New Sperm Morphology Analysis in Equids: Trumorph®Vs Eosin-Nigrosin Stain.

Sabrina Gacem,Jordi Miró,Iván Yánez-Ortiz,Jaime Catalán,Carles Soler

doi:10.3390/vetsci8050079

Sabrina Gacem, Jordi Miró + Show 3 more

Open Access

https://doi.org/10.3390/vetsci8050079

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Abstract

The evaluation of the male fertility potential is based on the analysis of the basic spermatic characteristics of concentration, motility and morphology. Thus, the study of sperm morphology is a fundamental element in the seminal analysis, but its real meaning has been biased by the techniques used for its evaluation. These techniques involve dehydration phases and subsequent staining, which involves the production of artifacts. The aim of the study is to compare two methods for equid semen morphology evaluation, Trumorph® using living sperm vs. eosin-nigrosine stain. A total of 49 ejaculates from stallions and donkeys were used. After semen collection and dilution, an aliquot was placed on the slide and introduced in the Trumorph® device. Then observation was made with a 40x objective and negative phase-contrast microscope. Another aliquot was stained using eosin-nigrosine stain and viewed using 100× magnification. Well-formed sperm were observed, and different abnormalities were identified using Trumorph®. The use of eosin-nigrosin staining method and Trumorph® led to the same results and both techniques can be used for stallion and donkey sperm morphological analysis. However, considering the fact that Trumorph® uses living sperm helps prevent sperm cell alteration during sample preparation. Therefore, Trumorph® can be a good alternative to the conventional staining method, which provides a quick test on live sperm.

Highlights

The common use of spermatozoa in assisted reproduction is expanding especially for freezing or FIV, leading to a steady strengthened interest intrigued with the distinguishing sub-fertile or infertile males [1,2,3,4,5]
The study of sperm morphology is a fundamental element in the seminal analysis, but its real meaning has been biased by the techniques used for its evaluation
The use of eosin-nigrosin staining method and Trumorph® led to the same results and both techniques can be used for stallion and donkey sperm morphological analysis

Summary

Introduction

The common use of spermatozoa in assisted reproduction is expanding especially for freezing or FIV, leading to a steady strengthened interest intrigued with the distinguishing sub-fertile or infertile males [1,2,3,4,5]. Morphology data are for high predictive value for male fertility potential [6,7,8,9,10,11] and are critical for selection using the assisted reproductive techniques treatment. Different stains and herbal dyes have been used for morphology analysis, resulting in ambiguous outcomes [13,14,15,16]. In this regard, eosinnigrosin staining has remained the most commonly used technique in equids allowing detection of morphological abnormalities and determination of viability [16]. It is important to mention that the staining technique passes through three steps (staining, smearing, drying), and each one critically affecting sperm morphology analysis

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