Characterization of NDM-1-producing carbapenemase in Acinetobacter spp. and E. coli isolates from diseased pigs

Abstract

In recent years, the mobile metallo-β-lactamase (MBL) genes have been found to correspond to one of the most important resistance characters identified in Gram-negative bacteria, severely affecting clinical chemotherapy and threatening public health. The prevalence of mobile MBL genes and their flanking regions in Gram-negative bacteria from diseased pigs in China was investigated. A total of 334 lung samples from diseased pigs were screened for Gram-negative bacteria classified as non-susceptible to meropenem (MIC≥4 mg·L⁻¹). Six isolates, including three Escherichia coli, two Acinetobacter baumanii and one A. calcoaeticus, exhibited MBL production and carried the bla_NDM-1 gene. S1-PFGE and Southern blot analysis showed that the bla_NDM-1 gene was located on the chromosome of one A. baumanii isolate and on plasmids of various sizes in the other five isolates. MIC testing using broth microdilution revealed that all bla_NDM-1-carrying isolates and some of their transconjugants exhibited resistance to almost all β-lactams tested. Whole genome sequencing revealed that the flanking region of the bla_NDM-1 gene from all porcine isolates had high levels of similarity with the corresponding regions in human isolates. One porcine E. coli isolate carrying bla_NDM-1 was typed as ST48, a common sequence type in human E. coli isolates. These results suggest the possibility of human-to-food animal transfer of bla_NDM-1-producing E. coli, highlighting the need for surveillance of carbapenemase producers among bacteria from food animals. In addition, the prudent use of antimicrobial agents to decrease the opportunities for co-selection of carbapenemase genes in food animals is also urgently needed.

Keywords

carbapenemase / NDM-1 / ISAba125 / Enterobacteriaceae / food safety

References

[1] Cornaglia G, Giamarellou H, Rossolini G M. Metallo-β-lactamases: a last frontier for β-lactams? The Lancet Infectious Diseases, 2011, 11(5): 381–393

[2] Yong D, Toleman M A, Giske C G, Cho H S, Sundman K, Lee K, Walsh T R. Characterization of a new metallo-β-lactamase gene, bla_NDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrobial Agents and Chemotherapy, 2009, 53(12): 5046–5054

[3] Satlin M J, Calfee D P, Chen L, Fauntleroy K A, Wilson S J, Jenkins S G, Feldman E J, Roboz G J, Shore T B, Helfgott D C, Soave R, Kreiswirth B N, Walsh T J. Emergence of carbapenem-resistant Enterobacteriaceae as causes of bloodstream infections in patients with hematologic malignancies. Leukemia & Lymphoma, 2013, 54(4): 799–806

[4] Dortet L, Poirel L, Nordmann P. Worldwide dissemination of the NDM-type carbapenemases in Gram-negative bacteria. BioMed Research International, 2014: 249856

[5] Walsh T R, Weeks J, Livermore D M, Toleman M A. Dissemination of NDM-1 positive bacteria in the New Delhi environment and its implications for human health: an environmental point prevalence study. The Lancet Infectious Diseases, 2011, 11(5): 355–362

[6] Isozumi R, Yoshimatsu K, Yamashiro T, Hasebe F, Nguyen B M, Ngo T C, Yasuda S P, Koma T, Shimizu K, Arikawa J. bla_NDM-1-positive Klebsiella pneumoniae from environment, Vietnam. Emerging Infectious Diseases, 2012, 18(8): 1383–1385

[7] Zhang C, Qiu S, Wang Y, Qi L, Hao R, Liu X, Shi Y, Hu X, An D, Li Z, Li P, Wang L, Cui J, Wang P, Huang L, Klena J D, Song H. Higher isolation of NDM-1 producing Acinetobacter baumannii from the sewage of the hospitals in Beijing. PLoS ONE, 2014, 8(6): e64857

[8] Wang  B,  Sun  D.  Detection of  NDM-1  carbapenemase-producing Acinetobacter calcoaceticus  and  Acinetobacter junii  in environmental samples from livestock farms. Journal of Antimicrobial Chemotherapy, 2015, 70(2): 611–613

[9] Zhang  W  J,  Lu  Z,  Schwarz  S,  Zhang  R  M,  Wang  X  M,  Si  W, Yu S, Chen L, Liu S. Complete sequence of the bla_NDM-1-carrying plasmid pNDM-AB from Acinetobacter baumannii of food animal origin. Journal of Antimicrobial Chemotherapy, 2013, 68(7): 1681–1682

