Skip to main content
SpringerLink
Log in

Antibiotic Resistance Genes and Antibiotic Susceptibility of Oral Enterococcus faecalis Isolates Compared to Isolates from Hospitalized Patients and Food

  • Chapter
  • First Online:
Advances in Microbiology, Infectious Diseases and Public Health

Part of the book series: Advances in Experimental Medicine and Biology ((AMIDPH,volume 1057))

Abstract

Enterococcus faecalis, a commensal of the intestinal tract of humans and animals is of great significance as leading opportunistic pathogen, and also prevalent in oral diseases, such as endodontic infections, as well as the healthy oral cavity. To investigate the potential of oral E. faecalis to constitute a reservoir of antibiotic resistance, isolates from supragingival plaque/saliva and from endodontic infections were screened regarding their resistance to selected antibiotics in comparison to nosocomial and food isolates.

70 E. faecalis isolates were analyzed with PCR regarding their equipment with the resistance genes tetM, tetO, ermB, ermC, vanA, vanB and blaTEM. Additionally, they were tested for their phenotypic resistance to doxycycline, azithromycin, rifampicin, amoxicillin and streptomycin using the Etest.

High percentages of the plaque/saliva, nosocomial and food isolates were resistant to doxycycline and azithromycin, particularly plaque/saliva isolates (81%) and nosocomial isolates (73.3%) showed resistance to doxycycline, significantly more than among the food and endodontic isolates. Rifampicin resistance was widespread among isolates from plaque/saliva (52.4%), endodontic infections (50%) and nosocomial infections (40%); all isolates were susceptible to amoxicillin and all oral isolates to high-level streptomycin. TetM genes were detected in the majority of all isolates and ermB genes were present in many nosocomial and plaque/saliva isolates. Thirty percent of the endodontic isolates and 53% of the nosocomial isolates were equipped with blaTEM genes.

The results suggest that the oral cavity can harbor E. faecalis strains with multiple resistances against different antibiotics and thus be regarded as a potential source of resistance traits.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Institutional subscriptions

References

  • Al-Ahmad A, Maier J, Follo M, Spitzmuller B, Wittmer A, Hellwig E et al (2010) Food-borne enterococci integrate into oral biofilm: an in vivo study. J Endod 36(11):1812–1819. doi:10.1016/j.joen.2010.08.011

    Article  PubMed  Google Scholar 

  • Al-Ahmad A, Ameen H, Pelz K, Karygianni L, Wittmer A, Anderson AC et al (2014) Antibiotic resistance and capacity for biofilm formation of different bacteria isolated from endodontic infections associated with root-filled teeth. J Endod 40(2):223–230. doi:10.1016/j.joen.2013.07.023

    Article  PubMed  Google Scholar 

  • Amsler K, Santoro C, Foleno B, Bush K, Flamm R (2010) Comparison of broth microdilution, agar dilution, and Etest for susceptibility testing of doripenem against gram-negative and gram-positive pathogens. J Clin Microbiol 48(9):3353–3357. doi:10.1128/jcm.00494-10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Anderson AC, Al-Ahmad A, Elamin F, Jonas D, Mirghani Y, Schilhabel M et al (2013) Comparison of the bacterial composition and structure in symptomatic and asymptomatic endodontic infections associated with root-filled teeth using pyrosequencing. PLoS One 8(12):e84960. doi:10.1371/journal.pone.0084960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Anderson AC, Jonas D, Huber I, Karygianni L, Wolber J, Hellwig E et al (2015) Enterococcus faecalis from food, clinical specimens, and oral sites: prevalence of virulence factors in association with biofilm formation. Front Microbiol 6:1534. doi:10.3389/fmicb.2015.01534

    Article  PubMed  Google Scholar 

  • Arias CA, Murray BE (2012) The rise of the Enterococcus: beyond vancomycin resistance. Nat Rev Microbiol 10(4):266–278. doi:10.1038/nrmicro2761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Barbosa-Ribeiro M, De-Jesus-Soares A, Zaia AA, Ferraz CC, Almeida JF, Gomes BP (2016) Antimicrobial susceptibility and characterization of virulence genes of Enterococcus faecalis isolates from teeth with failure of the endodontic treatment. J Endod. doi:10.1016/j.joen.2016.03.015

    Article  PubMed  Google Scholar 

  • Call DR, Bakko MK, Krug MJ, Roberts MC (2003) Identifying antimicrobial resistance genes with DNA microarrays. Antimicrob Agents Chemother 47(10):3290–3295

    Article  CAS  Google Scholar 

  • CLSI (2013) CLSI. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Third Informational Supplement. CLSI document M100-S23. Clinical and Laboratory Standards Institute, Wayne, PA

