Authors:
E. SapiUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by E. Sapi in
Current site
Google Scholar
PubMed
Close
,
K. BalasubramanianUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by K. Balasubramanian in
Current site
Google Scholar
PubMed
Close
,
A. PoruriUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by A. Poruri in
Current site
Google Scholar
PubMed
Close
,
J. S. MaghsoudlouUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by J. S. Maghsoudlou in
Current site
Google Scholar
PubMed
Close
,
K. M. SocarrasUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by K. M. Socarras in
Current site
Google Scholar
PubMed
Close
,
A. V. TimmarajuUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by A. V. Timmaraju in
Current site
Google Scholar
PubMed
Close
,
K. R. FilushUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by K. R. Filush in
Current site
Google Scholar
PubMed
Close
,
K. GuptaUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by K. Gupta in
Current site
Google Scholar
PubMed
Close
,
S. ShaikhUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by S. Shaikh in
Current site
Google Scholar
PubMed
Close
,
P. A. S. TheophilusUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by P. A. S. Theophilus in
Current site
Google Scholar
PubMed
Close
,
D. F. LueckeUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by D. F. Luecke in
Current site
Google Scholar
PubMed
Close
,
A. MacDonaldUniversity of New Haven, West Haven, CT 06516, USA

Search for other papers by A. MacDonald in
Current site
Google Scholar
PubMed
Close
, and
B. ZelgerMedical University Innsbruck, Innsbruck, Austria

Search for other papers by B. Zelger in
Current site
Google Scholar
PubMed
Close
View More View Less
Open access

Lyme borreliosis, caused by the spirochete Borrelia burgdorferi sensu lato, has grown into a major public health problem. We recently identified a novel morphological form of B. burgdorferi, called biofilm, a structure that is well known to be highly resistant to antibiotics. However, there is no evidence of the existence of Borrelia biofilm in vivo; therefore, the main goal of this study was to determine the presence of Borrelia biofilm in infected human skin tissues. Archived skin biopsy tissues from borrelial lymphocytomas (BL) were reexamined for the presence of B. burgdorferi sensu lato using Borrelia-specific immunohistochemical staining (IHC), fluorescent in situ hybridization, combined fluorescent in situ hybridization (FISH)–IHC, polymerase chain reaction (PCR), and fluorescent and atomic force microscopy methods. Our morphological and histological analyses showed that significant amounts of Borrelia-positive spirochetes and aggregates exist in the BL tissues. Analyzing structures positive for Borrelia showed that aggregates, but not spirochetes, expressed biofilm markers such as protective layers of different mucopolysaccharides, especially alginate. Atomic force microscopy revealed additional hallmark biofilm features of the Borrelia/alginate-positive aggregates such as inside channels and surface protrusions. In summary, this is the first study that demonstrates the presence of Borrelia biofilm in human infected skin tissues.

  • 1.

    Barbour AG , Hayes SF: Biology of Borrelia species. Microbiol Rev 50, 381400 (1986)

  • 2.

    Mead, PS : Epidemiology of Lyme Disease. Infect Dis Clin North Am 29, 187210 (2015)

  • 3.

    Liegner KB , Shapiro JR, Ramsay D, Halperin AJ, Hogrefe W, Kong L: Recurrent erythema migrans despite extended antibiotic treatment with minocycline in a patient with persisting Borrelia burgdorferi infection. J Am Acad Dermatol 28, 312314 (1993)

    • Search Google Scholar
    • Export Citation
  • 4.

    Dumler JS : Molecular diagnosis of Lyme disease: review and meta-analysis. Mol Diagn 6, 111 (2001)

  • 5.

    Klempner MS , Baker PJ, Shapiro ED, Marques A, Dattwyler RJ, Halperin JJ, Wormser GP: Treatment trials for postlyme disease symptoms revisited. Am J Med 126, 665669 (2013)

    • Search Google Scholar
    • Export Citation
  • 6.

    Steere AC , Angelis, SM: Therapy for Lyme arthritis: Strategies for the treatment of antibiotic-refractory arthritis. Arthritis Rheum 54, 30793086 (2006)

    • Search Google Scholar
    • Export Citation
  • 7.

    Berndtson K : Review of evidence for immune evasion and persistent infection in Lyme disease. Int J Gen Med 6, 291306 (2013)

  • 8.

