scholarly journals Genetic Characterization of mcr-1-Positive Multidrug-Resistant Salmonella enterica Serotype Typhimurium Isolated From Intestinal Infection in Children and Pork Offal in China

2022 ◽  
Vol 12 ◽  
Author(s):  
Haoran Zhang ◽  
Ying Xiang ◽  
Yong Huang ◽  
Beibei Liang ◽  
Xuebin Xu ◽  
...  

With the rapid emergence of plasmid-mediated colistin resistance gene mcr-1, the increased resistance of Salmonella has attracted extensive attention. This study reports on 11 multidrug-resistant Salmonella enterica serovar Typhimurium strains harboring mcr-1 in China. They all presented resistance to colistin, and additionally, one that was isolated from a child’s stool sample was also resistant to ceftriaxone and azithromycin. We screened 1454 strains of Salmonella for mcr-1 gene through PCR, and these strains are all preserved in our laboratory. Antimicrobial sensitivity analysis was carried out for the screened mcr-1 positive strains. Genetic polymorphism analysis of S. Typhimurium was performed by using the Pulsed-Field Gel Electrophoresis (PFGE). The plasmids harboring mcr-1 were identified by S1-PFGE and southern blotting. Plasmid conjugation assays were used to analyze the transferability of colistin resistance. The plasmids harboring mcr-1 were characterized by sequencing and bioinformatic analysis. Eleven S. Typhimurium strains harboring mcr-1 with colistin resistance (MICs 4μg/ml) were detected, which were isolated from children and pig offal in China. All of them were multidrug-resistant strains. PFGE results revealed that the strains isolated from different samples or locations have identical genotypes. S1-PFGE and southern blotting experiments showed that three plasmids of different sizes (33, 60, and 250 kb) all carried the mcr-1 gene. The plasmid conjugation assays revealed that Salmonella acquired mcr-1 harboring plasmids by horizontal transfer. Sequencing and plasmid type analysis revealed that these plasmids were types IncX4, IncI2, and IncHI2. Among them, IncX4 and IncI2 plasmids had extremely similar backbones and contained one resistant gene mcr-1. IncHI2 plasmid contained multiple resistant genes including blaCTX–M, oqxB, sul, aph, aadA, and blaTEM. We identified 11 mcr-1 harboring S. Typhimurium strains in China and described their characteristics. Our findings indicate that the mcr-1 gene can effectively spread among intestinal bacteria by horizontal transfer of three types of plasmids. Moreover, the IncHI2 plasmid can also mediate the transfer of other drug resistance genes. These results reveal that constant surveillance of mcr-1 harboring S Typhimurium is imperative to prevent the spread of colistin resistance.

2019 ◽  
Author(s):  
Laura M Carroll ◽  
Ahmed Gaballa ◽  
Claudia Guldimann ◽  
Genevieve Sullivan ◽  
Lory O Henderson ◽  
...  

Mobilized colistin resistance (mcr) genes are plasmid-borne genes that confer resistance to colistin, an antibiotic used to treat severe bacterial infections. To date, eight known mcr homologues have been described (mcr-1 to -8). Here, we describe mcr-9, a novel mcr homologue, detected in a Salmonella enterica serotype Typhimurium (S. Typhimurium) genome using an in silico approach, followed by experimental functional analysis. The amino acid sequence of mcr-9, detected in a multidrug resistant (MDR) S. Typhimurium strain isolated from a human patient in Washington State in 2010, most closely resembled mcr-3, aligning with 64.5% amino acid identity and 99.5% coverage using translated nucleotide blast. The S. Typhimurium strain was tested for phenotypic resistance to colistin and was found to be sensitive at the 2 mg/L European Committee on Antimicrobial Susceptibility Testing breakpoint under the tested conditions. To determine whether it was capable of conferring resistance to colistin when expressed in a heterologous host, mcr-9 was cloned in colistin-susceptible Escherichia coli NEB5α under an IPTG-induced promoter. Expression of mcr-9 conferred resistance to colistin in E. coli NEB5α at 1, 2, and 2.5 mg/L colistin, albeit at a lower level when compared to mcr-3. Pairwise comparisons of the predicted protein structures associated with all nine mcr homologues (Mcr-1 to -9) revealed that Mcr-9, Mcr-3, and Mcr-7 share a high degree of similarity at the structural level. The results of our approach indicate that mcr-9 is capable of conferring phenotypic resistance to colistin in Enterobacteriaceae and should be immediately considered when monitoring plasmid-mediated colistin resistance. Importance: Colistin is a last-resort antibiotic that is used to treat severe infections caused by MDR and extensively drug resistant (XDR) bacteria. The World Health Organization (WHO) has designated colistin as a Highest Priority Critically Important Antimicrobial for human medicine (WHO, Critically Important Antimicrobials for Human Medicine, 5th Revision, 2017), as it is often one of the only therapies available for treating serious bacterial infections in critically ill patients. Plasmid-borne mcr genes that confer resistance to colistin pose a threat to public health at an international scale, as they can be transmitted via horizontal gene transfer and have the potential to spread globally. Therefore, the establishment of a complete reference of mcr genes that can be used to screen for plasmid-mediated colistin resistance is essential for developing effective control strategies.


