Volume 5, Issue 3, September 2020, Page: 120-130
Detection of Biofilm Formation and Antibiotic Resistance in Klebsiella Oxytoca and Klebsiella Pneumoniae from Animal Origin Foods
Gedif Meseret Abebe, Department of Biology, College of Natural and Computational Science, Wolaita Sodo University, Wolaita Sodo, Ethiopia; Department of Biology, School of Natural and Applied Science, Gazi University, Ankara, Turkey
Received: May 17, 2020;       Accepted: May 29, 2020;       Published: Jul. 4, 2020
DOI: 10.11648/j.ijmb.20200503.17      View  207      Downloads  132
Abstract
Biofilms are surface-attached microbial communities with distinct properties, which have a tremendous impact on our health and food safety. The study was aimed to detect biofilm formation and antibiotic resistance by Klebsiella oxytoca and Klebsiella pneumoniae from animal origin foods. In this study100 food samples were examined for the presence of Klebsiella oxytoca, Klebsiella pneumoniae, and other Enterobacteriaceae family members. In this study, 19 Klebsiella oxytoca and 5 Klebsiella pneumoniae isolates were isolated from cheese and minced meat samples using standard biochemical tests and identification kit. Biofilm formation in these isolates was detected by using microplate, Congo red agar, and tube adherence methods. Antibiotic susceptibility testing was performed using the disk diffusion method on Muller-Hinton agar. Using a microplate method strong biofilm formation was observed in 16 (84%) and 5 (100%) isolates of Klebsiella oxytoca and Klebsiella pneumoniae after 24 hours of incubation on Tryptic Soy Broth medium containing 2% of glucose respectively. After 24 and 48 hours of incubation on Tryptic Soy Broth without glucose strong biofilm formation was detected in 10 (52.6%) and 2 (40%) isolates of Klebsiella oxytoca and Klebsiella pneumoniae respectively. After 24 hours of incubation on Congo red agar 14 (73.7%) and 3 (60%), isolates of Klebsiella oxytoca and Klebsiella pneumoniae were slime factor positive respectively. In tube adherence method 13 (68.4%) and 4 (80%), isolates of Klebsiella oxytoca and Klebsiella pneumoniae were seen to have adhered strongly after 24 hours of incubation on Tryptic Soy Broth medium containing 2% of glucose. In general, strong biofilm formation by these strains was seen on Tryptic Soy Broth medium when supplemented with glucose. Among all the Klebsiella oxytoca, the highest rates of susceptibility were seen toward Trimethoprim-Sulfamethoxazole (100%) and Imipenem (94.7%) followed by Chloramphenicol (73.7%) and gentamicin (68.4%). Among 19 Klebsiella oxytoca isolates, the highest rates of resistance were seen in streptomycin (73.7%) and Kanamycin (73.7%) followed by ampicillin (63.2%). The majority of Klebsiella pneumoniae isolates were resistant to Kanamycin (80%) and Streptomycin (80%) followed by Amikacin (60%). On the other hand, 80% of Klebsiella pneumoniae were susceptible to imipenem, chloramphenicol, Trimethoprim-Sulfamethoxazole, and cefotaxime. Generally, majority of Klebsiella oxytoca, and Klebsiella pneumoniae isolates showed strong biofilm production on different growth conditions and majority of the isolates were also resistance for antibiotics. Therefore, biofilm production by these nosocomial bacteria has an implication public health and pave the way for increased resistance of biofilm-associated organisms to antimicrobial agents.
Keywords
Klebsiella oxytoca, Klebsiella pneumoniae, Biofilm, Antibiotic Resistance
To cite this article
Gedif Meseret Abebe, Detection of Biofilm Formation and Antibiotic Resistance in Klebsiella Oxytoca and Klebsiella Pneumoniae from Animal Origin Foods, International Journal of Microbiology and Biotechnology. Vol. 5, No. 3, 2020, pp. 120-130. doi: 10.11648/j.ijmb.20200503.17
Copyright
Copyright © 2020 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Podschun R, Ullmann U (1998) Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev, 11: 589-603.
