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    Modeling co-infection of Ixodes tick-borne pathogens.
    Math Biosci Eng 2017 Oct/Dec 1;14(5-6):1301-1316
    Mathematics and Science College, Shanghai Normal University, Shanghai 200234, China
    Ticks, including the Ixodes ricinus and Ixodes scapularis hard tick species, are regarded as the most common arthropod vectors of both human and animal diseases in Europe and the United States capable of transmitting a large number of bacteria, viruses and parasites. Since ticks in larval and nymphal stages share the same host community which can harbor multiple pathogens, they may be co-infected with two or more pathogens, with a subsequent high likelihood of co-transmission to humans or animals. This paper is devoted to the modeling of co-infection of tick-borne pathogens, with special focus on the co-infection of Borrelia burgdorferi (agent of Lyme disease) and Babesia microti (agent of human babesiosis). Read More

    Characterization of Stress and Innate Immunity Resistance of Wild-type and Δp66 Borrelia burgdorferi.
    Infect Immun 2017 Nov 20. Epub 2017 Nov 20.
    Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Borrelia burgdorferi is a causative agent of Lyme disease, the most common arthropod-borne disease in the United States. B. burgdorferi evades host immune defenses to establish a persistent, disseminated infection. Read More

    Borrelia burgdorferi glycosaminoglycan binding protein, Bgp in the B31 strain is not essential for infectivity despite facilitating adherence and tissue colonization.
    Infect Immun 2017 Nov 20. Epub 2017 Nov 20.
    Department of Microbiology, Biochemistry and Molecular Genetics, Rutgers New Jersey Medical School, Newark, New Jersey, USA
    Lyme disease causing Borrelia burgdorferi is transmitted into the mammalian host by an infected tick bite. Successful infection relies on the ability of this extracellular pathogen to persist and colonize different tissues. B. Read More

    Unraveling Diagnostic Uncertainty Surrounding Lyme Disease in Children with Neuropsychiatric Illness.
    Child Adolesc Psychiatr Clin N Am 2018 Jan 21;27(1):27-36. Epub 2017 Oct 21.
    Division of Pediatric Emergency Medicine, Alpert Medical School of Brown University, Rhode Island Hospital - Hasbro Children's Hospital, 125 Whipple Street, UEMF Suite-3rd Floor, Providence, RI 02908, USA.
    Lyme disease is endemic in parts of the United States, including New England, the Atlantic seaboard, and Great Lakes region. The presentation has various manifestations, many of which can mimic psychiatric diseases in children. Distinguishing manifestations of Lyme disease from those of psychiatric illnesses is complicated by inexact diagnostic tests and misuse of these tests when they are not clinically indicated. Read More

    Ticks and the city - are there any differences between city parks and natural forests in terms of tick abundance and prevalence of spirochaetes?
    Parasit Vectors 2017 Nov 21;10(1):573. Epub 2017 Nov 21.
    Department of Parasitology, Institute of Zoology, Faculty of Biology, University of Warsaw, 1 Miecznikowa Street, 02-096, Warsaw, Poland.
    Background: Ixodes ricinus ticks are commonly encountered in either natural or urban areas, contributing to Lyme disease agents Borreliella [(Borrelia burgdorferi (sensu lato)] spp. and Borrelia miyamotoi enzootic cycles in cities. It is an actual problem whether urbanization affects pathogen circulation and therefore risk of infection. Read More

    The tick biocontrol agent Metarhizium brunneum (= M. anisopliae) (strain F52) does not reduce non-target arthropods.
    PLoS One 2017 20;12(11):e0187675. Epub 2017 Nov 20.
    Cary Institute of Ecosystem Studies, Sharon Turnpike, Millbrook, NY, United States of America.
    Previous studies have found that Met52®, which contains the entomopathogenic fungus Metarhizium brunneum, is effective in reducing the abundance of Ixodes scapularis, the tick vector for the bacterium causing Lyme disease and for other tick-borne pathogens. Given widespread interest in effective, safe methods for controlling ticks, Met52 has the potential to be used at increasing scales. The non-target impacts of Met52, as applied for tick control, have not yet been assessed. Read More

