Potential of bacteriophage ΦAB2 as an environmental biocontrol agent for the control of multidrug-resistant Acinetobacter baumannii

BMC Microbiology

Chun-Chieh Tseng²  Meng-Jiun Lai⁴  Nien-Tsung Lin³  Kai-Chih Chang⁴  Anren Hu⁴  Yu-Lin Liu²  Li-Kuang Chen¹ 

[1] Department of Laboratory Medicine, Clinical Pathology, Emerging Infectious Pathogen Research Laboratory, Buddhist Tzu Chi General Hospital, Hualien, Taiwan;Department and Graduate Institute of Public Health, Tzu Chi University, Hualien, Taiwan;Master Program, Microbiology, Immunology, and Biochemistry, School of Medicine, Tzu Chi University, Hualien, Taiwan;Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, Taiwan

关键词: MDRAB;    Biocontrol;    Bacteriophage;   

Others  :  1143500
DOI  :  10.1186/1471-2180-13-154

 received in 2013-04-10, accepted in 2013-07-02,  发布年份 2013

【 摘 要 】

Background

Multidrug-resistant Acinetobacter baumannii (MDRAB) is associated with nosocomial infections worldwide. To date, the use of a phage to prevent infections caused by MDRAB has not been demonstrated.

Results

The MDRAB-specific phage ϕAB2 was stable at 4°C and pH 7 in 0.5% chloroform solution, and showed a slight decrease in plaque-forming units (PFU)/ml of 0.3–0.9 log after 330 days of storage. The addition of ϕAB2 at a concentration of at least 10⁵ PFU/ml to an A. baumannii M3237 suspension killed >99.9% of A. baumannii M3237 after 5 min, regardless of A. baumannii M3237 concentration (10⁴, 10⁵, or 10⁶ colony-forming units (CFU)/ml). The addition of ϕAB2 at a concentration of 10⁸ PFU/slide (>10⁷ PFU/cm²) to glass slides containing A. baumannii M3237 at 10⁴, 10⁵, or 10⁶ CFU/slide, significantly reduced bacterial numbers by 93%, 97%, and 99%, respectively. Thus, this concentration is recommended for decontamination of glass surfaces. Moreover, infusion of ϕAB2 into 10% glycerol exhibited strong anti-MDRAB activity (99.9% reduction), even after 90 days of storage. Treatment of a 10% paraffin oil-based lotion with ϕAB2 significantly reduced (99%) A. baumannii M3237 after 1 day of storage. However, ϕAB2 had no activity in the lotion after 1 month of storage.

Conclusions

Phages may be useful for reducing MDRAB contamination in liquid suspensions or on hard surfaces. Phages may also be inoculated into a solution to produce an antiseptic hand wash. However, the phage concentration and incubation time (the duration of phage contact with bacteria) should be carefully considered to reduce the risk of MDRAB contamination.