    Google Scholar 

  • Dahlen G, Samuelsson W, Molander A, Reit C (2000) Identification and antimicrobial susceptibility of enterococci isolated from the root canal. Oral Microbiol Immunol 15(5):309–312

    Article  CAS  Google Scholar 

  • Dahlen G, Blomqvist S, Almstahl A, Carlen A (2012) Virulence factors and antibiotic susceptibility in enterococci isolated from oral mucosal and deep infections. J Oral Microbiol:4. doi:10.3402/jom.v4i0.10855

    Article  CAS  Google Scholar 

  • Depardieu F, Kolbert M, Pruul H, Bell J, Courvalin P (2004) VanD-type vancomycin-resistant Enterococcus faecium and Enterococcus faecalis. Antimicrob Agents Chemother 48(10):3892–3904. doi:10.1128/aac.48.10.3892-3904.2004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Duggan JM, Sedgley CM (2007) Biofilm formation of oral and endodontic Enterococcus faecalis. J Endod 33(7):815–818. doi:10.1016/j.joen.2007.02.016

    Article  PubMed  Google Scholar 

  • Dutka-Malen S, Evers S, Courvalin P (1995) Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. J Clin Microbiol 33(1):24–27

    CAS  PubMed  PubMed Central  Google Scholar 

  • EUCAST (2016) The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 6.0, 2016 http://www.eucast.org. http://www.eucast.org/clinical_breakpoints/

  • Fass RJ (1993) Erythromycin, clarithromycin, and azithromycin: use of frequency distribution curves, scattergrams, and regression analyses to compare in vitro activities and describe cross-resistance. Antimicrob Agents Chemother 37(10):2080–2086

    Article  CAS  Google Scholar 

  • Fisher K, Phillips C (2009) The ecology, epidemiology and virulence of Enterococcus. Microbiology 155(Pt 6):1749–1757. doi:10.1099/mic.0.026385-0

    Article  CAS  PubMed  Google Scholar 

  • Franz CM, Huch M, Abriouel H, Holzapfel W, Galvez A (2011) Enterococci as probiotics and their implications in food safety. Int J Food Microbiol 151(2):125–140. doi:10.1016/j.ijfoodmicro.2011.08.014

    Article  CAS  PubMed  Google Scholar 

  • Gaetti-Jardim EC, Marqueti AC, Faverani LP, Gaetti-Jardim E Jr (2010) Antimicrobial resistance of aerobes and facultative anaerobes isolated from the oral cavity. J Appl Oral Sci 18(6):551–559

    Article  Google Scholar 

  • Hammerum AM (2012) Enterococci of animal origin and their significance for public health. Clin Microbiol Infect 18(7):619–625. doi:10.1111/j.1469-0691.2012.03829.x

    Article  CAS  PubMed  Google Scholar 

  • Jacinto RC, Gomes BP, Ferraz CC, Zaia AA, Filho FJ (2003) Microbiological analysis of infected root canals from symptomatic and asymptomatic teeth with periapical periodontitis and the antimicrobial susceptibility of some isolated anaerobic bacteria. Oral Microbiol Immunol 18(5):285–292

    Article  CAS  Google Scholar 

  • Jamet E, Akary E, Poisson MA, Chamba JF, Bertrand X, Serror P (2012) Prevalence and characterization of antibiotic resistant Enterococcus faecalis in French cheeses. Food Microbiol 31(2):191–198. doi:10.1016/j.fm.2012.03.009

    Article  CAS  PubMed  Google Scholar 

  • Jorgensen JH, Ferraro MJ (2009) Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clin Infect Dis 49(11):1749–1755. doi:10.1086/647952

    Article  CAS  PubMed  Google Scholar 

  • Jungermann GB, Burns K, Nandakumar R, Tolba M, Venezia RA, Fouad AF (2011) Antibiotic resistance in primary and persistent endodontic infections. J Endod 37(10):1337–1344. doi:10.1016/j.joen.2011.06.028

    Article  PubMed  PubMed Central  Google Scholar 

  • Komiyama EY, Lepesqueur LS, Yassuda CG, Samaranayake LP, Parahitiyawa NB, Balducci I et al (2016) Enterococcus species in the oral cavity: prevalence, virulence factors and antimicrobial susceptibility. PLoS One 11(9):e0163001. doi:10.1371/journal.pone.0163001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kouidhi B, Zmantar T, Mahdouani K, Hentati H, Bakhrouf A (2011) Antibiotic resistance and adhesion properties of oral Enterococci associated to dental caries. BMC Microbiol 11:155. doi:10.1186/1471-2180-11-155

