Phenotypic detection of Carbapenamase among Klebsiella pneumoniae isolated from clinical samples using modified Hodged test

Carbapenemases are microbial enzymes that confer resistance to virtually all available beta-lactam antibiotics and the most frequent carbapenemases are the Klebsiella pneumoniae Carbapenamase (KPC). Detection of carbapenemases is a significant infection control strategy as the enzymes are often associated with extensive antimicrobial resistance, therapeutic failures and mortality associated with infectious diseases. A total of 400 clinical samples were collected from different groups of patients in Abubakar Tafawa Balewa University Teaching Hospital, Bauchi, Nigeria and 118 K. pneumoniae were isolated using standard microbiological techniques. The isolates were subjected to antibiotic susceptibility testing by Kirby-Bauer disc diffusion method, then screened for Carbapenamase production using modified Hodge test. The results indicated that the isolates were resistant to Ampicillin (61.9%), Ceftriaxone (50.8%) and Ceftazidime (50.8%), then Ciprofloxacin (54.2%), but predominantly sensitive to Imipenem (66.9%), Eterpenem (60.2%) and Meropenem (65.3%). It was found that 38 (32.2%) of the isolates phenotypically shows the presence of Carbapenamase, with highest frequency of (40.7%) among patients, mainly adult females with cases of Urinary Tract Infections (UTIs) and the least from wound (11.8%).This study revealed that the isolates produced other beta-lactamases than KPC or variants of Carbapenamase that cannot be detected by modified Hodge test, thus shows low resistance to carbapenems. Therefore further studies is needed to genotypically confirm the presence of KPC in these isolates.

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Phenotypic detection of beta-lactamases production in enterobacteriaceae

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh1408457c ◽

2014 ◽

Vol 142 (7-8) ◽

pp. 457-463 ◽

Cited By ~ 2

Author(s):

Ivana Cirkovic ◽

Ljiljana Pavlovic ◽

Neda Konstantinovic ◽

Katarina Kostic ◽

Snezana Jovanovic ◽

...

Keyword(s):

Susceptibility Testing ◽

Automated System ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Disk Diffusion Method ◽

Beta Lactamases ◽

Beta Lactam Antibiotics ◽

Clinical Center ◽

Different Types

Introduction. Beta-lactam antibiotics are the most commonly used antibacterial drugs. However, many bacteria have developed resistance to these antibiotics, and the most common form of resistance is the production of beta-lactamase enzymes. Many members of the Enterobacteriaceae family produce different types of these enzymes. Objective. The aim of this study was to perform phenotypic detection of production and identification of beta-lactamase type in Enterobacteriaceae isolated from different clinical specimens from patients hospitalized in the Clinical Center of Serbia. Methods. The strains of Enterobacteriaceae were collected between November 2011 and January 2012 in the laboratory of the Clinical Center of Serbia. The isolates were identified according to the standard microbiology procedures and confirmed by the Vitek2 automated system. Antimicrobial susceptibility testing was performed by the disk diffusion method, and the phenotypic detection of production and identification of betalactamases was performed according to previously described methodologies. Results. In this study, a total of 172 Enterobacteriaceae strains were isolated. Further testing was performed on 54/145 (37.2%) strains showing decreased susceptibility to beta-lactam antibiotics: 13/85 (15.3%) Escherichia coli, 31/46 (67.4%) Klebsiella pneumoniae and 10/14 (71.4%) Proteus mirabilis. Among them, 40/145 (27.6%) strains produced extended spectrum betalactamases (ESBLs), 9/145 (6.2%) - AmpC, 1/145 (0.7%) - K1 beta-lactamase and 4/145 (2.8%) - carbapenemases. Carbapenemases were predominantly detected in K. pneumoniae (75%). Conclusion. Enterobacteriaceae produce different types of betalactamases, and the most common type in our study was ESBLs. Production of carbapenemases detected in Enterobacteriaceae is also an associated problem.

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Oxacillin (Methicillin) Resistant Staphylococci in Domestic Animals in the Czech Republic

Pathogens ◽

10.3390/pathogens10121585 ◽

2021 ◽

Vol 10 (12) ◽

pp. 1585

Author(s):

Jaroslav Bzdil ◽

Monika Zouharova ◽

Katerina Nedbalcova ◽

Vladimir Sladecek ◽

David Senk ◽

...

