Philosophiae Doctor - PhD / Numerous studies on mammalian spermatozoa have reported large variations in the dimensions of the main sperm structural components, namely the head, midpiece and flagellum. These variations in sperm architecture are believed to be adaptations for functioning of spermatozoa in complex environments outside the male reproductive system. The midpiece of the mammalian permatozoon contains a varied number of mitochondria, but the reason for the marked difference in the size and structure of this sperm component is not clear. This study confirmed the variations in the sperm morphometry of seven selected mammalian species and revealed unique features of the sperm midpiece and sperm mitochondria of these seven species. Evaluation of several sperm kinematic parameters revealed the unique swimming characteristics of the different spermatozoa. The importance of using standardized motility parameters was highlighted as well as the assessment of different subpopulations of spermatozoa in order to produce more reliable and comparable data. Investigating the role of sperm mitochondria in human sperm metabolism indicated that these organelles are related to sperm function in terms of sperm motility. Furthermore, it was suggested that glycolysis and mitochondrial respiration are linked processes and that both are important for the maintenance of human sperm motility. By optimizing and employing standardized experimental procedures and analysis techniques, this study was able to confirm the species specificity of almost all the sperm parameters evaluated, while also elucidating the phylogenetic relatedness of the non-human primate species. In conclusion, the present study has confirmed that the various midpiece morphometry parameters are related to the remaining sperm morphometry parameters as well as to the sperm kinematic parameters. These proposed associations between the various sperm parameters were used to explain the sperm velocity of two hypothetical and morphologically different sperm structures. Therefore, the results of the current study support the idea of co-evolution between sperm components in mammalian spermatozoa and propose that the midpiece morphometry parameters that are selected for in these spermatozoa are midpiece volume, total number of mitochondrial gyres, thickness of the mitochondrial sheath and mitochondrial height. / South Africa

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Magister Scientiae (Medical Bioscience) - MSc(MBS) / Passage of spermatozoa through the female reproductive tract is essential for the regulation of fertilization, ensuring that healthy sperm reach the oocyte. Previous studies were devoted to morphological selection of sperm cells by the cervical mucus. However, research prove that the loss of integrity of the sperm plasma membrane is associated with infertile men, irrespective of their normal semen parameters. This indicates that the sperm plasma membrane plays an important role in fertilization. Further studies indicated that sperm surface proteins assist penetration through the female reproductive tract and would therefore provide useful insight in understanding other factors associated with male infertility. The aim of this project was to determine if there are any differences between sperm surface proteins of fertile donor samples in relation to infertile patient samples using different separation techniques and different detergents. Three different sperm separation techniques were employed, including wash, swim-up (SU) and Percoll density gradient centrifugation (DGC).Parallel to this, the deoxy-ribose nucleic acid (DNA) fragmentation of these cells were analysed for comparison of the extent of DNA damage induced due to different separation techniques used. This provided evidence that the best separation technique is the DGC as it minimises the amount of DNA fragmentation caused. Four different detergents were used in the process of extracting the membrane proteins from spermatozoa, namely sodium dodecyl sulphate (SDS), saponin,cetyl-trimethyl-ammonium bromide (CTAB), and TWEEN-20. The membrane proteins were then separated on a12% SDS poly-acrylamide gel electrophoresis (PAGE), and analysed by Coomassie blue and silver staining techniques as well as densitometry. Due to the different chemical nature of the detergents that extracted different surface proteins, CTAB (cationic) and SDS (anionic) extracted the most because of its strong solubilising abilities as non-ionic detergents. Common proteins that were extracted in donor samples included; 115, 92.5, 89, 61, 55.5, 51.5, 47, 44.5, 43, 38.5, 34 and 28 kDa proteins. In patients, commonly occurring proteins included; 92.5, 74.5, 70, 60.5, 51.5, 50, 44.5, 43, 36, 29.5, and 25.5 kDa proteins. Marked differences were found between membrane proteins extracted from donor samples in comparison to patient samples. Identification of these proteins was done using the SwissProt database and a literature search. Mostly non-genomic progesterone receptors were identified; others included oestrogen receptor, a phosphotyrosyl protein, P34H, equatorial segment protein, mannose lectin receptor, human guanylylcyclase receptor, epididymal protease inhibitor receptor, PH30 and estradiol binding protein. The function of the membrane surface proteins identified in this study plays a vital role in fertilization. A few of these functions include sperm attachment and binding to the oocyte as well as penetration thereof. Others play a role in signalling events such as capacitation, hyperactivation and acrosome reaction. The absence of these proteins in patient sperm possibly accounts for the functional inability to successfully achieve fertilization suggesting that this provides molecular insight to reasons for infertility amongst men. In addition to this, proteins presented by patient samples that were absent in healthy donors may too account for their infertility status. Estradiol binding protein and PH30 are two proteins presented only in patient samples. Their function plays a role in the inhibition of the acrosome reaction and sperm-egg fusion, respectively. In conclusion, these differences in protein expression between fertile donors and patients may form the molecular basis of infertility amongst men and indicates possibilities for novel proteonomic approaches to improve andrological diagnosis in future.

