Demographic and Life History Characteristics of Black Bullheads Ameiurus melas in a North Temperate USA Lake

Black bullheads Ameiurus melas are an environmentally tolerant omnivorous fish species that are found throughout much of North America and parts of Europe. Despite their prevalence, black bullheads are an infrequently studied species making their biology, ecology, and life history poorly understood. Although limited information has been published on black bullheads, evidence suggests that bullheads can dominate the fish biomass and have profound influences on the fish community in some north temperate USA lakes. The goal of our study was to provide additional information on black bullhead population demographics, growth rates, life history characteristics, and seasonal diet preferences in a northern Wisconsin lake. Using common fish collection gears (fyke netting, electrofishing), fish aging protocols, fecundity assessments, and diet indices, our results suggested that black bullheads exhibited relatively fast growth rates, early ages at maturity, moderate fecundity, and a diverse omnivorous diet. Due to these demographic and life history characteristics, black bullheads have the potential to dominate fish community biomass in their native and introduced range. Results from our study may inform the management of black bullhead as native and invasive species.

The interface between invasive species science and legal regulation, using Hymenopteran species and their pathogens as a model system

<p>Biological invasions are one of the major causes of biodiversity decline on the planet. The key driver of the global movement of invasive species is international trade. As a response to trade driven invasive species risk, international and domestic regulations have been promulgated with the goal of managing the spread and impact of non-native species. My aims in this thesis were twofold. First, my goal was to review a subset of international and domestic regulations with a view to commenting on their fitness for purpose and suggesting potential improvements. Second, I used the example of non-native and invasive Hymenoptera, as well as their pathogens, to illustrate the risks posed by invasive species and gaps in their management. In order to assess international and domestic regulations, I reviewed the World Trade Organization’s (WTO) Agreement on Sanitary and Phytosanitary Measures, as well as associated disputes. I argue that the WTO’s regulatory system does, for the most part, allow domestic regulators to manage invasive species risk as they see fit. Subsequently, the focus of the thesis narrows to investigate New Zealand’s pre- and post-border regime managing invasive species. I argue that New Zealand’s pre-border approach represents international best practice, but the post-border management of species is fragmented. The power to manage invasive species has been delegated to sub-national and regional bodies, which typically approach invasive species management in different ways. This variation has led to regulatory inconsistencies in pests managed and funding allocated. There appears to be a substantial lack of planning in some spaces, such as the risk of aquatic invasions. I make recommendations to ameliorate these inconsistencies. My second aim involved the study of non-native and invasive Hymenoptera in New Zealand, as well as the pathogens they carry, in order to illustrate the risks posed by invasive species and gaps in their management. I show that the globally widespread invasive Argentine ant (Linepithema humile) may play a role in the pathogen dynamics and mortality of honey bee hives where the species occur sympatrically. Hives in the presence of Argentine ants suffered significantly higher mortality rates relative to hives without ants and always had higher levels of a honey bee pathogen Deformed wing virus. I demonstrate that honey bee pathogens are found in a range of invasive Hymenoptera in New Zealand. I amplify entire genomes of the honey bee virus Kashmir bee virus (KBV) from three species of non-native or invasive Hymenoptera (Argentine ants, common wasps and honey bees). I show that there is KBV strain variability within and between regions, but more between regions. Further, I demonstrate the result that as sampled KBV sequence length increases, so too does sampled diversity. These results highlight how ‘an’ invasive species is typically not alone: they carry a range of diseases that are almost always not considered in international and regional management plans. Patterns of non-native Hymenoptera carrying honey bee diseases were not restricted to New Zealand. I used mitochondrial DNA to find the likely origin of invasive populations of the globally distributed invasive German wasp. I demonstrate that German wasps show reduced genetic diversity in the invaded range compared to the native range. Populations in the introduced range are likely to have arrived from different source populations. In some regions there were likely multiple introductions. Other regions are genetically homogenous and represent potential areas for use of gene drive technologies. All four different honey bee pathogens assayed for were found in German wasp populations worldwide. These results highlight how the introduction of one exotic species likely brings a range of pathogens. This example of pathogens in Hymenoptera is likely to be true for nearly all non-native introductions. Many of the impacts of biological invasions, such as predation and competition, are relatively obvious and are frequently studied. However some, such as the impact of pathogens, are unseen and poorly understood. Legal regulation is often a post-hoc response implemented once a problem has already arisen. At a global level regulatory regimes operate relatively effectively. As the focus becomes more granular, such as the case of pathogens of Hymenoptera, fewer controls exists. This thesis helps to reduce uncertainty in this area as well as makes recommendations as to how these risks may be managed.</p>

