@article{chen_ponnusamy_mouhamadou_fodjo_sadia_affoue_deguenon_roe_2022, title={Internal and external microbiota of home-caught Anopheles coluzzii (Diptera: Culicidae) from Cote d'Ivoire, Africa: Mosquitoes are filthy}, volume={17}, ISSN={["1932-6203"]}, url={https://doi.org/10.1371/journal.pone.0278912}, DOI={10.1371/journal.pone.0278912}, abstractNote={Over the past 10 years, studies using high-throughput 16S rRNA gene sequencing have shown that mosquitoes harbor diverse bacterial communities in their digestive system. However, no previous research has examined the total bacteria community inside versus outside of mosquitoes and whether bacteria found on the outside could represent a potential health threat through mechanical transfer. We examined the bacterial community of the external surface and internal body of female Anopheles coluzzii adults collected from homes in Côte d’Ivoire, Africa, by Illumina sequencing of the V3 to V4 region of 16S rRNA gene. Anopheles coluzzii is in the Anopheles gambiae sensu lato (s.l.) species complex and important in the transmission of malaria. The total 16S rRNA reads were assigned to 34 phyla, 73 orders, 325 families, and 700 genera. At the genus level, the most abundant genera inside and outside combined were Bacillus, Staphylococcus, Enterobacter, Corynebacterium, Kocuria, Providencia, and Sphingomonas. Mosquitoes had a greater diversity of bacterial taxa internally compared to the outside. The internal bacterial communities were similar between homes, while the external body samples were significantly different between homes. The bacteria on the external body were associated with plants, human and animal skin, and human and animal infections. Internally, Rickettsia bellii and Rickettsia typhi were found, potentially of importance, since this genus is associated with human diseases. Based on these findings, further research is warranted to assess the potential mechanical transmission of bacteria by mosquitoes moving into homes and the importance of the internal mosquito microbiota in human health.}, number={12}, journal={PLOS ONE}, author={Chen, Kaiying and Ponnusamy, Loganathan and Mouhamadou, Chouaibou S. and Fodjo, Behi Kouadio and Sadia, Gba Christabelle and Affoue, France Paraudie Kouadio and Deguenon, Jean M. and Roe, R. Michael}, editor={Terenius, OlleEditor}, year={2022}, month={Dec} } @article{richardson_ponnusamy_roe_2022, title={Mechanical Acaricides Active against the Blacklegged Tick, Ixodes scapularis}, volume={13}, ISSN={["2075-4450"]}, url={https://www.mdpi.com/2075-4450/13/8/672}, DOI={10.3390/insects13080672}, abstractNote={Cases of Lyme disease in humans are on the rise in the United States and Canada. The vector of the bacteria that causes this disease is the blacklegged tick, Ixodes scapularis. Current control methods for I. scapularis mainly involve chemical acaricides. Unfortunately, ticks are developing resistance to these chemicals, and more and more, the public prefers non-toxic alternatives to chemical pesticides. We discovered that volcanic glass, ImergardTM WP, and other industrial minerals such as Celite 610 were efficacious mechanical insecticides against mosquitoes, filth flies, and agricultural pests. In this report, when 6–10- and 50–70-day old unfed I. scapularis nymphs were dipped for 1–2 s into Celite, the time to 50% mortality (LT50) was 66.8 and 81.7 min, respectively, at 30 °C and 50% relative humidity (RH). The LT50 was actually shorter at a higher 70% RH, 43.8 min. Scanning electron microscopy showed that the ticks were coated over most of their body surface, including partial to almost total coverage of the opening to their respiratory system. The other mechanical insecticide, Imergard, had similar efficacy against blacklegged unfed nymphs with an LT50 at 30 °C and 50% RH of 70.4 min. Although more research is needed, this study suggests that industrial minerals could be used as an alternative to chemical pesticides to control ticks and Lyme disease.}, number={8}, journal={INSECTS}, author={Richardson, Elise A. and Ponnusamy, Loganathan and Roe, R. Michael}, year={2022}, month={Aug} } @article{deguenon_dhammi_ponnusamy_travanty_cave_lawrie_mott_reisig_kurtz_roe_2021, title={Bacterial Microbiota of Field-Collected Helicoverpa zea (Lepidoptera: Noctuidae) from Transgenic Bt and Non-Bt Cotton}, volume={9}, ISSN={["2076-2607"]}, url={https://www.mdpi.com/2076-2607/9/4/878}, DOI={10.3390/microorganisms9040878}, abstractNote={The bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is an important agricultural pest in U.S. cotton and is managed using transgenic hybrids that produce insecticidal proteins from the bacterium, Bacillus thuringiensis (Bt). The reduced efficacy against H. zea caterpillars of Bt plants expressing