Keyword(s):

Czech Republic ◽

Target Genes ◽

Susceptibility Testing ◽

Domestic Animals ◽

University Teaching ◽

Diffusion Method ◽

Clinical Samples ◽

Beta Lactam ◽

The Czech Republic ◽

Methicillin Resistant

The aim of this study was to describe the prevalence of different Staphylococcus species isolated from pathological processes and lesions in domestic animals in the Czech Republic and to detect and describe oxacillin (methicillin)-resistant strains (MRS). During the years 2019–2020, a total of 5218 veterinary clinical samples from the Czech Republic were tested. Testing was performed by culture methods and typing by molecular phenotypic methods MALDI-TOF MS and PCR. Antimicrobial susceptibility testing of the strains was performed by the disk diffusion method. A total of 854 staphylococci strains were identified (16.37% prevalence), out of which 43 strains of 6 species of staphylococci were MRS (n = 43; 0.82% prevalence). Of the MRS strains, the most prevalent species were Staphylococcus pseudintermedius (n = 24; 0.46% prevalence) and Staphylococcus aureus (n = 7; 0.13% prevalence). Susceptibility testing showed resistance to beta-lactam antibiotics and, depending on the species, also to trimethoprim/sulfamethoxazole, gentamicin, tetracycline, erythromycin, clindamycin, and enrofloxacin. For further characterization of MRS, PCR assay for virulence factor genes was performed. Seven of the 14 target genes were observed only in S. aureus, except for the eno gene encoding laminin-binding protein, which was also detected in other staphylococci. It is necessary to emphasize the issue of correct using of antimicrobials in practice and antibiotic policy in university teaching and to create stricter legislation that would prevent the widespread use of antimicrobials in veterinary medicine, especially in livestock to reduce the emergence and spread of antimicrobial resistance.

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Multicenter Evaluation of the Etest Gradient Diffusion Method for Ceftolozane-Tazobactam Susceptibility Testing of Enterobacteriaceae and Pseudomonas aeruginosa

Journal of Clinical Microbiology ◽

10.1128/jcm.00717-18 ◽

2018 ◽

Vol 56 (9) ◽

Cited By ~ 2

Author(s):

Adam L. Bailey ◽

Tom Armstrong ◽

Hari-Prakash Dwivedi ◽

Gerald A. Denys ◽

Janet Hindler ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Susceptibility Testing ◽

Clinical Isolates ◽

Diffusion Method ◽

Beta Lactam ◽

Content Type ◽

Gradient Diffusion ◽

Tract Infections ◽

Abdominal Infections ◽

Inhibitor Combination

ABSTRACT Ceftolozane-tazobactam (C/T) is a novel beta-lactam–beta-lactamase inhibitor combination antibiotic approved by the U.S. Food and Drug Administration in 2014 for the treatment of complicated intra-abdominal infections (in combination with metronidazole) and complicated urinary tract infections. In this study, we evaluated the performance of the C/T Etest, a gradient diffusion method. C/T Etest was compared to broth microdilution (BMD) for 51 Enterobacteriaceae challenge isolates and 39 Pseudomonas aeruginosa challenge isolates at three clinical sites. Essential agreement (EA) between the methods ranged from 47 to 49/51 (92.2 to 96.1%) for the Enterobacteriaceae, and categorical agreement (CA) ranged from 49 to 51/51 (96.1 to 100.0%). EA and CA for P. aeruginosa were 100% at all sites. The C/T Etest was also compared to BMD for susceptibility testing on 966 clinical isolates (793 Enterobacteriaceae, including 167 Klebsiella pneumoniae and 159 Escherichia coli isolates, in addition to 173 P. aeruginosa isolates) collected at four clinical sites. EA between Etest and BMD was 96.9% for Enterobacteriaceae isolates and 98.8% for P. aeruginosa isolates. Within the Enterobacteriaceae, isolates from each species examined had >96% CA. For the clinical isolates, no very major errors were identified but two major errors were found (one for K. pneumoniae and one for Providencia rettgeri). By BMD, 47.0% of Enterobacteriaceae and 46.2% of P. aeruginosa challenge strains were nonsusceptible to C/T by CLSI breakpoint criteria; 8.2% of clinical Enterobacteriaceae isolates and 12.1% of clinical P. aeruginosa isolates were nonsusceptible to C/T by CLSI breakpoint criteria. In conclusion, Etest is accurate and reproducible for C/T susceptibility testing of Enterobacteriaceae and P. aeruginosa.