    Population-based Borrelia burgdorferi sensu lato seroprevalence and associated risk factors in Finland.
    Ticks Tick Borne Dis 2017 Nov 11. Epub 2017 Nov 11.
    Department of Health Security, National Institute for Health and Welfare, Helsinki, Finland.
    Lyme borreliosis (LB) is caused by Borrelia burgdorferi sensu lato (Bb-sl) and is the most common vector-borne disease in Europe. The objectives of this study were to determine the Bb-sl seroprevalence among the general Finnish adult population and to identify risk factors associated with Bb-sl-seropositive status. Two thousand sera from a nationwide health survey from 2011 were tested by whole-cell sonicate IgG ELISA, C6 peptide ELISA, and recomBead IgG 2. Read More

    Meningoencephalitis in a Royal Marine after skinning reindeer in Norway.
    J R Army Med Corps 2017 Nov 18. Epub 2017 Nov 18.
    Department of Anaesthesia and Intensive Care Medicine, Royal Centre for Defence Medicine, Birmingham, UK.
    Meningoencephalitis presenting in service personnel overseas may present a diagnostic challenge due to the broad range of potential differential diagnosis as well as the requirement for rapid assessment and treatment. A 25-year-old Royal Marine was evacuated to the Royal Centre for Defence Medicine in Birmingham, UK, with a history of rash consistent with erythema chronicum migrans, a seizure, and lymphocytic pleocytosis after skinning reindeer in Norway. Neuroborreliosis was suspected and empirical antibiotics were administered. Read More

    Vector-borne diseases and climate change: a European perspective.
    FEMS Microbiol Lett 2017 Nov 15. Epub 2017 Nov 15.
    European Centre for Disease Prevention and Control.
    Climate change has already impacted the transmission of a wide-range of vector-borne diseases in Europe, and it will continue to do so in the coming decades. Climate change has been implicated in the observed shift of ticks to elevated altitudes and latitudes, notably including the Ixodes ricinus tick species which is a vector for Lyme borreliosis and tick-borne encephalitis. Climate change is also thought to have been a factor in the expansion of other important disease vectors in Europe: Aedes albopictus (the Asian tiger mosquito), which transmits diseases such as Zika, dengue, and chikungunya, and Phlebotomus sandfly species, which transmits diseases including Leishmaniasis. Read More

    Seroreactivity to the C6 Peptide in Borrelia Miyamotoi Infections Occurring in the Northeastern United States.
    Clin Infect Dis 2017 Nov 15. Epub 2017 Nov 15.
    Division of Infectious Diseases, New York Medical College, Valhalla, NY 10595.
    Background: There are no Food and Drug Administration (FDA) approved diagnostic tests for Borrelia miyamotoi infection, an emerging tick-borne illness in the United States. The purpose of this study was to evaluate whether the FDA-approved C6 peptide ELISA currently used to diagnose Lyme disease may potentially serve as a diagnostic test for B. miyamotoi infections. Read More

    Impact of patient age on clinical features, serologic test reactivity and long-term outcome of culture-confirmed early Lyme disease.
    Diagn Microbiol Infect Dis 2017 Dec 19;89(4):300-302. Epub 2017 Sep 19.
    Division of Infectious Diseases, New York Medical College, Valhalla, NY 10595. Electronic address:
    Whether age at time of diagnosis impacts the clinical presentation or long-term outcome of early Lyme disease is unknown. Subjects enrolled in this study had culture-confirmed early Lyme disease and were followed up annually in a long-term prospective study. Subjects ≤50years of age were compared with those who were ≥51years of age at time of study entry. Read More

    A tiny tick can cause a big health problem.
    Indian J Ophthalmol 2017 Nov;65(11):1228-1232
    Former Professor, Electron Microscopy Centre, Plymouth University, England, United Kingdom.
    Ticks are tiny crawling bugs in the spider family that feed by sucking blood from animals. They are second only to mosquitoes as vectors of human disease, both infectious and toxic. Infected ticks spread over a hundred diseases, some of which are fatal if undetected. Read More