    Article  PubMed  PubMed Central  Google Scholar 

  • Kristich CJ, Rice LB, Arias CA (2014) Enterococcal infection-treatment and antibiotic resistance. In: Gilmore MS, Clewell DB, Ike Y, Shankar N (eds) Enterococci: from commensals to leading causes of drug resistant infection. Massachusetts Eye and Ear Infirmary, Boston

    Google Scholar 

  • Kuch A, Willems RJ, Werner G, Coque TM, Hammerum AM, Sundsfjord A et al (2012) Insight into antimicrobial susceptibility and population structure of contemporary human Enterococcus faecalis isolates from Europe. J Antimicrob Chemother 67(3):551–558. doi:10.1093/jac/dkr544

    Article  CAS  PubMed  Google Scholar 

  • Lietzau S, Hoewner M, von Baum H, Marre R, Brenner H (2006) Antibiotic resistant fecal isolates of Enterococci among unselected patients outside the clinical sector: an epidemiological study from Southern Germany. Pharmacoepidemiol Drug Saf 15(4):275–277. doi:10.1002/pds.1167

    Article  CAS  PubMed  Google Scholar 

  • Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG et al (2012) Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 18(3):268–281. doi:10.1111/j.1469-0691.2011.03570.x

    Article  CAS  PubMed  Google Scholar 

  • Manson JM, Hancock LE, Gilmore MS (2010) Mechanism of chromosomal transfer of Enterococcus faecalis pathogenicity island, capsule, antimicrobial resistance, and other traits. Proc Natl Acad Sci U S A 107(27):12269–12274. doi:10.1073/pnas.1000139107

    Article  PubMed  PubMed Central  Google Scholar 

  • McBride SM, Fischetti VA, Leblanc DJ, Moellering RC Jr, Gilmore MS (2007) Genetic diversity among Enterococcus faecalis. PLoS One 2(7):e582. doi:10.1371/journal.pone.0000582

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Murray BE (1990) The life and times of the Enterococcus. Clin Microbiol Rev 3(1):46–65

    Article  CAS  Google Scholar 

  • Paganelli FL, Willems RJ, Leavis HL (2012) Optimizing future treatment of enterococcal infections: attacking the biofilm? Trends Microbiol 20(1):40–49. doi:10.1016/j.tim.2011.11.001

    Article  CAS  PubMed  Google Scholar 

  • Palmer KL, Kos VN, Gilmore MS (2010) Horizontal gene transfer and the genomics of enterococcal antibiotic resistance. Curr Opin Microbiol 13(5):632–639. doi:10.1016/j.mib.2010.08.004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Paulsen IT, Banerjei L, Myers GS, Nelson KE, Seshadri R, Read TD et al (2003) Role of mobile DNA in the evolution of vancomycin-resistant Enterococcus faecalis. Science 299(5615):2071–2074. doi:10.1126/science.1080613

    Article  CAS  PubMed  Google Scholar 

  • Perez-Trallero E, Montes M, Orden B, Tamayo E, Garcia-Arenzana JM, Marimon JM (2007) Phenotypic and genotypic characterization of Streptococcus pyogenes isolates displaying the MLSB phenotype of macrolide resistance in Spain, 1999 to 2005. Antimicrob Agents Chemother 51(4):1228–1233. doi:10.1128/aac.01054-06

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Perreten V, Vorlet-Fawer L, Slickers P, Ehricht R, Kuhnert P, Frey J (2005) Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria. J Clin Microbiol 43(5):2291–2302. doi:10.1128/jcm.43.5.2291-2302.2005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pinheiro ET, Gomes BP, Drucker DB, Zaia AA, Ferraz CC, Souza-Filho FJ (2004) Antimicrobial susceptibility of Enterococcus faecalis isolated from canals of root filled teeth with periapical lesions. Int Endod J 37(11):756–763. doi:10.1111/j.1365-2591.2004.00865.x

    Article  CAS  PubMed  Google Scholar 

  • Poeta P, Igrejas G, Goncalves A, Martins E, Araujo C, Carvalho C et al (2009) Influence of oral hygiene in patients with fixed appliances in the oral carriage of antimicrobial-resistant Escherichia coli and Enterococcus isolates. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108(4):557–564. doi:10.1016/j.tripleo.2009.06.002

    Article  PubMed  Google Scholar 

  • Portenier I, Waltimo TMT, Haapasalo M (2003) Enterococcus faecalis– the root canal survivor and ‘star’ in post-treatment disease. Endod Top 6(1):135–159. doi:10.1111/j.1601-1546.2003.00040.x