[10] Wang Y, Wu C, Zhang Q, Qi J, Liu H, Wang Y, He T, Ma L, Lai J, Shen Z, Liu Y, Shen J. Identification of New Delhi metallo-β-lactamase 1 in Acinetobacter lwoffii of food animal origin. PLoS ONE, 2012, 7(5): e37152

[11] Shaheen B W, Nayak R, Boothe D M. Emergence of a New Delhi metallo-β-lactamase (NDM-1)-encoding gene in clinical Escherichia coli isolates recovered from companion animals in the United States. Antimicrobial Agents and Chemotherapy, 2013, 57(6): 2902–2903

[12] Wang Y, He T, Schwarz S, Zhao Q, Shen Z, Wu C, Shen J. Multidrug resistance gene cfr in methicillin-resistant coagulase-negative staphylococci from chickens, ducks, and pigs in China. International Journal of Medical Microbiology, 2013, 303(2): 84–87

[13] CLSI document M100-S25. Performance standards for antimicrobial susceptibility testing; twenty-fifth informational supplement. Panama: Clinical and Laboratory Standards Institute, 2015

[14] Wang Y, Wang X, Schwarz S, Zhang R, Lei L, Liu X, Lin D, Shen J. IMP-45-producing multidrug-resistant Pseudomonas aeruginosa of canine origin. Journal of Antimicrobial Chemotherapy, 2014, 69(9): 2579–2581

[15] Benson D A, Cavanaugh M, Clark K, Karsch-Mizrachi I, Lipman D J, Ostell J, Sayers E W. GenBank. Nucleic Acids Research, 2013, 41(D1): D36–D42

[16] Liu Y, Wang Y, Schwarz S, Li Y, Shen Z, Zhang Q, Wu C, Shen J. Transferable multiresistance plasmids carrying cfr in Enterococcus spp. from swine and farm environment. Antimicrobial Agents and Chemotherapy, 2013, 57(1): 42–48

[17] Smet A, Martel A, Persoons D, Dewulf J, Heyndrickx M, Claeys G, Lontie M, Van Meensel B, Herman L, Haesebrouck F, Butaye P. Characterization of extended-spectrum β-lactamases produced by Escherichia coli isolated from hospitalized and nonhospitalized patients: emergence of CTX-M-15-producing strains causing urinary tract infections. Microbial Drug Resistance, 2010, 16(2): 129–134

[18] Ben Sallem R, Ben Slama K, Estepa V, Jouini A, Gharsa H, Klibi N, Sáenz Y, Ruiz-Larrea F, Boudabous A, Torres C. Prevalence and characterisation of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates in healthy volunteers in Tunisia. European Journal of Clinical Microbiology & Infectious Diseases, 2012, 31(7): 1511–1516

[19] Philippon A, Arlet G, Jacoby G A. Plasmid-determined AmpC-type β-lactamases. Antimicrobial Agents and Chemotherapy, 2002, 46(1): 1–11

[20] Hu H, Hu Y, Pan Y, Liang H, Wang H, Wang X, Hao Q, Yang X, Yang X, Xiao X, Luan C, Yang Y, Cui Y, Yang R, Gao G F, Song Y, Zhu B. Novel plasmid and its variant harboring both a bla_NDM-1 gene and type IV secretion system in clinical isolates of Acinetobacter lwoffii. Antimicrobial Agents and Chemotherapy, 2012, 56(4): 1698–1702

[21] Wang X, Xu X, Li Z, Chen H, Wang Q, Yang P, Zhao C, Ni M, Wang H. An outbreak of a nosocomial NDM-1-producing Klebsiella pneumoniae ST147 at a teaching hospital in mainland China. Microbial Drug Resistance, 2014, 20(2): 144–149

[22] Bonnin R A, Poirel L, Carattoli A, Nordmann P. Characterization of an IncFII plasmid encoding NDM-1 from Escherichia coli ST131. PLoS ONE, 2012, 7(4): e34752

Acknowledgements

This study was supported by grants from the National Basic Research Program of China (2013CB127200), and the National Natural Science Foundation of China (31422055).

Rongmin Zhang, Yang Wang, Zhihai Liu, Jiyun Li, Wenjuan Yin, Lei Lei, Congming Wu, and Jianzhong Shen declare that they have no conflict of interest or financial conflicts to disclose.

All applicable institutional and national guidelines for the care and use of animals were followed.

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg