A Blue-tongued skink ( Tiliqua gigas) is a rare exotic species which is very popular between breeders. Nevertheless, the social structure of this charismatic skink remains almost unknown. Because of its high aggressive- ness and difficulties connected with study in its natural habitat, experiments based on the odor discrimination looks like a good alternative for its bet- ter understanding. Our results revealed that the reaction on the odor is generally poor and is very individual. We observed the highest response of interest on odors produced throught dorsal parts of skin. The reaction on conspecific's odor did not lead to escape behavior. Behavioral analysis was complemented by morphological analysis of heads in T. gigas and T.scincoides. These species are divided into separate subspecies, which are formed by geographically distinct populations. We wondered whether it is possible to separate these populations by geometric morphometrics and whether this separation is consistent with phylogenetic studies. We found out that if the populations are united in bigger groups based on their congeniality they seem to be distinct according to the head shape. Our results also revealed the monomorphy in head shape. Unfortu- nately, it is very difficult to distinguish between sexes due to the absence of sexual dimorphism....

Thesis(M. A. Agricultural Management (Animal Production)) -- University of Limpopo, 2019 / The study was conducted to assess phenotypic and genotypic diversity in resistance to gastro-intestinal nematode (GIN) within communal goats in different agro-ecological zones of Limpopo province. Eighty goats were randomly sampled from Mopani = 20, Blouburg = 20, Vhembe = 20 and Capricorn = 20, representing four agro-ecological zones of Limpopo province. Frequencies and percentages of occurrence of qualitative traits and least square means (LSM) of quantitative traits were computed. Goats were characterized as multi-coloured (67.7%), black dominant coat colour (40%), flat face profile (73.8%), erect ears (68.75%), slopping rump (47.5%), no toggles (83.8%), horned (86.3%), while 71.0% and 59.4% were straight shaped and orientated backward respectively, in all agro-ecological zones. Faecal and blood samples were collected to assess the prevalence of GIN and genetic diversity of goats. The highest prevalence and abundance GIN parasites were Haemonchus contortus (nematode = 357.42) followed by Fasciola hapatica (trematode = 163) and Moniezia (cestodes = 121.50). The patterns of GIN prevalence varied (P < 0.05) across agro-ecological zones and seasons. Prevalence of Moniezia nematode varied (P < 0.05) amongst goat sexes, whilst prevalence of other GIN’s did not vary (P>0.05). For genetic variation in GIN resistance, goats were genotyped at 15 microsatellite markers recommended by the International Society of Animal Genetics. Expected heterozygosity (HE) ranged from 0.69 in arid zone goat population to 0.76 in sub-humid zone goat population, while the observed heterozygosity (HO) ranged from 0.53 in arid zone goat population to 0.60 in sub-humid zone goat population. Mean number of alleles (MNA) per population ranged from 6.13 to 7.40. Gene differentiation (FST) among populations was low (3.6%). The results revealed that genetic relationships between populations do not reflect their geographical proximity as revealed by the Nei’s genetic distance results. Low heterozygosity resulted in reducing some fitness traits for GIN parasite resistance. Inbreed goats showed low internal parasite resistance across all agro-ecological zones.

<p>Down syndrome (DS) is a genetic condition caused by the triplication of human chromosome  21 and presents with many phenotypes including decreased brain size, hypocellularity in the brain,  and assorted ocular phenotypes. Some of the ocular phenotypes seen are increased risk of cataracts,  accommodation difficulties, increased risk of refractive errors, and increased retinal thickness. The  Ts65Dn mouse model of DS is a classically used mouse model as it presents a number of  phenotypes also seen in those with DS. Some of these phenotypes include decreased brain volume,  abnormal synaptic plasticity, and ocular phenotypes. These ocular phenotypes include decreased  visual acuity, cataracts, and increased retinal thickness. The Ts65Dn mouse model is trisomic for <em>Dyrk1a</em>, a gene of interest in DS research. We hypothesize that there will be a genotypic and sex effect of retinal thickness and retinal ganglion cell (RGC) count at postnatal day 15 in the Ts65Dn  mouse model of DS. Retinal slices were taken from male and female trisomic and euploid Ts65Dn  mice at P15 and fluorescently labeled for RGCs and bipolar cells via immunohistochemistry. The  retinas were measured for total retinal thickness and RNA-binding protein (RBPMS) positive cells in the RGC layer were counted. There was no genotypic or sex effect when comparing retinal  thickness in trisomic mice as compared to euploid mice. There was a genotypic effect of RBPMS  positive cell count in which the trisomic mice had a higher number of RBPMS positive cells than  euploid mice. Increased retinal thickness along with increased RGC number have both been  implicated with decreased apoptosis in the retina. In the Ts65Dn mouse model along with in  individuals with DS, this could be due to an increase in DYRK1A protein levels reducing apoptosis.  In future studies, determining DYRK1A’s influence in retinal thickness and RGC number could  result in a treatment for overactive <em>DYRK1A</em> that could normalize retinal thickness and RGC  number in those with DS.</p>

