Technique comparison to assess sperm DNA fragmentation in hair ram

• Soza-Mata, María F. ^[1] ; Domínguez-Rebolledo, Álvaro E. ^[3] ; Baeza-Rodríguez, Juan J. ^[3] ; Loeza-Concha, Henry ^[2] ; Ramón-Ugalde, Julio P. ^[1]
  1. [1] Tecnológico Nacional de México

     Tecnológico Nacional de México

     México

     
  2. [2] Colegio de Postgraduados

     Colegio de Postgraduados

     México

     
  3. [3] Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias

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• Localización: Agro Productividad, ISSN-e 2594-0252, Vol. 17, Nº. 9, 2024 (Ejemplar dedicado a: Septiembre), págs. 91-100
• Idioma: inglés
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  • Objective: To evaluate 5 DNA fragmentation techniques in thawed sperm samples from hair ram subjected to fragmentation with H2O2.Design/methodology/approach: Samples were 36 straws from 4 Blackbelly rams that were thawed, mixed (pool), diluted in PBS to a concentration of  30X106 sperm/mL and divided into three treatments: T0: sample without oxidant (considered as 0% damaged DNA), T100: sample incubated with 300 M H2O2 for 24 hours (induction of DNA fragmentation (100%)). Subsequently, half of T0 and T100 were mixed to obtain a proportion of 50% sperm with fragmented DNA (T50). The samples were analyzed with different techniques: Aniline Blue (AB), Toluidine Blue (TB), Acridine Orange (AO), Chromomycin A3 (CMA3) and Sperm Chromatin Dispersion (SCD).Results: In the linear regression, all the techniques presented a significance level of less than 5%, as well as a significant correlation (r=0.962, P<0.01). However, between treatments, it was observed that the AO technique (34.82 31 ± 3.00%) at T50 and the AB technique (9.55 ± 1.45% at T100 were the least sensitive in detecting DNA damage compared to the other techniques.Limitations on study/implications: New techniques are increasingly. Findings/conclusions: The techniques that best evaluate the DNAFI of sperm in hair ram are CMA3, SCD and TB.