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The South African isolate of the Black Queen-Cell Virus (BQCV), a honeybee virus, was previously found to have an 8550 nucleotide genome excluding the poly(A) tail. Its genome contained two ORFs, a 5'-proximal ORF encoding a putative replicase protein and a 3'-proximal ORF encoding a capsid polyprotein.<br /> <br /> A reverse transcriptase PCR (RT -PCR) assay was developed for the detection of BQCV and acute bee-paralysis virus (ABPV). Complete genomes sequences w ere used to design unique PCR primers within a l-kb region from the 3' end of both genomes to amplify a fragment of 70.0 bp from BQCV and 900 bp from ABPV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing BQCV and ABPV. Sensitivities were of the order of 130 genome equivalents of purified BQCV and 1600 genome equivalents of ABPV.

The risks posed by rapidly evolving RNA viruses to human and animal health are well recognized. Epidemics in managed and wildlife populations can lead to considerable economic and biodiversity losses. Yet, we lack understanding of the ecological and evolutionary factors that promote disease emergence. Host-switching viruses may be a particular threat to species important for human welfare, such as pollinating bees. Both honeybees and wild bumblebees have faced sharp declines in the last decades, with high winter mortality seen in honeybees. Infectious and emerging diseases are considered one of the key drivers of declines, acting in synergy with habitat loss and pesticide use. Here I focus on multihost viruses that pose a risk to wild bumblebees. I first identify the risk factors driving viral spillover and emergence from managed honeybees to wild bumblebees, by synthesising current data and literature. Biological factors (i.e. the nature of RNA viruses and ecology of social bees) play a clear role in increasing the risk of disease emergence, but anthropogenic factors (trade and transportation of commercial honeybees and bumblebees) creates the greatest risk of viral spillover to wild bees. Basic knowledge of the pathogenic effect of many common pollinator viruses on hosts other than A. mellifera is currently lacking, yet vital for understanding the wider impacts of infection at a population level. Here, I provide evidence that a common bumblebee virus, Slow bee paralysis virus (SBPV), reduces the longevity of Bombus terrestris under conditions of nutrition stress. The invasion of Varroa destructor as an ectoparasitic viral vector in European honeybees has dramatically altered viral dynamics in honeybees. I test how this specialist honeybee vector affects multi-host pathogens that can infect and be transmitted by both honeybees and wild bumblebees. I sampled across three host species (A. mellifera, B. terrestris and B. pascuorum) from Varroa-free and Varroa-present locations. Using a combination of molecular and phylogenetic techniques I find that this specialist honeybee vector increases the prevalence of four multi-host viruses (deformed wing virus (type A and B), SBPV and black queen cell virus) in sympatric wild bumblebees. Furthermore, wild bumblebees are currently experiencing a DWV epidemic driven by the presence of virus-vectoring Varroa in A. mellifera. Overall this thesis demonstrates that wild bumblebees are at high risk of viral disease emergence. My research adds to the ever-expanding body of evidence indicating that stronger disease controls on commercial bee operations are crucial to protect our wild bumblebees.

Biogenic amines are important messenger substances in the central nervous system and in peripheral organs of vertebrates and of invertebrates. The honeybee, Apis mellifera, is excellently suited to uncover the functions of biogenic amines in behaviour, because it has an extensive behavioural repertoire, with a number of biogenic amine receptors characterised in this insect. In the honeybee, the biogenic amines dopamine, octopamine, serotonin and tyramine modulate neuronal functions in various ways. Dopamine and serotonin are present in high concentrations in the bee brain, whereas octopamine and tyramine are less abundant. Octopamine is a key molecule for the control of honeybee behaviour. It generally has an arousing effect and leads to higher sensitivity for sensory inputs, better learning performance and increased foraging behaviour. Tyramine has been suggested to act antagonistically to octopamine, but only few experimental data are available for this amine. Dopamine and serotonin often have antagonistic or inhibitory effects as compared to octopamine. Biogenic amines bind to membrane receptors that primarily belong to the large gene-family of GTP-binding (G) protein coupled receptors. Receptor activation leads to transient changes in concentrations of intracellular second messengers such as cAMP, IP3 and/or Ca2+. Although several biogenic amine receptors from the honeybee have been cloned and characterised more recently, many genes still remain to be identified. The availability of the completely sequenced genome of Apis mellifera will contribute substantially to closing this gap. In this review, we will discuss the present knowledge on how biogenic amines and their receptor-mediated cellular responses modulate different behaviours of honeybees including learning processes and division of labour.

No description available.

