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First reported New Delhi metallo-β-lactamase-1-producing Cedecea lapagei
ARTICLE IN PRESS International Journal of Antimicrobial Agents ■■ (2016) ■■–■■
Contents lists available at ScienceDirect
International Journal of Antimicrobial Agents j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / i j a n t i m i c a g
1
Q2 Letter to the Editor
2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54
First reported New Delhi metallo-β-lactamase1-producing Cedecea lapagei Sir, Cedecea lapagei is a Gram-negative, facultative anaerobic, nonspore-forming bacterium belonging to the family Enterobacteriaceae, first isolated by the US Centers for Disease Control and Prevention (CDC) laboratories in 1981. It has been reported as a pathogen in a few cases of bacterial peritonitis, wound infection, chemical burns and pneumonia [1]. New Delhi metallo-β-lactamase-1 (NDM-1) was first reported in 2009 in Klebsiella pneumoniae and Escherichia coli isolated from a patient in Sweden who had received medical care in New Delhi, India [2]. Here we report the presence of a clinically significant NDM-1producing C. lapagei in a 26-day-old female preterm baby admitted to the paediatric intensive care unit (ICU) of a 1300-bed tertiary care hospital in Aligarh, India. The patient was diagnosed with lateonset sepsis, apnoea and hypocalcaemia and was treated with cefotaxime and phenytoin (Epsolin) with no recovery observed after the first week. Amikacin was also added to the treatment along with cefotaxime for another week. The baby started recovering after 14 days. An NDM-1-producing C. lapagei isolate was detected in a blood sample, which to the best of our knowledge is the first report of NDM-1-producing C. lapagei. The identity of the isolated strain (AK68) was confirmed by BD PhoenixTM 100 Automated Microbiology System using panel NMIC/ ID-55 (Gram-negative susceptibility card) followed by 16S rDNA sequencing. Furthermore, antimicrobial susceptibility was determined by the standard disk diffusion method using Mueller– Hinton agar as per Clinical and Laboratory Standards Institute (CLSI) guidelines. The strain was found to be resistant to imipenem, meropenem, aztreonam, ceftazidime, cefotaxime, cefoxitin, cefepime and cefoperazone/sulbactam. Moreover, detection of metallo-βlactamase activity was performed using two imipenem disks (10 μg), one containing 10 μL of 0.1 M anhydrous ethylene diamine tetraacetic acid (EDTA). The disks were placed 25 mm apart on Mueller– Hinton agar plates [3]. Minimum inhibitory concentrations (MICs) were determined following CLSI guidelines and the results are given in Table 1. MIC data showed high resistance to β-lactams. PCR amplification of whole DNA from strain AK68 using previously described primers revealed the presence of blaNDM [4]. Following purification, the amplified product was sent for sequencing (SciGenom Labs Pvt. Ltd., Kerala, India). The obtained sequence was identified as Q3 blaNDM-1 by sequence analysis using BLAST (http://www.ncbi.nlm.nih .gov). The presence of other markers (blaVIM, blaOXA-1, blaOXA-9, blaCMY, blaSHV, blaCTX-M, blaTEM, blaIMP and blaKPC) was also checked for in association with blaNDM-1. Only blaCTX-M, blaSHV and blaTEM were found in coexistence with blaNDM-1. The genetic environment of blaNDM-1 was analysed for the presence of an insertion sequence (IS) known to be associated with the blaNDM-1 in Enterobacteriaceae [5]. Primers targeting the IS element Aba125 identified a complete ISAba125
Table 1 Minimum inhibitory concentration (MIC) and genetic profile of NDM-1-producing Cedecea lapagei strain AK68. MIC (μg/mL) IPM MEM ATM
Genetic environment of blaNDM-1 CAZ
CTX
FOX
CPM SCP Upstream Downstream
512 >256 >1024 >1024 >1024 >1024 128
64
Complete bleMBL absent ISAba125
MIC, minimum inhibitory concentration; IPM, imipenem; MEM, meropenem; ATM, aztreonam; CAZ, ceftazidime; CTX, cefotaxime; FOX, cefoxitin; CPM, cefepime; SCP, cefoperazone/sulbactam.
