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Assessment of stallion semen morphology using two different staining methods, microscopic techniques, and sample sizes
2016
Łącka Katarzyna | Kondracki Stanisław | Iwanina Maria | Wysokińska Anna
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.
Show more [+] Less [-]Assessment of stallion semen morphology using two different staining methods, microscopic techniques, and sample sizes
2016
Łącka, Katarzyna | Kondracki, Stanisław | Iwanina, Maria | Wysokińska, Anna
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.
Show more [+] Less [-]Single and mixed interaction of experimental Trypanosoma brucei brucei and Trypanosoma evansi on the semen collection reaction time and spermatozoa morphology of Yankasa rams
2016
Yunusa A. Wada | Sonnie J. Oniye | Peter I. Rekwot | Oluyinka O. Okubanjo
Objective: This study was conducted to evaluate the pathological effects of experimental trypanosomosis on the semen collection reaction time and spermatozoa morphology of Yankasa rams. Materials and Methods: Twelve apparently healthy Yankasa rams aging 24-30 months and weighing 22-25 Kg were randomly selected and were distributed into four (4) groups. Groups I and II were challenged with experimental Trypanosoma brucei brucei (Federe strain) and T. evansi (Sokoto strain) respectively, while group III was challenged with both T. brucei brucei and T. evansi parasites. Group IV was left as uninfected control. Each infected ram received 2 mL of the infected blood containing 2x106 trypomastigotes via the jugular vein. The animals were examined for clinical observations, reaction time for semen collection and abnormalities in the morphology of the spermatozoa.Results: Infection of rams with trypanosomes showed scrotal edema, scrotal atrophy, loss of libido, increased semen collection reaction time, and significant increase of spermatozoa morphological abnormalities in all the infected rams. The rams especially in groups I and III were all deemed unfit for breeding by the end of the 98 days post infection, while the uninfected rams remained as healthy and had normal values of sperm morphology throughout the study period. Conclusion: Single or mixed interaction with T. brucei brucei or T. evansi is capable of causing infertility and reproductive failure in Yankasa rams. [J Adv Vet Anim Res 2016; 3(4.000): 360-367]
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