Phenotypic and Molecular Detection of Extended Spectrum Beta-Lactamases Producing-Bacteria Isolated From Pregnant Women With Genital Tract Infection in Duhok Governorate
DOI:
https://doi.org/10.25271/sjuoz.2021.9.2.784Keywords:
Vaginal Infection, Pregnant Women, MDR, XDR, ESBL, IraqAbstract
Beta-lactamase producing bacteria have a worldwide distribution with a high degree of prevalence in both community and hospital. Furthermore, multidrug resistant (MDR) and extended spectrum β-lactamases (ESBL) producing bacterial isolates from women patients may limit treatment options available. This study was designed to determine the frequency of bacterial isolates associated with genital tract infection in pregnant women and their antimicrobial resistance profile and to assess the prevalence of extended spectrum β-lactamases producing bacteria. Demonstrating the β-lactamase genes (blaTEM, blaSHV and blaCTX-M) by using polymerase chain reaction (PCR) assay with specific primers, was carried out on patients who were admitted to Maternity and Obstetric Hospital in Duhok city from November 2018 to October 2019. A total of 100 high vaginal swabs were collected from pregnant women patients between the ages 18-45 years. All clinical samples were cultured and standard microbiological methods were used to identify bacterial isolates, then confirmed by Vitek®2 compact automated system. All gram negative bacterial isolates were studied phenotypically and genotypically for extended spectrum β-lactamases-production. Out of 100 vaginal swabs, 88% confirmed positive culture; 90.9% of which were bacterial isolates. From the total bacterial isolates, 38.8% were gram negative bacteria, with a predominant 54.8% Klebsiella pneumoniae followed by Escherichia coli 35.5%. 54.8% of the isolates were characterized as multidrug resistance isolates, 29% isolates were extensive drug resistance, and no pan drug resistance were detected. Among these, the commonest extended spectrum β-lactamases producing isolates were Escherichia coli 81.8% followed by, Klebsiella pneumoniae 58.8%. Extended spectrum β-lactamases-producing isolates have showed significantly higher resistance than non- extended spectrum β-lactamases producing isolates to third and fourth generation cephalosporins. CTX-M was the most common β-lactamase gene 73.7% among extended spectrum β-lactamase producing strains, followed by blaSHV, 57.9% and blaTEM 52.6%, 21.1% had combination of all bla genes, 15.8% had CTX-M only and combination of blaCTX-M with blaSHV and blaTEM. 10.5% among extended spectrum β-lactamases producing isolates carried SHV type only and in combination with TEM type while TEM gene were observed in 5.3%. We concluded that the drug resistant isolates were common, worryingly high and it may limit treatment options available. In this study a high level of the blaCTX-M gene was demonstrated among extended spectrum β-lactamases producing isolates.
References
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Ahmad, S.S. and Ali, F.A., (2015). Microbiological Study of Pathogens Isolated From Women with Genital Tract Infection. International Journal of MediPharm Research. Vol.01, No.02, pp 95-105.
Al-Hilali, S. A. M. H. (2010). Occurrence and molecular characterization of enteropathogenic Escherichia coli (EPEC) serotypes isolated from children with diarrhea in Najaf (Doctoral dissertation, M. Sc. thesis: Kufa University, College of Medicine, Iraq).
Al-Mayahie, S. M. (2013). Phenotypic and genotypic comparison of ESBL production by vaginal Escherichia coli isolates from pregnant and non-pregnant women. Annals of Clinical Microbiology and Antimicrobials, 12(1), 7.
Bali, E. B., Accedil, L., & Sultan, N. (2010). Phenotypic and molecular characterization of SHV, TEM, CTX-M and extended-spectrum-lactamase produced by Escherichia coli, Acinobacter baumannii and Klebsiella isolates in a Turkish hospital. African Journal of Microbiology Research, 4(8), 650-654.
Bindayna, K., Khanfar, H. S., Senok, A. C., & Botta, G. A. (2010). Predominance of CTX-M genotype among extended spectrum beta lactamase isolates in a tertiary hospital in Saudi Arabia. Saudi Med J, 31(8), 859-863.
