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Updated: May 25, 2025

Sperm Collection of Differential Quality Using Density Gradient Centrifugation
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Methods Used to Assess Bull Sperm Chromatin Integrity and Its Correlation with In Vitro Embryo Production Efficiency.

Matheus Vicente Silva1, Lays Oliveira Rocha2, Bruno Augusto Nassif Travençolo3

  • 1Faculdade de Medicina Veterinária, Universidade Federal de Uberlândia, Uberlândia 38400-902, Brazil.

Veterinary Sciences
|February 26, 2025
PubMed
Summary

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Sperm chromatin integrity analysis is vital for bull fertility. Acridine orange (AO) and transmission electron microscopy (TEM) effectively identified alterations impacting in vitro embryo production (IVEP) success.

Area of Science:

  • Reproductive biology
  • Animal science
  • Veterinary medicine

Background:

  • Sperm chromatin integrity is critical for male fertility and successful in vitro embryo production (IVEP).
  • Accurate and efficient methods are needed to assess sperm chromatin alterations in bulls.

Purpose of the Study:

  • To evaluate and compare various low-cost, efficient methods for detecting sperm chromatin integrity alterations.
  • To correlate these methods with in vitro embryo production (IVEP) outcomes in bulls.

Main Methods:

  • Semen samples from four bulls were analyzed using toluidine blue, Feulgen reaction (FR), acridine orange (AO) staining, and transmission electron microscopy (TEM).
  • Methods included visual and computational staining intensity assessments, epifluorescence and confocal microscopy, and TEM.
Keywords:
Feulgen reactionacridine orangefertilitytoluidine bluetransmission electron microscopy

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  • Samples were used for in vitro embryo production (IVEP) to assess cleavage and embryonic development rates.

    • Main Results:

    • Acridine orange (AO) and transmission electron microscopy (TEM) methods showed strong negative correlations with cleavage and embryonic development rates, respectively (r = -1.00).
    • Both AO and TEM proved effective in identifying chromatin alterations that negatively impact IVEP efficiency.
    • Toluidine blue and FR methods showed less consistent correlations with IVEP outcomes.

    Conclusions:

    • Transmission electron microscopy (TEM) and acridine orange (AO) staining are the most effective methods for assessing sperm chromatin integrity related to bull fertility and IVEP.
    • These methods provide valuable insights into factors limiting successful assisted reproductive technologies.