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Katarzyna Łącka, Stanisław Kondracki, Maria Iwanina and Anna Wysokińska

Abstract

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.

Open access

Krzysztof Górski, Stanisław Kondracki, Anna Wysokińska and Maria Iwanina

Abstract

Introduction: Determination of sperm concentration and morphology in ejaculate is essential in evaluating fertility. Sperm shape and dimensions may depend on their concentration in ejaculate. The aim of the study was to evaluate the dependence of Hypor boar morphological and morphometric sperm characteristics on concentration in ejaculate.

Material and Methods: The study was conducted on 120 ejaculates from 12 Hypor boars. Depending on sperm concentration, they were put into three groups: low, medium, and high. Ejaculate sperm concentration was determined with the photometric method. Slides were prepared from semen samples which were evaluated for the morphology of spermatozoa. The preparations for morphological analyses were by the eosin-gentian staining method.

Results: As the sperm concentration in the ejaculate increased, the length, perimeter, and area of the sperm heads also did. In the ejaculates with higher sperm concentration the sperm heads were rounder. The ratios of head dimensions to tail length or total length were the highest in the sperm from ejaculates with the highest concentrations. The highest percentage of morphologically abnormal sperm was noted in ejaculates with low concentrations.

Conclusion: The dimensions and shape of sperm depend on the sperm concentration in the ejaculate. In ejaculates with the highest concentrations, the sperm have larger dimensions. Sperm concentration affects the frequency of morphological anomalies in the spermatozoa.

Open access

Krzysztof Górski, Stanisław Kondracki and Anna Wysokińska

Abstract

Introduction: The aim of this study was to evaluate the dependence between ejaculate traits, sperm morphology, and ejaculate volume in Duroc boars.

Material and Methods: The analysis involved 121 ejaculates collected from 12 Duroc boars kept in three artificial insemination centres located in central Poland. Ejaculates were collected manually at one-month intervals, over a period of 10 months. At least 10 ejaculates were collected from each boar. The material was divided by ejaculate volume and each ejaculate was assigned to one of three volume groups: 160 mL and lower, 161–200 mL, and 201 mL and higher. The ejaculates were assessed to identify the basic physical traits and determine the incidence of morphological abnormalities in the spermatozoa, specifying major and minor abnormalities. Furthermore, the morphological structure indices for the spermatozoa were also calculated.

Results: In large-volume ejaculates, spermatozoa were more elongated in shape, their heads were more elongated and had the largest flagella. With an increase in the ejaculate volume, sperm concentration in the ejaculate decreased. Moreover, while the total number of sperm in the ejaculate increased, the number of insemination doses obtained from a single ejaculate were higher.

Conclusion: The volume of ejaculate has little impact on the occurrence of morphological abnormalities and the size of sperm cells. Ejaculate volume is important for the shape of the sperm cells.

Open access

Stanisław Kondracki, Anna Wysokińska, Magdalena Kania and Krzysztof Górski

Abstract

Introduction: The effect of two smear staining methods on the dimensions and shape of sperm cells in the semen of domestic pigs was evaluated. Material and Methods: The studies were carried out on 30 ejaculates collected from 15 boars, which included five Duroc boars, five Pietrain boars, and five hybrid Duroc × Pietrain boars. Each ejaculate was next sampled to make two microscopic slides, of which one was stained with eosin-nigrosin and the other with eosin-gentian dye. In total, 600 measurements of sperm cells were made. Each sperm was measured for the following morphometric parameters: head length, head width, head area, head perimeter, tail length, and the total sperm length. Results: Sperms measured on slides stained with eosin-nigrosin showed lower dimensions as compared with those stained with the eosin-gentian dye method. Sperm stained with eosin-nigrosin had shorter and narrower heads than sperm stained with eosin-gentian dye. The method of staining, therefore, affected not only the dimensions of the sperm, but also the proportions of the dimensions defining the shape of the sperm. Conclusions: The size and shape parameters in porcine sperm may take on different values depending on the method of semen staining. Sperm cells stained with eosin-nigrosin are smaller than the sperm stained with eosin-gentian dye. The sensitivity of the sperm to the type of dye used for the fixation may be associated with genetic factors.

Open access

Krzysztof Górski, Stanisław Kondracki, Karolina Strachocka and Anna Wysokińska

Abstract

Reproductive efficiency in pigs is largely dependent on the fertility of the boar. Boars used at insemination stations should produce a large amount of semen with high fertilization capacity. The sperm count influences the conception rate and the number of insemination doses produced. The aim of this study was to evaluate the morphological and morphometric characteristics of boars of the Hypor breed in relation to the total sperm count in the ejaculate. An analysis was performed of 120 ejaculates collected from 12 Hypor boars used at three insemination stations. The ejaculate sperm count was found to affect the morphometric characteristics of spermatozoa from boars. In the ejaculates with the lowest sperm count the spermatozoa heads were significantly shorter and narrower and had a smaller surface area. The sperm from ejaculates with the lowest sperm count had relatively small head dimensions in relation to the tail length and total sperm length. In the ejaculates with the most sperm cells, the proportion of spermatozoa with major defects was significantly greater than in the ejaculates from other groups. In the ejaculates with the lowest sperm count the frequency of sperm with progressive motility was significantly lower. The fewest insemination doses can be prepared from these ejaculates.

Open access

Joanna Luszczak, Michal Bartosik, Jolanta Rzymowska, Agnieszka Sochaczewska-Dolecka, Ewa Tomaszek, Joanna Wysokinska-Miszczuk and Anna Bogucka-Kocka

Abstract

According to some studies, the Entamoeba gingivalis colonizing the gingival tissue is an important agent in bringing about periodontitis. Other studies, however, deem it an opportunist that is able to survive in the medium induced by periodontal disease. The aim of this study was to investigate the prevalence of Entamoeba gingivalis infection in patients from the Department of Periodontology, and compare this population with that of healthy people, so as to analyze the relationships between infection and patient sex and age. The result of this work is that in both groups, a correlation (p = 0,19) has been noted between the occurrence of amoebae and other diseases in the oral cavity. Indeed, 81,4% of all patients with some periodontal disease showed the presence of amoeba. Among those who are not afflicted with oral diseases, the presence of amoeba was indicated in 62,5% of the total. In addition, a correlation between the person's age and the presence of protozoa (p = 0,15) was strongly marked among women (p = 0,19). In the three age groups of women in this study (40-49, 60-69, and above 80 years), we observed a 100% presence of protozoa.

Our study leads us to the conclusion that infections with Entamoeba gingivalis should be regarded as an factor that is associated with the pathological changes occurring in patients with periodontal diseases.