    Does more favourable handling of the cerebrospinal fluid increase the diagnostic sensitivity of Borrelia burgdorferi sensu lato-specific PCR in Lyme neuroborreliosis?
    Infect Dis (Lond) 2017 Nov 10:1-6. Epub 2017 Nov 10.
    f Department of Medical Microbiology , Sørlandet Hospital Trust , Kristiansand , Norway.
    Background: Tests for direct detection of Borrelia burgdorferi sensu lato (Bb) in Lyme neuroborreliosis (LNB) are needed. Detection of Bb DNA using PCR is promising, but clinical utility is hampered by low diagnostic sensitivity. We aimed to examine whether diagnostic sensitivity can be improved by the use of larger cerebrospinal fluid (CSF) volumes and faster handling of samples. Read More

    Surveillance for Lyme Disease - United States, 2008-2015.
    MMWR Surveill Summ 2017 Nov 10;66(22):1-12. Epub 2017 Nov 10.
    Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, CDC.
    Problem/condition: Lyme disease is the most commonly reported vectorborne disease in the United States but is geographically focal. The majority of Lyme disease cases occur in the Northeast, mid-Atlantic, and upper Midwest regions. Lyme disease can cause varied clinical manifestations, including erythema migrans, arthritis, facial palsy, and carditis. Read More

    Borrelia burgdorferi glycosaminoglycan-binding proteins: a potential target for new therapeutics against Lyme disease.
    Microbiology 2017 Nov 8. Epub 2017 Nov 8.
    5​Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
    The spirochete bacterium Borrelia burgdorferisensu lato is the causative agent of Lyme disease, the most common vector-borne disease in Europe and the United States. The spirochetes can be transmitted to humans via ticks, and then spread to different tissues, leading to arthritis, carditis and neuroborreliosis. Although antibiotics have commonly been used to treat infected individuals, some treated patients do not respond to antibiotics and experience persistent, long-term arthritis. Read More

    Multiple legumain isoenzymes in ticks.
    Int J Parasitol 2017 Nov 4. Epub 2017 Nov 4.
    Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice CZ-370 05, Czech Republic. Electronic address:
    By searching nucleotide databases for the North American Lyme disease vector, Ixodes scapularis, we have complemented the previously characterized European Ixodes ricinus legumain IrAE1 with a full set of nine analogous genes (isae1-9). Six of these were PCR confirmed as genes present in all tick genomes tested. The absolute mRNA copy number examined by quantitative (q)PCR enabled expression profiling and an absolute comparison of mRNA levels for individual I. Read More

    Health Information-Seeking Patterns of the General Public and Indications for Disease Surveillance: Register-Based Study Using Lyme Disease.
    JMIR Public Health Surveill 2017 Nov 6;3(4):e86. Epub 2017 Nov 6.
    National Institute for Health and Welfare, Helsinki, Finland.
    Background: People using the Internet to find information on health issues, such as specific diseases, usually start their search from a general search engine, for example, Google. Internet searches such as these may yield results and data of questionable quality and reliability. Health Library is a free-of-charge medical portal on the Internet providing medical information for the general public. Read More

    Analysis of recombinational switching at the antigenic variation locus of the Lyme spirochete using a novel PacBio sequencing pipeline.
    Mol Microbiol 2017 Nov 4. Epub 2017 Nov 4.
    Department of Biochemistry and Molecular Biology, Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
    The Lyme disease spirochete evades the host immune system by combinatorial variation of VlsE, a surface antigen. Antigenic variation occurs via segmental gene conversion from contiguous silent cassettes into the vlsE locus. Because of the high degree of similarity between switch variants and the size of vlsE, short-read NGS technologies have been unsuitable for sequencing vlsE populations. Read More

    Early Disseminated Lyme Disease Masquerading as Mononucleosis: A Case Report.
    J Emerg Med 2017 Oct 25. Epub 2017 Oct 25.
    Cooper University Hospital, Camden, New Jersey.
    Background: Disseminated Lyme disease can be difficult to diagnose, as it begins with nonspecific signs and symptoms, which, if not treated correctly, can lead to atrioventricular conduction blocks and meningitis. In addition, the diagnosis can be further complicated by potentially false-positive test results.

