Entomology Theses and Dissertations

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Quantifying the relative contribution and furthering qualitative understanding of ftz cis-regulatory elements in Drosophila melanogaster
(2022) Fischer, Matthew Douglas; Pick, Leslie; Entomology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Embryonic development is coordinated by interactions within gene regulatory networks. This process is orchestrated at the level of transcription through the regulatory properties of enhancers, which direct spatiotemporal expression patterns when bound by specific trans-acting factors. Though enhancers can act upon promoters located at great distances irrespective of orientation, the contributions from these cis-regulatory elements (CREs) are limited by insulators and/or tethering elements that organize chromatin architecture. Much research has been conducted towards understanding the coordination of the segmentation genes that pattern the basic body plan of the fruit fly, Drosophila melanogaster, during embryogenesis. The pair-rule genes (PRGs) of this pathway, such as fushi tarazu (ftz), are expressed in seven alternating stripes across the embryo. These PRGs are required for the development of body segments, and the mis-regulation of a single transcriptional domain can result in the loss of a segment. Here, I have investigated the ftz CREs to more precisely determine their sufficiency to direct expression within ftz stripe domains and their necessity for doing so in the native context of the gene. To investigate the sufficiency, I have generated 36 standardized reporter transgenes from 18 CREs, tested in both forward and reverse orientations. All CREs examined have been inserted into the same XbaI site of the reporter plasmid, and the transgenes have been inserted into the same genomic region. Through in situ hybridization experiments, I have determined that the qualitative patterns conferred by every CRE is orientation-dependent, and I have identified two putative insulators and/or tethering elements, proposed to explain this observation. To investigate their necessity, I targeted four genomic regulatory regions for precise deletion using the CRISPR/Cas9 system to generate seven deletion mutants. Though deletions were expected to cause lethality, most of the mutants are homozygous viable and fertile; only a mutant simultaneously removing two seven-stripe CREs was homozygous lethal. Quantitative gene expression analysis by fluorescent in situ hybridization chain reaction revealed that there is a critical threshold of ftz abundance required in each stripe for segmentation to proceed. In conclusion, I have determined that the ftz CREs are redundant and function together in a non-additive manner.
DEVELOPING A PERRENIAL LIVING MULCH SYSTEM FOR MANAGING PESTS AND AUGMENTING NATURAL BIOCONTROL IN MARYLAND CANTALOUPE SYSTEMS
(2022) Nunez, Demian Antonio; Hooks, Cerruti RR; Entomology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
This study investigated how alsike clover (Trifolium hybridum) and Virginia wildrye (Elymus virginicus), when interplanted as a living mulch with cantaloupe, (Cucumis melo var. cantalupensis) would impact herbivorous and beneficial arthropod numbers. An additional objective was to determine how these living mulches would impact fruit yield and quality. It was hypothesized that there would be a reduction of cantaloupe pest herbivores and increase in natural enemy abundances in the interplanting compared to monoculture cantaloupe system. Some arthropods conformed to these expectations. However, most had a neutral or inconsistent response to the living mulches. Striped cucumber beetles (Acalymma vitattum), a major pest, were unaffected by the living mulches on most sampling dates. During several periods in both study years, leaf piercing herbivores including aphids were found in greater numbers on cantaloupe interplanted with clover than wildrye and/or monoculture. Spiders were found in greater abundance in cantaloupe interplanted with clover than wildrye or monoculture plantings during several sampling periods. Other natural enemy guilds such as parasitic wasps and piercing predators were inconsistently influenced by living mulch types. Yield was highest in the monoculture plots and living mulch was correlated with changes in fruit texture and color.