In this project I studied the importance of pollinators in the reproduction of Gastrodia ‘long column’ and the preferences of New Zealand’s native and exotic insect pollinators. This was done in order to determine the specialisation of insect pollination in urban Christchurch. This knowledge can contribute to crop pollination, conservation efforts, and an understanding of the main drivers of the common floral traits (small, white flowers) in New Zealand. The strength of the relationship between the common traits of the New Zealand flora and the preferences of the native invertebrate fauna is not known. Traditionally it had been thought that New Zealand’s insects lack strong preferences, however recent work has not supported this. Changing landscapes worldwide have led to declines in pollinator numbers. Additionally, in New Zealand Apis mellifera numbers have declined as a result of the arrival of the varroa mite and it is important that alternative pollinating species for wild and agricultural pollination are identified. To address these needs I examined the abundance and preferences of insect pollinators in modified landscapes in and around Christchurch testing: the reliance upon pollinators by the undescribed native orchid Gastrodia ‘long column’, the preferences of New Zealand’s native and introduced insect pollinators in an extensive observational study, the results of which I further tested using a subsequent manipulative experiment of petal colour (according to human vision) at the Christchurch Botanical Gardens Sampling at the Christchurch Botanical Gardens, University of Canterbury, and Port Hills across a four month observation period (January – April 2012), revealed that native pollinators preferred white native flowers and exotic pollinators preferred not-white exotic flowers when data were grouped according to insect provenance. A more detailed visit-level analysis found that two native bees, Hylaeus and Leioproctus, showed a significant preference for native flowers; the exotic bees, Apis mellifera and Bombus species, preferred purple over white flowers; and Melangyna novaezelandiae (a native hoverfly) preferred white over yellow. However, a series of experimental arrays to present controlled choices between pairs of flower types at the Christchurch Botanical Gardens (14 December 2012 – 22 January 2013) did not find significant preferences by native or exotic insect pollinators between white and yellow flowers. This may have been a result of the plant species chosen, as a correlation between pollinator preference and plant species has been shown elsewhere. Visitation to experimental arrays was both low and highly skewed, with over half of all visits made by Lasioglossum bees and 615 of the 669 visits made by native species. This may have meant that lack of significant results were representing the choices of native insects generally and Lasioglossum bees specifically. A bagging experiment from 30 January 2012 – 16 February 2012 demonstrated the dependence of the undescribed native orchid species, Gastrodia ‘long column’, upon pollinating animals. Fruit set of most plants worldwide depends upon pollination (by wind or animal-transfer of pollen). In this case study final fruit sets were significantly reduced on bagged inflorescences, while open flowers had surprisingly high natural fruit sets (>75%). A week of observations (29 January 2012 – 6 February 2012) revealed that Gastrodia ‘long column’ was predominantly visited by Lasioglossum bees, and remarkably bee numbers were high enough in a residential property in the middle of Christchurch city for high fruit set on unmanipulated plants. The results of my thesis indicate that pollinators are important in the reproductive system of Gastrodia ‘long column’, suggesting that other previously overlooked plant species may also rely upon insect pollinators. Furthermore, the relative importance of native pollinating insects is high for native plants even when examined in an urban setting. New Zealand pollinators have preferences for certain floral traits which show trends when grouped broadly, but vary when considered at the insect species level. This contrasts with traditional views of unspecialised insect pollinators which lack preferences in regards to the plants they visit. Further work which serves to increase current understanding of the underlying mechanisms of pollination specialisation in New Zealand may wish to focus on single insect species. By identifying particular preferences of pollinators and the underlying ‘native’ traits selected for, alternative options to crop pollination may be found, targeted management strategies implemented, and the strength of the relationship between pollinator preferences and the traits of the plants they visit determined.

The South African isolate of the Black Queen-Cell Virus (BQCV), a honeybee virus, was previously found to have an 8550 nucleotide genome excluding the poly(A) tail. Its genome contained two ORFs, a 5'-proximal ORF encoding a putative replicase protein and a 3'-proximal ORF encoding a capsid polyprotein.<br /> <br /> A reverse transcriptase PCR (RT -PCR) assay was developed for the detection of BQCV and acute bee-paralysis virus (ABPV). Complete genomes sequences w ere used to design unique PCR primers within a l-kb region from the 3' end of both genomes to amplify a fragment of 70.0 bp from BQCV and 900 bp from ABPV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing BQCV and ABPV. Sensitivities were of the order of 130 genome equivalents of purified BQCV and 1600 genome equivalents of ABPV.

Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (leaves 185-189).

Thesis (M.Sc.)--Acadia University, 1999. / Includes bibliographical references (leaves 46-48). Also available on the Internet via the World Wide Web.

Thesis (M.Sc.)--Acadia University, 1999. / Includes bibliographical references (leaves 46-48). Also available on the Internet via the World Wide Web.