Correlation between biofilm formation and carbapenem resistance among clinical isolates of Klebsiella pneumoniae

Background: Klebsiella pneumoniae is a Gram-negative enteric bacterium that causes nosocomial infections; this bacterium has survived from harsh condition using biofilm formation in hospital equipment and cause severe infection. In the other hand, the emergence and extension of carbapenem resistance burden among K. pneumonia producing biofilm is the current concern of public health services. There are controversial findings about this subject. The aim of this study was to evaluate the correlation between biofilm formation and resistance to carbapenem among clinical isolates of K. pneumoniae.Methods: A total of 160 K. pneumoniae isolates were collected from various infections of hospitalized patients. The Carba NP test and molecular methods were used for detection of carbapenem resistance isolates of K. pneumonia. Subsequently, the ability for biofilm production was performed from all isolates. Finally, Correlation of biofilm formation among carbapenem resistant isolates was calculated using χ2 and Fisher’s exact tests.Results: Among K. pneumoniae isolates 42.5% have carbapenemase activity by Carba NP test, while carbapenemase genes were detected in 35.6% of isolates in amplification assay. Moreover, there are 52.5% (n= 84) of all isolates were formed a strong biofilm, while 38.1% (n= 61) and 9.3% (n= 15) of isolates were middle and weak biofilm producer, respectively. Among carbapenem resistant cases (n= 68), there are 77.9% (n= 53) and 22% (n= 15) of isolates were reported as strong and middle biofilm producer, respectively. We see a significant correlation was seen between biofilm formation ability and carbapenem resistant isolates (p-value < 0.00001).Conclusion: The increase of carbapenem resistance burden in biofilm producing isolates of K. pneumoniae is considered as serious alert and the basic measures to combat this phenomenon is imperative.

Download Full-text

Detection and Characterization of Carbapenem resistant Gram-negative bacilli isolates recovered from hospitalized patients at Soba University Hospital, Sudan

10.21203/rs.3.rs-31964/v1 ◽

2020 ◽

Author(s):

Hana S. Elbadawi ◽

Kamal M. Elhag ◽

Elsheikh Mahgoub ◽

Hisham N Altayb ◽

Francine Ntoumi ◽

...

Keyword(s):

Resistance Genes ◽

Nosocomial Infections ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

University Hospital ◽

P Value ◽

Relative Distribution ◽

Gram Negative ◽

Carbapenem Resistant ◽

Carbapenemase Gene

Abstract Background:Antimicrobial resistance (AMR) poses a threat to global health security. Whilst over the past decade, there has been an increase in reports of nosocomial infections globally caused by carbapenem resistant Gram-negative bacilli (GNB), data from Africa have been scanty. We performed a study of carbapenem resistance genes among GNB isolated from patients treated in hospitals in Khartoum state, Sudan.Methods:A cross-sectional study was conducted at Soba University Hospital (SUH) and Institute of Endemic Diseases, University of Khartoum for the period October 2016 to February 2017. A total of 206 GNB isolates from different clinical specimens were analyzed for carbapenem resistance genes using phenotypic tests and affirmed by genes detection. Multiplex PCR was performed for each strain to detect the carbapenemase genes, including the blaNDM, blaVIM, blaIMP, blaKPC, and blaOXA-48. In addition to blaCTXM, blaTEM and blaSHV. DNA sequencing and bioinformatics analysis were used to detect genes subtypes.Findings:Of 206 isolates, 171 (83%) were confirmed resistant phenotypically and 121 (58.7%) isolates were positive for the presence of one or more carbapenemase gene. New Delhi metallo-β-lactamase (NDM) types were the most predominant genes, blaNDM 107(88.4%). Others included blaIMP 7 (5.7%), blaOXA-48 5(4.1%), blaVIM 2 (1.6%) and blaKPC 0 (0%). Co- resistance genes with NDM producing GNB were detected in 87 (81.3%) of all blaNDM positive isolates. A significant association between phenotypic and genotypic resistance was observed (P- value < 0.001). NDM-1 was the most sub type was observed in 75 isolates (70 %), other subtypes were NDM- 5 and NDM-6. Infections due to Carbapenem resistant GNB are increasing at SUH, with the blaNDM being the prevalent genes among clinical isolates and belong to the Indian lineage.Conclusions:The frequency of carbapenemase producing bacilli was found to be improperly high in Khartoum hospitals. NDM was found to be the most prevalent carbapenemase gene among clinical isolates. Close surveillance across all hospitals in Sudan is required. The relative distribution of Carbapenemase genes among GNB in nosocomial infections in Africa needs to be defined.