Cry toxins is increasing in the field. In a first step towards understanding Bt cotton–bollworm–microbiota interactions, we investigated the internal bacterial microbiota of second–third stadium H. zea collected in the field from non-Bt versus Bt (WideStrike) cotton in close proximity (in North Carolina, USA). The bacterial populations were analyzed using culture-dependent and -independent molecular approaches. We found that WideStrike samples had a higher bacterial density and diversity per larva than insects collected from non-Bt cotton over two field seasons: 8.42 ± 0.23 and 5.36 ± 0.75 (log10 colony forming units per insect) for WideStrike compared to 6.82 ± 0.20 and 4.30 ± 0.56 for non-Bt cotton for seasons 1 and 2, respectively. Fifteen phyla, 103 families, and 229 genera were identified after performing Illumina sequencing of the 16S rRNA. At the family level, Enterobacteriaceae and Enterococcaceae were the most abundant taxa. The Enterococcaceae family was comprised mostly of Enterococcus species (E. casseliflavus and another Enterococcus sp.). Members of the Enterococcus genus can acidify their environment and can potentially reduce the alkaline activation of some Bt toxins. These findings argue for more research to better understand the role of cotton–bollworm–bacteria interactions and the impact on Bt toxin caterpillar susceptibility.}, number={4}, journal={MICROORGANISMS}, publisher={MDPI AG}, author={Deguenon, Jean M. and Dhammi, Anirudh and Ponnusamy, Loganathan and Travanty, Nicholas V and Cave, Grayson and Lawrie, Roger and Mott, Dan and Reisig, Dominic and Kurtz, Ryan and Roe, R. Michael}, year={2021}, month={Apr} } @article{lawrie_mitchell_deguenon_ponnusamy_reisig_pozo-valdivia_kurtz_roe_2022, title={Characterization of Long Non-Coding RNAs in the Bollworm, Helicoverpa zea, and Their Possible Role in Cry1Ac-Resistance}, volume={13}, ISSN={["2075-4450"]}, url={https://www.mdpi.com/2075-4450/13/1/12}, DOI={10.3390/insects13010012}, abstractNote={Multiple insect pest species have developed field resistance to Bt-transgenic crops. There has been a significant amount of research on protein-coding genes that contribute to resistance, such as the up-regulation of protease activity or altered receptors. However, our understanding of the role of non-protein-coding mechanisms in Bt-resistance is minimal, as is also the case for resistance to chemical pesticides. To address this problem relative to Bt, RNA-seq was used to examine statistically significant, differential gene expression between a Cry1Ac-resistant (~100-fold resistant) and Cry1Ac-susceptible strain of Helicoverpa zea, a prevalent caterpillar pest in the USA. Significant differential expression of putative long non-coding RNAs (lncRNAs) was found in the Cry1Ac-resistant strain (58 up- and 24 down-regulated gene transcripts with an additional 10 found only in resistant and four only in susceptible caterpillars). These lncRNAs were examined as potential pseudogenes and for their genomic proximity to coding genes, both of which can be indicative of regulatory relationships between a lncRNA and coding gene expression. A possible pseudogenic lncRNA was found with similarities to a cadherin. In addition, putative lncRNAs were found significantly proximal to a serine protease, ABC transporter, and CYP coding genes, potentially involved in the mechanism of Bt and/or chemical insecticide resistance. Characterization of non-coding genetic mechanisms in Helicoverpa zea will improve the understanding of the genomic evolution of insect resistance, improve the identification of specific regulators of coding genes in general (some of which could be important in resistance), and is the first step for potentially targeting these regulators for pest control and resistance management (using molecular approaches, such as RNAi and others).}, number={1}, journal={INSECTS}, author={Lawrie, Roger D. and Mitchell, Robert D. and Deguenon, Jean Marcel and Ponnusamy, Loganathan and Reisig, Dominic and Pozo-Valdivia, Alejandro Del and Kurtz, Ryan W. and Roe, Richard Michael}, year={2022}, month={Jan} } @article{luan_west_mccord_denhartog_shi_bettermann_li_travanty_mitchell_cave_et al._2021, title={Mosquito-Textile Physics: A Mathematical Roadmap to Insecticide-Free, Bite-Proof Clothing for Everyday Life}, volume={12}, ISSN={2075-4450}, url={http://dx.doi.org/10.3390/insects12070636}, DOI={10.3390/insects12070636}, abstractNote={Garments treated with chemical insecticides are commonly used to prevent mosquito bites. Resistance to insecticides, however, is threatening the efficacy of this technology, and people are increasingly concerned about the potential health impacts of wearing insecticide-treated clothing. Here, we report a mathematical model for fabric barriers