Relationship between Antibiotic Resistance, Biofilm Formation, and Biofilm-Specific Resistance in Acinetobacter baumannii

In this study, we aimed to examine the relationships between antibiotic resistance, biofilm formation, and biofilm-specific resistance in clinical isolates of Acinetobacter baumannii. The tested 272 isolates were collected from several hospitals in China during 2010–2013. Biofilm-forming capacities were evaluated using the crystal violet staining method. Antibiotic resistance/susceptibility profiles to 21 antibiotics were assessed using VITEK 2 system, broth microdilution method or the Kirby-Bauer disc diffusion method. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) to cefotaxime, imipenem, and ciprofloxacin were evaluated using micro dilution assays. Genetic relatedness of the isolates was also analyzed by pulsed-field gel electrophoresis (PFGE) and plasmid profile. Among all the 272 isolates, 31 were multidrug-resistant (MDR), and 166 were extensively drug-resistant (XDR). PFGE typing revealed 167 pattern types and 103 clusters with a similarity of 80%. MDR and XDR isolates built up the main prevalent genotypes. Most of the non-MDR isolates were distributed in a scattered pattern. Additionally, 249 isolates exhibited biofilm formation, among which 63 were stronger biofilm formers than type strain ATCC19606. Population that exhibited more robust biofilm formation likely contained larger proportion of non-MDR isolates. Isolates with higher level of resistance tended to form weaker biofilms. The MBECs for cefotaxime, imipenem, and ciprofloxacin showed a positive correlation with corresponding MICs, while the enhancement in resistance occurred independent of the quantity of biofilm biomass produced. Results from this study imply that biofilm acts as a mechanism for bacteria to get a better survival, especially in isolates with resistance level not high enough. Moreover, even though biofilms formed by isolates with high level of resistance are always weak, they could still provide similar level of protection for the isolates. Further explorations genetically would improve our understanding of these processes and provide novel insights in the therapeutics and prevention against A. baumannii biofilm-related infections.

[1]  Relationship between multiple drug resistance and biofilm formation in Staphylococcus aureus isolated from medical and non-medical personnel in Yaounde, Cameroon , 2014, The Pan African medical journal.

[2]  L. Kotra,et al.  Common beta-lactamases inhibit bacterial biofilm formation. , 2005, Molecular microbiology.

[3]  Costerton Jw,et al.  Bacterial resistance to antibiotics: the role of biofilms. , 1991 .

[4]  J. Vila,et al.  Biofilm formation in Acinetobacter baumannii: associated features and clinical implications. , 2008, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[5]  G. O’Toole Microtiter dish biofilm formation assay. , 2011, Journal of visualized experiments : JoVE.

[6]  L. Dijkshoorn,et al.  Standardization and Interlaboratory Reproducibility Assessment of Pulsed-Field Gel Electrophoresis-Generated Fingerprints of Acinetobacter baumannii , 2005, Journal of Clinical Microbiology.

[7]  Naohisa Matsunaga,et al.  [Acinetobacter spp]. , 2012, Nihon rinsho. Japanese journal of clinical medicine.

[8]  D. Robinson,et al.  Fnbpb Fibronectin-binding Proteins, Fnbpa and Phenotype Mediated by the Biofilm Staphylococcus Aureus a Novel , 2008 .

[9]  J. Wu,et al.  Heteroresistance to Cephalosporins and Penicillins in Acinetobacter baumannii , 2011, Journal of Clinical Microbiology.

[10]  K. Lewis,et al.  Biofilms and Planktonic Cells of Pseudomonas aeruginosa Have Similar Resistance to Killing by Antimicrobials , 2001, Journal of bacteriology.

[11]  J. Kaplan Antibiotic-Induced Biofilm Formation , 2011, The International journal of artificial organs.

[12]  R. Goyal,et al.  A Study on Device-Related Infections with Special Reference to Biofilm Production and Antibiotic Resistance , 2012, Journal of global infectious diseases.

[13]  Qiwen Yang,et al.  Molecular Epidemiology of Clinical Isolates of Carbapenem-Resistant Acinetobacter spp. from Chinese Hospitals , 2007, Antimicrobial Agents and Chemotherapy.