Biswas, S. M., Mia, M. R. A., Ara, N., Ibrahim, M., Nasir, T. A., & Yunus, S. (2013). Comparison of Three Dimensional Test and Double Disc Synergy Test for detection of Extended Spectrum β-Lactamase (ESBL) producing Gram negative bacteria. Pulse, 6(1-2), 12-19.
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Dagi, H. T., Al Dulaimi, D. A. A., Kus, H., Seyhan, T., Findik, D., Tuncer, I., & Arslan, U. (2015). Genotype distribution of extended Spectrum β-Lactamase producing Escherichia coli and Klebsiella pneumoniae. Biomedical Research, 26(2), 235-238.
Divya, G. A., & Karthika, J. (2015). Study to Detect ESBL producing Escherichia coli Isolates from Women with Genital Tract Infection. Indian Journal of Applied Research, 5(8).
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Donders, G. G., Vereecken, A., Bosmans, E., Dekeersmaecker, A., Salembier, G., & Spitz, B. (2002). Definition of a type of abnormal vaginal flora that is distinct from bacterial vaginosis: aerobic vaginitis. BJOG: an international journal of obstetrics and gynaecology, 109(1), 34-43.
Ensor, V. M., Jamal, W., Rotimi, V. O., Evans, J. T., & Hawkey, P. M. (2009). Predominance of CTX-M-15 extended spectrum β-lactamases in diverse Escherichia coli and Klebsiella pneumoniae from hospital and community patients in Kuwait. International journal of antimicrobial agents, 33(5), 487-489.
Fernando, M. M. P. S. C., Luke, W. A. N. V., Miththinda, J. K. N. D., Wickramasinghe, R. D. S. S., Sebastiampillai, B. S., Gunathilake, M. P. M. L., ... & Premaratna, R. (2017). Extended spectrum beta lactamase producing organisms causing urinary tract infections in Sri Lanka and their antibiotic susceptibility pattern–a hospital based cross sectional study. BMC infectious diseases, 17(1), 138.
Gaddar, N., Anastasiadis, E., Halimeh, R., Ghaddar, A., Matar, G. M., Abou Fayad, A., & El Chaar, M. (2020). Phenotypic and Genotypic Characterization of Extended-Spectrum Beta-Lactamases Produced by Escherichia coli Colonizing Pregnant Women. Infectious Diseases in Obstetrics and Gynecology.2020.1-7
Gazin, M., Paasch, F., Goossens, H., & Malhotra-Kumar, S. (2012). Current trends in culture-based and molecular detection of extended-spectrum-β-lactamase-harboring and carbapenem-resistant Enterobacteriaceae. Journal of clinical microbiology, 50(4), 1140-1146.
Gorish, B.M.T.(2019). Phenotypic Detection of Extended-Spectrum B-Lactamase Producing Escherichia coli and Klebsiella pneumoniae from a Vaginal Swab of Pregnant women in Khartoum State. Acta Scientific Women's Health, 1:13-17
Harada Y, Morinaga Y, Yamada K, Migiyama Y, Nagaoka K. (2013). Clinical and molecular epidemiology of extended-spectrum β-lactamase-producing Klebsiella pneumoniae and Escherichia coli in a Japanese tertiary hospital. J Med Microb Diagn. 2:2161-0703.
Hassan, M. I., Alkharsah, K. R., Alzahrani, A. J., Obeid, O. E., Khamis, A. H., & Diab, A. (2013). Detection of extended spectrum beta-lactamases-producing isolates and effect of AmpC overlapping. The Journal of Infection in Developing Countries, 7(08), 618-629.
Hawkey, P. M. (2008). Prevalence and clonality of extended-spectrum beta-lactamases in Asia. Clin. Microbiol. Infect., 14: 159-165.
Jabbar, A. D. (2013). Phenotypic and Genotypic Detection of Extended-Spectrum β-Lactamases (ESBL) among Escherichia coli Isolated from Symptomatic Female's Genital Tract Infection. Journal Of Wassit For Science & Medicine, 6(1), 59-67.