For the present study 106 strandings of Kogia breviceps and 85 strandings of K. sima along the South African coastline between 1880 and 1995 were analysed in order to examine the age and growth, male and female reproduction, diet, stranding patterns, and population genetic structure of both species. Length and weight at birth were about 120cm and 53kg for K. breviceps and about 103cm and 14kg for K. sima. Von Bertalanffy growth curves were fitted to the data and indicated that physical maturity was reached at around 15 years in both sexes of K. breviceps and at 13 years in female and 15 years in male K. sima. Asymptotic length was reached at 306.0 and 286.1cm in female and male K. breviceps and 249.14 and 263.75cm in female and male K. sima, respectively. Maximum ages were16 years for male K. breviceps and 23 years for females and 17 years for male K. sima and 22 years for females. Reversed sexual size dimorphism was suggested for K. breviceps, while in K. sima males were larger than females. Attainment of sexual maturity in males occurred at between 2.5 and 5 years of age in K. breviceps and 2.6 and 3 years in K. sima, corresponding to 241-242cm and 197cm body length, respectively. The maximum combined testis weight comprised 1.04% and 2.00% for K. breviceps and K. sima, respectively, and a polygynous mating system with a roving male strategy was proposed for both species. The sperm morphology for both Kogia species was described and is characterised by 20-25 spherical mitochondria arranged in rows around the midpiece. Attainment of sexual maturity in females occurred at 5 years in both Kogia species, and at 262cm and 215cm body length in K. breviceps and K. sima, respectively. The ovulation rates were 0.9 and 0.7 per year for K. breviceps and K. sima, respectively. In K. breviceps conceptions occurred from April to September and births from March to August, while in K. sima both conceptions and births occurred from December to March. Annual reproduction and a post-partum oestrus was suggested for both Kogia species. The diet of K. breviceps comprised 50 different cephalopod species from 22 families and 17 other prey species, while K. sima fed on 32 cephalopod species from 17 families and six others. Although niche overlap indices between the two species and between groups within each species were high, some differences in diet could be determined, which allow these two sympatrically occurring species to share the same ecological niche off the coast of Southern Africa. An analysis of the stranding patterns revealed that K. sima has a closer affinity to the Agulhas current and to higher water temperatures than K. breviceps, which is supported by differences in the size of the appendages between the two species. The population genetic analysis revealed a high haplotype and nucleotide diversity for K. breviceps in the Southern hemisphere, but a lack of significant phylogeographic structure, indicating substantial gene flow among populations and inhibiting genetic differentiation of local populations, although the South African population was somewhat isolated from others in the Southern Hemisphere. In contrast the data on the phylogeographic structure of K. sima were somewhat restrictive as the majority of the samples originated from South Africa. Nevertheless, both nucleotide and haplotype diversities were markedly lower than in K. breviceps and more similar to those for other small cetacean populations, suggesting a smaller population size for K. sima than for K. breviceps. Although both Kogia species belong to the medium to larger-sized odontocetes their life histories are located near the fast end of the slow-fast continuum of life histories of marine mammals, indicating high mortality rates. The “false-gill” marking and the ability to squirt ink are thought to reflect adaptations to predator mimicry and avoidance.

Philosophiae Doctor - PhD / 2022-04-30

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Apis mellifera (honey bees) are essential pollinators and thus considered a keystone species. Reproductive success is therefore important to the survival of the species, with drone sperm quality playing a vital role in fertilization success and colony health. Drone sperm quality is determined by multiple aspects including both structural and functional parameters. Particularly, motility, a functional parameter of sperm, plays an important role in reproductive success by determining the ability of sperm to move in the reproductive tract of the queen and to participate in fertilization. However, for the South African honey bee subspecies, the Cape honey bee (Apis mellifera capensis Escholtz), data on sperm quality parameters, are lacking. / 2023

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published_or_final_version / Obstetrics and Gynaecology / Doctoral / Doctor of Philosophy

No description available.

No description available.