The interface between invasive species science and legal regulation, using Hymenopteran species and their pathogens as a model system

<p>Biological invasions are one of the major causes of biodiversity decline on the planet. The key driver of the global movement of invasive species is international trade. As a response to trade driven invasive species risk, international and domestic regulations have been promulgated with the goal of managing the spread and impact of non-native species. My aims in this thesis were twofold. First, my goal was to review a subset of international and domestic regulations with a view to commenting on their fitness for purpose and suggesting potential improvements. Second, I used the example of non-native and invasive Hymenoptera, as well as their pathogens, to illustrate the risks posed by invasive species and gaps in their management. In order to assess international and domestic regulations, I reviewed the World Trade Organization’s (WTO) Agreement on Sanitary and Phytosanitary Measures, as well as associated disputes. I argue that the WTO’s regulatory system does, for the most part, allow domestic regulators to manage invasive species risk as they see fit. Subsequently, the focus of the thesis narrows to investigate New Zealand’s pre- and post-border regime managing invasive species. I argue that New Zealand’s pre-border approach represents international best practice, but the post-border management of species is fragmented. The power to manage invasive species has been delegated to sub-national and regional bodies, which typically approach invasive species management in different ways. This variation has led to regulatory inconsistencies in pests managed and funding allocated. There appears to be a substantial lack of planning in some spaces, such as the risk of aquatic invasions. I make recommendations to ameliorate these inconsistencies. My second aim involved the study of non-native and invasive Hymenoptera in New Zealand, as well as the pathogens they carry, in order to illustrate the risks posed by invasive species and gaps in their management. I show that the globally widespread invasive Argentine ant (Linepithema humile) may play a role in the pathogen dynamics and mortality of honey bee hives where the species occur sympatrically. Hives in the presence of Argentine ants suffered significantly higher mortality rates relative to hives without ants and always had higher levels of a honey bee pathogen Deformed wing virus. I demonstrate that honey bee pathogens are found in a range of invasive Hymenoptera in New Zealand. I amplify entire genomes of the honey bee virus Kashmir bee virus (KBV) from three species of non-native or invasive Hymenoptera (Argentine ants, common wasps and honey bees). I show that there is KBV strain variability within and between regions, but more between regions. Further, I demonstrate the result that as sampled KBV sequence length increases, so too does sampled diversity. These results highlight how ‘an’ invasive species is typically not alone: they carry a range of diseases that are almost always not considered in international and regional management plans. Patterns of non-native Hymenoptera carrying honey bee diseases were not restricted to New Zealand. I used mitochondrial DNA to find the likely origin of invasive populations of the globally distributed invasive German wasp. I demonstrate that German wasps show reduced genetic diversity in the invaded range compared to the native range. Populations in the introduced range are likely to have arrived from different source populations. In some regions there were likely multiple introductions. Other regions are genetically homogenous and represent potential areas for use of gene drive technologies. All four different honey bee pathogens assayed for were found in German wasp populations worldwide. These results highlight how the introduction of one exotic species likely brings a range of pathogens. This example of pathogens in Hymenoptera is likely to be true for nearly all non-native introductions. Many of the impacts of biological invasions, such as predation and competition, are relatively obvious and are frequently studied. However some, such as the impact of pathogens, are unseen and poorly understood. Legal regulation is often a post-hoc response implemented once a problem has already arisen. At a global level regulatory regimes operate relatively effectively. As the focus becomes more granular, such as the case of pathogens of Hymenoptera, fewer controls exists. This thesis helps to reduce uncertainty in this area as well as makes recommendations as to how these risks may be managed.</p>

Biological Control of Weeds: Is it Safe?