2021 ◽  
Author(s):  
Zhang Haoran ◽  
Huang Yong ◽  
Liang Beibei ◽  
Xie Jing ◽  
Xiang Ying ◽  
...  

Abstract Background: With the rapid emergence of plasmid mediated polymyxin resistance gene mcr-1, multidrug-resistant Salmonella caused great troubles in clinical treatment and attracted extensive attention. Here we report multidrug-resistant Salmonella strains harboring mcr-1 in China, which are resistant to both polymyxin, traditional antibiotics and even clinically wide-used antibiotics. Methods: We screened 1454 strains of Salmonella collected in our laboratory from 2006 to 2018 from 3 provinces or regions for mcr-1 by PCR. Antimicrobial susceptibility testing was determined. Plasmid conjugation assays were carried out to analysis the transferability of polymyxin resistance. Genetic polymorphism analysis of Salmonella was performed using the PFGE, and the plasmid profiles were characterized by S1-PFGE and southern blotting. The plasmids harboring mcr-1 were sequenced and compared. Results: Eleven S. Typhimurium isolates harboring mcr-1 with polymyxin resistance (MICs 4μg/ml) were identified from intestinal infections and foods in China. All S. Typhimurium isolates were multidrug-resistant to traditional antibiotics and even clinically wide-used antibiotics. Three types of plasmids harboring mcr-1 were recovered (IncHI2, IncX4 and IncI2). Compared with the reference plasmid, IncX4 and IncI2 plasmids had extremely similar typical backbone, and contain only mcr-1 resistance gene. However, IncHI2 were the most diverse type of plasmid due to containing a large MDR region, including blaCTX-M, oqxB, sul, aph, aadA and blaTEM. IncHI2 plasmids were observed to contain only one or no insertion sequence ISApl1 around mcr-1, without forming a circular intermediate. Conclusion: With the horizontal transfer of different types of plasmids, mcr-1 is widely spread worldwide. These prevalent plasmids are responsible for resistance to polymyxin, traditional antibiotics and even clinically wide-used antibiotics resulting from transmission of mcr-1 and other resistance genes. Our studying emphasizes the necessity to jointly monitor its international epidemic and preemptive further upgrade.


2022 ◽  
Vol 12 ◽  
Author(s):  
Xenia Vázquez ◽  
Vanesa García ◽  
Javier Fernández ◽  
Margarita Bances ◽  
María de Toro ◽  
...  

Colistin is a last-resort antibiotic in fighting severe infections caused by multidrug resistant Gram negative pathogens in hospitals. Zoonotic bacteria acquire colistin resistance in animal reservoirs and mediate its spread along the food chain. This is the case of non-typhoid serovars of Salmonella enterica. Colistin-resistant S. enterica in foods represents a threat to human health. Here, we assessed the prevalence of colistin-resistance in food-borne isolates of S. enterica (2014–2019; Asturias, Spain), and established the genetic basis and transferability of this resistance. Five out of 231 isolates tested (2.2%) were resistant to colistin. Four of them, belonging to the European monophasic ST34 clone of S. Typhimurium, were characterized in the present study. They were collected from pork or pork and beef meat-derived products, either in 2015 (three isolates) or 2019 (one isolate). Molecular typing with XbaI-PFGE and plasmid profiling revealed distinct patterns for each isolate, even though two of the 2015 isolates derived from the same sample. The MICs of colistin ranged from 8 to 16 mg/L. All isolates carried the mcr-1.1 gene located on conjugative plasmids of the incompatibility groups IncX4 (2015 isolates) or IncHI2 (2019 isolate). Apart from colistin resistance, the four isolates carried chromosomal genes conferring resistance to ampicillin, streptomycin, sulfonamides and tetracycline [blaTEM–1, strA-strB, sul2, and tet(B)] and heavy metals, including copper and silver (silESRCFBAGP and pcoGE1ABCDRSE2), arsenic (arsRSD2A2BCA1D1) ± mercury (merEDACPTR), which are characteristically associated with the European ST34 monophasic clone. The 2019 isolate was also resistant to other antibiotics, comprising third generation cephalosporins and cephamycins. The latter phenotype was conferred by the blaCMY–2 gene located on an IncI1-I(α)-ST2 plasmid. Results in the present study identified meat-derived products as a reservoir of a highly successful clone harboring transferable plasmids which confer resistance to colistin and other clinically important antibiotics. An important reduction in the number of food-borne S. enterica detected during the period of the study, together with the low frequency of colistin resistance, underlines the success of One Health initiatives, such as those implemented at the UE, to control zoonotic bacteria along the food chain and to halt the spread of antimicrobial resistance.


