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Poster Session 2 12:00 - 14:00 Tuesday, 11Th June, 2019 Zia Ballroom Presentation Type Poster

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Poster Session 2 12:00 - 14:00 Tuesday, 11Th June, 2019 Zia Ballroom Presentation Type Poster Poster Session 2 12:00 - 14:00 Tuesday, 11th June, 2019 Zia Ballroom Presentation type Poster 111 Molecular Detection of Rickettsia in American Dog Ticks Collected Along the Platte River in South Central Nebraska Brandon Luedtke1, Julie Shaffer1, Estrella Monrroy1, Corey Willicott1, Travis Bourret2 1University of Nebraska-Kearney, Kearney, USA. 2Creighton University School of Medicine, Omaha, USA Abstract Dermacentor variabilis is the predominant tick species in Nebraska and is presumed to be the primary vector of Rickettsia rickettsii associated with Nebraskans that have contracted Rocky Mountain spotted fever. Interestingly, cases of Rocky Mountain spotted fever in Nebraska have increased on a year over year basis, yet the prevalence of D. variabilis vectoring R. rickettsii has not been established for Nebraska. Here we sought to set a baseline for the prevalence of D. variabilis vectoring R. rickettsii and other spotted fever group (SFG) rickettsiae. Over a 3 year period, D. variabilis were collected along the Platte River in south central Nebraska. Individual tick DNA was analyzed using endpoint PCR to identify ticks carrying SFG rickettsiae. A total of 927 D. variabilis were analyzed by PCR and 38 (4.1%) ticks tested positive for SFG rickettsiae. Presumptive positives were sequenced to identify the Rickettsia species, of which 29 (76%) were R. montanensis, 5 (13%) were R. amblyommatis, 4 (11%) were R. bellii, and R. rickettsii was not detected. These data indicate that R. rickettsii is likely at a low prevalence in south central Nebraska and spillover of R. amblyommatis into D. variabilis is occurring likely due to the invasive lone star tick (Amblyoma americanum). In addition, our data suggest that R. montanensis and R. amblyommatis could be associated with the increase in SFG rickettsiosis in Nebraska. This information will be of value to clinicians and the general public for evaluating diagnosis of disease and risk associated environmental exposure, respectively. 101 Dynamic Gene Expression of Cat Flea-Derived Salivary Gland-Secreted Factors during Rickettsia felis Infection Monika Danchenko, Hanna Laukaitis, Kevin Macaluso Vector-borne Disease Laboratories, Department of Pathobiological Sciences, Louisiana State University, Baton Rouge, USA Abstract Hematophagous insect saliva contains multiple biomolecules with anticlotting, vasodilatory, and immunomodulatory activities. Moreover, while feeding on host organism, bloodsucking arthropods also secrete proteins to facilitate microbial survival, growth, and transmission. The cat flea, Ctenocephalides felis, is a recognized arthropod vector transmitting several human pathogens, including Rickettsia spp. The sialotranscriptome of the cat flea was already characterized; however, the exact role of salivary factors and the molecular interaction between flea-borne rickettsiae and their insect host are still largely unknown. We hypothesize that Rickettsia felis modulates gene expression in cat fleas, thus regulating biomolecules essential for the successful horizontal bacterial transmission. Therefore, we infected cat fleas with R. felis to analyze differential transcription of selected salivary gland-derived factors. In our study, newly emerged adult fleas were fed bovine blood for 14 days. For infection assay, R. felis (strain LSU) was propagated in cell culture. Female and male salivary glands were microdissected separately from both control and infected groups at different time points upon inoculation. Subsequently, total RNA was extracted from salivary glands and the transcripts obtained by cDNA synthesis were subjected to real-time qPCR for gene expression analysis. The relative abundance of transcripts, coding putative salivary secreted factors in the infected cat fleas, was calculated using 2-ΔΔCt method with elongation factor 1-α as a reference gene and compared to the uninfected flea group. Based on the expression profiles, we suggest there are flea-specific compounds responsive to rickettsial infection. Their role in R. felis transmission is under current study. 