Kaambo, E., Africa, C., Chambuso, R., & Passmore, J. A. S. (2018). Vaginal microbiomes associated with aerobic vaginitis and bacterial vaginosis. Frontiers in Public Health, 6, 78.
Kaftandzieva A, Trajkovska-Dokic E, Panovski N (2011) Prevalence and molecular characterization of Extended spectrum beta lactamase producing Escherichia coli and Klebsilla pneumonia. Contributions, Sec. Biol. Med. Sci., 32 (2),129-141
Khamees, S., 2012. Characterization of vaginal discharge among women complaining of genital tract infection. Int. J. of Pharm. & Life Sci.(IJPLS) Oct, 1(3),10.
Lamichhane, P., Joshi, D. R., Subedi, Y. P., Thapa, R., Acharya, G. P., & Lamsal, A. (2014). Study on types of vaginitis and association between bacterial vaginosis and urinary tract infection in pregnant women. IJBAR, 5(06), 305-307.
Leylabadlo, H. E., Pourlak, T., Aghazadeh, M., Asgharzadeh, M., & Kafil, H. S. (2017). Extended-spectrum beta-lactamase producing gram negative bacteria In Iran: A review. African journal of infectious diseases, 11(2), 39-53.
Livermore, D. M. (2012). Current epidemiology and growing resistance of gram-negative pathogens. The Korean journal of internal medicine, 27(2), 128.
Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., and Paterson, D. L. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical microbiology and infection, 18(3), 268-281.
Manoharan, A., Premalatha, K., Chatterjee, S., Mathai, D., & SARI Study Group. (2011). Correlation of TEM, SHV and CTX-M extended-spectrum beta lactamases among Enterobacteriaceae with their in vitro antimicrobial susceptibility. Indian journal of medical microbiology, 29(2), 161-164.
Mawlood, A. H., Omer, S. A., Jalal, S. T., & Haji, S. H. (2018). Molecular detection of SHV-Type ESBL in E. coli and K. pneumoniae and their antimicrobial resistance profile. Zanco Journal of Medical Sciences, 22(2), 262-272.
Mumtaz, S., Ahmad, M., Aftab, I., Akhtar, N., ul Hassan, M., & Hamid, A. (2008). Aerobic vaginal pathogens and their sensitivity pattern. J Ayub Med Coll Abbottabad, 20(1), 113-7.
Ogefere, H. O., Aigbiremwen, P. A., & Omoregie, R. (2015). Extended-spectrum beta-lactamase (ESBL)–producing Gram-negative isolates from urine and wound specimens in a tertiary health facility in southern Nigeria. Tropical Journal of Pharmaceutical Research, 14(6), 1089-1094.
Oksüz, L., & Gürler, N. (2009). Typing of extended-spectrum beta-lactamases in Escherichia coli and Klebsiella spp. strains and analysis of plasmid profiles. Mikrobiyoloji bulteni, 43(2), 183-194.
Pal, K., Sidhu, S. K., Deiv, P., Malhotra, S., Malhotra, A., & Soneja, S. (2017). Etiology of vaginal infections and antimicrobial resistance pattern of aerobic bacterial isolates in women of reproductive age group attending a tertiary care hospital. APJHS, 4(4), 15-18.
Pincus, D.H., (2010). Microbial identification using the bioMérieux Vitek® 2 system. Encyclopedia of rapid microbiological methods, 1, 1-32.
Ravishankar, N., & Prakash, M. (2017). Antibiogram of Bacterial Isolates from High Vaginal Swabs of Pregnant Women from Tertiary Care Hospital in Puducherry, India. Int. J. Curr. Microbiol. App. Sci, 6(1), 964-972.
Razzak M S A, Al-Charrakh A H and Al-Greitty B H. (2011). Relationship between lactobacilli and opportunistic bacterial pathogens associated with vaginittis. North American Journal of Medical Sciences; 3 (4): 185-192.
Sangeetha KT, Saroj Golia, Vasudha C. L. A (2015). A study of aerobic bacterial pathogens associated with vaginitis in reproductive age group women (15-45 years) and their sensitivity pattern. International Journal of Research in Medical Sciences, 3(9), 2268-2273.