55 56 57 58 59 60 61 62 63 64 65 66
upstream of blaNDM-1 in strain AK68. The bleomycin resistance gene bleMBL was not identified downstream of blaNDM-1, which is an unusual feature compared with earlier known species carrying blaNDM-1 [3] and needs to be further characterised. The plasmid incompatibility group was determined by PCRbased replicon typing (PBRT). The strain was found to be untypeable. Conjugal transfer was performed using C. lapagei AK68 as donor and azide-resistant E. coli J53 strain as recipient on selection medium with cefoxitin (10 μg/mL) and sodium azide (100 μg/mL). Conjugation confirmed the plasmid location of blaNDM-1. This is the first case of an NDM-1-producing C. lapagei strain isolated from a neonate admitted to the paediatric ICU of a north Indian hospital. Hence, there is an urgent need to perform a proper surveillance study to control further spread of NDM-1 in evolving new species. Nucleotide sequence accession number: The sequence of blaNDM-1 determined in this strain was submitted to GenBank with the accession no. KX231908. Funding: This study was supported by grants DBT Award BT/ HRD/NBA/34/01/2012, BT/PR8281/BID/7/448/2013 and ICMR grant AMR/5/2011-ECD-1 to AUK. Competing interests: None declared. Ethical approval: Clearance from the institution ethical committee was received for the whole study [no. 151/201517/PDFWM2015-2017-UTT-31140 (SAII)].
67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92
References [1] Salazar G, Almeida A, Gómez M. Cedecea lapagei traumatic wound infection: case report and literature review. Rev Chilena Infectol 2013;30:86–9, [in Spanish]. [2] Yong D, Toleman MA, Giske CG, Cho HS, Sundman K, Lee K, et al. Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother 2009;53:5046–54. [3] Khan AU, Parvez S. Detection of blaNDM-4 in Escherichia coli from hospital sewage. J Med Microbiol 2014;63:1404–6. [4] Khan AU, Nordmann P. NDM-1-producing Enterobacter cloacae and Klebsiella pneumoniae from diabetic foot ulcers in India. J Med Microbiol 2012;61:454–6.
http://dx.doi.org/10.1016/j.ijantimicag.2016.10.001 0924-8579/© 2016 Published by Elsevier B.V.
Please cite this article in press as: Nayeem Ahmad, Syed Manazir Ali, Asad U. Khan, First reported New Delhi metallo-β-lactamase-1-producing Cedecea lapagei, International Journal of Antimicrobial Agents (2016), doi: 10.1016/j.ijantimicag.2016.10.001
93 94 95 96 97 98 99 100 101 102 103 104
ARTICLE IN PRESS 2
Letter to the Editor / International Journal of Antimicrobial Agents ■■ (2016) ■■–■■
105 [5] Poirel L, Dortet L, Bernabeu S, Nordmann P. Genetic features of blaNDM-1-positive 106 Enterobacteriaceae. Antimicrob Agents Chemother 2011;55:5403–7. 107 Nayeem Ahmad 108 Q1 Interdisciplinary Biotechnology Unit, 109 Aligarh Muslim University, 110 Aligarh 202002, India 111 112 113 114 115 116
Syed Manazir Ali Pediatrics Department, Jawaharlal Nehru Medical College and Hospital, Aligarh Muslim University, Aligarh 202002, India
Asad U. Khan * Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh 202002, India * Corresponding author. Interdisciplinary Biotechnology Unit, Q4 Aligarh Muslim University, Aligarh 202002, India. E-mail address: [email protected] (A.U. Khan) 27 September 2016 2 October 2016
Please cite this article in press as: Nayeem Ahmad, Syed Manazir Ali, Asad U. Khan, First reported New Delhi metallo-β-lactamase-1-producing Cedecea lapagei, International Journal of Antimicrobial Agents (2016), doi: 10.1016/j.ijantimicag.2016.10.001
117 118 119 120 121 122 123 124 125 126 127 128
Contents lists available at ScienceDirect
International Journal of Antimicrobial Agents j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / i j a n t i m i c a g
