Effect of method and clinician on stallion sperm morphology evaluation

Brito, Leonardo F.C.; Greene, Lauren M.; Kelleman, Audrey A.; Knobbe, Marc; Turner, Regina M.

doi:10.1016/j.theriogenology.2011.04.007

Cited by 56 publications

(56 citation statements)

References 16 publications

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“…Sperm morphology was examined by light microscopy (Eclipse E600; Nikon) of smears stained with eosin and nigrosine (Brito et al 2011). At least 200 sperm per slide were counted to determine the percentage of morphologically normal sperm.…”

Section: Methodsmentioning

confidence: 99%

Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions - A pilot study

Šichtař¹,

Nehasilová²,

Šimoník³

et al. 2017

CZECH J ANIM SCI

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ABSTRACTŠichtař J., Nehasilová A., Šimoník O., Bubeníčková F. (2017): Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions -A pilot study. Czech J. Anim. Sci., 62, 227−233.The aim of the study was to evaluate the effect of two different extenders on sperm characteristic before equilibration and post-thaw in the endangered Old Kladruber stallions. Also, the response of individual stallions to the extenders used was tested. Semen was collected from six stallions every other day within one week. After centrifugation of the collected sperm-rich fraction, the supernatant was removed and sperm pellets were divided to two aliquots; these were diluted either with Gent (Minitube, Germany) or privately manufactured lactose-EDTA-egg yolk extender (Lact). Three cryopreserved insemination doses (IDs) from each extender (Gent and Lact) were prepared for each stallion from one collection (108 samples from six stallions in total). As a parameter of quality, the motility (computer assisted sperm analysis), viability (fluorescence staining), and morphology (eosin/nigrosine staining) were evaluated after dilution with freezing extenders (fresh) and after thawing (frozen-thawed). The different effects of chosen extenders on the quality of fresh semen were only manifested in higher kinematic parameters of sperm when the Lact extender was used. However, in frozen-thawed samples, the Gent extender yielded significantly better results in all of the evaluated parameters. The representation of sperm subpopulation was significantly influenced by extender in fresh as well as frozen-thawed samples; moreover, we found a significant effect of freezing on the distribution of these subpopulations. The response of individual stallions to chosen extenders was evident in the different quality of fresh as well as frozen-thawed IDs; Gent extender yielded better frozen-thawed IDs. Based on our results, among others describing quality parameters of ejaculate in endangered Old Kladruber stallions, we can recommend using Gent extender for the production of frozen-thawed IDs.

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Section: Methodsmentioning

confidence: 99%

Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions - A pilot study

Šichtař¹,

Nehasilová²,

Šimoník³

et al. 2017

CZECH J ANIM SCI

View full text Add to dashboard Cite

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“…Many authors using different staining methods describe various morphologic abnormalities of sperm in bulls (Revay et al 2009, Freneau et al 2010, Mandal et al 2010, Enciso et al 2011, stallions (Kavak et al 2004, Morrell et al 2008, Einarsson et al 2009, Brito et al 2011, Love 2011, rams (Lambrechts et al 2000, Janett et al 2001, Sweeney et al 2007, Lopez Armengol et al 2012) and dogs (Kawakami et al 2005, Nxñ ez-Martinez et al 2005, Peñ a et al 2007, Lange-Consiglio et al 2010). But we did not find concrete references what is the morphologically normal sperm cell in domestic animals.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of morphological criteria of sperm quality before in vitro fertilization and intracytoplasmic sperm injection

Lasienë

Gedrimas

Vitkus

et al. 2013

Polish Journal of Veterinary Sciences

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The quality of sperm has a direct influence on the fertilization and developmental competence of embryos. In the literature we did not find defined criteria for evaluation of normal sperm parameters in various species of domestic mammals. Therefore we attempted to review evaluation of criteria of morphologically normal human sperm and their abnormalities. All sperm cells observed in the stained sample are classified as normal or abnormal. Any abnormalities in morphology of sperm have a negative effect on the outcome in in vitro fertilization and intracytoplasmic sperm injection. Abnormal sperm are categorized into subgroups according to the observed defects (concerning the head and/or midpiece and/or tail). Most morphologically abnormal sperm have multiple defects. This article can be considered as guideline for the manual of sperm quality evaluation in different species of domestic mammals.

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“…Brito et al (6) showed that eosin-nigrosin staining method can give similar results in stallion sperm morphology analyses to those obtained with Papanicolaou staining. Similar observations are also reported by other researchers (26).…”

Section: Discussionmentioning

confidence: 82%

Assessment of stallion semen morphology using two different staining methods, microscopic techniques, and sample sizes

Lacka

Kondracki

Iwanina

et al. 2016

Journal of Veterinary Research

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Introduction: The aim of this study was to propose the optimal methodology for stallion semen morphology analysis while taking into consideration the staining method, the microscopic techniques, and the workload generated by a number of samples. Material and Methods:Ejaculates from eight pure-bred Arabian horses were tested microscopically for the incidence of morphological defects in the spermatozoa. Two different staining methods (eosin-nigrosin and eosin-gentian dye), two different techniques of microscopic analysis (1000× and 400× magnifications), and two sample sizes (200 and 500 spermatozoa) were used. Results: Well-formed spermatozoa and those with major and minor defects according to Blom's classification were identified. The applied staining methods gave similar results and could be used in stallion sperm morphology analysis. However, the eosin-nigrosin method was more recommendable, because it allowed to limit the number of visible artefacts without hindering the identification of protoplasm drops and enables the differentiation of living and dead spermatozoa. Conclusion: The applied microscopic techniques proved to be equally efficacious. Therefore, it is practically possible to opt for the simpler and faster 400x technique of analysing sperm morphology to examine stallion semen. We also found that the number of spermatozoa clearly affects the results of sperm morphology evaluation. Reducing the number of spermatozoa from 500 to 200 causes a decrease in the percentage of spermatozoa identified as normal and an increase in the percentage of spermatozoa determined as morphologically defective.

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Effect of method and clinician on stallion sperm morphology evaluation

Cited by 56 publications

References 16 publications

Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions - A pilot study

Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions - A pilot study

Evaluation of morphological criteria of sperm quality before in vitro fertilization and intracytoplasmic sperm injection

Assessment of stallion semen morphology using two different staining methods, microscopic techniques, and sample sizes

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