mBio ◽  
2019 ◽  
Vol 10 (3) ◽  
Author(s):  
Laura M. Carroll ◽  
Ahmed Gaballa ◽  
Claudia Guldimann ◽  
Genevieve Sullivan ◽  
Lory O. Henderson ◽  
...  

ABSTRACT Mobilized colistin resistance (mcr) genes are plasmid-borne genes that confer resistance to colistin, an antibiotic used to treat severe bacterial infections. To date, eight known mcr homologues have been described (mcr-1 to -8). Here, we describe mcr-9, a novel mcr homologue detected during routine in silico screening of sequenced Salmonella genomes for antimicrobial resistance genes. The amino acid sequence of mcr-9, detected in a multidrug-resistant (MDR) Salmonella enterica serotype Typhimurium (S. Typhimurium) strain isolated from a human patient in Washington State in 2010, most closely resembled mcr-3, aligning with 64.5% amino acid identity and 99.5% coverage using Translated Nucleotide BLAST (tblastn). The S. Typhimurium strain was tested for phenotypic resistance to colistin and was found to be sensitive at the 2-mg/liter European Committee on Antimicrobial Susceptibility Testing breakpoint under the tested conditions. mcr-9 was cloned in colistin-susceptible Escherichia coli NEB5α under an IPTG (isopropyl-β-d-thiogalactopyranoside)-induced promoter to determine whether it was capable of conferring resistance to colistin when expressed in a heterologous host. Expression of mcr-9 conferred resistance to colistin in E. coli NEB5α at 1, 2, and 2.5 mg/liter colistin, albeit at a lower level than mcr-3. Pairwise comparisons of the predicted protein structures associated with all nine mcr homologues (Mcr-1 to -9) revealed that Mcr-9, Mcr-3, Mcr-4, and Mcr-7 share a high degree of similarity at the structural level. Our results indicate that mcr-9 is capable of conferring phenotypic resistance to colistin in Enterobacteriaceae and should be immediately considered when monitoring plasmid-mediated colistin resistance. IMPORTANCE Colistin is a last-resort antibiotic that is used to treat severe infections caused by MDR and extensively drug-resistant (XDR) bacteria. The World Health Organization (WHO) has designated colistin as a “highest priority critically important antimicrobial for human medicine” (WHO, Critically Important Antimicrobials for Human Medicine, 5th revision, 2017, https://www.who.int/foodsafety/publications/antimicrobials-fifth/en/), as it is often one of the only therapies available for treating serious bacterial infections in critically ill patients. Plasmid-borne mcr genes that confer resistance to colistin pose a threat to public health at an international scale, as they can be transmitted via horizontal gene transfer and have the potential to spread globally. Therefore, the establishment of a complete reference of mcr genes that can be used to screen for plasmid-mediated colistin resistance is essential for developing effective control strategies.


Author(s):  
Dr. Manish Kulshrestha ◽  
Dr. Anjali Kulshrestha

INTRODUCTION: Enteric fever includes typhoid and paratyphoid fever. Peak incidence is seen in children 5–15 years of age; but in regions where the disease is highly endemic, as in India, children younger than 5 years of age may have the highest infection rates. There are about 22 million new typhoid cases occur each year. Young children in poor, resource limited areas, who make up the majority of the new cases and there is a mortality figures of 215,000 deaths annually. A sharp decline in the rates of complications and mortality due to typhoid fever is observed as a result of introduction of effective antibiotic therapy since 1950s. MDR-ST became endemic in many areas of Asia, including India soon after multidrug-resistant strains of Salmonella enterica serotype typhi (MDR-ST) that were resistant to all the three first-line drugs then in use, namely chloramphenicol, amoxycillin and co-trimoxazole emerged in early 1990s. MATERIAL AND METHODS: Only blood culture or bone marrow culture positive cases were included. The patients with culture isolated enteric fever were included in the study. Antimicrobial susceptibility testing was carried out by disk diffusion method using antibiotic discs. The analysis of the antimicrobial susceptibility was carried out as per CLSI interpretative guidelines. RESULTS: A total of 82 culture positive cases were included in the present study. 80 culture isolates were from blood culture and 2 from the bone marrow culture. Salmonella entericasubspecies enterica serovartyphi (S typhi) was isolated from 67 (81.70%) patients while Salmonella enterica subspecies entericaserovarparatyphi (S paratyphi A) was isolated from 13 (15.85%) cases and 2 (2.44%) were Salmonella enterica subspecies entericaserovarschottmuelleri (S paratyphi B). Of the 82 cases 65(79.3%) isolates were resistant to ciprofloxacin, 17 (20.7%) were resistant to nalidixic acid, one (1.2%) case each was resistant to Cefotaxime and ceftriaxone, 2 (2.4%) were resistant to chloramphenicol, 10 (12.2%) were resistant and to cotrimoxazole 3 (3.7%) were resistant. CONCLUSION: In a culture positive cases 65(79.3%) isolates were resistant to ciprofloxacin and 17 (20.7%) were resistant to nalidixic acid. Multidrug resistant isolates were 65(79.3%).