Download Full-text

Molecular Detection of bla OXA-48 Gene Encoding Carbapenem Resistance Pseudomonas aeruginosa Clinical Isolates from Khartoum State Hospitals, Sudan

10.1101/2020.06.22.20137034 ◽

2020 ◽

Author(s):

Doha Omer Ali ◽

Mohamed M.A. Nagla

Keyword(s):

Pseudomonas Aeruginosa ◽

Teaching Hospital ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Clinical Samples ◽

P Value ◽

Gene Encoding ◽

State Hospitals ◽

Carbapenem Resistant ◽

Different Strains

AbatractCarbapenem resistance in Pseudomonas.aeruginosa is particularly worrisome because this class of β-lactam represents the last therapeutic resource for control of bacterial infection.So this study aimed to detect the frequency of bla OXA-48 resistance gene among Pseudomonas aeruginosa clinical isolates during the period from November 2018 to November 2019.Hundred Pseudomonas aeruginosa clinical isolates, 81 carbapenems (imipenem meropenem) resistant and 19 carbapenems sensitive were collected from Omdurman Teaching Hospital, Fedail Hospital and Soba Teaching Hospital in Khartoum State-Sudan. All isolates were re-identified using conventional bacteriological techniques, their susceptibility to carbapenems were tested using Kirby-Bauer method for confirmation and investigated for the presence of the bla OXA-48 gene using conventional PCR technique.60 (60.0%) out of 100 Pseudomonas aeruginosa clinical isolates were positive for blaOXA-48 gene. Out of 81 carbapenem resistant isolates 54(66.7%) were positive for bla OXA-48 gene, while among the (19) carbapenem sensitive isolates 6 (31.6%) were positive for blaOXA-48 gene. There was statistically significant association between carbapenem resistant isolates and the presence of blaOXA-48 gene (P-value = 0.006).Wound swabs were the predominant clinical samples detected harboring bla OXA-48 gene both among the sensitive 5 (83.3%) and carbapenem resistant isolates 29(53.7) (P.value> 0.05).Our findings revealed high frequency of bla OXA-48 among carbapenem resistant isolates so identification of bla OXA-48 producing strains and taking efforts to reduce the rate of transferring these gene between the different strains is essential for optimization of therapy and improves of patients outcomes.

Download Full-text

Polyphasic characterization of carbapenem-resistant Klebsiella pneumoniae clinical isolates suggests vertical transmission of the blaKPC-3 gene

PLoS ONE ◽

10.1371/journal.pone.0247058 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0247058

Author(s):

Catarina Ferreira ◽

Santosh K. Bikkarolla ◽

Karolin Frykholm ◽

Saga Pohjanen ◽

Margarida Brito ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Vertical Transmission ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Host Bacterium ◽