[2]
Darby A, Lertpiriyapong K, Sarkar U, Seneviratne U, Park DS, et al. (2014). Cytotoxic and Pathogenic Properties of Klebsiella oxytoca Isolated from Laboratory Animals. PLoS ONE 9 (7): e100542. doi: 10.1371/journal.pone.0100542.
[3]
Singh C. L, Cariappa C. M. P., Kaur L. C. M. (2016). Klebsiella oxytoca: An emerging pathogen? Medical journal armed forces india 72, 5 9–61.
[4]
Zollner-Schwetz I, Genauer C H, Joainig M, Weberhofer P, Gorkiewicz G, Valentin T, Hinterleitner TA, and Krause R (2008) Role of Klebsiella oxytoca in Antibiotic-Associated Diarrhea, Clinical Infectious Diseases 2008; 47: e74–8.
[5]
Wei J, Wenjie Y, Ping L, Na W, Haixia R, Xuequn Z (2018). Antibiotic resistance of Klebsiella pneumoniae through β-arrestin recruitment-induced β-lactamase signaling pathway, Experimental and therapeutic medicine 15: 2247-2254.
[6]
Ferreira RL, da Silva BCM, Rezende GS, Nakamura-Silva R, Pitondo-Silva A, Campanini EB, Brito MCA, da Silva EML, Freire CCM, Cunha AF and Pranchevicius MC (2019) High Prevalence of Multidrug-Resistant Klebsiella pneumoniae Harboring Several Virulence and β-Lactamase Encoding Genes in a Brazilian Intensive Care Unit. Front. Microbiol. 9: 3198. doi: 10.3389/fmicb.2018.03198.
[7]
Brisse S, Fevre C, Passet V, Issenhuth-Jeanjean S, Tournebize R, et al. (2009) Virulent Clones of Klebsiella pneumoniae: Identification and Evolutionary Scenario Based on Genomic and Phenotypic Characterization. PLoS ONE 4 (3): e4982. doi: 10.1371/journal.pone.0004982.
[8]
Haryani Y, Noorzaleha A. S, Fatimah, A. B., Noorjahan, B. A., Patrick, G. B., Shamsinar, A. T., et al. (2007). Incidence of Klebsiella pneumoniae in street foods sold in Malaysia and their characterization by antibiotic resistance, plasmid profiling, and RAPD–PCR analysis. Food Control 18, 847–853. doi: 10.1016/j.foodcont.2006.04.009.
[9]
Chenga F, Li Z, Lana S, Liua W, Li X (2018). Characterization of Klebsiella pneumoniae associated with cattle infections in southwest China using multi-locus sequence typing (MLST), antibiotic resistance and virulence-associated gene profile analysis, Brazilian journal of microbiology 49S, 93–100, https://doi.org/10.1016/j.bjm.2018.06.004.
[10]
Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15: 167-193.
[11]
Shi X, Zhu X (2009) Biofilm formation and food safety in food industries, Review 20, 407-413.
[12]
Singh S, Singh S K, Chowdhury I, Singh R (2017) Understanding the Mechanism of Bacterial Biofilms Resistance to Antimicrobial Agents, The Open Microbiology Journal, 11, 53-62.
[13]
Schroll C, Barken K. B, Krogfelt K. A and Struve C., (2010) Research article Role of type 1 and type 3 fimbriae in Klebsiella pneumoniae biofilm formation, BMC Microbiology, 10: 179 http://www.biomedcentral.com/1471-2180/10/179.
[14]
Zheng J, Lin Z, Chen C, Chen Z, Lin F, Wu Y, Yang S, Sun X, Yao W, Li D, Yu Z, Jin J, Qu D and Deng Q (2018) Biofilm Formation in Klebsiella pneumoniae Bacteremia Strains Was Found to be Associated with CC23 and the Presence of wcaG. Front. Cell. Infect. Microbiol. 8: 21. doi: 10.3389/fcimb.2018.00021.
