Sammanfattning

Single-layer centrifugation (SLC) with a low-density colloid is an efficient method for removing contaminating microorganisms from boar semen while recovering most spermatozoa from the original sample. This study tested the performance of this technique, using 50-ml tubes, by spiking commercial semen doses prepared without antibiotics with selected bacterial species followed by storage at 17 degrees C. The doses were spiked up to 102/ml CFU (colony forming units) of the bacteria Burkholderia ambifaria, Pseudomonas aeruginosa, and Staphylococcus sim-ulans. The semen was processed by SLC (15 ml of sample and 15 ml of colloid) with the colloid Porcicoll at 20% (P20) and 30% (P30), with a spiked control (CTL) and an unspiked control (CTL0), analyzing microbiology and sperm quality on days 0, 3 and 7. SLC completely removed B. ambifaria and S. simulans, considerably reducing P. aeruginosa and overall contamination (especially P30, similar to 104 CFU/ml of total contamination on day 7, median). Sperm viability was lower in P20 and P30 samples at day 0, with higher cytoplasmic ROS. Still, results were similar in all groups on day 3 and reversed on day 7, indicating a protective effect of SLC (possibly directly by removal of damaged sperm and indirectly because of lower bacterial contamination). Sperm chromatin was affected by the treatment (lower DNA fragmentation and chromatin decondensation) and storage (higher overall condensation on day 7 as per chromomycin A3 and monobromobimane staining). In conclusion, SLC with low-density colloids can remove most bacteria in a controlled contamination design while potentially improving sperm quality and long-term storage at practical temperatures.

Nyckelord

bacteria; Single-layer centrifugation; Silane-coated silica colloids; Environmental contaminants; Boar semen

Publicerad i

Research in Veterinary Science
2023, Volym: 158, sidor: 215-225
Utgivare: ELSEVIER SCI LTD

SLU författare

• Morrell, Jane

  • Institutionen för kliniska vetenskaper, Sveriges lantbruksuniversitet
    

UKÄ forskningsämne

Klinisk vetenskap


Publikationens identifierare

DOI: https://doi.org/10.1016/j.rvsc.2023.03.024

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/122272