Whole Genome ◽

Healthcare Facilities ◽

Genetic Elements ◽

Dna Mapping ◽

Carbapenem Resistant

Carbapenem-resistant Klebsiella pneumoniae are a major global threat in healthcare facilities. The propagation of carbapenem resistance determinants can occur through vertical transmission, with genetic elements being transmitted by the host bacterium, or by horizontal transmission, with the same genetic elements being transferred among distinct bacterial hosts. This work aimed to track carbapenem resistance transmission by K. pneumoniae in a healthcare facility. The study involved a polyphasic approach based on conjugation assays, resistance phenotype and genotype analyses, whole genome sequencing, and plasmid characterization by pulsed field gel electrophoresis and optical DNA mapping. Out of 40 K. pneumoniae clinical isolates recovered over two years, five were carbapenem- and multidrug-resistant and belonged to multilocus sequence type ST147. These isolates harboured the carbapenemase encoding blaKPC-3 gene, integrated in conjugative plasmids of 140 kbp or 55 kbp, belonging to replicon types incFIA/incFIIK or incN/incFIIK, respectively. The two distinct plasmids encoding the blaKPC-3 gene were associated with distinct genetic lineages, as confirmed by optical DNA mapping and whole genome sequence analyses. These results suggested vertical (bacterial strain-based) transmission of the carbapenem-resistance genetic elements. Determination of the mode of transmission of antibiotic resistance in healthcare facilities, only possible based on polyphasic approaches as described here, is essential to control resistance propagation.

Download Full-text

Synergistic Combination of AS101 and Azidothymidine against Clinical Isolates of Carbapenem-Resistant Klebsiella pneumoniae

Pathogens ◽

10.3390/pathogens10121552 ◽

2021 ◽

Vol 10 (12) ◽

pp. 1552

Author(s):

Chung-Lin Sung ◽

Wei-Chun Hung ◽

Po-Liang Lu ◽

Lin Lin ◽

Liang-Chun Wang ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Carbapenem Resistance ◽

Antibacterial Activities ◽

Clinical Isolates ◽

World Health ◽

Synergistic Activity ◽

Antibiotic Development ◽

Carbapenem Resistant ◽

Health Organization ◽

Carbapenem Resistant Enterobacteriaceae

Owing to the over usage of carbapenems, carbapenem resistance has become a vital threat worldwide, and, thus, the World Health Organization announced the carbapenem-resistant Enterobacteriaceae (CRE) as the critical priority for antibiotic development in 2017. In the current situation, combination therapy would be one solution against CRE. Azidothymidine (AZT), a thymidine analog, has demonstrated its synergistically antibacterial activities with other antibiotics. The unexpected antimicrobial activity of the immunomodulator ammonium trichloro(dioxoethylene-o,o’)tellurate (AS101) has been reported against carbapenem-resistant Klebsiella pneumoniae (CRKP). Here, we sought to investigate the synergistic activity between AS101 and AZT against 12 CRKP clinical isolates. According to the gene detection results, the blaOXA-1 (7/12, 58.3%), blaDHA (7/12, 58.3%), and blaKPC (7/12, 58.3%) genes were the most prevalent ESBL, AmpC, and carbapenemase genes, respectively. The checkerboard analysis demonstrated the remarkable synergism between AS101 and AZT, with the observable decrease in the MIC value for two agents and the fractional inhibitory concentration (FIC) index ≤0.5 in all strains. Hence, the combination of AS101 and azidothymidine could be a potential treatment option against CRKP for drug development.

Download Full-text

High-Level Carbapenem Resistance in a Klebsiella pneumoniae Clinical Isolate Is Due to the Combination of blaACT-1 β-Lactamase Production, Porin OmpK35/36 Insertional Inactivation, and Down-Regulation of the Phosphate Transport Porin PhoE

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00285-06 ◽

2006 ◽

Vol 50 (10) ◽

pp. 3396-3406 ◽

Cited By ~ 141

Author(s):

Frank M. Kaczmarek ◽

Fadia Dib-Hajj ◽

Wenchi Shang ◽

Thomas D. Gootz

Keyword(s):