【 授权许可】

   
2013 Chen et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Bergogne-Berezin E, Towner KJ: Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features. Clin Microbiol Rev 1996, 9:148-165.
• [2]Villegas MV, Hartstein AI: Acinetobacter outbreaks, 1977-2000. Infect Control Hosp Epidemiol 2003, 24:284-295.
• [3]Okpara AU, Maswoswe JJ: Emergence of multidrug-resistant isolates of Acinetobacter baumannii. Am J Hosp Pharm 1994, 51:2671-2675.
• [4]Gaynes R, Edwards JR: Overview of nosocomial infections caused by gram-negative bacilli. Clin Infect Dis 2005, 41:848-854.
• [5]Meric M, Kasap M, Gacar G, Budak F, Dundar D, Kolayli F, Eroglu C, Vahaboglu H: Emergence and spread of carbapenem-resistant Acinetobacter baumannii in a tertiary care hospital in Turkey. FEMS Microbiol Lett 2008, 282:214-218.
• [6]Hsueh PR, Teng LJ, Chen CY, Chen WH, Yu CJ, Ho SW, Luh KT: Pandrug-resistant Acinetobacter baumannii causing nosocomial infections in a university hospital, Taiwan. Emerg Infect Dis 2002, 8:827-832.
• [7]Annual Report of Nosocomial Infections Surveillance System: Annual Report of Nosocomial Infections Surveillance System. Taiwan: Center for Disease Control; 2009. http://www.cdc.gov.tw/english/ webcite
• [8]Dijkshoorn L, Nemec A, Seifert H: An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nat Rev Microbiol 2007, 5:939-951.
• [9]Chang HL, Tang CH, Hsu YM, Wan L, Chang YF, Lin CT, Tseng YR, Lin YJ, Sheu JJ, Lin CW, et al.: Nosocomial outbreak of infection with multidrug-resistant Acinetobacter baumannii in a medical center in Taiwan. Infect Control Hosp Epidemiol 2009, 30:34-38.
• [10]Sengstock DM, Thyagarajan R, Apalara J, Mira A, Chopra T, Kaye KS: Multidrug-resistant Acinetobacter baumannii: an emerging pathogen among older adults in community hospitals and nursing homes. Clin Infect Dis 2010, 50:1611-1616.
• [11]Joseph NM, Sistla S, Dutta TK, Badhe AS, Rasitha D, Parija SC: Role of intensive care unit environment and health-care workers in transmission of ventilator-associated pneumonia. J Infect Dev Ctries 2010, 4:282-291.
• [12]Wang CY, Wu HD, Lee LN, Chang HT, Hsu YL, Yu CJ, Yang PC, Hsueh PR: Pasteurization is effective against multidrug-resistant bacteria. Am J Infect Control 2006, 34:320-322.
• [13]Rastogi VK, Wallace L, Smith LS: Disinfection of Acinetobacter baumannii-contaminated surfaces relevant to medical treatment facilities with ultraviolet C light. Mil Med 2007, 172:1166-1169.
• [14]Doidge M, Allworth AM, Woods M, Marshall P, Terry M, O’Brien K, Goh HM, George N, Nimmo GR, Schembri MA, et al.: Control of an outbreak of carbapenem-resistant Acinetobacter baumannii in Australia after introduction of environmental cleaning with a commercial oxidizing disinfectant. Infect Control Hosp Epidemiol 2010, 31:418-420.
• [15]Donahue M, Watson LR, Torress-Cook A, Watson PA: Novel use of antimicrobial hand sanitizer in treatment of nosocomial Acinetobacter infection. Orthopedics 2009, 32:58.
• [16]Martro E, Hernandez A, Ariza J, Dominguez MA, Matas L, Argerich MJ, Martin R, Ausina V: Assessment of Acinetobacter baumannii susceptibility to antiseptics and disinfectants. J Hosp Infect 2003, 55:39-46.
• [17]Sharma M, Hudson JB: Ozone gas is an effective and practical antibacterial agent. Am J Infect Control 2008, 36:559-563.
• [18]Wong MS, Sun DS, Chang HH: Bactericidal performance of visible-light responsive titania photocatalyst with silver nanostructures. PLoS One 2010, 5:e10394.
• [19]Wisplinghoff H, Schmitt R, Wohrmann A, Stefanik D, Seifert H: Resistance to disinfectants in epidemiologically defined clinical isolates of Acinetobacter baumannii. J Hosp Infect 2007, 66:174-181.
• [20]Edwards J, Patel G, Wareham DW: Low concentrations of commercial alcohol hand rubs facilitate growth of and secretion of extracellular proteins by multidrug-resistant strains of Acinetobacter baumannii. J Med Microbiol 2007, 56:1595-1599.
• [21]Hudson JA, Billington C, Carey-Smith G, Greening G: Bacteriophages as biocontrol agents in food. J Food Prot 2005, 68:426-437.
• [22]Bigwood T, Hudson JA, Billington C: Influence of host and bacteriophage concentrations on the inactivation of food-borne pathogenic bacteria by two phages. FEMS Microbiol Lett 2009, 291:59-64.
• [23]Guenther S, Huwyler D, Richard S, Loessner MJ: Virulent bacteriophage for efficient biocontrol of Listeria monocytogenes in ready-to-eat foods. Appl Environ Microbiol 2009, 75:93-100.