Talaiekhozani, A., Alaee, S., & Ponraj, M. (2015). Guidelines for quick application of biochemical tests to identify unknown bacteria. AOBR, 2(2), 65-82.
Tang, Y., Yu, F., Hu, Z., Peng, L., & Jiang, Y. (2020). Characterization of aerobic vaginitis in late pregnancy in a Chinese population: A STROBE-compliant study. Medicine, 99(25).
Tariq, N., Jaffery, T., Ayub, R., Alam, A. Y., Javid, M. H., & Shafique, S. (2006). Frequency and antimicrobial susceptibility of aerobic bacterial vaginal isolates. Journal of the College of Physicians and Surgeons--Pakistan: JCPSP, 16(3), 196-199.
Wax, R. G., Lewis, K., Salyers, A. A., & Taber, H.. (2008). Bacterial resistance to antimicrobials. 2nd ed.). NW. Taylor & Francis Group: CRC Press.
Ahmad, S.S. and Ali, F.A., (2015). Microbiological Study of Pathogens Isolated From Women with Genital Tract Infection. International Journal of MediPharm Research. Vol.01, No.02, pp 95-105.
Al-Hilali, S. A. M. H. (2010). Occurrence and molecular characterization of enteropathogenic Escherichia coli (EPEC) serotypes isolated from children with diarrhea in Najaf (Doctoral dissertation, M. Sc. thesis: Kufa University, College of Medicine, Iraq).
Al-Mayahie, S. M. (2013). Phenotypic and genotypic comparison of ESBL production by vaginal Escherichia coli isolates from pregnant and non-pregnant women. Annals of Clinical Microbiology and Antimicrobials, 12(1), 7.
Bali, E. B., Accedil, L., & Sultan, N. (2010). Phenotypic and molecular characterization of SHV, TEM, CTX-M and extended-spectrum-lactamase produced by Escherichia coli, Acinobacter baumannii and Klebsiella isolates in a Turkish hospital. African Journal of Microbiology Research, 4(8), 650-654.
Bindayna, K., Khanfar, H. S., Senok, A. C., & Botta, G. A. (2010). Predominance of CTX-M genotype among extended spectrum beta lactamase isolates in a tertiary hospital in Saudi Arabia. Saudi Med J, 31(8), 859-863.
Biswas, S. M., Mia, M. R. A., Ara, N., Ibrahim, M., Nasir, T. A., & Yunus, S. (2013). Comparison of Three Dimensional Test and Double Disc Synergy Test for detection of Extended Spectrum β-Lactamase (ESBL) producing Gram negative bacteria. Pulse, 6(1-2), 12-19.
Clinical and Laboratory Standard Institute. (2018). M100: Performance standards for antimicrobial susceptibility testing. 28th edition .Clinical Lab Standards Institute
Coque, T. M., Baquero, F., & Canton, R. (2008). Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe. Eurosur eillance, 13(47), 19044.
Dagi, H. T., Al Dulaimi, D. A. A., Kus, H., Seyhan, T., Findik, D., Tuncer, I., & Arslan, U. (2015). Genotype distribution of extended Spectrum β-Lactamase producing Escherichia coli and Klebsiella pneumoniae. Biomedical Research, 26(2), 235-238.
Divya, G. A., & Karthika, J. (2015). Study to Detect ESBL producing Escherichia coli Isolates from Women with Genital Tract Infection. Indian Journal of Applied Research, 5(8).
Donders, G. G. G., Bellen, G., & Rezeberga, D. (2011). Aerobic vaginitis in pregnancy. BJOG: An International Journal of Obstetrics & Gynaecology, 118(10), 1163-1170.
Donders, G. G., Vereecken, A., Bosmans, E., Dekeersmaecker, A., Salembier, G., & Spitz, B. (2002). Definition of a type of abnormal vaginal flora that is distinct from bacterial vaginosis: aerobic vaginitis. BJOG: an international journal of obstetrics and gynaecology, 109(1), 34-43.