New York ◽

New York City ◽

Klebsiella Pneumoniae ◽

York City ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Wild Type ◽

Down Regulation ◽

Carbapenem Resistant ◽

Insertional Inactivation

ABSTRACT Clinical isolates of Klebsiella pneumoniae resistant to carbapenems and essentially all other antibiotics (multidrug resistant) are being isolated from some hospitals in New York City with increasing frequency. A highly related pair of K. pneumoniae strains isolated on the same day from one patient in a hospital in New York City were studied for antibiotic resistance. One (KP-2) was resistant to imipenem, meropenem, and sulopenem (MICs of 16 to 32 μg/ml) while the other (KP-1) was susceptible (MIC of 0.5 μg/ml); both contained the bla ACT-1, bla SHV-1, and bla TEM-1 β-lactamases. bla ACT-1 in both strains was encoded on a large ∼150-kb plasmid. Both isolates contained an identical class 1 integron encoding resistance to aminoglycosides and chloramphenicol. They each had identical insertions in ompK35 and ompK36, resulting in disruption of these key porin genes. The carbapenem-resistant and -susceptible isolates were extensively studied for differences in the structural and regulatory genes for the operons acrRAB, marORAB, romA-ramA, soxRS, micF, micC, phoE, phoBR, rpoS, and hfq. No changes were detected between the isolates except for a significant down-regulation of ompK37, phoB, and phoE in KP-2 as deduced from reverse transcription-PCR analysis of mRNA and polyacrylamide gel electrophoresis separation of outer membrane proteins. Backcross analysis was conducted using the wild-type phoE gene cloned into the vector pGEM under regulation of its native promoter as well as the lacZ promoter following transformation into the resistant KP-2 isolate. The wild-type gene reversed carbapenem resistance only when under control of the heterologous lacZ promoter. In the background of ompK35-ompK36 gene disruption, the up-regulation of phoE in KP-1 apparently compensated for porin loss and conferred carbapenem susceptibility. Down-regulation of phoE in KP-2 may represent the normal state of this gene, or it may have been selected from KP-1 in vivo under antibiotic pressure, generating the carbapenem-resistant clone. This is the first study in the Enterobacteriaceae where expression of the phosphate-regulated PhoE porin has been associated with resistance to antimicrobials. Our results with this pair of Klebsiella clinical isolates highlight the complex and evolving nature of multiple drug resistance in this species.

Download Full-text

Network Integrative Genomic and Transcriptomic Analysis of Carbapenem-ResistantKlebsiella pneumoniaeStrains Identifies Genes for Antibiotic Resistance and Virulence

mSystems ◽

10.1128/msystems.00202-19 ◽

2019 ◽

Vol 4 (4) ◽

Cited By ~ 5

Author(s):

Muyoung Lee ◽

Naina Adren Pinto ◽

Chan Yeong Kim ◽

Sunmo Yang ◽

Roshan D’Souza ◽

...

Keyword(s):

Antibiotic Resistance ◽

Klebsiella Pneumoniae ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Functional Modules ◽

Antibiotic Resistant ◽

Content Type ◽

Functional Association ◽

Carbapenem Resistant ◽

Tract Infections

ABSTRACTGlobal increases in the use of carbapenems have resulted in several strains of Gram-negative bacteria acquiring carbapenem resistance, thereby limiting treatment options.Klebsiella pneumoniaeis a common carbapenem-resistant pathogenic bacterium that is widely studied to identify novel antibiotic resistance mechanisms and drug targets. Antibiotic-resistant clinical isolates generally harbor many genetic alterations, and the identification of responsible mutations would provide insights into the molecular mechanisms of antibiotic resistance. We propose a method to prioritize mutated genes responsible for antibiotic resistance on the basis of expression changes in their local subnetworks, hypothesizing that mutated genes that show significant expression changes among the corresponding functionally associated genes are more likely to be involved in the carbapenem resistance. For network-based gene prioritization, we developed KlebNet (www.inetbio.org/klebnet), a genome-scale cofunctional network ofK. pneumoniaegenes. Using KlebNet, we reconstructed the functional modules for carbapenem resistance and virulence and identified the functional association between antibiotic resistance and virulence. Using complementation assays with the top candidate genes, we were able to validate a novel gene that negatively regulated carbapenem resistance and four novel genes that positively regulated virulence inGalleria mellonellalarvae. Therefore, our study demonstrated the feasibility of network-based identification of genes required for antibiotic resistance and virulence of human-pathogenic bacteria.IMPORTANCEKlebsiella pneumoniaeis a major bacterial pathogen that causes pneumonia and urinary tract infections in human.K. pneumoniaeinfections are treated with carbapenem, but carbapenem-resistantK. pneumoniaehas been spreading worldwide. We are able to identify antimicrobial-resistant genes among mutated genes of the antibiotic-resistant clinical isolates. However, they usually harbor many mutated genes, including those that cause weak or neutral functional effects. Therefore, we need to prioritize the mutated genes to identify the more likely candidates for the follow-up functional analysis. For this study, we present a functional network ofK. pneumoniaegenes and propose a network-based method of prioritizing the mutated genes of the resistant clinical isolates. We also reconstructed the network-based functional modules for carbapenem resistance and virulence and retrieved the functional association between antibiotic resistance and virulence. This study demonstrated the feasibility of network-based analysis of clinical genomics data for the study ofK. pneumoniaeinfection.

