Dairy Science 81, 3321-3330.
| Crossref | Google Scholar | PubMed |

Maya-Soriano MJ, Taberner E, Sabés-Alsina M, López-Béjar M (2013) Retinol might stabilize sperm acrosomal membrane in situations of oxidative stress because of high temperatures. Theriogenology 79, 367-373.
| Crossref | Google Scholar | PubMed |

Melean M, Herrera C, Siuda M, Bollwein H, Malama E (2022) 120 Effect of bull exposure to high temperature-humidity index levels on the quality of sperm selected through density gradient centrifugation for in vitro fertilisation. Reproduction, Fertility and Development 34, 297.
| Crossref | Google Scholar |

Morrell JM, Nongbua T, Valeanu S, Lima Verde I, Lundstedt-Enkel K, Edman A, Johannisson A (2017) Sperm quality variables as indicators of bull fertility may be breed dependent. Animal Reproduction Science 185, 42-52.
| Crossref | Google Scholar | PubMed |

Murphy EM, Kelly AK, O’Meara C, Eivers B, Lonergan P, Fair S (2018a) Influence of bull age, ejaculate number, and season of collection on semen production and sperm motility parameters in Holstein Friesian bulls in a commercial artificial insemination centre. Journal of Animal Science 96, 2408-2418.
| Crossref | Google Scholar | PubMed |

Murphy EM, Eivers B, O’Meara CM, Lonergan P, Fair S (2018b) Effect of increasing equilibration time of diluted bull semen up to 72 h prior to freezing on sperm quality parameters and calving rate following artificial insemination. Theriogenology 108, 217-222.
| Crossref | Google Scholar | PubMed |

Nagy S, Johannisson A, Wahlsten T, Ijäs R, Andersson M, Rodriguez-Martinez H (2013) Sperm chromatin structure and sperm morphology: their association with fertility in AI-dairy Ayrshire sires. Theriogenology 79, 1153-1161.
| Crossref | Google Scholar | PubMed |

NOAA National Centers for Environmental Information (2021) State of the climate: global climate report for August 2021. Available at https://www.ncdc.noaa.gov/sotc/global/202108

Ollech D (2022) seastests: seasonality tests. R package version 0.15.4. Available at https://cran.r-project.org/package=seastests

Orgal S, Zeron Y, Elior N, Biran D, Friedman E, Druker S, Roth Z (2012) Season-induced changes in bovine sperm motility following a freeze-thaw procedure. Journal of Reproduction and Development 58, 212-218.
| Crossref | Google Scholar | PubMed |

Papa PM, Maziero RD, Guasti PN, Junqueira CR, Freitas-Dell’Aqua CP, Papa FO, Vianna FP, Alvarenga MA, Crespilho AM, Dell’Aqua JA, Jr. (2015) Effect of glycerol on the viability and fertility of cooled bovine semen. Theriogenology 83(1), 107-113.
| Crossref | Google Scholar | PubMed |

Pardede BP, Supriatna I, Yudi Y, Agil M (2020) Decreased bull fertility: age-related changes in sperm motility and DNA fragmentation. E3S Web of Conferences 151, 01010.
| Crossref | Google Scholar |

Pieper L, Meschede T, Jung M, Janowitz U, Schulze M (2023) Influence of equilibration time and bull-specific extender for cryopreservation on semen quality and fertility in German Holstein Friesian bulls: a controlled field trial. Animals 13, 2285.
| Crossref | Google Scholar | PubMed |

Pinheiro J, Bates D, DebRoy S, Sarkar D, The R Development Core Team (2015) nlme: linear and nonlinear mixed effects models. R package version 3.1-120. Available at http://CRAN.R-project.org/package=nlme

Puglisi R, Pozzi A, Foglio L, Spanò M, Eleuteri P, Grollino MG, Bongioni G, Galli A (2012) The usefulness of combining traditional sperm assessments with in vitro heterospermic insemination to identify bulls of low fertility as estimated in vivo. Animal Reproduction Science 132, 17-28.
| Crossref | Google Scholar | PubMed |

Rahman MB, Kamal MM, Rijsselaere T, Vandaele L, Shamsuddin M, Van Soom A (2014) Altered chromatin condensation of heat-stressed spermatozoa perturbs the dynamics of DNA methylation reprogramming in the paternal genome after in vitro fertilisation in cattle. Reproduction, Fertility and Development 26, 1107-1116.
| Crossref | Google Scholar | PubMed |

Sabés-Alsina M, Lundeheim N, Johannisson A, López-Béjar M, Morrell JM (2019) Relationships between climate and sperm quality in dairy bull semen: a retrospective analysis. Journal of Dairy Science 102, 5623-5633.
| Crossref | Google Scholar |