Invasive plants are non-native plant species that cause harm in their introduced range. Classical biological control of invasive plants is the use of natural enemies, imported insects and mites or pathogens, to control the target plants. This publication explains the strategies and rules in place to ensure that organisms released for the biological control of weeds are safe and effective.

Variation in behavioural traits and their molecular basis in the globally invasive Argentine ant

<p>A salient question in invasion biology is to elucidate what traits make some species invasive and not others. Recently, the introduction process has been hypothesised to function as a filter, selecting for traits that promote invasiveness in introduced populations. Behaviour has been increasingly recognised as a significant driver of animal invasions, but few studies focus on how the introduction process may affect behavioural variation across an invader’s range. This thesis aims at investigating the consequences of the introduction process on behavioural variation along an introduction pathway in the globally widespread Argentine ant. In Chapter I, I investigated transcriptome-wide variations associated with morphological caste as well as worker foraging propensity. I found a number of transcripts showed caste and behaviour-specific expression profiles, and that foraging workers exhibit higher expression of genes related to chemosensory receptors and neurological pathways known to be associated with foraging behaviour and aggression. In chapter II, I investigated octopamine-mediated modulation of foraging behaviour and aggression using pharmacological manipulations using synthetic octopamine (OA) and one of its antagonists. I found that OA was associated with colony-level foraging activity, but did not appear to similarly drive changes in individual behaviour. However, my results suggest that OA is associated with individual aggressiveness, and I also found an association between OA and foraging propensity. In chapter III, I tested the hypothesis that the introduction process selected for increased risk-taking behaviour along the Argentine ant introduction pathway. I used behavioural assays combined with OA diet supplementation in four regions along the introduction pathway, and showed that both behavioural variation and plasticity were consistent across the range, not in support of our hypothesis. Finally, in chapter IV I investigated transcriptome-wide expression profiles in workers along the introduction pathway and discovered distinct transcriptomic signatures associated with range. I found that specific immune and neural pathways were differentially regulated in the introduced range compared to the native range. Overall, my results contribute to a better understanding of caste differentiation and neuromodulation of key behaviours in invasive ants. While I did not find behavioural differentiation between native and invasive Argentine ant populations, I show that they are characterised by specific transcriptomic signatures perhaps underlying the ant’s invasion success. This is one of the first global and integrative studies to focus on changes associated with the introduction of invasive species, and may have important implications for the understanding of biological invasions.</p>

Microbiota of an Invasive Wasp Vespula vulgaris and Hymenopteran relatives: Interpreting the microbiome

<p>Invasive species represent a critical threat to ecosystems and ecological communities, causing changes through overwhelming predation as well as competing with native species for resources. Understanding the mechanisms behind invasive success is essential for understanding why they invade and the consequences of their invasions. Furthermore, invasive species, like all macroscopic organisms, harbour symbiotic and pathogenic microbes that constitute their microbiomes, which could explain invasive success. The complex ecological interaction networks within the microbiome can have a positive or negative impact on host abundance and dominance. These interactions may be significant for invasive species, where microbial influences acting on an exotic host can potentially drive the ecological success of an invasive population to the detriment of recipient communities. This thesis explores the microbiota of one of the most globally invasive species, the common wasp Vespula vulgaris, with the overall aim to investigate and characterise the microbiome of V. vulgaris, using metagenomics, bioinformatics and molecular techniques. The initial comparative microbiota study focused on three distinct life stages (larvae, worker and queen), from two ranges. This analysis revealed a core bacteriome community present in V. vulgaris. There was evidence of higher microbial diversity in wasp larvae compared with workers and queens. The Queen (gyne) microbiome revealed a more specific microbiome with absences of certain microbiota found in larvae and workers from the same nest, indicating a more distinctive microbiome. Interestingly, analysis of life stages between ranges showed significant dissimilarity in microbiomes, with microbiota loses, and acquisitions in the introduced New Zealand range. Using the same techniques, the microbiota of V. vulgaris and four hymenopteran hosts (Apis mellifera, Bombus terrestris, Vespula germanica and Linepithema humile), were comparatively analysed. The analysis investigated both shared microbiota and host specific microbiota. This analysis indicated the polyphagous V. vulgaris as having a diverse microbiome varying between nests and sites, indicating less specific microbiota in comparison to other hymenopteran hosts in this study. Vespid wasp colonies are known to occasionally crash in the presence of diseases; however, there is a lack of reliable evidence indicating pathogenic micro-organisms play an essential role in wasp colony dynamics. Incorporating knowledge gained in previous analyses, the next aim was to investigate V. vulgaris nests symptomatic of an infectious agent to discover the cause of pathology. Through molecular techniques, such as Illumina RNA-Seq, PCR and Sanger sequencing, the potential cause of infection and decline of diseased nests was examined. The metatranscriptomic comparison of diseased and healthy larvae highlighted five putative infectious agents. The bacteria Moellerella wisconsensis, Moku virus, Kashmir Bee Virus, Aspergillus and the microsporidian Vavraia culicis floridensis found in infected larvae, potentially causing pathology in the host. The first known instance of Moku virus, and potentially V. culicis floridensis and M. wisconsensis was documented as potential pathogens of V. vulgaris present in New Zealand. To test for potential virulence of these putative infectious agents, an infection study was carried out. Vespula vulgaris nests and larvae were orally infected in the lab using homogenised infected larvae. Subsequently, test and control larvae were sampled to conduct and quantify a time series analysis of infection using RT-qPCR using designed primers. This dissertation provided the first insight into the microbiome of V. vulgaris in the native and introduced range providing a baseline for further research. This analysis and the subsequent microbiota identified may play a role in wasp population dynamics, giving a better understanding of the observed thriving V. vulgaris population dynamics in New Zealand.</p>

