Thousands of sexual assault cases in the United States are backlogged. This has been a growing issue for years that has increased the difficulty of solving these cases and providing closure to the victims. The analysis process for each case includes the identification of body fluids, presumptive testing, confirmatory testing, and DNA extraction. The only confirmatory method for semen identification is a microscopic visualization of sperm cells. The time spent on microscopic analysis varies depending on the complexity of the samples and the skills of the analyst. While the identification of sperm cells is informative, it can be very time-consuming and labor intensive. Some forensic laboratories choose to skip this step and submit samples directly for DNA analysis. Conducting DNA analysis on unscreened samples can increase the cost of testing when negative samples are analyzed as well as the time it takes to process each case. Automated microscopy has been available for decades and more recently has been paired with artificial intelligence to detect sperm cells on microscope slides. In this research, the MetaSystems automated microscope was used to analyze slides that mimic forensic sexual assault samples. Slides were also examined using traditional microscopy. The automated system quickly provided an accurate quantification of the number of sperm cells present in a sample, which can inform downstream DNA testing. The software was successful in identifying sperm cells treated with Christmas tree and hematoxylin and eosin stains, even among epithelial cells and various contaminants. Results demonstrated that an artificial intelligence-driven forensic sperm cell detection microscope can significantly reduce the time it takes to locate and identify sperm cells and estimate sperm cell quantity compared to a lengthier and more tedious manual search. Drawbacks to the system include the relatively high cost and reduced ability to accurately detect sperm cells amid contaminants that are of similar morphology.

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This study aimed at complementing studies of sperm whale social and vocal behaviour that were restricted to the Pacific Ocean. The characteristic multi-pulsed structure of sperm whale clicks allows for estimation of whales' size from measurements of the inter-pulse intervals (IPI). I have developed two new automatic methods for IPI estimation from clicks recorded during foraging dives. When compared to other previously developed methods, the newly developed method that averages several clicks' autocorrelation function showed the best performance amongst the automatic methods. Previous studies did not support individual identity advertisement among social unit members as the function for the sperm whale communication signals called codas. I tested within coda type variation for individual specific patterns and found that, while some coda types do not allow for individual discrimination, one did so. This variation suggests that different coda types may have distinct functions. Analysis of social structure in the Azores found that, similar to the Eastern Tropical Pacific, sperm whales form long term social units of about 12 individuals. Unlike the Pacific Ocean, Azorean social units do not form temporary groups with other units, suggesting differences in the costs and benefits of group formation. I argue that these are due to differences in terms of predation pressure and intraspecific competition between the Azores and the Pacific study sites. The variation of coda repertoires in the Atlantic also showed a pattern dissimilar to that previously documented in the Eastern Tropical Pacific. In the North Atlantic, coda repertoire variation is mostly geographic, which is parsimoniously explained by random drift of culturally transmitted coda repertoires. No sympatric vocal clans with distinct dialects were found as has been noted in the Pacific. Drawing upon the differences found in social structure I argue that selection for maximization of differences between units with similar foraging strategies may have led to the Pacific vocal clans. The differences between oceans suggest that sperm whales may adaptively adjust their behaviour according to experienced ecological conditions.

Sperm phenotype is an essential indicator of the male ejaculate quality and may have a significant impact on male reproductive success. Sperm phenotypes are considerably variable across species but variation is also found among males within species. This thesis examines (1) variation in sperm phenotypes among males in barn swallows (Hirundo rustica), (2) changes in male ejaculate quality with age, (3) relationships between sperm morphology and motility, (4) effects of sperm phenotypes (morphology and motility) on male fertilization success, using a large dataset of 174 observation for 130 males, and (5) the influence of female reproductive environment on sperm motility. From the tested variables, only midpiece length correlated with male age. Older males had shorter midpiece but no relationship between male age and reproductive success was found. Sperm length negatively affected sperm motility and, simultaneosly, relative midpiece length posively correlated with sperm motility. No correlation was found between the male reproductive success and sperm motility, presence of abnormalities, length of outermost tail feathers or age. Males with shorter relative midpiece were more successful in within-pair paternity, and males with shorter sperms but longer relative midpiece were more successful in...