    Kurtti TJ , Munderloh, UG, Johnson RC, Ahlstrand GG: Colony formation and morphology in Borrelia burgdorferi. J Clin Microbiol 25, 20542058 (1987)

    • Search Google Scholar
    • Export Citation
  • 9.

    Mursic VP , Wanner G, Reinhardt S, Wilske B, Busch U, Marget W: Formation and cultivation of Borrelia burgdorferi spheroplast-L-form variants. Infection 24, 218226 (1996)

    • Search Google Scholar
    • Export Citation
  • 10.

    Brorson O , Brorson SH: In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium. Infection 26, 144150 (1998)

    • Search Google Scholar
    • Export Citation
  • 11.

    Hampp EG : Further studies on the significance of spirochetal granules. J Bacteriol 62, 347349 (1951)

  • 12.

    Alban PS , Johnson PW, Nelson DR: Serum-starvation-induced changes in protein synthesis and morphology of Borrelia burgdorferi. Microbiology 146, 119127 (2000)

    • Search Google Scholar
    • Export Citation
  • 13.

    Gruntar I , Malovrh T, Murgia R, Cinco M: Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. APMIS 109, 383388 (2001)

    • Search Google Scholar
    • Export Citation
  • 14.

    Murgia R , Cinco M: Induction of cystic forms by different stress conditions in Borrelia burgdorferi. APMIS 112, 5762 (2004)

  • 15.

    Brorson Ø , Brorson SH: An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole. Int Microbiol 7, 13942 (2004)

    • Search Google Scholar
    • Export Citation
  • 16.

    MacDonald AB : Spirochetal cyst forms in neurodegenerative disorders,…hiding in plain sight. Med Hypotheses 67, 819832 (2006)

  • 17.

    Miklossy J , Kasas S, Zurn AD, McCall S, Yu S, McGeer PL: Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis. J Neuroinflammation 5, 118 (2008)

    • Search Google Scholar
    • Export Citation
  • 18.

    Kersten A , Poitschek C, Rauch S, Aberer E: Effects of penicillin, ceftriaxone, and doxycycline on morphology of Borrelia burgdorferi. Antimicrob Agents Chemother 39, 11271133 (1995)

    • Search Google Scholar
    • Export Citation
  • 19.

    Brorson Ø , Brorson S-H, S cythes J, MacAllister J, Wier A, Margulis L: Destruction of spirochete Borrelia burgdorferi round-body propagules (RBs) by the antibiotic tigecycline. Proc Natl Acad Sci 106, 1865618661 (2009)

    • Search Google Scholar
    • Export Citation
  • 20.

    Sapi E , Kaur N, Anyanwu S, Luecke DF, Datar A, Patel S, Rossi M, Stricker RB: Evaluation of in vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi. Infect Drug Resist 4, 97113 (2011)

    • Search Google Scholar
    • Export Citation
  • 21.

    Feng J , Wang T, Shi W, Zhang S, Sullivan D, Auwaerter PG Zhang Y: Identification of novel activity against Borrelia burgdorferi persisters using an FDA approved drug library. Emerg Microbes Infect 3, e49 (2014)

    • Search Google Scholar
    • Export Citation
  • 22.

    Feng J , Auwaerter PG, Zhang Y: Drug combinations against Borrelia burgdorferi persisters in vitro: eradication achieved by using daptomycin, cefoperazone and doxycycline. PLoS One 10, 115 (2015)

    • Search Google Scholar
    • Export Citation
  • 23.

    Straubinger RK , Summers, BA, Chang YF, Appel MJ: Persistence of Borrelia burgdorferi in experimentally infected dogs after antibiotic treatment. J Clin Microbiol 35, 111116 (1997)

    • Search Google Scholar
    • Export Citation
  • 24.

    Hodzic E , Feng S, Holden K, Freet KJ, Barthold SW: Persistence of Borrelia burgdorferi following antibiotic treatment in mice. Antimicrob Agents Chemother 52, 17281736 (2008)

    • Search Google Scholar
    • Export Citation
  • 25.

    Barthold SW , Hodzic E, Imai DM, Feng S, Yang S, Luft BJ: Ineffectiveness of tigecycline against persistent Borrelia burgdorferi. Antimicrob Agents Chemother 54, 643651 (2010)

    • Search Google Scholar
    • Export Citation
  • 26.