1
Q2 Letter to the Editor
2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54
First reported New Delhi metallo-β-lactamase1-producing Cedecea lapagei Sir, Cedecea lapagei is a Gram-negative, facultative anaerobic, nonspore-forming bacterium belonging to the family Enterobacteriaceae, first isolated by the US Centers for Disease Control and Prevention (CDC) laboratories in 1981. It has been reported as a pathogen in a few cases of bacterial peritonitis, wound infection, chemical burns and pneumonia [1]. New Delhi metallo-β-lactamase-1 (NDM-1) was first reported in 2009 in Klebsiella pneumoniae and Escherichia coli isolated from a patient in Sweden who had received medical care in New Delhi, India [2]. Here we report the presence of a clinically significant NDM-1producing C. lapagei in a 26-day-old female preterm baby admitted to the paediatric intensive care unit (ICU) of a 1300-bed tertiary care hospital in Aligarh, India. The patient was diagnosed with lateonset sepsis, apnoea and hypocalcaemia and was treated with cefotaxime and phenytoin (Epsolin) with no recovery observed after the first week. Amikacin was also added to the treatment along with cefotaxime for another week. The baby started recovering after 14 days. An NDM-1-producing C. lapagei isolate was detected in a blood sample, which to the best of our knowledge is the first report of NDM-1-producing C. lapagei. The identity of the isolated strain (AK68) was confirmed by BD PhoenixTM 100 Automated Microbiology System using panel NMIC/ ID-55 (Gram-negative susceptibility card) followed by 16S rDNA sequencing. Furthermore, antimicrobial susceptibility was determined by the standard disk diffusion method using Mueller– Hinton agar as per Clinical and Laboratory Standards Institute (CLSI) guidelines. The strain was found to be resistant to imipenem, meropenem, aztreonam, ceftazidime, cefotaxime, cefoxitin, cefepime and cefoperazone/sulbactam. Moreover, detection of metallo-βlactamase activity was performed using two imipenem disks (10 μg), one containing 10 μL of 0.1 M anhydrous ethylene diamine tetraacetic acid (EDTA). The disks were placed 25 mm apart on Mueller– Hinton agar plates [3]. Minimum inhibitory concentrations (MICs) were determined following CLSI guidelines and the results are given in Table 1. MIC data showed high resistance to β-lactams. PCR amplification of whole DNA from strain AK68 using previously described primers revealed the presence of blaNDM [4]. Following purification, the amplified product was sent for sequencing (SciGenom Labs Pvt. Ltd., Kerala, India). The obtained sequence was identified as Q3 blaNDM-1 by sequence analysis using BLAST (http://www.ncbi.nlm.nih .gov). The presence of other markers (blaVIM, blaOXA-1, blaOXA-9, blaCMY, blaSHV, blaCTX-M, blaTEM, blaIMP and blaKPC) was also checked for in association with blaNDM-1. Only blaCTX-M, blaSHV and blaTEM were found in coexistence with blaNDM-1. The genetic environment of blaNDM-1 was analysed for the presence of an insertion sequence (IS) known to be associated with the blaNDM-1 in Enterobacteriaceae [5]. Primers targeting the IS element Aba125 identified a complete ISAba125
Table 1 Minimum inhibitory concentration (MIC) and genetic profile of NDM-1-producing Cedecea lapagei strain AK68. MIC (μg/mL) IPM MEM ATM
Genetic environment of blaNDM-1 CAZ
CTX
FOX
CPM SCP Upstream Downstream
512 >256 >1024 >1024 >1024 >1024 128
64
Complete bleMBL absent ISAba125
MIC, minimum inhibitory concentration; IPM, imipenem; MEM, meropenem; ATM, aztreonam; CAZ, ceftazidime; CTX, cefotaxime; FOX, cefoxitin; CPM, cefepime; SCP, cefoperazone/sulbactam.
