Flies are diverse models for the study of the evolution of the parasitic habit. Multiple repeated origins of specialized feeding in flies can reveal the molecular mechanisms underlying adaptive changes that both produce and sustain the parasitic lifestyle. With 1525 described species the blow flies (family Calliphoridae), are an ideal candidate for comparative study of the genetic and behavioral determinants of feeding adaptations. In this family of flies, different species have been recorded feeding on living tissues of a vertebrates, living tissues of invertebrates; decaying organic matter-- especially carrion or wound tissue; and blood. Considering the feeding habits of their larvae, blow flies are classified in: necrosaprophagous species, which feed on decomposing tissue; facultative ectoparasites, which feed on dead organic matter (necrophagous), or infest necrotic tissues of living vertebrates; and obligate parasites, which feed only on the living tissues of their hosts. For this reason, they are important as indicator organisms in forensic cases, are major agricultural pests of livestock, and they play a major role in nearly all terrestrial ecosystems as decomposers of organic material. The origin and evolutionary history of diverse specialized feeding habits in Calliphoridae are still vastly understudied, and are limited by a lack of phylogenetic, genetic and ecological information. With this project, an established team of evolutionary, ecological and genomic scientists will collaboratively investigate the causes and consequences of trophic specialization as a driver of species diversity across three integrated dimensions of blow fly research: (1) phylogenetic: reconstructing the history of blow fly evolutionary relationships and mapping the transitions of feeding habits that occurred repeatedly in the family; (2) functional, quantifying the index of preference for different food sources, and testing the effects of selection on feeding preferences; and investigating the role of the blow fly gut microbiome in mediating diet adaptations and maintaining diverse biotic interactions; and (3) genetic/genomic, identifying genes and gene regions associated with specific specialized feeding habits and using an established CRISPR Cas9 gene editing system to test hypotheses about the lability, maintenance, and control of feeding strategies in diverse blow fly models. The project includes a novel cross laboratory training and data collection for graduate students and postdocs between NCSU and Sao Paulo, Brazil, the development of an on-line data resource of genomic, taxonomic and ecological information on blow flies, and participation in multi-institutional training and research opportunities that include resources and outreach to individuals and communities that are underrepresented in science.

Project is in support of PSI. Drs. Huerta, Baltzegar, and Gold are pleased to submit this internal pre-proposal to NC State in support of their commitment and desire to write and submit a full-length proposal to the USDA-NIFA Equipment Grant Program, where they will lead a team of 26 other faculty requesting funds to acquire a Whole Genome Sequel IIe System sequencer from PabBio. The PacBio Sequel IIe is the current state-of-the-art technology for high-quality sequencing of genomes, transcriptomes, and epigenomes. If successful, this would be the first Sequel IIe in the University and possibly the state. The PacBio Sequel IIe system (product code 101-986-400) is priced at $525,000.00; The on-site system training (product code 100-125- 100) is $12,500; and the necessary uninterrupted power supply (UPS) (product code 100-609-000) is $7,000. The total for the instrument, installation and training for the instrument is $544,500.00. This sum is well above the funding limit for the USDA-NIFA EGP. However, Dr. Huerta has been in conversation with Dr. Nicole Newell, Sequencing Application Specialist at PacBio, to negotiate a 10% discount on the above listed sale price to bring the price down to $490,050 (see quote attached and letter of support from PacBio���s willingness to collaborate with us to make this grant and purchase a reality).

The Mikaelyan lab and Elysia Creative Biology are collaborating on a project to mitigate atmospheric methane. The lab will leverage its expertise in methanogenesis and methanotrophy in insects and other environments to aid in construct design. Additionally, they will conduct enzyme discovery and perform analytical chemistry analyses.

Methane, a greenhouse gas 84 times more potent than carbon dioxide. Livestock digestion is the predominant single contributor to the methane budget, contributing a remarkable 5% to the total greenhouse gas budget. Feed additives like bromoform from red seaweeds offer solutions, but their application been stymied by issues such as high costs and complex production needs. Our innovative approach synthesizes bromoform in terrestrial plant seeds, making methane reduction in livestock feasible and cost-effective. By infusing this technology into staple crops like corn, we leverage existing agricultural systems for scalability and affordability. The heart of our innovation is a transgenic pathway that allows land plants to produce bromoform. Preliminary tests on the model plant Arabidopsis have shown these genetically modified plants accumulate bromoform at levels capable of offsetting 98% of methane emissions from cattle. Ensuring safety, bromoform production is localized within seed tissues, suggesting exciting revelations about plant metabolism. This breakthrough, besides its commercial viability, also expands our understanding of plant molecular biology. Importantly, our technology addresses the urgent issue of climate change. If adopted globally, it could substantially diminish livestock methane emissions, which currently eclipse emissions from air travel, shipping, and deforestation combined. With many institutions pledging methane reductions, our innovation not only presents environmental solutions but also taps into a combined market potential exceeding $100 billion, aligning economic growth with environmental stewardship.

Flies carry an extensive range of pathogenic microorganisms, both internally and externally, that are critically important to public health, veterinary medicine, and food production. By feeding on and breeding in decaying organic matter such as human food, feces, and animal carcasses, flesh flies, house flies, and blowflies are frequently implicated as vectors of clinically relevant viral and bacterial pathogens, including those carrying genes encoding resistance to clinically important antibiotics. However, very few studies have examined the role of the microbiomes associated with these so called "filth flies" in the transmission of potential human and veterinary pathogens. Our study will draw on principles of entomology, disease ecology, forensics, bacteriology and virology to address the question of disease transmission by blowflies. Using sea gull carcasses, our experiments will track the incorporation and maintenance of potential pathogens (bacteria and viruses) in microbiomes of filth flies in natural environments.