Antibiotics ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 80 ◽  
Author(s):  
Silpak Biswas ◽  
Mohammed Elbediwi ◽  
Guimin Gu ◽  
Min Yue

Colistin is considered to be a ‘last-resort’ antimicrobial for the treatment of multidrug-resistant Gram-negative bacterial infections. Identification of Enterobacteriaceae, carrying the transferable colistin resistance gene mcr-1, has recently provoked a global health concern. This report presents the first detection of a hydrogen sulfide (H2S)-producing Escherichia coli variant isolated from a human in China, with multidrug resistance (MDR) properties, including colistin resistance by the mcr-1 gene, which could have great implications for the treatment of human infections.


Author(s):  
Girija Roka ◽  
Subash Pandaya ◽  
Md. Reyad-ul Ferdous ◽  
Manson Pandey ◽  
Naba Raj Pokhrel ◽  
...  

The present study determined the susceptibility to ciprofloxacin of nalidixic acid resistant <em>Salmonella</em> (NARS) isolated from enteric fever patients at Scheer Memorial Hospital, Banepa, Nepal, from June 2012 to December 2012. The antimicrobial sensitivity to nalidixic acid and ciprofloxacin was determined using modified Kirby-Bauer disc diffusion and broth dilution method according to the guidelines of the Clinical and Laboratory Standard Institute. <em>Salmonella</em> was isolated from 34 out of 992 (3.43%) blood cultures collected during the study period, and 10 (29.4%) isolates were identified as <em>Salmonella enterica</em> serotype Typhi, while 24 (70.6%) were identified as <em>Salmonella enterica</em> serotype Paratyphi. Out of the total isolates, 31 (91.2%) were nalidixic acid-resistant <em>Salmonella</em> (NARS). Among NARS, the minimum inhibitory concentration values for ciprofloxacin ranged from 0.25 to 2 mg/L and were constantly higher than those shown by the nalidixic acid-susceptible <em>Salmonella</em>. Therefore, in typhoid <em>Salmonella</em> nalidixic acid resistance may be the indicator of decreased susceptibility to ciprofloxacin.


2011 ◽  
Vol 55 (11) ◽  
pp. 5262-5266 ◽  
Author(s):  
Sophie A. Granier ◽  
Laura Hidalgo ◽  
Alvaro San Millan ◽  
Jose Antonio Escudero ◽  
Belen Gutierrez ◽  
...  

ABSTRACTThe 16S rRNA methyltransferase ArmA is a worldwide emerging determinant that confers high-level resistance to most clinically relevant aminoglycosides. We report here the identification and characterization of a multidrug-resistantSalmonella entericasubspecies I.4,12:i:− isolate recovered from chicken meat sampled in a supermarket on February 2009 in La Reunion, a French island in the Indian Ocean. Susceptibility testing showed an unusually high-level resistance to gentamicin, as well as to ampicillin, expanded-spectrum cephalosporins and amoxicillin-clavulanate. Molecular analysis of the 16S rRNA methyltransferases revealed presence of thearmAgene, together withblaTEM-1,blaCMY-2, andblaCTX-M-3. All of these genes could be transferreden blocthrough conjugation intoEscherichia coliat a frequency of 10−5CFU/donor. Replicon typing and S1 pulsed-field gel electrophoresis revealed that thearmAgene was borne on an ∼150-kb broad-host-range IncP plasmid, pB1010. To elucidate howarmAhad integrated in pB1010, a PCR mapping strategy was developed for Tn1548, the genetic platform forarmA.The gene was embedded in a Tn1548-like structure, albeit with a deletion of the macrolide resistance genes, and an IS26was inserted within themelgene. To our knowledge, this is the first report of ArmA methyltransferase in food, showing a novel route of transmission for this resistance determinant. Further surveillance in food-borne bacteria will be crucial to determine the role of food in the spread of 16S rRNA methyltransferase genes worldwide.


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