    Case Report: We report a case of early-disseminated Lyme disease presenting with Borrelia meningitis and concomitant Lyme carditis, which was misdiagnosed as mononucleosis. Read More

    Transcriptome Assessment of Erythema Migrans Skin Lesions in Patients with Early Lyme Disease Reveals Predominant Interferon Signaling.
    J Infect Dis 2017 Nov 1. Epub 2017 Nov 1.
    Department of Microbiology and Immunology, New York Medical College, Valhalla, New York, USA.
    Background: The most common clinical manifestation of early Lyme disease is the erythema migrans (EM) skin lesion that develops at the tick bite site typically between 7 and 14 days following infection with Borreliella burgdorferi. The host-pathogen interactions that occur in the skin may have a critical role in determining outcome of infection.

    Methods: Gene arrays were utilized to characterize the global transcriptional alterations in skin biopsy samples of EM lesions from untreated adult patients with Lyme disease in comparison to controls. Read More

    Rhode Island Lyme Disease Surveillance Summary 2014-2016.
    R I Med J (2013) 2017 Nov 1;100(11):41-44. Epub 2017 Nov 1.
    State Epidemiologist and Medical/Division Director, Division of Preparedness, Response, Infectious Disease, and Emergency Medical Services, and Clinical Assistant Professor, Department of Health Services, Policy, and Practice, Brown School of Public Health.
    [Full article available at http://rimed.org/rimedicaljournal-2017-11.asp]. Read More

    Biochemical characterization of Borrelia burgdorferi's RecA protein.
    PLoS One 2017 31;12(10):e0187382. Epub 2017 Oct 31.
    Department of Microbiology & Immunology, College of Medicine, University of Saskatchewan, Academic Health Sciences Building, Saskatoon, Saskatchewan, Canada.
    RecA plays key roles in DNA recombination, replication and repair. Mutation of recA in the Lyme disease spirochete, Borrelia burgdorferi, fails to produce some of the phenotypes expected from study of recA mutation in other organisms. 'Missing' recA phenotypes include a lack of growth or viability effects, including in the presence of DNA damage, and a lack of a role in vlsE antigenic variation and infectivity. Read More

    Assessing diagnostic coding practices among a sample of healthcare facilities in Lyme disease endemic areas: Maryland and New York - A Brief Report.
    Zoonoses Public Health 2017 Oct 30. Epub 2017 Oct 30.
    New York State Department of Health, Albany, NY, USA.
    The value of using diagnostic codes in Lyme disease (LD) surveillance in highly endemic states has not been well studied. Surveys of healthcare facilities in Maryland (MD) and New York (NY) regarding coding practices were conducted to evaluate the feasibility of using diagnostic codes as a potential method for LD surveillance. Most respondents indicated that their practice utilized electronic medical records (53%) and processed medical/billing claims electronically (74%). Read More

    To test or not to test? Laboratory support for the diagnosis of Lyme borreliosis.
    Clin Microbiol Infect 2017 Oct 24. Epub 2017 Oct 24.
    Department of Infectious Diseases, University Medical Centre Ljubljana, Ljubljana, Slovenia.
    As stated in the recent review antibodies are expected to develop in almost all patients within 6-8 weeks and the negative predictive value is very high. This is well supported in the literature. Thus, testing for borrelia specific antibodies may, as explained in detail in the review, effectively rule out Lyme borreliosis, and other diagnostic possibilities should be considered. Read More

    Selective Essential Oils from Spice or Culinary Herbs Have High Activity against Stationary Phase and Biofilm Borrelia burgdorferi.
    Front Med (Lausanne) 2017 11;4:169. Epub 2017 Oct 11.
    Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, United States.
    Although the majority of patients with acute Lyme disease can be cured with the standard 2-4 week antibiotic treatment, about 10-20% of patients continue suffering from chronic symptoms described as posttreatment Lyme disease syndrome. While the cause for this is debated, one possibility is that persister bacteria are not killed by the current Lyme antibiotics and remain active in the system. It has been reported that essential oils have antimicrobial activities and some have been used by patients with persisting Lyme disease symptoms. Read More