    Embers ME , Barthold SW, Borda JT, Bowers L, Doyle L, Hodzic E, Jacobs MB, Hasenkampf NR, Martin DS, Narasimhan S, Phillippi-Falkenstein KM, Purcell JE, Ratterree MS, Philipp MT: Persistence of Borrelia burgdorferi in rhesus macaques following antibiotic treatment of disseminated infection. PLoS One 7, e29914 (2012)

    • Search Google Scholar
    • Export Citation
  • 27.

    Hodzic E , Imai D, Feng S, Barthold SW: Resurgence of persisting non-cultivable Borrelia burgdorferi following antibiotic treatment in mice. PLoS One 23, 111 (2014)

    • Search Google Scholar
    • Export Citation
  • 28.

    Costerton, JW , Stewart, PS, Greenberg, EP: Bacterial biofilms: a common cause of persistent infections. Science 284, 13181322 (1999)

  • 29.

    Donlan RM , Costerton JW: Biofilms: Survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15, 16793 (2002)

  • 30.

    Sapi E , Bastian SL, Mpoy CM, Scott S, Rattelle A, Pabbati N, Poruri A, Burugu D, Theophilus PA, Pham TV, Datar A, Dhaliwal NK, MacDonald A, Rossi MJ, Sinha SK, Luecke DF: Characterization of biofilm formation by Borrelia burgdorferiin vitro. PLoS One 7, e48277 (2012)

    • Search Google Scholar
    • Export Citation
  • 31.

    Fitzpatrick F , Humphreys H, O’Gara JP: Environmental regulation of biofilm development in methicillin-resistant and methicillin-susceptible Staphylococcus aureus clinical isolates. J Hosp Infect 62, 120122 (2006)

    • Search Google Scholar
    • Export Citation
  • 32.

    Dordel J , Kim C, Chung M, Pardos de la Gándara M, Holden MT, Parkhill J, de Lencastre H, Bentley SD, Tomasz A: Novel determinants of antibiotic resistance: identification of mutated loci in highly methicillin-resistant subpopulations of methicillin-resistant Staphylococcus aureus. MBio 5, e01000 (2014)

    • Search Google Scholar
    • Export Citation
  • 33.

    Clementi, F : Alginate production by Azotobacter vinelandii. Crit Rev Biotechnol 17, 327361 (1997)

  • 34.

    Hentzer M , Teitzel GM, Balzer GJ, Heydorn A, Molin S, Givskov M, Parsek MR: Alginate overproduction affects Pseudomonas aeruginosa biofilm structure and function. J Bacteriol 183, 53955401 (2001)

    • Search Google Scholar
    • Export Citation
  • 35.

    Branda SS , Vik Å, Friedman L, Kolter R: Biofilms: the matrix revisited. Trends Microbiol 13, 2026 (2005)

  • 36.

    Allesen-Holm M , Barken KB, Yang L, Klausen M, Webb JS, Kjelleber S, Molin S, Givskov M, Tolker-Nielsen T: A characterization of DNA release in Pseudomonas aeruginosa cultures and biofilms. Mol Microbiol 59, 11141128 (2006)

    • Search Google Scholar
    • Export Citation
  • 37.

    Ristow P , Bourhy P, Kerneis S, Schmitt C, Prevost MC, Lilenbaum W, Picardeau M: Biofilm formation by saprophytic and pathogenic leptospires. Microbiology 154, 13091317 (2008)

    • Search Google Scholar
    • Export Citation
  • 38.

    Timmaraju VA , Theophilus PAS, Balasubramanian K, Shakih S, Luecke DF, Sapi E: Biofilm formation by Borrelia sensu lato. FEMS Microbiol Lett 362, fnv120 (2015)

    • Search Google Scholar
    • Export Citation
  • 39.

    Remminghorst U , Rehm BHA: Bacterial alginates: from biosynthesis to applications. Biotechnol Lett 28, 17011712 (2006)

  • 40.

    Sharma B , Brown A V., Matluck NE, Hu LT, Lewis K: Borrelia burgdorferi, the causative agent of Lyme disease, forms drug-tolerant persister cells. Antimicrob Agents Chemother 59, 46164624 (2015)

    • Search Google Scholar
    • Export Citation
  • 41.

