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Christopher J. Pappas, Ph.D. - Manhattanville College. Purchase, NY, US

Christopher J. Pappas, Ph.D. Christopher J. Pappas, Ph.D.

Associate Professor and Director of Laboratory Safety, Biology | Manhattanville College

Purchase, NY, UNITED STATES

An expert in Borrelia burgdorferi, the etiologic agent of Lyme disease.

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Biography

Classically trained as a molecular microbiologist, Dr. Pappas teaches students in lectures and laboratories within the disciplines of general biology, parasitology, genetics, molecular biology, biochemistry, and nutrition. His research has included studies involving Borrelia burgdorferi, the etiologic agent of Lyme disease, in which he focused specifically on the the survival of the bacteria when inside the tick. His current research interests include genetic mechanisms of survival within saprophytic Leptospira, as well as other vector-borne and enzootic diseases. Outside of the laboratory and classroom, Dr. Pappas is involved in community activities such as judging at annual science fairs and working within Manhattanville College’s Organic Community Garden.

Areas of Expertise (4)

Lyme Disease

Ticks and Tick Borne Diseases

Leptospirosis

Lab Safety / OSHA Regulations

Accomplishments (5)

Faculty Member of the Year Award.

2013 Awarded by the Student Government Association, Manhattanville College

Martha Lucas Pate, Ph.D. Memorial Award.

2011 For demonstration of academic excellence and leadership in social and humane concerns in medicine, science, and health

1st Place, Poster Presentation.

2010 “Borrelia burgdorferi Glycerol-3-Phosphate Dehydrogenase (GlpD) is Important for Spirochete Maintenance in the Tick.”, New York Medical College 22nd Annual Graduate Student Research Forum, Valhalla, NY

1 st Place, Oral Presentation

2009 “Borrelia burgdorferi Glycerol-3-Phosphate Dehydrogenase (GlpD) Is Important for Spirochete Maintenance in the Tick.”, New York Medical College 21st Annual Graduate Student Research Forum, Valhalla, NY

2nd Place, Poster Presentation.

2008 “Borrelia burgdorferi Glycerol-3-Phosphate Dehydrogenase (GlpD) is Involved in Temperature-Dependent Stationary Phase Adaptation.” New York Medical College 20th Annual Graduate Student Research Forum, Valhalla, NY

Education (3)

New York Medical College: Ph.D., Microbiology & Immunology 2011

New York Medical College: M.S., Microbiology & Immunology 2006

Syracuse University: B.S., Psychology, Biology and Neuroscience 2002

Affiliations (7)

  • Member, Council on Undergraduate Research
  • Full Member, Sigma Xi Scientific Research Society
  • Member, American Society for Microbiology, New York City Branch
  • Member, American Society for Microbiology
  • Member, New York Academy of Sciences
  • Chair, Faculty Budget Committee
  • Founder and Chair, Laboratory Safety Committee, Manhattanville College

Selected Event Appearances (5)

Identification of Molecular Mechanisms of Adaptation to Osmotic Stress in Leptospira biflexa

Gordon Research Conference: Biology of Spirochetes, January, 2018  Ventura, CA

An Enhancer Binding Protein Affects RpoN Regulated Gene Expression in Leptospira interrogans

Gordon Research Conference: Biology of Spirochetes, January, 2016  Ventura, CA

Development of Genetic Manipulation Tools for Use in Leptospira spp. has provided Insight into the Biology of Pathogenic Leptospires

Gordon Research Seminar: Biology of Spirochetes, January, 2016  Ventura, CA

Development of Genetic Manipulation Tools for Use in Leptospira spp. has Provided Insight into the Biology of Pathogenic Leptospires

6th Annual Young Researchers in Life Sciences Conference, May, 2015  Paris, France

Formate Hydrogenlyase Activator (FhlA) Enhances RpoN Regulated Gene Expression in Leptospira interrogans

7th International Conference on Emerging Zoonosis, October, 2014.  Berlin, Germany

Selected Articles (5)

A novel ex vivo biosassay suggests DEET is an effective repellent of Rhipicephalus sanguineus Journal of Agricultural and Urban Entomology