    Regulation of Gene and Protein Expression in the Lyme Disease Spirochete.
    Curr Top Microbiol Immunol 2017 Oct 25. Epub 2017 Oct 25.
    Department of Biology, University of Texas at San Antonio, 78249, San Antonio, TX, USA.
    The infectious cycle of Borrelia burgdorferi necessitates persistent infection of both vertebrates and ticks, and efficient means of transmission between those two very different types of hosts. The Lyme disease spirochete has evolved mechanisms to sense its location in the infectious cycle, and use that information to control production of the proteins and other factors required for each step. Numerous components of borrelial regulatory pathways have been characterized to date. Read More

    IrC2/Bf - A yeast and Borrelia responsive component of the complement system from the hard tick Ixodes ricinus.
    Dev Comp Immunol 2018 Feb 20;79:86-94. Epub 2017 Oct 20.
    Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice CZ-370 05, Czech Republic. Electronic address:
    Ticks possess components of a primordial complement system that presumably play a role in the interaction of the tick immune system with tick-borne pathogens and affect their transmission. Here we characterized a novel complement component, tagged as IrC2/Bf, from the hard tick Ixodes ricinus, the principal vector of Lyme disease in Europe. IrC2/Bf is a multi-domain molecule composed of 5-7 CCP modules, varied by alternative splicing, followed by a von Willebrand factor A domain and a C-terminal trypsin-like domain. Read More

    Parallelisms and Contrasts in the Diverse Ecologies of the Anaplasma phagocytophilum and Borrelia burgdorferi Complexes of Bacteria in the Far Western United States.
    Vet Sci 2016 Sep 22;3(4). Epub 2016 Sep 22.
    School of Veterinary Medicine, Medicine and Epidemiology, University of California, Davis, CA 95616, USA.
    Anaplasma phagocytophilum and Borrelia burgdorferi are two tick-borne bacteria that cause disease in people and animals. For each of these bacteria, there is a complex of closely related genospecies and/or strains that are genetically distinct and have been shown through both observational and experimental studies to have different host tropisms. In this review we compare the known ecologies of these two bacterial complexes in the far western USA and find remarkable similarities, which will help us understand evolutionary histories and coadaptation among vertebrate host, tick vector, and bacteria. Read More

    Selective intrathecal enrichment of G1m1-positive B cells in multiple sclerosis.
    Ann Clin Transl Neurol 2017 Oct 27;4(10):756-761. Epub 2017 Sep 27.
    Department of Immunology and Transfusion MedicineFaculty of MedicineUniversity of Oslo and Oslo University Hospital RikshospitaletOsloNorway.
    Immunoglobulin gamma (IgG) heavy chain genes are associated with susceptibility to multiple sclerosis (MS) and IgG levels in the cerebrospinal fluid (CSF). However, how these variants are implicated in disease mechanisms remains unknown. Here, we show that proliferating plasmablasts expressing the G1m1 allotype of IgG1 are selectively enriched in CSF of G1m1/G1m3 heterozygous MS patients, whereas plasmablasts expressing either G1m1 or G1m3 are evenly distributed in blood. Read More

    Variable clinical presentations of babesiosis: A case series.
    Nurse Pract 2017 Nov;42(11):1-7
    Pamela Paparone is an NP at the Atlantic County Health Department, Northfield, N.J. Philip W. Paparone is a communicable disease physician at the Atlantic County Health Department, Northfield, N.J.
    Human babesiosis continues to spread in multiple regions of the United States. It is transmitted by Ixodes species ticks, as are Lyme disease and anaplasmosis. Its variable clinical presentations, together with serologic detection limitations, require that a high index of clinical suspicion be present for prompt diagnosis. Read More

    Weak Organic Acids Decrease Borrelia burgdorferi Cytoplasmic pH, Eliciting an Acid Stress Response and Impacting RpoN- and RpoS-Dependent Gene Expression.
    Front Microbiol 2017 29;8:1734. Epub 2017 Sep 29.
    Laboratory of Zoonotic Pathogens, Gene Regulation Section, Division of Intramural Research, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
    The spirochete Borrelia burgdorferi survives in its tick vector, Ixodes scapularis, or within various hosts. To transition between and survive in these distinct niches, B. burgdorferi changes its gene expression in response to environmental cues, both biochemical and physiological. Read More