Aside from few model organisms, little is known about early embryonic development or sex determination in insects, in particular mosquitoes which are major vectors of worldwide disease. The goals of this work were to investigate a mosquito-specific transcription factor and its intronic miRNA cluster and characterize a novel Y chromosome gene in An. stephensi. The aims of these experiments were to expand on the knowledge of genes involved in embryonic development and sex determination with potential application in vector control strategies. In Ae. aegypti a mosquito-specific bZIP1 transcription factor was demonstrated to be conserved among divergent mosquito species. It was maternally and zygotically-expressed and knock-down of bZIP1 mRNA via siRNA microinjection in the embryo resulted in embryonic death. The expression profile of this gene was determined through the use of RT-PCR and qRT-PCR. Additionally, this gene contains a miRNA cluster that is also relatively conserved amongst members of the Culicidae family suggesting its evolutionary importance. The miRNAs are also maternally and zygotically expressed and are the most abundant embryonic miRNAs as determined by small RNA sequencing and Northern analysis. Promoter activity for bZIP1 was characterized and the promoter was used to direct maternal and zygotic transgene expression in An. stephensi. Y chromosome genes were successfully identified in An. stephensi from Illumina sequencing data. This work focused on a gene unique to the Y 1 (GUY1). It was shown that GUY1 was male specific and linked to the Y chromosome. RT-PCR and single embryo analysis suggested that GUY1 was expressed during the maternal to zygotic transition and was only expressed in male embryos. It was shown in multiple transient and transgenic assays that the ectopic expression of GUY1 can influence the sex of subjected individuals and skew sex distribution to a male bias. There is still much to be investigated before a GUY1-based transgenic line can be tested and implemented for use in vector population control. However, the work in this dissertation represents a major step towards novel mosquito control strategies based on the manipulation of Y chromosome genes. / Ph. D.

Sex determination is a key developmental process in several species regulated by sexrelated transcription factors. In many species a gene called doublesex and mab-3 related transcription factor 1 (dmrt1), plays an important role in sexual differentiation. I used African clawed frogs (Xenopus) to examine function of dmrt1 in two species: a diploid species, X. tropicalis, and an allotetraploid species, X. laevis. In both species, dmrt1 is an autosomal gene; Xenopus tropicalis has one copy of dmrt1 and X. laevis has two homeologous copies that by definition are derived from whole genome duplication: dmrt1.L and dmrt1.S in X. laevis. We generated knockouts of each of these genes to further examine their function in sexual differentiation. Histological examination showed testicular dysgenesis in X. tropicalis dmrt1 and X. laevis dmrt1.L null males whereas dmrt1.S null males presented no obvious difference in sperm density compared to wildtype males. X. tropicalis dmrt1 and X. laevis dmrt1.L null females were found to completely lack reproductive organs and are infertile whereas dmrt1.S null females appeared unaffected. The contrasting results between dmrt1.L and dmrt1.S in X. laevis provides evidence of both neofunctionalization and subfunctionalization following gene duplication and suggest that gene duplication is a major contributor to evolutionary change. Additional investigation of the transcriptome of these frogs and the role of dmrt1 in the secondary sex characteristic vocalization provides further evidence of the role of dmrt1 in development. Comprehensively, this investigation provides further knowledge of the role of dmrt1 and homeologs of this gene in sexual differentiation and introduces a novel aspect of this gene in female development. Future efforts are focused on generating double knockouts for dmrt1.L and dmrt1.S, further examining the role of dmrt1.S in somatic cell function and developing additional mutant lines in other Xenopus for comparative analysis. / Thesis / Master of Science (MSc) / In many species sexual differentiation is a crucial developmental event. Surprisingly, however, the systems orchestrating sexual differentiation are highly variable among species. The doublesex and mab-3 related transcription factor 1 (dmrt1) gene plays a role in sexual differentiation in many groups, but its specific roles in this process are incompletely characterized and potentially diverse. We used genetic engineering in two species of African clawed frog (Xenopus) to disable function of dmrt1 in order to explore effects on gonadal development and the development of secondary sex characteristics. We found that dmrt1 is required for normal ovary or testis development in both Xenopus species, and that functional divergence occurred following duplication of dmrt1 by whole genome duplication. Taken together, these findings identify previously uncharacterized roles of dmrt1 in Xenopus and provide evidence of dynamic functional evolution of this important gene.