Sexual selection plays an important role in the evolution of animals. Today we already know that it takes place not only before copulation (precopulatory sexual selection), but also after copulation. This type of sexual selection is called postcopulatory sexual selection, and occurs mainly in promiscuous species, where females mate with multiple males. In this case, sperm competition occurs in the female reproductive tract. To increase the likelihood of their reproductive success, males began to develop surprisingly diverse sperm adaptations at the morphological, physiological, or behavioural levels. These adaptations often affect sperm velocity (motility), which is a key factor for successful egg fertilization. However, the result of reproductive success can also be influenced by females, who may prefer sperm with a certain phenotype in the process of cryptic female choice, and thus, for example, obtain better genes for offspring. In some species, females even can have the ability to sort and store sperm in specialized organs in which the sperms are nourished for some time, and then used to fertilize the egg. Even in this case, the storage of sperm is often affected by sperm morphology. Due to these mechanisms of postcopulatory sexual selection, sperm are under strong selection pressure, which can...

BACKGROUND:The SPE-8 group gene products transduce the signal for spermatid activation initiated by extracellular zinc in C. elegans. Mutations in the spe-8 group genes result in hermaphrodite-derived spermatids that cannot activate to crawling spermatozoa, although spermatids from mutant males activate through a pathway induced by extracellular TRY-5 protease present in male seminal fluid.RESULTS:Here, we identify SPE-8 as a member of a large family of sperm-expressed non-receptor-like protein-tyrosine kinases. A rescuing SPE-8::GFP translational fusion reporter localizes to the plasma membrane in all spermatogenic cells from the primary spermatocyte stage through spermatids. Once spermatids become activated to spermatozoa, the reporter moves from the plasma membrane to the cytoplasm. Mutations in the spe-8 group genes spe-12, spe-19, and spe-27 disrupt localization of the reporter to the plasma membrane, while localization appears near normal in a spe-29 mutant background.CONCLUSIONS:These results suggest that the SPE-8 group proteins form a functional complex localized at the plasma membrane, and that SPE-8 is correctly positioned only when all members of the SPE-8 group are present, with the possible exception of SPE-29. Further, SPE-8 is released from the membrane when the activation signal is transduced into the spermatid.