[14]  P. Stewart,et al.  A genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance , 2003, Nature.

[15]  M. Falagas,et al.  Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. , 2012, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[16]  Diarmaid Hughes,et al.  Antibiotic resistance and its cost: is it possible to reverse resistance? , 2010, Nature Reviews Microbiology.

[17]  A. Peleg,et al.  Insights into Acinetobacter baumannii pathogenicity , 2011, IUBMB life.

[18]  G. Donelli,et al.  Biofilm formation in Acinetobacter baumannii. , 2014, The new microbiologica.

[19]  Paul Stoodley,et al.  Bacterial biofilms: from the Natural environment to infectious diseases , 2004, Nature Reviews Microbiology.

[20]  M. Levine,et al.  A plasmid of enterohemorrhagic Escherichia coli O157:H7 is required for expression of a new fimbrial antigen and for adhesion to epithelial cells , 1987, Infection and immunity.

[21]  H. Ceri,et al.  The MBEC Assay System: multiple equivalent biofilms for antibiotic and biocide susceptibility testing. , 2001, Methods in enzymology.

[22]  J. Costerton,et al.  Bacterial resistance to antibiotics: the role of biofilms. , 1991, Progress in drug research. Fortschritte der Arzneimittelforschung. Progres des recherches pharmaceutiques.

[23]  S. Abidi,et al.  Drug resistance profile and biofilm forming potential of Pseudomonas aeruginosa isolated from contact lenses in Karachi-Pakistan , 2013, BMC Ophthalmology.

[24]  J. Bartlett,et al.  Bad bugs need drugs: an update on the development pipeline from the Antimicrobial Availability Task Force of the Infectious Diseases Society of America. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[25]  Michael J. MacCoss,et al.  Aminoglycoside antibiotics induce bacterial biofilm formation , 2005, Nature.

[26]  B. Levin,et al.  Non-inherited antibiotic resistance , 2006, Nature Reviews Microbiology.

[27]  E Bergogne-Bérézin,et al.  Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features , 1996, Clinical microbiology reviews.

[28]  J. Bryers Bacterial biofilms. , 1993, Current opinion in biotechnology.

[29]  Jung Mogg Kim,et al.  Capacity of multidrug-resistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces. , 2008, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[30]  Mary Jane Ferraro,et al.  Performance standards for antimicrobial susceptibility testing : twelfth informational supplement , 2002 .

[31]  L. Černohorská,et al.  Determination of minimal regrowth concentration (MRC) in clinical isolates of various biofilm-forming bacteria , 2008, Folia Microbiologica.

[32]  L. Kotra,et al.  Common β‐lactamases inhibit bacterial biofilm formation , 2005 .

[33]  A. Banik,et al.  Association of biofilm production with multidrug resistance among clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa from intensive care unit , 2013, Indian journal of critical care medicine : peer-reviewed, official publication of Indian Society of Critical Care Medicine.

[34]  P. Collignon,et al.  Evolution of multi-resistance plasmids in Australian clinical isolates of Escherichia coli. , 2004, Microbiology.

[35]  E. Pérez-Rodríguez,et al.  Influence of whole-body washing of critically ill patients with chlorhexidine on Acinetobacter baumannii isolates. , 2014, American journal of infection control.

[36]  Diane McDougald,et al.  Should we stay or should we go: mechanisms and ecological consequences for biofilm dispersal , 2011, Nature Reviews Microbiology.

[37]  L. Perez Acinetobacter baumannii displays inverse relationship between meropenem resistance and biofilm production , 2015, Journal of chemotherapy.

[38]  R. Bonomo,et al.  Why are we afraid of Acinetobacter baumannii? , 2008, Expert review of anti-infective therapy.

[39]  T. Mah,et al.  Identification of Genes Involved in Pseudomonas aeruginosa Biofilm-Specific Resistance to Antibiotics , 2013, PloS one.

[40]  M. Welsch [Bacterial resistance to antibiotics]. , 1955, Schweizerische medizinische Wochenschrift.