Theoretical research has demonstrated that ecological interactions in sympatry or parapatry can generate disruptive selection that in concert with assortative mating can lead to speciation. However, empirical examples are few and restricted to terrestrial and lacustrine systems. New Zealand triplefin fishes (Family Tripterygiidae) are an ideal model system to study speciation in the sea, as they conform to the criteria of an adaptive radiation, being philopatric, speciose and abundant, and having largely sympatric distributions. This thesis investigates two key aspects of the New Zealand triplefin radiation: 1) which ecological traits are under selection?; and 2) which traits are potentially available for the development of assortative mating? Habitat use was identified as a possible key trait for selection and investigated in detail in this thesis. Habitat use of the majority of New Zealand triplefin species was censused quantitatively throughout most of their latitudinal range and analysed using novel statistical methods. Analyses showed that habitat use was highly divergent between species and thus diversification in habitat may have been a major component in the evolution of this clade. The phylogenetic analysis of habitat characters confirmed that there has been rapid evolution in habitat use among species. Habitat selection at settlement was highly species-specific, indicating that interspecific differences in adult habitat use may be the outcome of active habitat choice established at settlement. These species-specific habitat associations showed no evidence for geographic variation in habitat use. Laboratory trials and field observations of the sister-species pair Ruanoho decemdigitatus and R. whero showed that competition was linked with body size, with R. decemdigitatus being the larger and consequently dominant species. The second part of this thesis investigated which traits may have contributed to prezygotic isolation, and thus to assortative mating. Little evidence was found for divergence in breeding season or male colour patterns. However, divergence in habitat affected breeding habitat choice, as triplefins court and mate in the same territory as that occupied year round. This suggests that assortative mating in New Zealand triplefin species could be the by-product of adaptation to habitat resources. Body size affected mate choice and time at first maturity in the Ruanoho sister-species pair, suggesting that size is important in the maintenance of reproductive isolation in these species. Differences in body size may have also lead to assortative mating in other New Zealand triplefin sister-species pairs, as all sister-species pairs differ in maximum body size. The findings of this thesis invoke a strong role for ecologically-based selection in speciation, and support the hypothesis that adaptation to habitat has been a major factor in speciation in this system. / This PhD was funded by the New Zealand Royal Society (Marsden Fund 02-UOA-005 to Kendall Clements), the Tertiary Education Commission (Top Achiever Doctoral Scholarship), and the University of Auckland (University of Auckland Doctoral Scholarship.

Theoretical research has demonstrated that ecological interactions in sympatry or parapatry can generate disruptive selection that in concert with assortative mating can lead to speciation. However, empirical examples are few and restricted to terrestrial and lacustrine systems. New Zealand triplefin fishes (Family Tripterygiidae) are an ideal model system to study speciation in the sea, as they conform to the criteria of an adaptive radiation, being philopatric, speciose and abundant, and having largely sympatric distributions. This thesis investigates two key aspects of the New Zealand triplefin radiation: 1) which ecological traits are under selection?; and 2) which traits are potentially available for the development of assortative mating? Habitat use was identified as a possible key trait for selection and investigated in detail in this thesis. Habitat use of the majority of New Zealand triplefin species was censused quantitatively throughout most of their latitudinal range and analysed using novel statistical methods. Analyses showed that habitat use was highly divergent between species and thus diversification in habitat may have been a major component in the evolution of this clade. The phylogenetic analysis of habitat characters confirmed that there has been rapid evolution in habitat use among species. Habitat selection at settlement was highly species-specific, indicating that interspecific differences in adult habitat use may be the outcome of active habitat choice established at settlement. These species-specific habitat associations showed no evidence for geographic variation in habitat use. Laboratory trials and field observations of the sister-species pair Ruanoho decemdigitatus and R. whero showed that competition was linked with body size, with R. decemdigitatus being the larger and consequently dominant species. The second part of this thesis investigated which traits may have contributed to prezygotic isolation, and thus to assortative mating. Little evidence was found for divergence in breeding season or male colour patterns. However, divergence in habitat affected breeding habitat choice, as triplefins court and mate in the same territory as that occupied year round. This suggests that assortative mating in New Zealand triplefin species could be the by-product of adaptation to habitat resources. Body size affected mate choice and time at first maturity in the Ruanoho sister-species pair, suggesting that size is important in the maintenance of reproductive isolation in these species. Differences in body size may have also lead to assortative mating in other New Zealand triplefin sister-species pairs, as all sister-species pairs differ in maximum body size. The findings of this thesis invoke a strong role for ecologically-based selection in speciation, and support the hypothesis that adaptation to habitat has been a major factor in speciation in this system. / This PhD was funded by the New Zealand Royal Society (Marsden Fund 02-UOA-005 to Kendall Clements), the Tertiary Education Commission (Top Achiever Doctoral Scholarship), and the University of Auckland (University of Auckland Doctoral Scholarship.

The microbiome plays a key symbiotic role in maintaining host health and aids in acquiring nutrients, supporting development and immune function, and modulating behavior. However, more research is needed to elucidate the potential impact of environmental pollutants on host microbial communities and how microbiomes can modulate the toxicity of contaminants to the host. Through a literature review of 18 studies that assessed the impacts of various anthropogenic chemicals on fish-associated microbiomes, we found that toxicants generally decrease microbial diversity, which could lead to long-term health impacts if chronically stressed, and can increase the host’s susceptibility to disease as well as the chemical resistance of certain microbes. These findings led us to explore the impacts of one of the reviewed contaminants, polycyclic aromatic hydrocarbons (PAHs), typically found in oil. The Deepwater Horizon disaster of April 2010 was the largest oil spill in U.S. history and had catastrophic effects on several ecologically important fish species in the Gulf of Mexico (GoM). This study tested the hypotheses that exposure to weathered oil would cause significant shifts in fish gut-associated microbial communities, with taxa known for hydrocarbon degradation increasing in abundance and that foraging behavior would decrease, potentially due to microbial dysbiosis via the gut-brain axis. We characterized the gut microbiome (with 16S rRNA gene sequencing) of a native GoM estuarine species, the sheepshead minnow (Cyprinodon variegatus). Fish were exposed to High Energy Water Accommodated Fractions (HEWAF; tPAH = 80.99 ± 12.5 μg/L) of oil over a 7-day period and whole gastrointestinal tracts were sampled for microbiome analyses. A foraging behavioral assay was used to determine feeding efficiency before and after oil exposure. The fish gut microbiome did not experience any significant changes in alpha or beta diversity but known hydrocarbon degrading taxa were noticeably present in oil-exposed communities and were absent in controls. We found the order Pseudomonadales, the family Paenibacillaceae, and Pseudomonas pachastrellae to be among these, with Pseudomonadales increasing in abundance. Foraging behavior was not significantly affected by oil exposure. This work highlights the need for further research to elucidate the functional metagenomic responses of the fish gut-microbiome under oil spill conditions.