55 56 57 58 59 60 61 62 63 64 65 66
upstream of blaNDM-1 in strain AK68. The bleomycin resistance gene bleMBL was not identified downstream of blaNDM-1, which is an unusual feature compared with earlier known species carrying blaNDM-1 [3] and needs to be further characterised. The plasmid incompatibility group was determined by PCRbased replicon typing (PBRT). The strain was found to be untypeable. Conjugal transfer was performed using C. lapagei AK68 as donor and azide-resistant E. coli J53 strain as recipient on selection medium with cefoxitin (10 μg/mL) and sodium azide (100 μg/mL). Conjugation confirmed the plasmid location of blaNDM-1. This is the first case of an NDM-1-producing C. lapagei strain isolated from a neonate admitted to the paediatric ICU of a north Indian hospital. Hence, there is an urgent need to perform a proper surveillance study to control further spread of NDM-1 in evolving new species. Nucleotide sequence accession number: The sequence of blaNDM-1 determined in this strain was submitted to GenBank with the accession no. KX231908. Funding: This study was supported by grants DBT Award BT/ HRD/NBA/34/01/2012, BT/PR8281/BID/7/448/2013 and ICMR grant AMR/5/2011-ECD-1 to AUK. Competing interests: None declared. Ethical approval: Clearance from the institution ethical committee was received for the whole study [no. 151/201517/PDFWM2015-2017-UTT-31140 (SAII)].
67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92
References [1] Salazar G, Almeida A, Gómez M. Cedecea lapagei traumatic wound infection: case report and literature review. Rev Chilena Infectol 2013;30:86–9, [in Spanish]. [2] Yong D, Toleman MA, Giske CG, Cho HS, Sundman K, Lee K, et al. Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother 2009;53:5046–54. [3] Khan AU, Parvez S. Detection of blaNDM-4 in Escherichia coli from hospital sewage. J Med Microbiol 2014;63:1404–6. [4] Khan AU, Nordmann P. NDM-1-producing Enterobacter cloacae and Klebsiella pneumoniae from diabetic foot ulcers in India. J Med Microbiol 2012;61:454–6.
http://dx.doi.org/10.1016/j.ijantimicag.2016.10.001 0924-8579/© 2016 Published by Elsevier B.V.
Please cite this article in press as: Nayeem Ahmad, Syed Manazir Ali, Asad U. Khan, First reported New Delhi metallo-β-lactamase-1-producing Cedecea lapagei, International Journal of Antimicrobial Agents (2016), doi: 10.1016/j.ijantimicag.2016.10.001
93 94 95 96 97 98 99 100 101 102 103 104
ARTICLE IN PRESS 2
Letter to the Editor / International Journal of Antimicrobial Agents ■■ (2016) ■■–■■
105 [5] Poirel L, Dortet L, Bernabeu S, Nordmann P. Genetic features of blaNDM-1-positive 106 Enterobacteriaceae. Antimicrob Agents Chemother 2011;55:5403–7. 107 Nayeem Ahmad 108 Q1 Interdisciplinary Biotechnology Unit, 109 Aligarh Muslim University, 110 Aligarh 202002, India 111 112 113 114 115 116
Syed Manazir Ali Pediatrics Department, Jawaharlal Nehru Medical College and Hospital, Aligarh Muslim University, Aligarh 202002, India
Asad U. Khan * Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh 202002, India * Corresponding author. Interdisciplinary Biotechnology Unit, Q4 Aligarh Muslim University, Aligarh 202002, India. E-mail address: [email protected] (A.U. Khan) 27 September 2016 2 October 2016
Please cite this article in press as: Nayeem Ahmad, Syed Manazir Ali, Asad U. Khan, First reported New Delhi metallo-β-lactamase-1-producing Cedecea lapagei, International Journal of Antimicrobial Agents (2016), doi: 10.1016/j.ijantimicag.2016.10.001
117 118 119 120 121 122 123 124 125 126 127 128