Salimiyekta Y, Jensen J, Su G, Gebreyesus G (2023) Age-dependent genetic and environmental variance of semen quality in Nordic Holstein bulls. Journal of Dairy Science 106, 2598-2612.
| Crossref | Google Scholar | PubMed |

Salman A, Fernández-Alegre E, Francisco-Vázquez R, Gómez-Martín R, Fernández-Fernández A, Areán-Dablanca H, Domínguez JC, González-Montaña JR, Caamaño JN, Martínez-Pastor F (2023) Extension of the equilibration period up to 24 h maintains the post-thawing quality of Holstein bull semen frozen with OPTIXcell®. Animal Reproduction Science 250, 107209.
| Crossref | Google Scholar |

Seifi-Jamadi A, Zhandi M, Kohram H, Luceño NL, Leemans B, Henrotte E, Latour C, Demeyere K, Meyer E, Van Soom A (2020) Influence of seasonal differences on semen quality and subsequent embryo development of Belgian Blue bulls. Theriogenology 158, 8-17.
| Crossref | Google Scholar | PubMed |

Sellem E, Broekhuijse MLWJ, Chevrier L, Camugli S, Schmitt E, Schibler L, Koenen EPC (2015) Use of combinations of in vitro quality assessments to predict fertility of bovine semen. Theriogenology 84, 1447-1454.e5.
| Crossref | Google Scholar | PubMed |

Simões R, Feitosa WB, Siqueira AFP, Nichi M, Paula-Lopes FF, Marques MG, Peres MA, Barnabe VH, Visintin JA, Assumpçaõ MEO (2013) Influence of bovine sperm DNA fragmentation and oxidative stress on early embryo in vitro development outcome. Reproduction 146, 433-441.
| Crossref | Google Scholar | PubMed |

Snoj T, Kobal S, Majdic G (2013) Effects of season, age, and breed on semen characteristics in different Bos taurus breeds in a 31-year retrospective study. Theriogenology 79, 847-852.
| Crossref | Google Scholar | PubMed |

Söderquist L, Janson L, Håård M, Einarsson S (1996) Influence of season, age, breed and some other factors on the variation in sperm morphological abnormalities in Swedish dairy A.I. bulls. Animal Reproduction Science 44, 91-98.
| Crossref | Google Scholar |

The R Development Core Team (2022) ‘R: a language and environment for statistical computing.’ (R Foundation for Statistical Computing). Available at http://www.r-project.org/

Trevizan JT, Carreira JT, Carvalho IR, Kipper BH, Nagata WB, Perri SHV, Franco Oliveira ME, Pierucci JC, Koivisto MBd (2018) Does lipid peroxidation and oxidative DNA damage differ in cryopreserved semen samples from young, adult and aged Nellore bulls? Animal Reproduction Science 195, 8-15.
| Crossref | Google Scholar | PubMed |

Valeanu S, Johannisson A, Lundeheim N, Morrell JM (2015) Seasonal variation in sperm quality parameters in Swedish red dairy bulls used for artificial insemination. Livestock Science 173, 111-118.
| Crossref | Google Scholar |

Vince S, Žura Žaja I, Samardžija M, Majić Balić I, Vilić M, Đuričić D, Valpotić H, Marković F, Milinković-Tur S (2018) Age-related differences of semen quality, seminal plasma, and spermatozoa antioxidative and oxidative stress variables in bulls during cold and warm periods of the year. Animal 12, 559-568.
| Crossref | Google Scholar | PubMed |

Vincent P, Underwood SL, Dolbec C, Bouchard N, Kroetsch T, Blondin P (2021) Bovine semen quality control in artificial insemination centers. In ‘Bovine Reproduction’. pp. 1019–1031. (John Wiley & Sons, Inc). doi:10.1002/9781119602484.ch81

Waterhouse KE, Haugan T, Kommisrud E, Tverdal A, Flatberg G, Farstad W, Evenson DP, De Angelis PM (2006) Sperm DNA damage is related to field fertility of semen from young Norwegian Red bulls. Reproduction, Fertility and Development 18, 781-788.
| Crossref | Google Scholar | PubMed |

Westfalewicz B, Słowińska M, Judycka S, Ciereszko A, Dietrich MA (2021) Comparative proteomic analysis of young and adult bull (Bos taurus) cryopreserved semen. Animals 11, 2013.
| Crossref | Google Scholar |

WMO (2022) Provisional state of the global climate in 2022. World Meteorological Organization, Geneva, Switzerland. Available at https://library.wmo.int/idurl/4/66214. ISBN: 978-92-63-11316-0

Yeste M, Estrada E, Rivera del Álamo M-M, Bonet S, Rigau T, Rodríguez-Gil J-E (2014) The increase in phosphorylation levels of serine residues of protein HSP70 during holding time at 17°C is concomitant with a higher cryotolerance of boar spermatozoa. PLoS ONE 9(3), e90887.
| Crossref | Google Scholar | PubMed |