Variation in behavioural traits and their molecular basis in the globally invasive Argentine ant

<p>A salient question in invasion biology is to elucidate what traits make some species invasive and not others. Recently, the introduction process has been hypothesised to function as a filter, selecting for traits that promote invasiveness in introduced populations. Behaviour has been increasingly recognised as a significant driver of animal invasions, but few studies focus on how the introduction process may affect behavioural variation across an invader’s range. This thesis aims at investigating the consequences of the introduction process on behavioural variation along an introduction pathway in the globally widespread Argentine ant. In Chapter I, I investigated transcriptome-wide variations associated with morphological caste as well as worker foraging propensity. I found a number of transcripts showed caste and behaviour-specific expression profiles, and that foraging workers exhibit higher expression of genes related to chemosensory receptors and neurological pathways known to be associated with foraging behaviour and aggression. In chapter II, I investigated octopamine-mediated modulation of foraging behaviour and aggression using pharmacological manipulations using synthetic octopamine (OA) and one of its antagonists. I found that OA was associated with colony-level foraging activity, but did not appear to similarly drive changes in individual behaviour. However, my results suggest that OA is associated with individual aggressiveness, and I also found an association between OA and foraging propensity. In chapter III, I tested the hypothesis that the introduction process selected for increased risk-taking behaviour along the Argentine ant introduction pathway. I used behavioural assays combined with OA diet supplementation in four regions along the introduction pathway, and showed that both behavioural variation and plasticity were consistent across the range, not in support of our hypothesis. Finally, in chapter IV I investigated transcriptome-wide expression profiles in workers along the introduction pathway and discovered distinct transcriptomic signatures associated with range. I found that specific immune and neural pathways were differentially regulated in the introduced range compared to the native range. Overall, my results contribute to a better understanding of caste differentiation and neuromodulation of key behaviours in invasive ants. While I did not find behavioural differentiation between native and invasive Argentine ant populations, I show that they are characterised by specific transcriptomic signatures perhaps underlying the ant’s invasion success. This is one of the first global and integrative studies to focus on changes associated with the introduction of invasive species, and may have important implications for the understanding of biological invasions.</p>

Microbiota of an Invasive Wasp Vespula vulgaris and Hymenopteran relatives: Interpreting the microbiome