Ensor, V. M., Jamal, W., Rotimi, V. O., Evans, J. T., & Hawkey, P. M. (2009). Predominance of CTX-M-15 extended spectrum β-lactamases in diverse Escherichia coli and Klebsiella pneumoniae from hospital and community patients in Kuwait. International journal of antimicrobial agents, 33(5), 487-489.
Fernando, M. M. P. S. C., Luke, W. A. N. V., Miththinda, J. K. N. D., Wickramasinghe, R. D. S. S., Sebastiampillai, B. S., Gunathilake, M. P. M. L., ... & Premaratna, R. (2017). Extended spectrum beta lactamase producing organisms causing urinary tract infections in Sri Lanka and their antibiotic susceptibility pattern–a hospital based cross sectional study. BMC infectious diseases, 17(1), 138.
Gaddar, N., Anastasiadis, E., Halimeh, R., Ghaddar, A., Matar, G. M., Abou Fayad, A., & El Chaar, M. (2020). Phenotypic and Genotypic Characterization of Extended-Spectrum Beta-Lactamases Produced by Escherichia coli Colonizing Pregnant Women. Infectious Diseases in Obstetrics and Gynecology.2020.1-7
Gazin, M., Paasch, F., Goossens, H., & Malhotra-Kumar, S. (2012). Current trends in culture-based and molecular detection of extended-spectrum-β-lactamase-harboring and carbapenem-resistant Enterobacteriaceae. Journal of clinical microbiology, 50(4), 1140-1146.
Gorish, B.M.T.(2019). Phenotypic Detection of Extended-Spectrum B-Lactamase Producing Escherichia coli and Klebsiella pneumoniae from a Vaginal Swab of Pregnant women in Khartoum State. Acta Scientific Women's Health, 1:13-17
Harada Y, Morinaga Y, Yamada K, Migiyama Y, Nagaoka K. (2013). Clinical and molecular epidemiology of extended-spectrum β-lactamase-producing Klebsiella pneumoniae and Escherichia coli in a Japanese tertiary hospital. J Med Microb Diagn. 2:2161-0703.
Hassan, M. I., Alkharsah, K. R., Alzahrani, A. J., Obeid, O. E., Khamis, A. H., & Diab, A. (2013). Detection of extended spectrum beta-lactamases-producing isolates and effect of AmpC overlapping. The Journal of Infection in Developing Countries, 7(08), 618-629.
Hawkey, P. M. (2008). Prevalence and clonality of extended-spectrum beta-lactamases in Asia. Clin. Microbiol. Infect., 14: 159-165.
Jabbar, A. D. (2013). Phenotypic and Genotypic Detection of Extended-Spectrum β-Lactamases (ESBL) among Escherichia coli Isolated from Symptomatic Female's Genital Tract Infection. Journal Of Wassit For Science & Medicine, 6(1), 59-67.
Kaambo, E., Africa, C., Chambuso, R., & Passmore, J. A. S. (2018). Vaginal microbiomes associated with aerobic vaginitis and bacterial vaginosis. Frontiers in Public Health, 6, 78.
Kaftandzieva A, Trajkovska-Dokic E, Panovski N (2011) Prevalence and molecular characterization of Extended spectrum beta lactamase producing Escherichia coli and Klebsilla pneumonia. Contributions, Sec. Biol. Med. Sci., 32 (2),129-141
Khamees, S., 2012. Characterization of vaginal discharge among women complaining of genital tract infection. Int. J. of Pharm. & Life Sci.(IJPLS) Oct, 1(3),10.
Lamichhane, P., Joshi, D. R., Subedi, Y. P., Thapa, R., Acharya, G. P., & Lamsal, A. (2014). Study on types of vaginitis and association between bacterial vaginosis and urinary tract infection in pregnant women. IJBAR, 5(06), 305-307.
Leylabadlo, H. E., Pourlak, T., Aghazadeh, M., Asgharzadeh, M., & Kafil, H. S. (2017). Extended-spectrum beta-lactamase producing gram negative bacteria In Iran: A review. African journal of infectious diseases, 11(2), 39-53.
Livermore, D. M. (2012). Current epidemiology and growing resistance of gram-negative pathogens. The Korean journal of internal medicine, 27(2), 128.
Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., and Paterson, D. L. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical microbiology and infection, 18(3), 268-281.
Manoharan, A., Premalatha, K., Chatterjee, S., Mathai, D., & SARI Study Group. (2011). Correlation of TEM, SHV and CTX-M extended-spectrum beta lactamases among Enterobacteriaceae with their in vitro antimicrobial susceptibility. Indian journal of medical microbiology, 29(2), 161-164.
Mawlood, A. H., Omer, S. A., Jalal, S. T., & Haji, S. H. (2018). Molecular detection of SHV-Type ESBL in E. coli and K. pneumoniae and their antimicrobial resistance profile. Zanco Journal of Medical Sciences, 22(2), 262-272.
Mumtaz, S., Ahmad, M., Aftab, I., Akhtar, N., ul Hassan, M., & Hamid, A. (2008). Aerobic vaginal pathogens and their sensitivity pattern. J Ayub Med Coll Abbottabad, 20(1), 113-7.
Ogefere, H. O., Aigbiremwen, P. A., & Omoregie, R. (2015). Extended-spectrum beta-lactamase (ESBL)–producing Gram-negative isolates from urine and wound specimens in a tertiary health facility in southern Nigeria. Tropical Journal of Pharmaceutical Research, 14(6), 1089-1094.
Oksüz, L., & Gürler, N. (2009). Typing of extended-spectrum beta-lactamases in Escherichia coli and Klebsiella spp. strains and analysis of plasmid profiles. Mikrobiyoloji bulteni, 43(2), 183-194.
Pal, K., Sidhu, S. K., Deiv, P., Malhotra, S., Malhotra, A., & Soneja, S. (2017). Etiology of vaginal infections and antimicrobial resistance pattern of aerobic bacterial isolates in women of reproductive age group attending a tertiary care hospital. APJHS, 4(4), 15-18.
Pincus, D.H., (2010). Microbial identification using the bioMérieux Vitek® 2 system. Encyclopedia of rapid microbiological methods, 1, 1-32.
Ravishankar, N., & Prakash, M. (2017). Antibiogram of Bacterial Isolates from High Vaginal Swabs of Pregnant Women from Tertiary Care Hospital in Puducherry, India. Int. J. Curr. Microbiol. App. Sci, 6(1), 964-972.
Razzak M S A, Al-Charrakh A H and Al-Greitty B H. (2011). Relationship between lactobacilli and opportunistic bacterial pathogens associated with vaginittis. North American Journal of Medical Sciences; 3 (4): 185-192.
Sangeetha KT, Saroj Golia, Vasudha C. L. A (2015). A study of aerobic bacterial pathogens associated with vaginitis in reproductive age group women (15-45 years) and their sensitivity pattern. International Journal of Research in Medical Sciences, 3(9), 2268-2273.
Talaiekhozani, A., Alaee, S., & Ponraj, M. (2015). Guidelines for quick application of biochemical tests to identify unknown bacteria. AOBR, 2(2), 65-82.
Tang, Y., Yu, F., Hu, Z., Peng, L., & Jiang, Y. (2020). Characterization of aerobic vaginitis in late pregnancy in a Chinese population: A STROBE-compliant study. Medicine, 99(25).
Tariq, N., Jaffery, T., Ayub, R., Alam, A. Y., Javid, M. H., & Shafique, S. (2006). Frequency and antimicrobial susceptibility of aerobic bacterial vaginal isolates. Journal of the College of Physicians and Surgeons--Pakistan: JCPSP, 16(3), 196-199.
Wax, R. G., Lewis, K., Salyers, A. A., & Taber, H.. (2008). Bacterial resistance to antimicrobials. 2nd ed.). NW. Taylor & Francis Group: CRC Press.
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2021-06-30
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Nerway, S. M., & Al-Delaimi, M. S. (2021). Phenotypic and Molecular Detection of Extended Spectrum Beta-Lactamases Producing-Bacteria Isolated From Pregnant Women With Genital Tract Infection in Duhok Governorate. Science Journal of University of Zakho, 9(2), 80–88. https://doi.org/10.25271/sjuoz.2021.9.2.784
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