that resist bites from Aedes aegypti mosquitoes based on textile physical structure and no insecticides. The model was derived from mosquito morphometrics and analysis of mosquito biting behavior. Woven filter fabrics, precision polypropylene plates, and knitted fabrics were used for model validation. Then, based on the model predictions, prototype knitted textiles and garments were developed that prevented mosquito biting, and comfort testing showed the garments to possess superior thermophysiological properties. Our fabrics provided a three-times greater bite resistance than the insecticide-treated cloth. Our predictive model can be used to develop additional textiles in the future for garments that are highly bite resistant to mosquitoes.}, number={7}, journal={Insects}, publisher={MDPI AG}, author={Luan, Kun and West, Andre J. and McCord, Marian G. and DenHartog, Emiel A. and Shi, Quan and Bettermann, Isa and Li, Jiayin and Travanty, Nicholas V. and Mitchell, Robert D., III and Cave, Grayson L. and et al.}, year={2021}, month={Jul}, pages={636} } @article{ponnusamy_sutton_mitchell_sonenshine_apperson_roe_2021, title={Tick Ecdysteroid Hormone, Global Microbiota/Rickettsia Signaling in the Ovary versus Carcass during Vitellogenesis in Part-Fed (Virgin) American Dog Ticks, Dermacentor variabilis}, volume={9}, ISSN={["2076-2607"]}, url={https://www.mdpi.com/2076-2607/9/6/1242}, DOI={10.3390/microorganisms9061242}, abstractNote={The transovarial transmission of tick-borne bacterial pathogens is an important mechanism for their maintenance in natural populations and transmission, causing disease in humans and animals. The mechanism for this transmission and the possible role of tick hormones facilitating this process have never been studied. Injections of physiological levels of the tick hormone, 20-hydroxyecdysone (20E), into part-fed (virgin) adult females of the American dog tick, Dermacentor variabilis, attached to the host caused a reduction in density of Rickettsia montanensis in the carcass and an increase in the ovaries compared to buffer-injected controls. This injection initiates yolk protein synthesis and uptake by the eggs but has no effect on blood feeding. Francisella sp. and R. montanensis were the predominant bacteria based on the proportionality in the carcass and ovary. The total bacteria load increased in the carcass and ovaries, and bacteria in the genus Pseudomonas increased in the carcass after the 20E injection. The mechanism of how the Rickettsia species respond to changes in tick hormonal regulation needs further investigation. Multiple possible mechanisms for the proliferation of R. montanensis in the ovaries are proposed.}, number={6}, journal={MICROORGANISMS}, author={Ponnusamy, Loganathan and Sutton, Haley and Mitchell, Robert D. and Sonenshine, Daniel E. and Apperson, Charles S. and Roe, Richard Michael}, year={2021}, month={Jun} } @article{deguenon_travanty_zhu_carr_denning_reiskind_watson_roe_ponnusamy_2019, title={Exogenous and endogenous microbiomes of wild-caught Phormia regina (Diptera: Calliphoridae) flies from a suburban farm by 16S rRNA gene sequencing}, volume={9}, ISSN={["2045-2322"]}, url={http://dx.doi.org/10.1038/s41598-019-56733-z}, DOI={10.1038/s41598-019-56733-z}, abstractNote={AbstractThe black blow fly,Phormia regina(Meigen) (Diptera: Calliphoridae) is one of the most abundant carrion flies in North America. Calliphorids are important in agriculture and animal production, veterinary sciences, forensics and medical entomology. While the role of flies in the epidemiology of human and animal diseases is an active area of research, little is known about the microorganisms associated with these insects. We examined the diversity of wild-caught black blow fly endogenous (internal body) and exogenous (external body) microbial communities using 16S rRNA gene sequencing. Overall, 27 phyla, 171 families and 533 genera were detected, and diversity was significantly higher (P< 0.05) on external body surfaces. At the genus level,Dysgonomonas,Ignatzschineria,Acinetobacter,Vagococcus,Myroides, andWohlfahrtiimonaswere predominant. Cloning and sequencing of nearly full-length fragments of the 16S rRNA gene showed that some of the species identified are known to be pathogenic to humans, animals, and plants.Myroides odoratimimusandAcinetobacter radioresistensare well-known, multi-drug resistant bacteria. These results provide a snapshot of the microbial communities harbored by adult black blow flies and call for more comprehensive studies to better characterize the role these flies may play in the transmission of pathogenic microorganisms.