    Recombinant E. coli Dualistic Role as an Antigen-adjuvant Delivery Vehicle for Oral Immunization.
    Methods Mol Biol 2018 ;1690:347-357
    US BIOLOGIC, Memphis, TN, 38103, USA.
    Escherichia coli is the mainstay tool for fundamental microbiology research due to its ease of cultivation and safety. Auxotrophic strains of the K-12 and B lineages of E. coli are the organisms of choice to produce recombinant proteins. Read More

    Xenodiagnosis Using Ixodes scapularis Larval Ticks in Humans.
    Methods Mol Biol 2018 ;1690:337-346
    Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 10 Center Drive BG 10 RM 12C118 MSC 1899, Bethesda, MD, 20892-1899, USA.
    Xenodiagnosis is the use of a natural vector to detect the presence of an organism, and xenodiagnosis using Ixodes ticks has long been used by entomologists in Lyme disease research to provide evidence of the host's infectious status with Borrelia burgdorferi. We developed the methodology and performed the first human research study using uninfected larval Ixodes scapularis ticks to assess evidence of B. burgdorferi infection. Read More

    Magnetic Isolation of Phagosomes Containing Toll-Like Receptor Ligands.
    Methods Mol Biol 2018 ;1690:329-336
    Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA, 02111, USA.
    Phagosomal compartments are critical in microbial defense as vesicles that degrade invading organisms. In a broader context, vesicular trafficking plays an important role in shuttling many different types of cargo that are critical for proper function of the cell. Endosomal and phagosomal vesicles are thus important locations for the assembly of intracellular signaling platforms that mediate host responses to invasive pathogens such as Borrelia burgdorferi. Read More

    In Vitro Models of Cutaneous Inflammation.
    Methods Mol Biol 2018 ;1690:319-327
    EA7290: Virulence Bactérienne Précoce: Groupe Borréliose de Lyme, Université de Strasbourg, 3 rue Koeberlé, 67000, Strasbourg, France.
    The skin plays an essential role in the transmission of Lyme borreliosis since it is the first interface between the Ixodes tick and the host during the inoculation of Borrelia burgdorferi sensu lato. A better understanding of the inflammatory reaction at this location is key to develop better strategies (e.g. Read More

    Measuring Borrelia burgdorferi Motility and Chemotaxis.
    Methods Mol Biol 2018 ;1690:313-317
    Department of Oral Biology, Microbiology and Immunology, State University of New York at Buffalo, 3435 Main Street, Buffalo, NY, USA.
    Swimming plate, cell motion tracking, and capillary tube assays are very useful tools to quantitatively measure bacterial motility and chemotaxis. These methods were modified and applied to study Borrelia burgdorferi motility and chemotaxis. By using these methods, numerous motility and chemotaxis mutants have been characterized and several chemoattractants were identified. Read More

    Phagocytosis Assays for Borrelia burgdorferi.
    Methods Mol Biol 2018 ;1690:301-312
    CIC bioGUNE, Macrophage and Tick Vaccine Laboratory, Parque Tenolágico de Bizkaia, 48160, Derio, Bizkaia, Spain.
    Phagocytosis of Borrelia burgdorferi, the causative agent of Lyme disease, is a poorly understood process, despite its importance during the host immune response to infection. Thus, macrophages infiltrate the infected tissues, including the base of the heart and phagocytose the spirochete, therefore contributing to their elimination from infected tissues and leading to inflammation. An impaired bacterial clearance will result in bacterial persistence that may interfere with normal physiology of the heart, such as electrical signals from the heart, resulting in an impaired coordination of the beating of the heart or "heart block. Read More

    Two Photon Intravital Microscopy of Lyme Borrelia in Mice.
    Methods Mol Biol 2018 ;1690:279-290
    Department of Internal Medicine/Section of Rheumatology, Yale University School of Medicine, New Haven, CT, 06520, USA.
    Two-photon intravital microscopy is a powerful tool that allows visualization of cells in intact tissues in a live animal in real time. In recent years, this advanced technology has been applied to understand pathogen-host interactions using fluorescently labeled bacteria. In particular, infectious fluorescent transformants of the Lyme disease spirochete Borrelia burgdorferi, an Ixodes tick-transmitted pathogen, have been imaged by two-photon intravital microscopy to study bacterial motility and interactions of the pathogen with feeding ticks and host tissues. Read More