    Article  Google Scholar 

  • Poveda Roda R, Bagan JV, Sanchis Bielsa JM, Carbonell Pastor E (2007) Antibiotic use in dental practice: a review. Med Oral Patol Oral Cir Bucal 12(3):E186–E192

    PubMed  Google Scholar 

  • Preshaw PM, Hefti AF, Jepsen S, Etienne D, Walker C, Bradshaw MH (2004) Subantimicrobial dose doxycycline as adjunctive treatment for periodontitis: a review. J Clin Periodontol 31(9):697–707. doi:10.1111/j.1600-051X.2004.00558.x

    Article  CAS  PubMed  Google Scholar 

  • Rams TE, Feik D, Mortensen JE, Degener JE, van Winkelhoff AJ (2013) Antibiotic susceptibility of periodontal Enterococcus faecalis. J Periodontol 84(7):1026–1033. doi:10.1902/jop.2012.120050

    Article  CAS  PubMed  Google Scholar 

  • Reinert RR, Filimonova OY, Al-Lahham A, Grudinina SA, Ilina EN, Weigel LM et al (2008) Mechanisms of macrolide resistance among Streptococcus pneumoniae isolates from Russia. Antimicrob Agents Chemother 52(6):2260–2262. doi:10.1128/aac.01270-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roberts AP, Mullany P (2010) Oral biofilms: a reservoir of transferable, bacterial, antimicrobial resistance. Expert Rev Anti-Infect Ther 8(12):1441–1450. doi:10.1586/eri.10.106

    Article  CAS  PubMed  Google Scholar 

  • Roberts AP, Mullany P (2011) Tn916-like genetic elements: a diverse group of modular mobile elements conferring antibiotic resistance. FEMS Microbiol Rev 35(5):856–871. doi:10.1111/j.1574-6976.2011.00283.x

    Article  CAS  PubMed  Google Scholar 

  • Roberts AP, Cheah G, Ready D, Pratten J, Wilson M, Mullany P (2001) Transfer of TN916-like elements in microcosm dental plaques. Antimicrob Agents Chemother 45(10):2943–2946. doi:10.1128/aac.45.10.2943-2946.2001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rocas IN, Siqueira JF Jr (2013) Detection of antibiotic resistance genes in samples from acute and chronic endodontic infections and after treatment. Arch Oral Biol 58(9):1123–1128. doi:10.1016/j.archoralbio.2013.03.010

    Article  CAS  PubMed  Google Scholar 

  • Ruiz-Garbajosa P, Canton R, Pintado V, Coque TM, Willems R, Baquero F et al (2006) Genetic and phenotypic differences among Enterococcus faecalis clones from intestinal colonisation and invasive disease. Clin Microbiol Infect 12(12):1193–1198. doi:10.1111/j.1469-0691.2006.01533.x

    Article  CAS  PubMed  Google Scholar 

  • Schirrmeister JF, Liebenow AL, Braun G, Wittmer A, Hellwig E, Al-Ahmad A (2007) Detection and eradication of microorganisms in root-filled teeth associated with periradicular lesions: an in vivo study. J Endod 33(5):536–540. doi:10.1016/j.joen.2007.01.012

    Article  PubMed  Google Scholar 

  • Schlegelova J, Babak V, Klimova E, Lukasova J, Navratilova P, Sustackova A et al (2002) Prevalence of and resistance to anti-microbial drugs in selected microbial species isolated from bulk milk samples. J Vet Med B Infect Dis Vet Public Health 49(5):216–225

    Article  CAS  Google Scholar 

  • Sedgley CM, Molander A, Flannagan SE, Nagel AC, Appelbe OK, Clewell DB et al (2005a) Virulence, phenotype and genotype characteristics of endodontic Enterococcus spp. Oral Microbiol Immunol 20(1):10–19. doi:10.1111/j.1399-302X.2004.00180.x

    Article  CAS  PubMed  Google Scholar 

  • Sedgley CM, Nagel AC, Shelburne CE, Clewell DB, Appelbe O, Molander A (2005b) Quantitative real-time PCR detection of oral Enterococcus faecalis in humans. Arch Oral Biol 50(6):575–583. doi:10.1016/j.archoralbio.2004.10.017

    Article  CAS  PubMed  Google Scholar 

  • Sedgley C, Buck G, Appelbe O (2006) Prevalence of Enterococcus faecalis at multiple oral sites in endodontic patients using culture and PCR. J Endod 32(2):104–109. doi:10.1016/j.joen.2005.10.022