Download Full-text

Co-existence of NDM-1 and OXA-48 genes in Carbapenem Resistant Klebsiella pneumoniae clinical isolates in Kafrelsheikh, Egypt

African Health Sciences ◽

10.4314/ahs.v21i2.2 ◽

2021 ◽

Vol 21 (2) ◽

pp. 489-496

Author(s):

Ramadan Ahmed El-Domany ◽

Tarek El-Banna ◽

Fatma Sonbol ◽

Samar Hamed Abu-Sayedahmed

Keyword(s):

Klebsiella Pneumoniae ◽

Carbapenem Resistance ◽

Multidrug Resistant ◽

Clinical Isolates ◽

Carbapenem Resistant ◽

High Incidence ◽

Synergy Test ◽

Objective Determination ◽

Mdr Genes

Background: The noteworthy spread of carbapenem-resistant K. pneumoniae (CR-KP) isolates represents a significant safety threat. Objective: Determination of the carbapenemase genes incidence among CR-KP clinical isolates in Kafrelsheikh, Egypt. Methods: A total of 230 K. pneumoniae isolates were recovered from four hospitals in Kafrelsheikh, Egypt. Susceptibility testing was conducted using Kirby-Bauer method and automated-Vitek2 system. CR-KP isolates were tested using modified Hodge test (MHT) and combined disk synergy test. PCR and DNA sequencing were conducted for CR-KP isolates to rec- ognize the included carbapenemase-genes. Results: Out of 230 K. pneumoniae isolates, 50 isolates presented resistance to carbapenem (meropenem). All 50 CR-KP iso- lates were multidrug-resistant (MDR). Genes like blaNDM-1 and blaOXA-48 were the only detected genes among CR-KP with an incidence of 70.0% and 52.0%, respectively. Up to 74.0% of the tested isolates carried at least one of the two record- ed genes, among them 48.0% co-harbored both blaNDM-1 and blaOXA-48 genes. The accession-numbers of sequenced blaNDM-1 and blaOXA-48 genes were MG594615 and MG594616, respectively. Conclusion: This study reported a high incidence of MDR profile with the emergence of blaNDM-1 and blaOXA-48 genes co-existence in CR-KP isolates in Kafrelsheikh, Egypt. Hence, more restrictions should be applied against the spread of such serious pathogens. Keywords: Klebsiella pneumoniae; Egypt; carbapenem resistance; MDR; PCR; blaNDM-1; blaOXA-48; sequencing.

Download Full-text

Genome Sequences of Five Clinical Isolates of Klebsiella pneumoniae

Genome Announcements ◽

10.1128/genomea.00040-16 ◽

2016 ◽

Vol 4 (2) ◽

Cited By ~ 3

Author(s):

L. Letti Lopez ◽

Brigida Rusconi ◽

Heidi Gildersleeve ◽

Chao Qi ◽

Milena McLaughlin ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Broad Spectrum ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Nosocomial Pathogen ◽

Genome Sequences ◽

Class A ◽

Carbapenem Resistant ◽

Broad Spectrum Antibiotics

Klebsiella pneumoniae is a nosocomial pathogen of emerging importance and displays resistance to broad-spectrum antibiotics, such as carbapenems. Here, we report the genome sequences of five clinical K. pneumoniae isolates , four of which are carbapenem resistant. Carbapenem resistance is conferred by hydrolyzing class A β-lactamases found adjacent to transposases.