[15]
Hall-Stoodley L, Costerton J W, Stoodley P. (2004) Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol. 2004 Feb; 2 (2): 95-108.
[16]
Ghasemian A, Mobarez A M., Peerayeh S N, Abadi A T B (2018). The association of surface adhesin genes and the biofilm formation among Klebsiella oxytoca clinical isolates, NMNI, 27, 36–39.
[17]
Ribeiro SM, de la Fuente-Núñez C, Baquir B, Faria-Junior C, Franco OL, Hancock REW (2015). Antibiofilm peptides increase the susceptibility of carbapenemase-producing Klebsiella pneumoniae clinical isolates to -lactam antibiotics. Antimicrob Agents Chemother 59: 3906 –3912.
[18]
Oh S, Chen P, Kang D (2007) Biofilm formation by Enterobacter sakazakii grown in artificial broth and infant formula on plastic surface. J Rapid Methods Automat Microbiol 15: 311-319.
[19]
Iversen C, Lane M. Forsythe S J (2004) The growth profilethermotolerance and biofilm formation of Enterobacter sakazakii grown in infant formula milk. Lett Appl Microbiol38: 378-382.
[20]
Kim H, Ryu J, Beuchat L (2006): Attachment of and biofilm formation by Enterobacter sakazakii on stainless steel and enteral feeding tubes: Appl Environ Microbiol 72 (9): 5846-5856.
[21]
Najjuka C F, Kateete DP, Kajumbula HM, Joloba ML, Essack SY (2016). Antimicrobial susceptibility profiles of Escherichia coli and Klebsiella pneumonia isolated from outpatients in urban and rural districts of Uganda, BMC Res Notes (2016) 9: 235, DOI 10.1186/s13104-016-2049-8.
[22]
Navon-Venezia S, Kondratyeva K, Carattoli A (2017) Klebsiella pneumoniae: a major worldwide source and shuttle for antibiotic resistance, Journal investing in science, FEMS Microbiology Reviews, 252–275.
[23]
Adamo R, Margarit I. (2018). Fighting antibiotic-resistant Klebsiella pneumoniae with “sweet” immune targets. mBio 9: e00874-18. https://doi.org/10.1128/mBio.00874-18.
[24]
Stepanovic S, Vukovic D, Hola V, Ventura GB, Djukic S, Irkovic Ruzika F (2007) “Quantification of biofilm in microtiter plates: Overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci “, Ap mis, 115: 891–9.
[25]
Nasr R A, Abu Shady H M, Hussein H S (2012) Biofilm formation and presence of icaAD gene in clinical isolates of staphylococci, The Egyptian Journal of Medical Human Genetics, 13: 269–274.
[26]
Christensen, G D, Simpson W A, Bisno A L, Beachey E H, Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces”, Infection and Immunity, 37: 318–26 (1982).
[27]
Freeman D J, Falkiner F R, Keane C T (1989). New method for detecting slime production by coagulase negative staphylococci, Journal of Clinical Pathology, 42: 872-4.
[28]
National Committee for Clinical Laboratory Standards (2003) Performance standards for antimicrobial susceptibility testing, 13th informational supplement, approved standard M100eS13 (M2). National Committee for Clinical Laboratory Standards, Wayne.
[29]
Seifi K, Kazemian H, Heidari H, Rezagholizadeh F, Saee Y, Shirvani F, and Houri H (2016) Evaluation of Biofilm Formation Among Klebsiella pneumoniae Isolates and Molecular Characterization by ERIC-PCR, Jundishapur J Microbiol, 9 (1).
[30]
Monirzadeh F, Shakibaie MR, Gholamrezazadeh M, Masoumi S (2018). Susceptibility of catheter-related Klebsiella pneumoniae strains to quaternary ammonium compounds under, Canadian Journal of Infection Control, 33 (4), 209-215.