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ESBL Producers in Gram Negative Isolates from Clinical Samples

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.14.3.42 ◽

2020 ◽

Vol 14 (3) ◽

pp. 2027-2032

Author(s):

Mita D. Wadekar ◽

J.V. Sathish ◽

C. Pooja ◽

S. Jayashree

Keyword(s):

Treatment Options ◽

Multidrug Resistant ◽

Diffusion Method ◽

Clinical Samples ◽

Beta Lactamase ◽

Beta Lactam ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Beta Lactam Antibiotics ◽

Esbl Producers

Resistance to beta lactam antibiotics is the most common cause for beta-lactamase production. Increasing number of extended spectrum beta-lactamase (ESBL) producers has reduced the treatment options which resulted in emergence of multidrug resistant strains, treatment failure and hence increased mortality. To detect phenotypically, ESBL producers in Gram negative isolates from different samples and to know their susceptibility pattern. A retrospective study of Gram negative isolates was conducted. Total of 521 isolates were isolated from various samples. They were processed and identified by standard procedures. The antibiotic susceptibility testing was performed by Kirby- Bauer disc diffusion method using CLSI guidelines. ESBL was detected by combination disk test. A total of 521 Gram negative isolates were isolated which included E. coli, Klebsiella pneumoniae, Citrobacter spp., Enterobacter spp., Proteus spp. and Acinetobacter spp. Pseudomonas aeruginosa. Of 521 isolates tested, ESBL was detected in 329 (63.1%) isolates. These isolates showed maximum susceptibility to piperacillin- tazobactam (86%) followed by imipenem (78.4%), amikacin (63.5%), cotrimoxazole (54.4%), ciprofloxacin (51%), amoxi-clav (44.9%), cefepime (44.1%), gentamicin (38.9%), cefoxitin (34.9%) and ampicillin (19.1%). ESBL producers which are resistant to beta lactam antibiotics have become a major problem. Detection of these beta-lactamase enzymes by simple disk method and its reporting will help clinicians in prescribing proper antibiotics.

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Investigation of Antibiotic Resistance and Biofilm Formation in Clinical Isolates of Klebsiella pneumoniae

International Journal of Microbiology ◽

10.1155/2021/5573388 ◽

2021 ◽

Vol 2021 ◽

pp. 1-6

Author(s):

Kiana Karimi ◽

Omid Zarei ◽

Parinaz Sedighi ◽

Mohammad Taheri ◽

Amin Doosti-Irani ◽

...

Keyword(s):

Antibiotic Resistance ◽

Klebsiella Pneumoniae ◽

Biofilm Formation ◽

Diffusion Method ◽

Clinical Samples ◽

Resistance Pattern ◽

Chi Square ◽

Disk Diffusion Method ◽

High Level ◽

Tract Infections

Aim. Klebsiella pneumoniae (K. pneumoniae) is an encapsulated Gram-negative bacterium that can lead to 14–20% of nosocomial infections. The ability of biofilm formation in this bacterium decreases the host immune response and antibiotic efficacy. This may impose a huge impact on patients and healthcare settings. This study aimed to evaluate the antibiotic resistance pattern and biofilm formation in K. pneumoniae strains isolated from two major Hamadan hospitals, west of Iran. Methods. A total of 83 K. pneumoniae strains were isolated from clinical samples of patients in different wards of Hamadan hospitals from September 2018 to March 2019. Determination of antimicrobial susceptibility was performed using the disk diffusion method. Biofilm formation was evaluated by the crystal violet method. Data were analyzed by the SPSS software and chi-square test. Results. The results showed that clinical samples included 18 urinary tract samples (22%), 6 wound samples (7%), 6 blood samples (7%), 17 tracheal tube aspiration samples (20%), 32 throat cultures (38%), 2 sputum samples (2.5%), and 2 abscess drain cultures (2.5%). High-level resistance to cefotaxime was detected in 92%, and all of isolates were susceptible to colistin. Biofilm formation was seen in 62 (75%) isolates. Strong biofilm formation was observed in 17 (20%) strains. A significant correlation was seen between biofilm formation and antibiotic resistance ( P value <0.05). Conclusion. Our findings emphasize the need for proper diagnosis, control, and treatment of infections caused by K. pneumoniae especially in respiratory tract infections due to the strong biofilm formation and high antibiotic resistance in these strains.

