Elsevier

Theriogenology

Volume 53, Issue 7, 15 April 2000, Pages 1489-1499
Theriogenology

Flow cytometric evaluation of antibiotic effects on viability and mitochondrial function of refrigerated spermatozoa of nile tilapia

https://doi.org/10.1016/S0093-691X(00)00291-0Get rights and content

Abstract

Improved techniques for storage and evaluation of fish sperm would enhance breeding programs around the world. The goal of this study was to test the effect of antibiotics on refrigerated sperm from Nile tilapia (Oreochromis niloticus) by use of flow cytometry with 2 dual-staining protocols for objective assessment of sperm quality. Concentrations of 1 × 109. sperm/mL were suspended in Ringer's buffer at 318 mOsmol/kg (pH 8.0). The fluorescent stains Sybr 14 (10 μM), propidium iodide (2.4 mM), and rhodamine 123 (0.13 μM) were used to assess cell viability and mitochondrial function. Three concentrations of ampicillin, gentamicin, and an antibiotic/antimycotic solution were added to fresh spermatozoa. Motility estimates and flow cytometry measurements were made daily during 7 d of refrigerated storage (4 °C). The highest concentrations of gentamicin and antibiotic/antimycotic and all 3 concentrations of ampicillin significantly reduced sperm viability. The highest of each of the 3 antibiotic concentrations significantly reduced mitochondrial function. This study demonstrates that objective sperm quality assessments can be made using flow cytometry and that addition of antibiotics at appropriate concentrations can lengthen refrigerated storage time for tilapia spermatozoa. With minor modifications, these protocols can be adapted for use with sperm from other species and with other tissue types.

References (24)

  • EvensonDP et al.

    Simultaneous measurement by flow cytometry of sperm cell viability and mitochondrial membrane potential related to cell motility

    J Histochem Cytochem

    (1982)
  • GarnerDL et al.

    Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analysis

    Biol Reprod

    (1986)
  • Cited by (0)

    View full text