Thesis (MMed)--Stellenbosch University, 2105. / ENGLISH ABSTRACT: Intracytoplasmic sperm injection (ICSI), as well as other micromanipulation assisted reproductive technology methods, such as physiologic ICSI (PICSI) and intracytoplasmic morphologically selected sperm injection (IMSI), are routinely used in many fertility laboratories around the world. An integral part of these methods is the manipulation of spermatozoa in preparation of the injection into the oocyte. It is common practice to place prepared spermatozoa in a viscous holding medium to facilitate the handling, manipulation and slowdown of spermatozoon movement during the immobilization and injection processes of ICSI. The possible effect of these holding mediums on basic semen parameters, as well as the sperm deoxyribonucleic acid (DNA) and structural integrity of spermatozoa, is of importance. Hamilton Thorne IVOS® developed an automated software solution for live sperm morphology evaluation under high magnification, called IMSI StrictTM. It combines Tygerberg Strict Criteria morphological classification of human spermatozoa with motile sperm organelle morphology examination (MSOME) and provides software-based categorization. The IMSI StrictTM software was developed to aid in the IMSI spermatozoon selection process that enables objective classification of spermatozoa to remove inter-technician variation. For good optics and spermatozoon evaluation in IMSI StrictTM, spermatozoa need to be moving very slowly or be immotile, but still viable. This can be achieved by placing spermatozoa in a viscous holding medium, either polyvinylpyrrolidone (PVP) or SpermSlowTM, sometimes for a substantial time period. Before marketing the clinical use of IMSI StrictTM, the possible toxicity or deleterious effect of PVP and SpermSlowTM on spermatozoa needs to be excluded. The primary objective of this study was to evaluate the effect of PVP and SpermSlowTM on human spermatozoa after different exposure times using a viability stain, CASA motility and kinetic parameters, chromatin packaging analysis (CMA3 staining analysis) and DNA fragmentation analysis (TUNEL analysis). The secondary objective was to evaluate the effect of PVP and SpermSlowTM on human spermatozoa‟s ultrastructure with Transmission Electron Microscopy. This prospective analytical study was conducted at Drs Aevitas Fertility Clinic (Vincent Pallotti Hospital, Cape Town, South Africa) as well as the Fertility Unit at Tygerberg Hospital (Cape Town, South Africa) between July 2013 and October 2014. A total of 90 separate (no duplication) semen samples were analysed for the quantitative analysis (primary objective) and 1 sample for the descriptive analysis (secondary objective). Results showed that although PVP and SpermSlowTM treated sperm outcomes often differed significantly after typical statistical analysis, clinically these two mediums were shown to be equivalent (using a specific statistical test for equivalence) for the tested outcomes. PVP and SpermSlowTM had no detrimental effect clinically on sperm viability, motility parameters, chromatin packaging and DNA fragmentation rate. The secondary investigation indicated that SpermSlowTM might exert a disintegrating effect on various sperm membranes, and as a secondary consequence of the eventual necrotic process, alteration of chromatin and cytoskeletal components. PVP medium on the other hand did not show these disintegrating effects. This finding needs to be further investigated since only one semen sample was evaluated. Based on this study‟s results, either PVP or SpermSlowTM can be used for IMSI StrictTM purposes. However, the study did not include the technical aspects of the usage of PVP and SpermSlowTM. / AFRIKAANSE OPSOMMING: Intrasitoplasmiese sperm inspuiting (ICSI), sowel as ander mikro-manipulasie voortplantings tegnieke, soos fisiologiese ICSI (PICSI) en intrasitoplasmiese morfologies geselekteerde sperm inspuiting (IMSI), word in baie fertiliteitsklinieke regoor die wêreld gebruik. 'n Integrale deel van hierdie metodes is die manipulasie van spermatosoa ter voorbereiding van die inspuitproses. Dit is algemeen om voorbereide spermatosoa in 'n viskose medium te plaas om die hantering, manipulasie en vertraging van spermatosoön beweging tydens die immobilisasie en inspuitproses van ICSI te fasiliteer. Die effek van hierdie mediums op basiese semenparameters, sowel as die sperm deoksiribonukleïensuur (DNS) en strukturele integriteit van spermatosoa, is van belang. Hamilton Thorne IVOS® het 'n sagteware oplossing, IMSI StrictTM, vir lewende sperm morfologie evaluering onder hoë vergroting ontwikkel. Hierdie sagteware bied sagteware-gebaseerde morfologiese klassifikasie deur die Tygerberg streng kriteria morfologiese klassifikasie met beweeglike spermorganel morfologie ondersoek (MSOME) te kombineer. Die IMSI StrictTM sagteware is ontwikkel om die objektiewe klassifikasie van spermatosoa vir IMSI spermatosoön seleksie moontlik te maak. Spermatosoa moet baie stadig beweeg of immotiel, maar steeds lewensvatbaar wees om goeie optika en spermatosoön evaluering vir IMSI StrictTM te verseker. Dit sal bereik kan word deur spermatosoa in 'n viskose medium, hetsy PVP (“polyvinylpyrrolidone”) of SpermSlowTM, vir 'n aansienlike tydperk te inkubeer. Voordat IMSI StrictTM vir kliniese gebruik bemark kan word moet die moontlike toksisiteit of nadelige effek van PVP en SpermSlowTM op spermatosoa uitgesluit word. Die primêre doel van hierdie studie was om die effek van PVP en SpermSlowTM op menslike spermatosoa na verskillende inkubasie tye te evalueer deur ʼn lewensvatbaarheid kleuring toets, twee sperm DNS toetse (CMA3 en TUNEL) en rekenaar geëvalueerde sperm beweeglikheid toetse te gebruik. Die sekondêre doel was om die effek van PVP en SpermSlowTM op menslike spermatosoa se ultrastruktuur deur middel van Transmissie Elektronmikroskopie te evalueer. Hierdie studie is by Drs Aevitas Fertiliteitskliniek (Vincent Pallotti Hospitaal, Kaapstad, Suid-Afrika) sowel as die Fertiliteitseenheid by Tygerberg Hospitaal (Kaapstad, Suid-Afrika) tussen Julie 2013 en Oktober 2014 uitgevoer. 'n Totaal van 90 semenmonsters vir die kwantitatiewe analise (primêre doel) en een vir die beskrywende analise (sekondêre doel) is ontleed. Resultate het getoon dat alhoewel PVP en SpermSlowTM geïnkubeerde spermuitkomste dikwels na ʼn tipiese statistiese analise betekenisvol verskil, hierdie twee mediums vir die geëvalueerde uitkomste klinies ekwivalent (bepaal deur middel van spesifieke statistiese toetse vir ekwivalensie) is. Die mediums het ook nie klinies 'n nadelige effek op sperm lewensvatbaarheid, beweeglikheid parameters, chromatien verpakking en DNS fragmentasie koers getoon nie. Die sekondêre ondersoek het getoon dat SpermSlowTM hoofsaaklik 'n effek van disintegrasie op verskeie spermmembrane getoon het. Hierdie nekrotiese proses kan lei tot verandering van chromatien en sitoskelet komponente. PVP medium het egter nie dieselfde disintegrerende effek getoon nie. Hierdie bevinding moet egter verder ondersoek word, aangesien slegs een semenmonster geëvalueer is. Alhoewel hierdie studie nie die tegniese aspekte van die gebruik van PVP en SpermSlowTM geëvalueer het nie, kan aanbeveel word dat óf PVP óf SpermSlowTM op grond van geëvalueerde uitkomste tydens die IMSI StrictTM sperm seleksie proses gebruik word.