Philip Meade, Alfa L. Abate, Jason Pavo, Anna K. Yeung-Cheung, Christopher J. Pappas

2017 Ticks are vectors that pose a threat to public health. N,N-diethyl-3-methylbenzamide (DEET) is commonly applied as a repellent to prevent attachment of ticks to humans and animals. Typical commercially available repellents contain between 5–100% DEET. Lower concentrations of DEET may be necessary to minimize potential health risks associated with DEET. To characterize the repellency of low concentrations of DEET, we performed an in vitro vertical bioassay, and developed a novel ex vivo vertical bioassay using porcine skin for use with the adult brown dog tick, Rhipicephalus sanguineus (Latreille) (Acari: Ixodidae). DEET applied at concentrations of 0.19% in vitro and 12.5% ex vivo immediately after application, and at 0.38% in vitro and 40% ex vivo at 4 h after application, repelled over 90% of ticks. In both in vitro and ex vivo assessments, and at both 0 and 4 h post application, the repellency against female ticks was similar to that against male ticks. This study demonstrates that concentrations of DEET lower than those in commercial repellents may provide sufficient repellency when potential tick exposure occurs shortly after application. Additionally, the development of a porcine ex vivo bioassay provides an alternative assessment tool for future repellency studies.

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Genome-Wide Transcriptional Start Site Mapping and sRNA Identification in the Pathogen Leptospira Interrogans Frontiers in Cellular and Infection Microbiology.

Christopher J. Pappas et al.

2017 Leptospira are emerging zoonotic pathogens transmitted from animals to humans typically through contaminated environmental sources of water and soil. Regulatory pathways of pathogenic Leptospira spp. underlying the adaptive response to different hosts and environmental conditions remains elusive. In this study, we provide the first global Transcriptional Start Site (TSS) map of a Leptospira species. RNA was obtained from the pathogen Leptospira interrogans grown at 30°C (optimal in vitro temperature) and 37°C (host temperature) and selectively enriched for 5′ ends of native transcripts. A total of 2865 and 2866 primary TSS (pTSS) were predicted in the genome of L. interrogans at 30 and 37°C, respectively. The majority of the pTSSs were located between 0 and 10 nucleotides from the translational start site, suggesting that leaderless transcripts are a common feature of the leptospiral translational landscape. Comparative differential RNA-sequencing (dRNA-seq) analysis revealed conservation of most pTSS at 30 and 37°C. Promoter prediction algorithms allow the identification of the binding sites of the alternative sigma factor sigma 54. However, other motifs were not identified indicating that Leptospira consensus promoter sequences are inherently different from the Escherichia coli model. RNA sequencing also identified 277 and 226 putative small regulatory RNAs (sRNAs) at 30 and 37°C, respectively, including eight validated sRNAs by Northern blots. These results provide the first global view of TSS and the repertoire of sRNAs in L. interrogans. These data will establish a foundation for future experimental work on gene regulation under various environmental conditions including those in the host.

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The EbpA-RpoN regulatory pathway of the pathogen Leptospira interrogans is essential for the survival in the environment Applied and Environmental Microbiology.

Christopher J. Pappas et al.

2017 Leptospira interrogans is the agent of leptospirosis, a reemerging zoonotic disease. It is transmitted to humans through environmental surface waters contaminated by the urine of mammals chronically infected by pathogenic strains able to survive in water for long periods. Little is known about the regulatory pathways underlying environmental sensing and host adaptation of L. interrogans during its enzootic cycle. This study identifies the EbpA-RpoN regulatory pathway in L. interrogans. In this pathway, EbpA, a σ54 activator and putative prokaryotic enhancer-binding protein (EBP), and the alternative sigma factor RpoN (σ54) control expression of at least three genes, encoding AmtB (an ammonium transport protein) and two proteins of unknown function. Electrophoresis mobility shift assay demonstrated that recombinant RpoN and EbpA bind to the promoter region and upstream of these three identified genes, respectively. Genetic disruption of ebpA in L. interrogans serovar Manilae virtually abolished expression of the three genes, including amtB in two independent ebpA mutants. Complementation of the ebpA mutant restored expression of these genes. Intraperitoneal inoculation of gerbils with the ebpA mutant did not affect mortality. However, the ebpA mutant had decreased cell length in vitro and had a significantly lowered cell density at stationary phase when grown with l-alanine as the sole nitrogen source. Furthermore, the ebpA mutant has dramatically reduced long-term survival ability in water. Together, these studies identify a regulatory pathway, the EbpA-RpoN pathway, that plays an important role in the zoonotic cycle of L. interrogans.