The Bee Louse, Braula coeca Nitzsch, its Distribution and Biology on Honey Bees
(1978) Smith, Irving Barton Jr.; Caron, Dewey M.; Entomology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md)
Bee lice were found in 28% of Maryland apiaries and 18% of the colonies examined. In apiaries with lice, 50% of the colonies contained lice. Laboratory tests demonstrated that bee lice had no preference between 1, 5, 15, and 30 day old honey bee (Apis mellifera L.) workers at 25° while there was a preference for 1 day old workers at 34° C. Lice preferred young drones over old drones and virgin and mated queens over young drones at 25 and 34° C. Lice preferred virgin queens over old drones at 25° C while no preference was observed at 24° C. Mated queens were preferred over old drones at 25 and 34° C. There was a preference of lice for foraging age workers over old drones at 25° C while there was no preference at 34° C. Lice preferred both virgin and mated queens over random age workers at 25 and 34° C. Louse larval tunnels were numerous in nucs (4 frame honey bee colonies) stocked with lice from May through August corresponding with periods of nectar flow when bees were capping honey. In field colonies, louse populations decreased in the late spring to a low in early June. During July and after, populations of lice rose with the emergence of new lice. Few immature and adult lice were observed in control nucs having similar populations of bees. In nucs, 1 or more lice were observed on 24% of the queens between August and December. Only 2% of the virgin queens contained lice during the same period. In field colonies, 62% of the queens examined from June through the rest of the season harbored lice; 58% of these lice were pale in color indicating they were less than 1 day old. One louse was observed on 98.6% of the workers with lice, while 1.2% harbored 2 lice and 0.2% had 3 lice; 4 .2% of the lice were on drones. A single bee louse was observed on 3,092 foraging honey bees sampled. One-hundred-seventeen lice were collected on 14,459 bees collected from the brood nest of the same hives. Control samples indicated a 14 to 15% loss of lice during sampling. Tests demonstrated that during visual observations of lice on bees only 49% of the lice present were observed. Fluctuation in louse population levels were similar to those found elsewhere in this study.
Feeding behavior and distribution of Varroa destructor on adult bees of Apis mellifera
(2022) Lamas, Zachary Siqueira; Hawthorne, David J; Entomology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Varroa destructor is a competent vector of honey bee viruses and the leading cause of colony losses worldwide. Much about its feeding behavior and distribution on adult bees remains unknown. This work shows that Varroa are promiscuous feeders of adult bees, actively switching from one host to another. Laboratory trials showed there is a large heterogeneity in the host switching rate with some Varroa switching infrequently while others switched at high rates. The consequences of Varroa feeding on adult bees is largely unknown because adult feeding has largely been overlooked. This work shows that there is a high relative risk of death from Varroa feedings. Adult workers die quickly without developing high levels of infection after being fed upon by an infectious Varroa, and confer lower risk to their non-parasitized nestmates than counterparts which were nestmates to longer lived parasitized bees. Further experiments showed communicable routes of virus transmission may explain these findings. Trophallaxis between adult workers allowed for the movement of the pathogen to naïve nestmates. These nestmates act as an infectious reservoir to naïve Varroa showing communicable transmission between hosts can influence the acquisition and subsequent vectoring of the same pathogen by the vector. Another social behavior, cannibalization, was shown to have the same influence on Varroa vectoring. Varroa were also shown to be susceptible to viral acquisition through shared feedings on adult bee and brood hosts. Naïve Varroa readily acquired and then transmitted deformed wing virus when sharing the same host with an infectious Varroa. Collectively this work exemplifies how host social behavior and Varroa-Varroa transmission routes can increase the risk of vectors becoming infectious. Varroa feedings and virus transmission on adult workers cannot describe one of the most glaring features of Varroa infestations. For a portion of the year Varroa aggregate predominantly on adult drones, largely ignoring the worker cohort. Parasite burden only shifts onto workers when drone production ceases.
A Survey of Bees in the University of Maryland Campus Area
(2021) Striegel, Theodore; Hawthorne, David; Entomology; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
I sampled the wild bee community in the University of Maryland campus area in College Park, Maryland for one season and combined those data with that from previous years. The protocol was modified to significantly expand the geographic scope of the survey, covering a much wider spatial range of the campus. This combined data was used to assess population trends, best practices, and determine whether or not the expanded protocol offers advantages in capturing local wild bee diversity and/or analytical benefits over the previous survey. New data allowed for characterization of the local wild bee community and discerned a landscape driver of spatial variance in local diversity. Appropriateness of different geographic parameters was found to depend on survey goals.

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