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<p>Sex chromosomes have evolved independently multiple times in plants and animals. Sex chromosome evolution theory predicts the gradual degeneration of the sex-specific sex chromosome due to suppression of recombination, which lowers the efficacy of natural selection. Suppressed recombination also acts to resolve sexual conflict and ensure proper segregation of sex-specific alleles. However, sex chromosome degeneration is not always the case, and evolutionarily old, and young, but homomorphic (nondegenerate) sex chromosomes have been observed. African clawed frogs (<em>Xenopus</em> and <em>Silurana</em>) have homomorphic sex chromosomes due to a recent turnover event. However, occasional recombination between the sex chromosomes may contribute to the maintenance of homomorphic sex chromosomes in African clawed frogs. Mechanisms that prevent divergence of sex chromosomes may be related to polyploidization, which is frequently observed in African clawed frogs. The studies herein construct a phylogenetic framework to test alternative hypotheses for selection on sex-linked and autosomal genes involved in sex determination, map sex chromosomes and compare sex chromosomes across African clawed frogs. I have also explored the relationship between phenomena like recent turnover events, recombination and polyploidization to sex chromosome degeneration (or lack thereof). In this dissertation, I have discussed the potential for multiple mechanisms of sex determination and the unique pseudoautosomal nature of sex chromosomes within this group of frogs. This body of work provides a comprehensive study of sex chromosomes in a group lacking phylogenetic resolution and sheds light on the origin and evolution of sex chromosomes in other organisms.</p> / Doctor of Philosophy (PhD)

In aquatic turtles, females select nest sites that have a high degree of solar exposure, and exploit recently tilled agricultural fields for nesting, presumably because of increased solar exposure and/or easier nest excavation, and the importance of incubation temperature on survival and offspring phenotype. These same disturbed sites are often contaminated by pollutants and turtles can incorporate high levels of pollutants into their eggs which negatively impact hatch success. For my M.S. research, I investigated turtle nest site selection in a system dominated by agricultural and industrial land use, the impact of crop growth on the thermal and hydric dynamics of turtle nests, and I used paired field and laboratory experiments to examine the individual and interactive impacts of agricultural land use and Hg contamination on hatch success and offspring phenotype in Chelydra serpentina. Of the 150 turtle nests found during this research, 84% were located in human-disturbed soils. Nest site characteristics were similar among nests found in Hg contaminated and reference areas. Agriculture and control nests did not differ in temperature at the time of nesting, but temperatures diverged as crops grew, with temperatures in nests in agricultural fields averaging 2.5 °C lower than control nests over the course of incubation. Similarly, despite no initial difference, nest moisture levels diverged throughout incubation and moisture averaged 107 kPa lower in agricultural than control soils throughout incubation. In my field and laboratory experiments, I found that in comparison to turtles from control incubation conditions (i.e., warmer), turtles incubated under agricultural thermal regimens (i.e., colder) took longer to hatch, hatched at smaller structural body sizes, lost more mass after hatching, had lower post-hatching structural growth rates, and were more likely to be male. Additionally, thermal conditions associated with agricultural land use interacted with high levels of mercury to impact hatching success and offspring sex ratios. My thesis research provides one of the first documentations of negative interactive effects of mercury pollution and habitat quality on early vertebrate development and highlights the importance of examining the combined influence of multiple global changes on biological systems. / Master of Science / In aquatic turtles, females select nest sites that have a high degree of solar exposure, and exploit recently tilled agricultural fields for nesting, presumably because of increased solar exposure and/or easier nest excavation, and the importance of incubation temperature on survival and offspring phenotype. These same disturbed sites are often contaminated by pollutants and turtles can incorporate high levels of pollutants into their eggs which negatively impact hatch success. For my M.S. research, I investigated turtle nest site selection in a system dominated by agricultural and industrial land use, the impact of crop growth on the thermal and hydric dynamics of turtle nests, and I used paired field and laboratory experiments to examine the individual and interactive impacts of agricultural land use and Hg contamination on hatch success and offspring phenotype in Chelydra serpentina. Of the 150 turtle nests found during this research, 84% were located in human-disturbed soils. Nest site characteristics were similar among nests found in Hg contaminated and reference areas. Agriculture and control nests did not differ in temperature at the time of nesting, but temperatures diverged as crops grew, with temperatures in nests in agricultural fields averaging 2.5 °C lower than control nests over the course of incubation. Similarly, despite no initial difference, nest moisture levels diverged throughout incubation and moisture averaged 107 kPa lower in agricultural than control soils throughout incubation. In my field and laboratory experiments, I found that in comparison to turtles from control incubation conditions (i.e., warmer), turtles incubated under agricultural thermal regimens (i.e., colder) took longer to hatch, hatched at smaller structural body sizes, lost more mass after hatching, had lower post-hatching structural growth rates, and were more likely to be male. Additionally, thermal conditions associated with agricultural land use interacted with high levels of mercury to impact hatching success and offspring sex ratios. My thesis research provides one of the first documentations of negative interactive effects of mercury pollution and habitat quality on early vertebrate development and highlights the importance of examining the combined influence of multiple global changes on biological systems.