<p>Invasive species represent a critical threat to ecosystems and ecological communities, causing changes through overwhelming predation as well as competing with native species for resources. Understanding the mechanisms behind invasive success is essential for understanding why they invade and the consequences of their invasions. Furthermore, invasive species, like all macroscopic organisms, harbour symbiotic and pathogenic microbes that constitute their microbiomes, which could explain invasive success. The complex ecological interaction networks within the microbiome can have a positive or negative impact on host abundance and dominance. These interactions may be significant for invasive species, where microbial influences acting on an exotic host can potentially drive the ecological success of an invasive population to the detriment of recipient communities. This thesis explores the microbiota of one of the most globally invasive species, the common wasp Vespula vulgaris, with the overall aim to investigate and characterise the microbiome of V. vulgaris, using metagenomics, bioinformatics and molecular techniques. The initial comparative microbiota study focused on three distinct life stages (larvae, worker and queen), from two ranges. This analysis revealed a core bacteriome community present in V. vulgaris. There was evidence of higher microbial diversity in wasp larvae compared with workers and queens. The Queen (gyne) microbiome revealed a more specific microbiome with absences of certain microbiota found in larvae and workers from the same nest, indicating a more distinctive microbiome. Interestingly, analysis of life stages between ranges showed significant dissimilarity in microbiomes, with microbiota loses, and acquisitions in the introduced New Zealand range. Using the same techniques, the microbiota of V. vulgaris and four hymenopteran hosts (Apis mellifera, Bombus terrestris, Vespula germanica and Linepithema humile), were comparatively analysed. The analysis investigated both shared microbiota and host specific microbiota. This analysis indicated the polyphagous V. vulgaris as having a diverse microbiome varying between nests and sites, indicating less specific microbiota in comparison to other hymenopteran hosts in this study. Vespid wasp colonies are known to occasionally crash in the presence of diseases; however, there is a lack of reliable evidence indicating pathogenic micro-organisms play an essential role in wasp colony dynamics. Incorporating knowledge gained in previous analyses, the next aim was to investigate V. vulgaris nests symptomatic of an infectious agent to discover the cause of pathology. Through molecular techniques, such as Illumina RNA-Seq, PCR and Sanger sequencing, the potential cause of infection and decline of diseased nests was examined. The metatranscriptomic comparison of diseased and healthy larvae highlighted five putative infectious agents. The bacteria Moellerella wisconsensis, Moku virus, Kashmir Bee Virus, Aspergillus and the microsporidian Vavraia culicis floridensis found in infected larvae, potentially causing pathology in the host. The first known instance of Moku virus, and potentially V. culicis floridensis and M. wisconsensis was documented as potential pathogens of V. vulgaris present in New Zealand. To test for potential virulence of these putative infectious agents, an infection study was carried out. Vespula vulgaris nests and larvae were orally infected in the lab using homogenised infected larvae. Subsequently, test and control larvae were sampled to conduct and quantify a time series analysis of infection using RT-qPCR using designed primers. This dissertation provided the first insight into the microbiome of V. vulgaris in the native and introduced range providing a baseline for further research. This analysis and the subsequent microbiota identified may play a role in wasp population dynamics, giving a better understanding of the observed thriving V. vulgaris population dynamics in New Zealand.</p>

The influence of semiochemicals on the co-occurrence patterns of a global invader and native species