• [24]Garcia P, Martinez B, Rodriguez L, Rodriguez A: Synergy between the phage endolysin LysH5 and nisin to kill Staphylococcus aureus in pasteurized milk. Int J Food Microbiol 2010, 141:151-155.
• [25]Kim KP, Klumpp J, Loessner MJ: Enterobacter sakazakii bacteriophages can prevent bacterial growth in reconstituted infant formula. Int J Food Microbiol 2007, 115:195-203.
• [26]Abuladze T, Li M, Menetrez MY, Dean T, Senecal A, Sulakvelidze A: Bacteriophages reduce experimental contamination of hard surfaces, tomato, spinach, broccoli, and ground beef by Escherichia coli O157:H7. Appl Environ Microbiol 2008, 74:6230-6238.
• [27]Kocharunchitt C, Ross T, McNeil DL: Use of bacteriophages as biocontrol agents to control Salmonella associated with seed sprouts. Int J Food Microbiol 2009, 128:453-459.
• [28]Obeso JM, Garcia P, Martinez B, Arroyo-Lopez FN, Garrido-Fernandez A, Rodriguez A: Use of logistic regression for prediction of the fate of Staphylococcus aureus in pasteurized milk in the presence of two lytic phages. Appl Environ Microbiol 2010, 76:6038-6046.
• [29]Garcia P, Madera C, Martinez B, Rodriguez A, Evaristo Suarez J: Prevalence of bacteriophages infecting Staphylococcus aureus in dairy samples and their potential as biocontrol agents. J Dairy Sci 2009, 92:3019-3026.
• [30]FDA: Food additives permitted for direct addition to food for human consumption; bacteriophage preparation. Fed Regist 2006, 71:47729-47732.
• [31]Barbolla RE, Centron D, Maimone S, Rospide F, Salgueira C, Altclas J, Catalano M: Molecular epidemiology of Acinetobacter baumannii spread in an adult intensive care unit under an endemic setting. Am J Infect Control 2008, 36:444-452.
• [32]Otter JA, Yezli S, French GL: The role played by contaminated surfaces in the transmission of nosocomial pathogens. Infect Control Hosp Epidemiol 2011, 32:687-699.
• [33]Monk AB, Rees CD, Barrow P, Hagens S, Harper DR: Bacteriophage applications: where are we now? Lett Appl Microbiol 2010, 51:363-369.
• [34]O’Flaherty S, Ross RP, Meaney W, Fitzgerald GF, Elbreki MF, Coffey A: Potential of the polyvalent anti-Staphylococcus bacteriophage K for control of antibiotic-resistant staphylococci from hospitals. Appl Environ Microbiol 2005, 71:1836-1842.
• [35]Lin NT, Chiou PY, Chang KC, Chen LK, Lai MJ: Isolation and characterization of phi AB2: a novel bacteriophage of Acinetobacter baumannii. Res Microbiol 2010, 161:308-314.
• [36]Hagens S, Loessner MJ: Application of bacteriophages for detection and control of foodborne pathogens. Appl Microbiol Biotechnol 2007, 76:513-519.
• [37]Iriarte FB, Balogh B, Momol MT, Smith LM, Wilson M, Jones JB: Factors affecting survival of bacteriophage on tomato leaf surfaces. Appl Environ Microbiol 2007, 73:1704-1711.
• [38]Chang KC, Lin NT, Hu A, Lin YS, Chen LK, Lai MJ: Genomic analysis of bacteriophage ϕAB1, a ϕKMV-like virus infecting multidrug-resistant Acinetobacter baumannii. Genomics 2011, 97:249-255.
• [39]McConnell MJ, Perez-Ordonez A, Perez-Romero P, Valencia R, Lepe JA, Vazquez-Barba I, Pachon J: Quantitative real-time PCR for detection of Acinetobacter baumannii colonization in the hospital environment. J Clin Microbiol 2012, 50:1412-1414.
• [40]Yang H, Liang L, Lin S, Jia S: Isolation and characterization of a virulent bacteriophage AB1 of Acinetobacter baumannii. BMC Microbiol 2010, 10:131. BioMed Central Full Text
• [41]Boyce JM, Kelliher S, Vallande N: Skin irritation and dryness associated with two hand-hygiene regimens: soap-and-water hand washing versus hand antisepsis with an alcoholic hand gel. Infect Control Hosp Epidemiol 2000, 21:442-448.
• [42]Goroncy-Bermes P, Schouten MA, Voss A: In vitro activity of a nonmedicated handwash product, chlorhexidine, and an alcohol-based hand disinfectant against multiply resistant gram-positive microorganisms. Infect Control Hosp Epidemiol 2001, 22:194-196.
• [43]Trick WE, Vernon MO, Hayes RA, Nathan C, Rice TW, Peterson BJ, Segreti J, Welbel SF, Solomon SL, Weinstein RA: Impact of ring wearing on hand contamination and comparison of hand hygiene agents in a hospital. Clin Infect Dis 2003, 36:1383-1390.
• [44]Mendez J, Jofre J, Lucena F, Contreras N, Mooijman K, Araujo R: Conservation of phage reference materials and water samples containing bacteriophages of enteric bacteria. J Virol Methods 2002, 106:215-224.
• [45]Adams M: Bacteriophages. Edited by Hershey AD. New York: Interscience; 1959:137-159.