Download Full-text

Contribution of β-Lactamases and Porin Proteins OmpK35 and OmpK36 to Carbapenem Resistance in Clinical Isolates of KPC-2-Producing Klebsiella pneumoniae

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02045-12 ◽

2013 ◽

Vol 58 (2) ◽

pp. 1214-1217 ◽

Cited By ~ 41

Author(s):

Ying Zhang ◽

Xiaofei Jiang ◽

Yanyan Wang ◽

Gang Li ◽

Yueru Tian ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Content Type ◽

Carbapenem Resistant ◽

Sds Page ◽

Porin Proteins ◽

A Minor ◽

Sequence Types ◽

M 14

ABSTRACTFifty-seven carbapenem-resistantKlebsiella pneumoniaeisolates belonging to ST11 (50 isolates), ST423 (5 isolates), and two other sequence types were studied. All were positive forblaKPC-2,blaTEM-1, andblaCTX-M-14. SDS-PAGE analysis of six representative isolates demonstrated varied porin expression. Nevertheless, whenblaKPC-2was deleted, carbapenem resistance was markedly reduced. Additionally, SHV-12, DHA-1, and/or VIM-1 appeared to contribute to accessory carbapenemase activity. In contrast, OmpK35 and/or OmpK36 deficiency seemed to serve only as a minor cooperative factor.

Download Full-text

Clonal spread of carbapenem-resistant Klebsiella pneumoniae among patients at admission and discharge at a Vietnamese neonatal intensive care unit

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-021-01033-3 ◽

2021 ◽

Vol 10 (1) ◽

Author(s):

Björn Berglund ◽

Ngoc Thi Bich Hoang ◽

Ludwig Lundberg ◽

Ngai Kien Le ◽

Maria Tärnberg ◽

...

Keyword(s):

Intensive Care Unit ◽

Intensive Care ◽

Klebsiella Pneumoniae ◽

Neonatal Intensive Care Unit ◽

Neonatal Intensive Care ◽

Carbapenem Resistance ◽

Clinical Isolates ◽

Whole Genome ◽

Carbapenem Resistant ◽

Increased Risk

Abstract Background The increasing prevalence of carbapenem-resistant Enterobacteriaceae (CRE) is a growing problem globally, particularly in low- to middle-income countries (LMICs). Previous studies have shown high rates of CRE colonisation among patients at hospitals in LMICs, with increased risk of hospital-acquired infections. Methods We isolated carbapenem-resistant Klebsiella pneumoniae (CRKP) from faecal samples collected in 2017 from patients at admission and discharge at a Vietnamese neonatal intensive care unit (NICU). 126 CRKP were whole-genome sequenced. The phylogenetic relationship between the isolates and between clinical CRKP isolates collected in 2012–2018 at the same hospital were investigated. Results NDM-type carbapenemase-(61%) and KPC-2-encoding genes (41%) were the most common carbapenem resistance genes observed among the admission and discharge isolates. Most isolates (56%) belonged to three distinct clonal clusters of ST15, carrying blaKPC-2, blaNDM-1 and blaNDM-4, respectively. Each cluster also comprised clinical isolates from blood collected at the study hospital. The most dominant ST15 clone was shown to be related to isolates collected from the same hospital as far back as in 2012. Conclusions Highly resistant CRKP were found colonising admission and discharge patients at a Vietnamese NICU, emphasising the importance of continued monitoring. Whole-genome sequencing revealed a population of CRKP consisting mostly of ST15 isolates in three clonally related clusters, each related to blood isolates collected from the same hospital. Furthermore, clinical isolates collected from previous years (dating back to 2012) were shown to likely be clonally descended from ST15 isolates in the largest cluster, suggesting a successful hospital strain which can colonise inpatients.

Download Full-text