[31]
Khodadadian R, Rahdar HA, Javadi A, Safari M, Khorshidi A (2018). Detection of VIM-1 and IMP-1 genes in Klebsiella pneumoniae and relationship with biofilm formation, Microbial Pathogenesis, 115, 25-30, https://doi.org/10.1016/j.micpath.2017.12.036.
[32]
Kokare C R, Chakraborty S, Khopade A N, Mahadik K R. (2009) Biofilm: importance and applications. Indian J Biotechnol.; 8: 159–168.
[33]
Osungunna M O, Onawunmi G O (2018), Antibiotic resistance profiles of biofilm-forming bacteria associated with urine and urinary catheters in a tertiary hospital in Ile-Ife, Nigeria, Southern African Journal of Infectious Diseases, 33 (3): 80–85.
[34]
Barati A, Ghaderpour A, Chew L. L, Bong C. W, Thong K. L, Chong V. C and Chai L. C (2016) Isolation and Characterization of Aquatic-Borne Klebsiella pneumoniae from Tropical Estuaries in Malaysia, Int. J. Environ. Res. Public Health, 13, 426.
[35]
Mathur T, Singhal S, Khan S, Upadhyay D J, Fatma T, Rattan A. (2006) Detection of biofilm formation among the clinical isolates of staphylococci: An evaluation of three different screening methods. Indian J Med Microbiol; 24: 25-9.
[36]
Kumar C G, Anand S K (1998). Significance of microbial biofilms in food industry: a review. International Journal of Food Microbiology, 42 (1), 9-27.
[37]
Frank JF, Ehlers J, Wicker L (2003) Removal of Listeria monocytogenes and poultry soil-containing biofilm using chemical cleaning and sanitizing agents under static conditions. Food Protection Trends23: 654-663.
[38]
Chakraborty S, Mohsına K, Sarker P K, Alam M. Z, Karım M I A, Sayem SM A (2016) Prevalence, antibiotic susceptibility profiles and ESBL production in Klebsiella pneumoniae and Klebsiella oxytoca among hospitalized patients, PERIODICUM BIOLOGORUM, 118, 1, 53–58.
[39]
Ghasemian A, Mobarez AM, Peerayeh SN, Abadi ATM, Khodaparast S, Mahmood SS (2019) Expression of adhesin genes and biofilm formation among Klebsiella oxytoca clinical isolates from patients with antibiotic-associated haemorrhagic colitis, Journal of Medical Microbiology, 68 (7), https://doi.org/10.1099/jmm.0.000965.
[40]
Zhang S, Yang G, Ye Q, Wu Q, Zhang J, Huang Y (2018) Phenotypic and Genotypic Characterization of Klebsiella pneumoniae Isolated From Retail Foods in China, Front. Microbiol. 9: 289. doi: 10.3389/fmicb.2018.00289.
[41]
Guo Y, Zhou H, Qin L, Pang Z, Qin T, Ren H, et al. (2016) Frequency, Antimicrobial Resistance and Genetic Diversity of Klebsiella pneumoniae in Food Samples. PLoS ONE 11 (4): e0153561. doi: 10.1371/journal.pone.0153561.
[42]
Khaertynov KS, Anokhin VA, Rizvanov AA, Davidyuk YN, Semyenova DR, Lubin SA and Skvortsova NN (2018) Virulence Factors and Antibiotic Resistance of Klebsiella pneumoniae Strains Isolated From Neonates With Sepsis. Front. Med. 5: 225. doi: 10.3389/fmed.2018.00225.
[43]
Kidd TJ, Mills G, Sá-Pessoa J, Dumigan A, Frank CG, Insua JL, Ingram R, Hobley L, Bengoechea JA (2017) A Klebsiella pneumoniae antibiotic resistance mechanism that subdues host defences and promotes virulence, EMBO Mol Med (2017) 9: 430–447, DOI 10.15252/emmm.201607336.
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