98 Role of Sca4 in the Dissemination and Transmission of Rickettsia parkeri in Amblyomma maculatum Chanakan Suwanbongkot1, Chanida Fongsaran1, Ingeborg Langohr2, Rebecca L. Lamason3, Kevin Macaluso1 1Vector-Borne Disease Laboratories, Department of Pathobiological Science, School of Veterinary Medicine, Louisiana State University, Baton Rouge, USA. 2Department of Pathobiological Science, School of Veterinary Medicine, Louisiana State University, Baton Rouge, USA. 3Department of Biology Massachusetts Institute of Technology, Massachusetts, USA Abstract Rickettsia parkeri, a member of spotted fever group Rickettsia, is believed to disseminate to host tissue by a cell-to-cell spread mechanism which involves actin base motility to provide a physical force to spread. More recently, it has been demonstrated that R. parkeri uses an alternate mechanism for cell-to-cell spread by secreting effector sca4 to modulate protrusion and engulfment by manipulating vinculin-dependent intercellular tension. However, the interactions of sca4 in the tick infection model are unknown. Using R. parkeri lacking functional Sca4 (R. parkeri sca4::tn), we compared replication, growth kinetics and cell-to-cell spread in vitro, in tick and mammalian cells. In order to assess the role of Sca4 in dissemination in the tick host and the transmission in vivo, Rickettsia-free Amblyomma maculatum, the natural vector of R. parkeri, was exposed to wild type or R. parkeri sca4::tn bacteria. Individual tick saliva and tissues including salivary glands, mid gut and ovaries, were analyzed by qPCR and IFA at 3 and 7 days post exposure after blood meal acquisition. A portion of the ticks were allowed to feed to repletion and subsamples of each stage of F1 from each cohort were assessed for transovarial and transstadial transmission by qPCR. Blood from vertebrate hosts was collected at the same time points the ticks were collected. Blood, testis, and skin were assessed for infection using ELISA and histopathology, respectively. The preliminary data suggest the absence of Sca4 gene reduces Rickettsia infection in the tick host. 94 Comparative proteomics of different developmental stages of the Rickettsiales Orientia tsutsugamushi Jantana Wongsantichon1, Radoslaw Mikolaj Sobota2, Jeanne Salje1,3,4 1Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. 2Institute of Molecular and Cell Biology, A*STAR, Singapore, Singapore. 3Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom. 4Public Health Research Institute, Rutgers Biomedical and Health Science, Newark, New Jersey, USA Abstract Orientia tsutsugamushi (Ot) is an obligate intracellular bacterium and the causative agent of the human disease scrub typhus. The cellular infection cycle of Ot lasts 5-7 days in adherent fibroblast cells, and we have recently described how the bacterium transitions through 5 distinct stages during this infection: early entry, pre-replicative, replicative, maturation and extracellular (described in a separate poster by Suparat Giengkam et. al.). Here, we have isolated Ot populations in three of these stages (replicative, maturation and extracellular) and compared their protein profiles using tandem mass spectrometry. We have identified proteins whose expression levels are correlated with specific stages, and these include specific surface autotransporter proteins and ankyrin repeat proteins upregulated in the maturation and extracellular stages, as well as a number of tRNA- and ribosome-related genes also upregulated in the stages. These findings provide testable hypotheses about the mechanisms that this obligate intracellular bacterium uses in transitioning to a dedicated extracellular state that is primed for infection of new host cells. 92 Anaplasma marginale infection dynamics in an endemic region Roberta Koku1,2, David Herndon3, Johannesty Avillan1,2, James Futse4,1, Susan M. Noh3,2,1 1Paul G. Allen School of Global Animal Health, Pullman, USA. 2College of Veterinary Microbiology and Pathology, Washington State University, Pullman, USA. 3United States Department of Agriculture, Pullman, USA. 4College of Basic and Applied Science, University of Ghana, Accra, Ghana Abstract Anaplasma marginale, is a tick-borne, bacterial pathogen of cattle, which causes significant losses to producers worldwide. Cattle in high prevalence endemic regions of Central America are persistently infected with multiple strains of A. marginale. Multi-strain infections are important because they can provide selective pressure for altered transmission and virulence phenotypes. Multi-strain infections occur via co-infection, in which multiple strains are acquired prior to the development of an adaptive immune response, or superinfection, in which a strain is acquired after the development of the adaptive immune response. The latter scenario requires greater genetic diversity. Under natural transmission conditions, it is unknown if strains are predominantly acquired through co-infection or super-infection. Due to the high genetic diversity among A. marginale strains, we hypothesize that A. marginale strains are primarily acquired by superinfection. We will address this question by tracking the acquisition of A. marginale strains over time under natural transmission conditions in a region of Ghana with high A. marginale prevalence and high transmission pressure. Twenty-five
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