    Detection of Bioluminescent Borrelia burgdorferi from In Vitro Cultivation and During Murine Infection.
    Methods Mol Biol 2018 ;1690:241-257
    Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, Riverside Pkwy, Bryan, Texas, 77807, USA.
    Borrelia burgdorferi, etiologic agent of Lyme disease, is the leading tick-borne disease in the United States with approximately 300,000 cases diagnosed annually. Disease occurs in stages beginning localized infection at the site of a tick bite and progresses to disseminated infection when antibiotic treatment is not administered in a timely manner. A multi-systemic infection develops following dissemination to numerous immunoprotective tissues, such as the heart, bladder, and joints, resulting in late Lyme disease. Read More

    Generation of Conditional Mutants in Borrelia burgdorferi.
    Methods Mol Biol 2018 ;1690:225-239
    Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, 4301 West Markham Street, Little Rock, AR, 72205, USA.
    Mutational studies aimed at characterizing the function(s) of bacterial genes required for growth or viability are constrained by the inability to generate deletion strains lacking the gene of interest. To circumvent this limitation, it is possible to generate conditional mutants in which a copy of the gene of interest is introduced into the bacteria to compensate for the loss of the native allele. Expression of the non-native copy of the target gene is typically under control of an inducible promoter, which allows for controllable and regulated production of the gene of interest. Read More

    Genome-Wide Mutagenesis in Borrelia burgdorferi.
    Methods Mol Biol 2018 ;1690:201-223
    Department of Pathology and Laboratory Medicine, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX, USA.
    Signature-tagged mutagenesis (STM) is a functional genomics approach to identify bacterial virulence determinants and virulence factors by simultaneously screening multiple mutants in a single host animal, and has been utilized extensively for the study of bacterial pathogenesis, host-pathogen interactions, and spirochete and tick biology. The signature-tagged transposon mutagenesis has been developed to investigate virulence determinants and pathogenesis of Borrelia burgdorferi. Mutants in genes important in virulence are identified by negative selection in which the mutants fail to colonize or disseminate in the animal host and tick vector. Read More

    Genetic Transformation and Complementation.
    Methods Mol Biol 2018 ;1690:183-200
    Division of Biological Sciences, University of Montana, Missoula, MT, USA.
    The disciplines of Borrelia (Borreliella) burgdorferi microbiology and Lyme disease pathogenesis have come to depend on the genetic manipulation of the spirochete. Generating mutants in these recalcitrant bacteria, while not straightforward, is routinely accomplished in numerous laboratories, although there are several crucial caveats to consider. This chapter describes the design of basic molecular genetic experiments as well as the detailed methodologies to prepare and transform competent cells, select for and isolate transformants, and complement or genetically restore mutants. Read More

    Selection of Borrelia burgdorferi Promoter Sequences Active During Mammalian Infection Using In Vivo Expression Technology.
    Methods Mol Biol 2018 ;1690:137-154
    Division of Immunity and Pathogenesis, Burnett School of Biomedical Sciences, University of Central Florida College of Medicine, 6900 Lake Nona Blvd., Orlando, FL, 32827, USA.
    In vivo expression technology (IVET) has been applied to a variety of organisms to identify active promoters in specific environments or growth conditions of interest. Here, we describe modifications to employ this genome-wide screening method for Borrelia burgdorferi, the Lyme disease spirochete, during an active murine infection. Utilization of this technique provides valuable insights into the B. Read More

    Artificial Infection of Ticks with Borrelia burgdorferi Using a Microinjection Method and Their Detection In Vivo Using Quantitative PCR Targeting flaB RNA.
    Methods Mol Biol 2018 ;1690:105-114
    Department of Veterinary Medicine, University of Maryland, College Park, MD, USA.
    Borrelia burgdorferi is maintained in nature by a tick-rodent infection cycle where it traverses and colonizes a variety of host and vector tissues. A tick-borne murine model has been developed to study Lyme disease in the laboratory, which has a substantial impact in advancing our knowledge of spirochete infectivity and pathogenesis. Here, we detail a microinjection-based method for rapid and efficient infection of ticks with B. Read More

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