Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

Waberski, Dagmar; Luther, Anne-Marie; Grünther, Benita; Jäkel, Helen; Henning, Heiko; Vogel, Charlotte; Peralta, W. A.; Weitze, K. F.

doi:10.1038/s41598-019-51319-1

Cited by 64 publications

(91 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In this context, it is evident that the temperature is an important factor that can interfere on bacterial dissemination during semen storage, particularly for stallion and boar cooled-semen technology, in which protocols highlight use of a relatively high temperature (15 to 17 °C) in a nutrient-rich extender that can favor bacterial growth. However, it was demonstrated that even for stallions (Price et al, 2008) and boars (Waberski et al, 2019), the hypothermic storage (5 °C) may reduce the use of antimicrobial drugs (Table 1).…”

Section: Use Of Antibacterial Substances In Semen Extendersmentioning

confidence: 99%

Current and alternative trends in antibacterial agents used in mammalian semen technology

Santos¹,

Silva²

2020

Anim Reprod

View full text Add to dashboard Cite

How to cite: Santos CS, Silva AR. Current and alternative trends in antibacterial agents used in mammalian semen technology. AbstractThe use of antibacterial substances as additives in extenders for ensuring the sanitary quality of the semen employed in reproductive biotechniques and preserving it from bacterial deterioration has been reported since the mid-twentieth century. However, the deleterious effects of these drugs on the sperm quality as well as their effectiveness in controlling bacterial growth in the preserved semen have been questioned. The aim of this review was to report the antimicrobials primarily used in the extenders added to the semen of mammals, and to present alternatives to their use. Among the various mammalian species, there is a large variation regarding the antimicrobial types added to semen extenders as cephalosporins (ceftiofur, cefdinir, eg) and quinolones (ofloxacin, ciprofloxacin), alone or in combination with large action spectra substances as penicillin-streptomycin and gentamicin-tylosin-lincomycin-spectinomycin. To combat problems related to bacterial resistance to these drugs, the emergence of alternatives is increasingly evident. Among these alternatives, use of physical methods as centrifugation and filtration, as well as the use of antimicrobial peptides and other substances from different origins have been highlighted for presenting antimicrobial potential.

show abstract

Section: Use Of Antibacterial Substances In Semen Extendersmentioning

confidence: 99%

Current and alternative trends in antibacterial agents used in mammalian semen technology

Santos¹,

Silva²

2020

Anim Reprod

View full text Add to dashboard Cite

show abstract

“…It is important to consider that the satisfactory in vivo results previously obtained 5 were from sows inseminated with heterospermic doses from pools of a total of 23 boars. Thus, the boar effect was masked to some extent.…”

Section: Discussionmentioning

confidence: 99%

“…For analyses performed during storage, a different semen dose tube was used at each time of evaluation (24, 72, and 120 hours of storage) to avoid the influence of manipulation. Total and progressive motility were assed using a CASA system (AndroVison ® version 1.1.4; Minitüb GmbH) under phase contrast microscopy (200× magnification, Axio Scope.A1, Zeiss ® ) 4,5 …”

Section: Methodsmentioning

confidence: 99%

“…Samples of raw and extended semen were fixed in a formalin‐citrate solution for sperm morphology and acrosome integrity analyses, 4,5 respectively. All samples were analyzed under a phase contrast microscope at 1000× magnification, and 200 spermatozoa were assessed for sperm morphology and classified as normal or abnormal 12 .…”

Section: Methodsmentioning

confidence: 99%

“…Aiming to promote the prudent use of antibiotics, a recent commercial antibiotic‐free boar semen extender has been tested under hypothermic storage 4,5 . Liquid‐extended boar semen without antibiotics preserved at 5°C or 10°C reduced bacterial load and maintained satisfactory sperm motility (>75%) and plasma membrane integrity (>80%) during 120 hours of storage 4 .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Differential seminal plasma proteome signatures of boars with high and low resistance to hypothermic semen preservation at 5°C