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Clinical relevance, Speciation, and Antibiogram of Non -Diphtherial Corynebacteria isolated from various clinical samples in a tertiary care hospital in Zagazig, Egypt

10.51429/ejmm30207 ◽

2021 ◽

Vol 30 (2) ◽

pp. 43-49

Author(s):

Shymaa Yahia ◽

May M. Sami

Keyword(s):

Tertiary Care ◽

Resistance Rate ◽

University Hospital ◽

British Society ◽

Diffusion Method ◽

Clinical Samples ◽

Care Hospital ◽

Beta Lactam ◽

Beta Lactam Antibiotics ◽

Isolated Species

Background: The non-diphtherial Corynebacteria, also called “Coryneforms” bacteria are a diversified group of gram positive non sporing bacilli belonging to the genus Corynebacteria. Such bacteria are considered members of human microbiota (skin, respiratory and genital mucus membranes). Coryneform bacteria's pathogenic capacity has been undervalued until recently. Despite of frequently deemed as contaminants, these bacteria have been correlated to diverse clinical infections recently. Objectives: To isolate, speciate, and determine antimicrobial susceptibility pattern of clinically relevant non-diphtherial Corynebacteria from various clinical samples. Methodology: Different clinical samples (blood, urine, sputum, wound swabs, pus) collected from hospitalized patients attending at Zagazig University Hospital. The samples were processed and cultured as per conventional bacteriological methods. A total of 75 clinically relevant corynebacterial isolates exhibited speciation utilizing matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis and their antibiogram was done by disc diffusion method by means of combined guidelines of Clinical and Laboratory Standards Institute (CLSI) as well as British Society for Antimicrobial Chemotherapy (BSAC) because of lack of definite CLSI guidelines. Results: The mean age of the studied patients was 64.6 ± 14.9 years, 60% were male and 40% female. A total 75 clinically relevant Corynebacteria species were obtained from different clinical samples, including wound swabs and pus (53%), sputum (20%), and blood (17%). Forty two percent were isolated from ICUs. The most prevalent isolated species was C.amycolatum (27%), C. striatum (20%), and C.jeikieum (16%). Beta lactam antibiotics showed least activity against Corynebacteria species with resistance rate against penicillin 76% and ceftriaxone 72%, while all isolates exhibited uniform sensitivity (100%) against vancomycin as well as linezolid. Conclusion: This study showed isolation of different clinically relevant non-diaphterial Corynebacteria from different clinical samples with pus and wound swabs as the most common samples from which Corynebacteria were isolated. In particular, C.amycolatum was the most common isolated species. Beta lactam antibiotics (penicillin, ceftriaxone) showed the least activity while vancomycin and linezolid were the most active agents against nondiapdhterial Corynebacteria isolates. Herein, we confirm diphtheroids’ clinical importance among different infections that necessitate evaluating their susceptibility patterns to some common antibacterial agents for guide the best antibiotic to treat infections caused by these species.

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Beta-lactamase inhibitors from laboratory to clinic.

Clinical Microbiology Reviews ◽

10.1128/cmr.1.1.109 ◽

1988 ◽

Vol 1 (1) ◽

pp. 109-123 ◽

Cited By ~ 109

Author(s):

K Bush

Keyword(s):

Side Effects ◽

Clavulanic Acid ◽

Broad Spectrum ◽

Pathogenic Bacteria ◽

Defense Mechanism ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactamases ◽

Beta Lactam Antibiotics ◽

Tract Infections

beta-Lactamases constitute the major defense mechanism of pathogenic bacteria against beta-lactam antibiotics. When the beta-lactam ring of this antibiotic class is hydrolyzed, antimicrobial activity is destroyed. Although beta-lactamases have been identified with clinical failures for over 40 years, enzymes with various abilities to hydrolyze specific penicillins or cephalosporins are appearing more frequently in clinical isolates. One approach to counteracting this resistance mechanism has been through the development of beta-lactamase inactivators. beta-Lactamase inhibitors include clavulanic acid and sulbactam, molecules with minimal antibiotic activity. However, when combined with safe and efficacious penicillins or cephalosporins, these inhibitors can serve to protect the familiar beta-lactam antibiotics from hydrolysis by penicillinases or broad-spectrum beta-lactamases. Both of these molecules eventually inactivate the target enzymes permanently. Although clavulanic acid exhibits more potent inhibitory activity than sulbactam, especially against the TEM-type broad-spectrum beta-lactamases, the spectrum of inhibitory activities are very similar. Neither of these inhibitors acts as a good inhibitor of the cephalosporinases. Clavulanic acid has been most frequently combined with amoxicillin in the orally active Augmentin and with ticarcillin in the parenteral beta-lactam combination Timentin. Sulbactam has been used primarily to protect ampicillin from enzymatic hydrolysis. Sulbactam has been used either in the orally absorbed prodrug form as sultamicillin or as the injectable combination ampicillin-sulbactam. Synergy has been demonstrated for these combinations for most members of the Enterobacteriaceae, although those organisms that produce cephalosporinases are not well inhibited. Synergy has also been observed for Neisseria gonorrhoeae, Haemophilus influenzae, penicillinase-producing Staphylococcus aureus, and anaerobic organisms. These antibiotic combinations have been used clinically to treat urinary tract infections, bone and soft-tissue infections, gonorrhea, respiratory infections, and otitis media. Gastrointestinal side effects have been reported for Augmentin and sultamicillin; most side effects with these agents have been mild. Although combination therapy with beta-lactamase inactivators has been used successfully, the problem of resistance development to two agents must be considered. Induction of cephalosporinases can occur with clavulanic acid. Permeability mutants could arise, especially with added pressure from a second beta-lactam.(ABSTRACT TRUNCATED AT 250 WORDS)