}, journal={SCIENTIFIC REPORTS}, author={Deguenon, Jean M. and Travanty, Nicholas and Zhu, Jiwei and Carr, Ann and Denning, Steven and Reiskind, Michael H. and Watson, David W. and Roe, R. Michael and Ponnusamy, Loganathan}, year={2019}, month={Dec} } @article{dhammi_krestchmar_ponnusamy_bacheler_reisig_herbert_del pozo-valdivia_roe_2016, title={Biology, pest status, microbiome and control of kudzu bug (Hemiptera: Heteroptera: Plataspidae): a new invasive pest in the U.S.}, volume={17}, ISSN={["1422-0067"]}, url={http://dx.doi.org/10.3390/ijms17091570}, DOI={10.3390/ijms17091570}, abstractNote={Soybean is an important food crop, and insect integrated pest management (IPM) is critical to the sustainability of this production system. In recent years, the introduction into the United States of the kudzu bug currently identified as Megacopta cribraria (F.), poses a threat to soybean production. The kudzu bug was first discovered in the state of Georgia, U.S. in 2009 and since then has spread to most of the southeastern states. Because it was not found in the North American subcontinent before this time, much of our knowledge of this insect comes from research done in its native habitat. However, since the U.S. introduction, studies have been undertaken to improve our understanding of the kudzu bug basic biology, microbiome, migration patterns, host selection and management in its expanding new range. Researchers are not only looking at developing IPM strategies for the kudzu bug in soybean, but also at its unique relationship with symbiotic bacteria. Adult females deposit bacterial packets with their eggs, and the neonates feed on these packets to acquire the bacteria, Candidatus Ishikawaella capsulata. The kudzu bug should be an informative model to study the co-evolution of insect function and behavior with that of a single bacteria species. We review kudzu bug trapping and survey methods, the development of bioassays for insecticide susceptibility, insecticide efficacy, host preferences, impact of the pest on urban environments, population expansion, and the occurrence of natural enemies. The identity of the kudzu bug in the U.S. is not clear. We propose that the kudzu bug currently accepted as M. cribraria in the U.S. is actually Megacopta punctatissima, with more work needed to confirm this hypothesis.}, number={9}, journal={Int. J. Mol. Sci}, publisher={MDPI AG}, author={Dhammi, Anirudh and Krestchmar, Jaap B. and Ponnusamy, Loganathan and Bacheler, Jack S. and Reisig, Dominic D. and Herbert, Ames and Del Pozo-Valdivia, Alejandro I. and Roe, R. Michael}, year={2016} } @article{mitchell_dhammi_wallace_hodgson_roe_2016, title={Impact of Environmental Chemicals on the Transcriptome of Primary Human Hepatocytes: Potential for Health Effects}, volume={30}, ISSN={1095-6670}, url={http://dx.doi.org/10.1002/JBT.21801}, DOI={10.1002/jbt.21801}, abstractNote={New paradigms for human health risk assessment of environmental chemicals emphasize the use of molecular methods and human‐derived cell lines. In this study, we examined the effects of the insect repellent DEET (N,N‐diethyl‐m‐toluamide) and the phenylpyrazole insecticide fipronil (fluocyanobenpyrazole) on transcript levels in primary human hepatocytes. These chemicals were tested individually and as a mixture. RNA‐Seq showed that 100 μM DEET significantly increased transcript levels (α = 0.05) for 108 genes and lowered transcript levels for 64 genes and fipronil at 10 μM increased the levels of 2246 transcripts and decreased the levels for 1428 transcripts. Fipronil was 21‐times more effective than DEET in eliciting changes, even though the treatment concentration was 10‐fold lower for fipronil versus DEET. The mixture of DEET and fipronil produced a more than additive effect (levels increased for 3017 transcripts and decreased for 2087 transcripts). The transcripts affected for all chemical treatments were classified by GO analysis and mapped to chromosomes. The overall treatment responses, specific pathways, and individual transcripts affected were discussed at different levels of fold‐change. Changes found in transcript levels in response to treatments will require further research to understand their importance in overall cellular, organ, and organismic function.}, number={8}, journal={Journal of Biochemical and Molecular Toxicology}, publisher={Wiley}, author={Mitchell, Robert D., III and Dhammi, Anirudh and Wallace, Andrew and Hodgson, Ernest and Roe, R. Michael}, year={2016}, month={Apr}, pages={375–395} } @article{fuente_waterhouse_sonenshine_roe_ribeiro_sattelle_hill_2016, title={Tick genome assembled: New opportunities for research on tick-host-pathogen interactions}, volume={6}, journal={Frontiers in Cellular and Infection Microbiology}, author={Fuente, J. and Waterhouse, R. M. and Sonenshine, D. E. and Roe, R. M. and Ribeiro, J. M. and Sattelle, D. B. and Hill, C. A.}, year={2016} }