et al. 2020

View full text Add to dashboard Cite

Background Hypothermic storage at 5°C has been investigated as an alternative to promote the prudent use of antibiotics for boar artificial insemination doses. However, this temperature is challenging for some ejaculates or boars. Objective The present study aimed to identify putative biomarkers for semen resistance to hypothermic storage at 5°C by comparing the seminal plasma proteomes of boars with high and low seminal resistance to preservation at 5°C. Materials and methods From an initial group of 34 boars, 15 were selected based on the following criteria: ejaculate with ≤20% abnormal spermatozoa and at least 70% progressive motility at 120 hours of storage at 17°C. Then, based on the response to semen hypothermic storage at 5°C, boars were classified into two categories: high resistance—progressive motility of >75% in the three collections (n = 3); and low resistance—progressive motility of <75% in the three collections (n = 3). Seminal plasma proteins were analyzed in pools, and differential proteomics was performed using Multidimensional Protein Identification Technology. Results Progressive motility was lower at 120 hours of storage in low resistance, compared to high resistance boars (P < .05). Acrosome and plasma membrane integrity were not affected by the boar category, storage time, or their interaction (P ≥ .104). Sixty‐five proteins were considered for differential proteomics. Among the differentially expressed and exclusive proteins, the identification of proteins such cathepsin B, legumain, and cystatin B suggests significant changes in key enzymes (eg, metalloproteinases) involved in spermatogenesis, sperm integrity, and fertilizing potential. Discussion and Conclusion Differences in the seminal plasma suggest that proteins involved in the proteolytic activation of metalloproteinases and proteins related to immune response modulation could disrupt key cellular pathways during spermatogenesis and epididymal maturation, resulting in altered resistance to chilling injury. Further in vivo studies focusing on the immunological crosstalk between epithelial cells and gametes might explain how the immune regulators influence sperm resistance to hipothermic storage.

show abstract

Assessment of chilling injury in hypothermic stored boar spermatozoa by multicolor flow cytometry

et al. 2021

Self Cite

View full text Add to dashboard Cite

Hypothermic storage of boar semen may allow antibiotic‐free semen preservation but is limited due to chilling sensitivity of boar spermatozoa. Progress in this area requires sensitive tools to detect chilling injury. Therefore, multiparameter flow cytometry panels were evaluated to ascertain whether they are useful tools for identifying sublethal damage of sperm function at a single cell level, thus considering the high intrinsic sperm heterogeneity in a sample. The first fluorochrome panel consisted of Hoechst 33342 to identify DNA‐containing events, Yo‐Pro 1 to detect viability, merocyanine 540 to describe membrane fluidity, and PNA‐Alexa Fluor™ 647 to identify acrosomic integrity. The second fluorochrome panel consisted of SiR700‐DNA to identify DNA‐containing events, JC‐1 to characterize the mitochondrial transmembrane potential (MMP), and Calbryte 630 to assess the intracellular calcium level. Extended boar semen was stored either at 17°C (control) or 5°C (chilled). It is shown that chilling increased membrane fluidity in the viable (Yo‐Pro 1 negative) sperm population at 24 h (p < 0.05). At 144 h, the viable, acrosomic intact sperm population with low membrane fluidity was similar for both storage temperatures. Moreover, chilling reduced the main sperm population with high MMP, medium fluorescence for JC‐1 monomer and low intracellular calcium level (p < 0.05). However, after in vitro sperm capacitation, this population did not differ between the two storage temperatures. Exemplary computational data visualization in t‐distributed stochastic neighbor embedding (t‐SNE) maps and moving radar plots revealed similar subpopulations as identified by three‐dimensional stacked bar charts. In conclusion, sperm surviving an initial chilling injury withstand long‐term storage and respond in a similar manner to capacitation conditions as sperm stored conventionally at 17°C. Multicolor flow cytometry is a valuable tool for detecting chilling‐induced alterations of cell function in sperm subpopulations.

show abstract

Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

Cited by 64 publications

References 48 publications

Current and alternative trends in antibacterial agents used in mammalian semen technology

Current and alternative trends in antibacterial agents used in mammalian semen technology

Differential seminal plasma proteome signatures of boars with high and low resistance to hypothermic semen preservation at 5°C

Assessment of chilling injury in hypothermic stored boar spermatozoa by multicolor flow cytometry

Contact Info

Product

Resources

About