    Eisendle K , Grabner T, Zelger B: Focus floating microscopy: “Gold Standard” for cutaneous Borreliosis? Am J Clin Pathol 127, 213222 (2007)

    • Search Google Scholar
    • Export Citation
  • 42.

    Eisendle K , Zelger B: The expanding spectrum of cutaneous borreliosis. G Ital Dermatol Venereol 144, 157171 (2009)

  • 43.

    Duray P , Kusnitz A, Ryan J: Demonstration of the Lyme disease spirochete by a modified Dieterle stain method. Lab Med 16: 685687 (1985)

    • Search Google Scholar
    • Export Citation
  • 44.

    Schüler W , Bunikis I, Weber-Lehman J, Comstedt P, Kutschan-Bunikis S, Stanek G, Huber J, Meinke A, Bergström S, Lundberg U: Complete genome sequence of Borrelia afzelii K78 and comparative genome analysis. PLoS One 10, e0120548 (2015)

    • Search Google Scholar
    • Export Citation
  • 45.

    Høiby N , Ciofu O, Bjarnsholt T: Pseudomonas aeruginosa biofilms in cystic fibrosis. Future Microbiol 5, 16631674 (2010)

  • 46.

    Stapper AP : Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation. J Med Microbiol 53, 679690. (2004)

    • Search Google Scholar
    • Export Citation
  • 47.

    Aspe M , Jensen L: The role of alginate and extracellular DNA in biofilm-meditated Pseudomonas aeruginosa gentamicin resistance. J Exp Microbiol Immunol 16, 4248. (2012)

    • Search Google Scholar
    • Export Citation
  • 48.

    Wormser JP , Nadelman RB, Schwartz I: The amber theory of Lyme arthritis: initial description and clinical implications. Clin Rheumatol 31, 989994 (2012)

    • Search Google Scholar
    • Export Citation
  • 49.

    Bockenstedt, LK, Gonzales DG, Haberman AM, Belperron AA: Spirochete antigens persist near cartilage after murine Lyme borreliosis therapy. J Clin Invest 122, 26522660 (2012)

    • Search Google Scholar
    • Export Citation
  • 50.

    Lynch JF , Lappin-Scott HM, Costerton JW (2003): Microbial Biofilms, Cambridge University Press, Cambridge, UK

  • 51.

    Ehrlich GD , DeMeo PJ, Costerton JW, Winkler H (eds.): Culture negative orthopedic biofilm infections. In: Springer Series on Biofilms, Springer-Yerlag, Berlin Heidelberg, pp. 128

    • Search Google Scholar
    • Export Citation
  • 52.

    Baddour LM , Freeman WK, Suri RM, Wilson WR (2014): Cardiovascular infections. In: Braunwald’s Heart Disease: A Textbook of Cardiovascular Medicine, eds. Mann DL, Zipes DP, Libby P, Bonow RO, Braunwald E, Elsevier Saunders: Philadelphia, PA, Chapter 64

    • Search Google Scholar
    • Export Citation
  • 53.

    Singh R , Stine OC, Smith DL, Spitznagel JK Labib ME, Williams HN: Microbial diversity of biofilms in dental unit water systems. Appl Environ Microbiol 69, 34123420 (2003)

    • Search Google Scholar
    • Export Citation
  • 54.

    Vesey PM : Genetic analysis of Treponema denticola ATCC 35405 biofilm formation. Microbiology 150, 24012407 (2004)

  • 55.

    Balasubramanian K : Evidence for the presence of Borrelia burgdorferi biofilm in infected human and mouse tissues. Master Thesis. University of New Haven, Department of Biology and Environmental Science (2015)

    • Search Google Scholar
    • Export Citation
  • 56.

    Stevenson B , Babb K: LuxS-mediated quorum sensing in Borrelia burgdorferi, the Lyme disease spirochete. Infect Immun 70, 40994105 (2002)

    • Search Google Scholar
    • Export Citation
  • 57.

    Mpoy CM : Expression profile of quorum sensing biomarkers during biofilm development of Borrelia burgdorferi. Master Thesis. University of New Haven, Department of Biology and Environmental Science (2012)

    • Search Google Scholar
    • Export Citation
  • Collapse
  • Expand
The author instruction is available in PDF.
Please, download the file from HERE.
 