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Dissemination and Molecular Epidemiology of KPC-Producing Klebsiella pneumoniae Collected in Puerto Rico Medical Center Hospitals during a 1-Year Period

Epidemiology Research International ◽

10.1155/2011/698705 ◽

2011 ◽

Vol 2011 ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Iraida E. Robledo ◽

Guillermo J. Vázquez ◽

Ellen S. Moland ◽

Edna E. Aquino ◽

Richard V. Goering ◽

...

Keyword(s):

Puerto Rico ◽

Klebsiella Pneumoniae ◽

Susceptibility Testing ◽

Genetic Relatedness ◽

Medical Center ◽

Pulsed Field Gel Electrophoresis ◽

Beta Lactam ◽

Beta Lactamases ◽

Surveillance Studies ◽

Pfge Profiles

During a 2003-2004 PCR-based surveillance study conducted in 6 Puerto Rico Medical Center hospitals, 27/92 multi-beta-lactam-resistant Klebsiella pneumoniae strains were identified as carbapenemase (KPC) positive in 4 hospitals. The objectives of this study were to identify the KPC variants, their genetic relatedness, and any other beta-lactamases present. Susceptibility testing, pulsed field gel electrophoresis (PFGE), isoelectric focusing, PCR, and DNA sequencing were performed. KPC variants -2, -3, -4, and -6 were identified. Additional beta-lactamases detected were TEM, DHA, OXA-9 and -30. Antimicrobial susceptibility to carbapenems varied depending on the KPC variant. Five PFGE genetically related groups were identified in 15 isolates and 12 unrelated types. PFGE profiles suggested that both clonal and horizontal transfer are contributing to the dissemination of these isolates among the various hospitals. Comparison of the 2003 and a 2009 surveillance studies showed a significant increase in the KPC-positive K. pneumoniae isolates in the latter.

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The Usage of Antibiotics by COVID-19 Patients with Comorbidities: The Risk of Increased Antimicrobial Resistance

Antibiotics ◽

10.3390/antibiotics11010035 ◽

2021 ◽

Vol 11 (1) ◽

pp. 35

Author(s):

Basit Zeshan ◽

Mohmed Isaqali Karobari ◽

Nadia Afzal ◽

Amer Siddiq ◽

Sakeenabi Basha ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Klebsiella Pneumoniae ◽

Bacterial Infections ◽

Tertiary Care ◽

Diffusion Method ◽

Clinical Samples ◽

Care Hospital ◽

Surgical Intensive Care ◽

E Coli ◽

Tract Infections

Antimicrobial resistance (AMR) is a global health issue that plays a significant role in morbidity and mortality, especially in immunocompromised patients. It also becomes a serious threat to the successful treatment of many bacterial infections. The widespread and irrelevant use of antibiotics in hospitals and local clinics is the leading cause of AMR. Under this scenario, the study was conducted in a tertiary care hospital in Lahore, Pakistan, from 2 August 2021 to 31 October 2021 to discover the prevalence of bacterial infections and AMR rates in COVID-19 patients admitted in surgical intensive care units (SICUs). Clinical samples were collected from the patients and we proceeded to identify bacterial isolates, followed by antibiotic susceptibility testing (AST) using the Kirby Bauer disk diffusion method and minimum inhibitory concentration (MIC). The data of other comorbidities were also collected from the patient’s medical record. The current study showed that the most common pathogens were E. coli (32%) and Klebsiella pneumoniae (17%). Most E. coli were resistant to ciprofloxacin (16.8%) and ampicillin (19.8%). Klebsiella pneumoniae were more resistant to ampicillin (13.3%) and amoxycillin (12.0%). The most common comorbidity was chronic kidney disease (CKD) and urinary tract infections (UTIs). Around 17 different types of antibiotic, the carbapenem, fluoroquinolones, aminoglycoside, and quinolones, were highly prevalent in ICU patients. The current study provides valuable data on the clinical implication of antibiotics consumed by COVID-19 patients in SICUs and the AMR rates, especially with different comorbidities.

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