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Control of Gene Expression in Leptospira spp. by Transcription Activator-Like Effectors Demonstrates a Potential Role for LigA and LigB in Leptospira interrogans virulence Applied and Environmental Microbiology.

Christopher J. Pappas et al.

2015 Leptospirosis is a zoonotic disease that affects ∼1 million people annually, with a mortality rate of >10%. Currently, there is an absence of effective genetic manipulation tools for targeted mutagenesis in pathogenic leptospires. Transcription activator-like effectors (TALEs) are a recently described group of repressors that modify transcriptional activity in prokaryotic and eukaryotic cells by directly binding to a targeted sequence within the host genome. To determine the applicability of TALEs within Leptospira spp., two TALE constructs were designed. First, a constitutively expressed TALE gene specific for the lacO-like region upstream of bgaL was trans inserted in the saprophyte Leptospira biflexa (the TALEβgal strain). Reverse transcriptase PCR (RT-PCR) analysis and enzymatic assays demonstrated that BgaL was not expressed in the TALEβgal strain. Second, to study the role of LigA and LigB in pathogenesis, a constitutively expressed TALE gene with specificity for the homologous promoter regions of ligA and ligB was cis inserted into the pathogen Leptospira interrogans (TALElig). LigA and LigB expression was studied by using three independent clones: TALElig1, TALElig2, and TALElig3. Immunoblot analysis of osmotically induced TALElig clones demonstrated 2- to 9-fold reductions in the expression levels of LigA and LigB, with the highest reductions being noted for TALElig1 and TALElig2, which were avirulent in vivo and nonrecoverable from animal tissues. This study reconfirms galactosidase activity in the saprophyte and suggests a role for LigA and LigB in pathogenesis. Collectively, this study demonstrates that TALEs are effective at reducing the expression of targeted genes within saprophytic and pathogenic strains of Leptospira spp., providing an additional genetic manipulation tool for this genus.

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A replicative plasmid vector Allows efficient complementation of pathogenic Leptospira strains Applied and Environmental Microbiology.

Christopher J. Pappas et al.

2015 Leptospirosis, an emerging zoonotic disease, remains poorly understood because of a lack of genetic manipulation tools available for pathogenic leptospires. Current genetic manipulation techniques include insertion of DNA by random transposon mutagenesis and homologous recombination via suicide vectors. This study describes the construction of a shuttle vector, pMaORI, that replicates within saprophytic, intermediate, and pathogenic leptospires. The shuttle vector was constructed by the insertion of a 2.9-kb DNA segment including the parA, parB, and rep genes into pMAT, a plasmid that cannot replicate in Leptospira spp. and contains a backbone consisting of an aadA cassette, ori R6K, and oriT RK2/RP4. The inserted DNA segment was isolated from a 52-kb region within Leptospira mayottensis strain 200901116 that is not found in the closely related strain L. mayottensis 200901122. Because of the size of this region and the presence of bacteriophage-like proteins, it is possible that this region is a result of a phage-related genomic island. The stability of the pMaORI plasmid within pathogenic strains was tested by passaging cultures 10 times without selection and confirming the presence of pMaORI. Concordantly, we report the use of trans complementation in the pathogen Leptospira interrogans. Transformation of a pMaORI vector carrying a functional copy of the perR gene in a null mutant background restores the expression of PerR and susceptibility to hydrogen peroxide comparable to that of wild-type cells. In conclusion, we demonstrate the replication of a stable plasmid vector in a large panel of Leptospira strains, including pathogens. The shuttle vector described will expand our ability to perform genetic manipulation of Leptospira spp.

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