    Article  PubMed  Google Scholar 

  • Sedgley CM, Lee EH, Martin MJ, Flannagan SE (2008) Antibiotic resistance gene transfer between Streptococcus gordonii and Enterococcus faecalis in root canals of teeth ex vivo. J Endod 34(5):570–574. doi:10.1016/j.joen.2008.02.014

    Article  PubMed  Google Scholar 

  • Siqueira JF Jr, Rocas IN (2004) Polymerase chain reaction-based analysis of microorganisms associated with failed endodontic treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 97(1):85–94. doi:10.1016/s1079210403003536

    Article  PubMed  Google Scholar 

  • Souto R, Colombo AP (2008) Prevalence of Enterococcus faecalis in subgingival biofilm and saliva of subjects with chronic periodontal infection. Arch Oral Biol 53(2):155–160. doi:10.1016/j.archoralbio.2007.08.004

    Article  CAS  PubMed  Google Scholar 

  • Sun J, Song X, Kristiansen BE, Kjaereng A, Willems RJ, Eriksen HM et al (2009) Occurrence, population structure, and antimicrobial resistance of enterococci in marginal and apical periodontitis. J Clin Microbiol 47(7):2218–2225. doi:10.1128/jcm.00388-09

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Templer SP, Baumgartner A (2007) Enterococci from Appenzeller and Schabziger raw milk cheese: antibiotic resistance, virulence factors, and persistence of particular strains in the products. J Food Prot 70(2):450–455

    Article  CAS  Google Scholar 

  • Tenover FC, Baker CN, Swenson JM (1996) Evaluation of commercial methods for determining antimicrobial susceptibility of Streptococcus Pneumoniae. J Clin Microbiol 34(1):10–14

    CAS  PubMed  PubMed Central  Google Scholar 

  • Thurnheer T, Belibasakis GN (2015) Integration of non-oral bacteria into in vitro oral biofilms. Virulence 6(3):258–264. doi:10.4161/21505594.2014.967608

    Article  CAS  PubMed  Google Scholar 

  • Van Tyne D, Gilmore MS (2014) Friend turned foe: evolution of enterococcal virulence and antibiotic resistance. Annu Rev Microbiol 68:337–356. doi:10.1146/annurev-micro-091213-113003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wenzler S, Schmidt-Eisenlohr E, Daschner F (2004) Comparative in vitro activities of three new quinolones and azithromycin against aerobic pathogens causing respiratory tract and abdominal wound infections. Chemotherapy 50(1):40–42. doi:10.1159/000077284

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank Bettina Spitzmüller, Kristina Kollmar and Annette Wittmer for excellent technical assistance and Nicole Arweiler, Daniel Jonas and Ingrid Huber for providing part of the isolates, as well as Grant Anderson for English language correction.

Conflicts of Interest

The authors deny any conflicts of interest related to this study.

Ethical Statement

All endodontic and clinical isolates were obtained after approval by the Ethics Committee (no. 140/09, University of Freiburg).

Author information

Authors and Affiliations

  1. Department of Operative Dentistry and Periodontology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Straße 55, 79106, Freiburg, Germany

    Annette Carola Anderson, Huria Andisha, Elmar Hellwig & Ali Al-Ahmad

  2. Institute for Environmental Health Sciences and Hospital Infection Control, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Breisacher Straße 115b, 79106, Freiburg, Germany

    Daniel Jonas

  3. Center for Medical Biometry and Medical Informatics, Institute for Medical Biometry and Statistics, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Stefan-Meier Straße 26, 79104, Freiburg, Germany

    Kirstin Vach

Authors
  1. Annette Carola Anderson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huria Andisha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elmar Hellwig
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniel Jonas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kirstin Vach
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ali Al-Ahmad
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Annette Carola Anderson .

Editor information

Editors and Affiliations

  1. Microbial Biofilm Laboratory, Fondazione Santa Lucia IRCCS, Rome, Italy

    Gianfranco Donelli

Supplementary Material

Supplementary Material

Table S2 Antibiotic susceptibility and presence of antibiotic resistance genes in 70 Enterococcus faecalis isolates from four different sources
Full size table

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer International Publishing AG

About this chapter

Cite this chapter

Anderson, A.C., Andisha, H., Hellwig, E., Jonas, D., Vach, K., Al-Ahmad, A. (2017). Antibiotic Resistance Genes and Antibiotic Susceptibility of Oral Enterococcus faecalis Isolates Compared to Isolates from Hospitalized Patients and Food. In: Donelli, G. (eds) Advances in Microbiology, Infectious Diseases and Public Health. Advances in Experimental Medicine and Biology(), vol 1057. Springer, Cham. https://doi.org/10.1007/5584_2017_53

Download citation

Publish with us

Policies and ethics

Search

Navigation