<p>The success of invasive species in their introduced range is often influenced by interactions with resident species communities. Chemical communication is one the factors which contributes to a variety of aspects of a species life cycle, ranging from mating, to food localization and interactions with members of the same and other species. In my thesis, I investigate the effects of venoms and semiochemicals on interactions between the invasive Argentine ant (Linepethima humile) with other resident ant species and demonstrate how pheromones can potentially be utilized as an area wide control mechanism of this species, by disrupting their foraging success. I studied the effects of venom composition, their toxicity and utilization on the outcome of aggressive interactions between the Argentine ant and the four Monomorium species in New Zealand occurring. The toxicity of the venom of the two species co-occurring with Argentine ants was significantly higher than the toxicity of the species which do not. However, no correlation between venom toxicity and Monomorium survival was found. For M. antipodum a significant relationship between venom utilization and its mortality was found, indicating that the way venom is used might be an important aspect of these interactions. Physical Aggression between Monomorium and Argentine ants also had strong effects on Monomorium worker mortality, which provided evidence that a variety of factors and strategies contribute to the ability of interacting organisms to withstand the pressure of a dominant invader at high abundance. I conducted bioassays with food sources and synthetic trail pheromones, providing a proof of concept on disrupting the foraging ability of Argentine ants. Other resident species benefited from the reduced success of Argentine ants, but to a varying degree. Behavioural variations between the resident species provided an explanation for observed differences in foraging success and how much each of these individual competitors was able to increase their foraging. The mechanism for the observed increase in resource acquisition of resident species appeared to be a decrease in aggressive behaviour displayed by Argentine ants. I expanded the usage of the synthetic pheromone to a commercial vineyard, were Argentine ants can have negative effects on crop development by dispersing and tending to homopteran pest species. Argentine ants’ access to the crop canopy could be significantly reduced by placing pheromone dispensers at the base of the vine plant, while dispensers in the plant canopy had little effect on Argentine ant numbers. Doubling the amount of pheromone did not result in an additional reduction of ant activity. Lastly incorporating the knowledge gained in the previous chapter, I extended the application of the pheromone to a large field trial over a three month period. Argentine ant activity and foraging success was significantly supressed compared to untreated control plots, providing evidence that this form of large scale application might be a possible way to control large invasive ant populations by disrupting their trail following and foraging behaviour for a prolonged period of time. While initial calculations have suggested that the treatment is currently not feasible (13.3 US$/mg/ha), I found a significant reduction in body fat in workers collected from treated plots compared with untreated plots, suggesting adverse effects on nest fitness. My findings provide new insights into chemical communication between invasive and resident species, support existing dominance hierarchy models in ant populations, and help to establish a target specific potential management technique of wide-spread invasive ant species.</p>

The influence of semiochemicals on the co-occurrence patterns of a global invader and native species

<p>The success of invasive species in their introduced range is often influenced by interactions with resident species communities. Chemical communication is one the factors which contributes to a variety of aspects of a species life cycle, ranging from mating, to food localization and interactions with members of the same and other species. In my thesis, I investigate the effects of venoms and semiochemicals on interactions between the invasive Argentine ant (Linepethima humile) with other resident ant species and demonstrate how pheromones can potentially be utilized as an area wide control mechanism of this species, by disrupting their foraging success. I studied the effects of venom composition, their toxicity and utilization on the outcome of aggressive interactions between the Argentine ant and the four Monomorium species in New Zealand occurring. The toxicity of the venom of the two species co-occurring with Argentine ants was significantly higher than the toxicity of the species which do not. However, no correlation between venom toxicity and Monomorium survival was found. For M. antipodum a significant relationship between venom utilization and its mortality was found, indicating that the way venom is used might be an important aspect of these interactions. Physical Aggression between Monomorium and Argentine ants also had strong effects on Monomorium worker mortality, which provided evidence that a variety of factors and strategies contribute to the ability of interacting organisms to withstand the pressure of a dominant invader at high abundance. I conducted bioassays with food sources and synthetic trail pheromones, providing a proof of concept on disrupting the foraging ability of Argentine ants. Other resident species benefited from the reduced success of Argentine ants, but to a varying degree. Behavioural variations between the resident species provided an explanation for observed differences in foraging success and how much each of these individual competitors was able to increase their foraging. The mechanism for the observed increase in resource acquisition of resident species appeared to be a decrease in aggressive behaviour displayed by Argentine ants. I expanded the usage of the synthetic pheromone to a commercial vineyard, were Argentine ants can have negative effects on crop development by dispersing and tending to homopteran pest species. Argentine ants’ access to the crop canopy could be significantly reduced by placing pheromone dispensers at the base of the vine plant, while dispensers in the plant canopy had little effect on Argentine ant numbers. Doubling the amount of pheromone did not result in an additional reduction of ant activity. Lastly incorporating the knowledge gained in the previous chapter, I extended the application of the pheromone to a large field trial over a three month period. Argentine ant activity and foraging success was significantly supressed compared to untreated control plots, providing evidence that this form of large scale application might be a possible way to control large invasive ant populations by disrupting their trail following and foraging behaviour for a prolonged period of time. While initial calculations have suggested that the treatment is currently not feasible (13.3 US$/mg/ha), I found a significant reduction in body fat in workers collected from treated plots compared with untreated plots, suggesting adverse effects on nest fitness. My findings provide new insights into chemical communication between invasive and resident species, support existing dominance hierarchy models in ant populations, and help to establish a target specific potential management technique of wide-spread invasive ant species.</p>