Senior editors

Editor(s)-in-Chief: Dunay, Ildiko Rita

Editor(s)-in-Chief: Heimesaat, Markus M.

Vice Editor(s)-in-Chief: Fuchs, Anja

Editorial Board

Chair of the Editorial Board:
Jeffrey S. Buguliskis (Thomas Jefferson University, USA)

  • Jörn Albring (University of Münster, Germany)
  • Stefan Bereswill (Charité - University Medicine Berlin, Germany)
  • Dunja Bruder (University of Megdeburg, Germany)
  • Jan Buer (University of Duisburg, Germany)
  • Jeff Buguliskis (Thomas Jefferson University, USA)
  • Edit Buzas (Semmelweis University, Hungary)
  • Charles Collyer (University of Sydney, Australia)
  • Renato Damatta (UENF, Brazil)
  • Ivelina Damjanova (Semmelweis University, Hungary)
  • Maria Deli (Biological Research Center, HAS, Hungary)
  • Olgica Djurković-Djaković (University of Belgrade, Serbia)
  • Jean-Dennis Docquier (University of Siena, Italy)
  • Anna Erdei (Eötvös Loránd University, Hungary)
  • Zsuzsanna Fabry (University of Washington, USA)
  • Beniam Ghebremedhin (Witten/Herdecke University, Germany)
  • Nancy Guillen (Institute Pasteur, France)
  • Georgina L. Hold (University of Aberdeen, United Kingdom)
  • Ralf Ignatius (Charité - University Medicine Berlin, Germany)
  • Zsuzsanna Izsvak (MDC-Berlin, Germany)
  • Achim Kaasch (University of Cologne, Germany)
  • Tamás Laskay (University of Lübeck, Germany)
  • Oliver Liesenfeld (Roche, USA)
  • Shreemanta Parida (Vaccine Grand Challenge Program, India)
  • Matyas Sandor (University of Wisconsin, USA)
  • Ulrich Steinhoff (University of Marburg, Germany)
  • Michal Toborek (University of Miami, USA)
  • Mary Jo Wick (University of Gothenburg, Sweden)
  • Susanne A. Wolf (MDC-Berlin, Germany)

 

Dr. Dunay, Ildiko Rita
Magdeburg, Germany
E-mail: ildikodunay@gmail.com

Indexing and Abstracting Services:

  • PubMed Central
  • Scopus
  • ESCI
  • CABI
  • CABELLS Journalytics

 

2021  
Web of Science  
Total Cites
WoS
790
Journal Impact Factor not applicable
Rank by Impact Factor not applicable
Impact Factor
without
Journal Self Cites
not applicable
5 Year
Impact Factor
not applicable
Journal Citation Indicator 0,64
Rank by Journal Citation Indicator Microbiology 81/157
Scimago  
Scimago
H-index
not indexed
Scimago
Journal Rank
not indexed
Scimago Quartile Score not indexed
Scopus  
Scopus
Cite Score
not indexed
Scopus
CIte Score Rank
  not indexed
Scopus
SNIP
not indexed

2020  
CrossRef Documents 23
WoS Cites 708
Wos H-index 27
Days from submission to acceptance 219
Days from acceptance to publication 176
Acceptance Rate 70%

2019  
WoS
Cites
558
CrossRef
Documents
24
Acceptance
Rate
92%

 

European Journal of Microbiology and Immunology
Publication Model Gold Open Access
Submission Fee none
Article Processing Charge 600 EUR/article
Regional discounts on country of the funding agency World Bank Lower-middle-income economies: 50%
World Bank Low-income economies: 100%
Further Discounts Editorial Board / Advisory Board members: 50%
Corresponding authors, affiliated to an EISZ member institution subscribing to the journal package of Akadémiai Kiadó: 100%
Subscription Information Gold Open Access
Purchase per Title  

European Journal of Microbiology and Immunology
Language English
Size A4
Year of
Foundation
2011
Volumes
per Year
1
Issues
per Year
4
Founder Akadémiai Kiadó
Founder's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 2062-509X (Print)
ISSN 2062-8633 (Online)

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Oct 2022 0 39 10
Nov 2022 0 58 31
Dec 2022 0 55 19
Jan 2023 0 86 32
Feb 2023 0 54 22
Mar 2023 0 116 33
Apr 2023 0 0 0