<p>The increasing number of patients undergoing treatment with assisted reproductive techniques (ART) during the past years have led to the need of developing different methods for separation of spermatozoa that can be used for different fertilisation procedures and for freezing. Cryopreservation of spermatozoa includes preparation, freezing, storage and thawing.</p><p>In this study two different cryomedia (Cryo Protec I and Cryo Protec II) for human spermatozoa were compared. The main outcome was spermsurvival rate for spermatozoa after freezing. Sperm viability was assessed using the Hypo-osmotic swelling test which is based on osmolality.</p><p>A total of 86 samples of semen were used in this study (Cryo Protec I=38, Cryo Protec II=48). The survival rate between the two cryomedia did not differ much but Cryo Protectant I showed a small increase in survival for the spermatozoa after freezing. The Hypo-osmotic swelling test also showed similar values of viable spermatozoa for the two cryomedia both before and after freezing.</p>

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Over the last century, sexual selection has grown from a controversial theory into a vast field of theoretical and empirical research. Although Darwin outlined two major mechanisms within his theory, male-male competition and female mate choice, the latter has promoted a wealth of research by virtue of its complexity. Despite decades of research into how female preferences and sexually selected traits have evolved, there is still little consensus as to why females prefer the males they do. Preferences are thought to evolve from either direct selection on the preference, as females themselves benefit directly from mating with a preferred male, or through indirect selection on the preference via offspring fitness. In all cases however, female preferences should compensate for the costs of discriminating between potential mates, if they are to remain overall beneficial. The fitness benefits of mating with preferred males were investigated here using the fruitfly Drosophila simulans, employing a range of behavioural, phenotypic and quantitative genetic approaches. The findings presented here indicate that female Drosophila simulans do not gain directly from mating with a preferred male. Multiple mating can increase fecundity, although costs from male harassment can reduce the net benefit. They also indicate that females may benefit indirectly from mating with attractive males as attractiveness is heritable and sons of preferred males are themselves preferred. There is also evidence that attractive males are successful in both the pre- and post-copulatory sense, as preferred males are better sperm competitors than less-preferred males. However, although there appear to be benefits from preferred males via their sons, there appear to be no benefits from males via their daughters’ fitness. These findings collectively indicate that female preferences in Drosophila simulans are driven by indirectly selected benefits (via Fisherian sons), and that females benefit directly from mating multiply.