Molecular analysis of clinical isolates of ceftazidime-avibactam-resistant Klebsiella pneumoniae

ObjectivesTo analyse the strains collected during a 1-year survey of ceftazidime-avibactam-resistant KPC-producing Klebsiella pneumoniae, in order to investigate the molecular mechanisms potentially responsible for their resistant phenotype.MethodsClinical KPC-producing K. pneumoniae isolates were collected from 31 patients in six different hospitals in Rome. For eight of the patients, an additional strain grown before the start of treatment was also available, bringing the total of isolates studied to 39. Antimicrobial susceptibility was determined by automated system, broth microdiluition and E-test as appropriate. In silico analysis of acquired resistance genes was achieved by whole-genome sequencing, while multilocus sequence typing and core genome multilocus sequence typing were employed for molecular typing. Mutations associated with ceftazidime-avibactam resistance were identified by Sanger sequencing of the blaKPC gene. Possible mutations in OmpK35 and OmpK36 outer membrane proteins were also investigated.ResultsMolecular analyses highlighted the circulation of the ST512, 101 and 307 high-risk clones; 26 of the 31 patients carried a mutated KPC variant, five had a wild-type KPC-3. Among the KPC variants detected, 11 were different mutations within the bla_KPC-3 gene, four of which were novel mutational changes.ConclusionsDifferent mutations including single amino-acid substitutions, insertions or deletions within the bla_KPC gene were found in 26/31 ceftazidime-avibactam-resistant KPC-producing K. pneumoniae strains belonging to high-risk clones circulating in Italy. Of note, in 14/31 cases the isolates displayed resistance to both ceftazidime-avibactam and carbapenems, raising concerns for the possible selection of a multidrug-resistant phenotype.     

Time-kill synergy tests of tigecycline combined with imipenem, amikacin, and ciprofloxacin against clinical isolates of multidrug-resistant Klebsiella pneumoniae and Escherichia coli

This study evaluated the activity of tigecycline combined with imipenem, amikacin, and ciprofloxacin against clinical isolates of multidrug-resistant Klebsiella pneumoniae and Escherichia coli co-producing extended-spectrum β-lactamases and acquired AmpC β-lactamases. Broth microdilution tests were performed for cefotaxime, ceftazidime, cefepime, imipenem, amikacin, ciprofloxacin, and tigecycline. Time-kill synergy studies were tested for tigecycline plus imipenem, tigecycline plus amikacin, and tigecycline plus ciprofloxacin. Imipenem (MIC(90) = 1 μg/ml for both K. pneumoniae and E. coli) and tigecycline (MIC(90) = 2 μg/ml for K. pneumoniae and 1 μg/ml for E. coli) were the most potent agents. Combination studies with tigecycline plus imipenem resulted in synergy against 18 K. pneumoniae and 3 E. coli isolates; tigecycline plus amikacin yielded synergy against 8 K. pneumoniae and 3 E. coli isolates; tigecycline plus ciprofloxacin yielded synergy against 7 K. pneumoniae and 2 E. coli isolates. No antagonism was observed with any combination. In the present study, imipenem, amikacin, and ciprofloxacin led to indifferent and some synergistic effects in combination with tigecycline, and none of them demonstrated antagonistic effects.     

Resistance to amikacin and gentamicin among Gram-negative bloodstream isolates in a university hospital between 1989 and 1994

Objective: To characterize antimicrobial resistance patterns to amikacin (AN) and gentamicin (GM) among Gramnegative bloodstream isolates and to determine the possible relationship between use of AN and GM and the occurrence of antibiotic resistance during a 6-year period.
Methods: Standard media and techniques of isolation and identification were used. Antimicrobial susceptibility testing was performed with the disk diffusion method and API rapid ATE E strips. Data on consumption of aminoglycosides were collected by the central hospital pharmacy and were expressed as daily defined doses.
Results: One thousand nine hundred and four bloodstream isolates were tested for AN and GM susceptibility between 1989 and 1994. Activities of AN and GM remained high during the study period against most isolates of Gram-negative bacteria. No relationship could be observed between the use of AN/GM and the rate of AN/GM resistance. Nosocomial Gram-negative No relationship could be bloodstream isolates showed a higher degree of resistance towards both AN (3.9% of all nosocomial iveisolates) and GM (7.9%) than community-acquired isolates (1.8% toward AN and 3.1% towards GM, respectively). There was a significant increase (7.9%)than(P= 0.004) in the risk of GM resistance in patients with nosocomial Gram-negative bacteremia detected more than 14 days after admission. The proportion of GM-susceptible Pseudornonas aeruginosa isolates detected decreased linearly from 97% for infections acquired between day 3 and day 10 following admission to 80% for bacteremia developing 30 days or more after admission (P= 0.008).
Conclusions: AN and GM remain highly active antimicrobial drugs for treatment of GNB in times of growing resistance to cephalosporins and fluoroquinolones.     

The effect of tigecycline and ertapenem against clinical isolates of Brucella melitensis detected by E-test on different media

In this study, in vitro activity of tigecycline (TIG) and ertapenem (ERT) against clinical isolates of Brucella melitensis and the effect of different media on in vitro test results were investigated. The in vitro effects of TIG and ERT to 38 B. melitensis isolates were comparatively investigated in brucella agar and 5% sheep blood agar. MIC value of ERT was 0.032 μg/mL in 23 of 38 and 20 of 38 isolates on blood and brucella agar, respectively. Minimum inhibitory concentration values of TIG were substantially different ranging between 0.064-0.25 μg/mL on blood agar. However, MIC values of TIG were similar on brucella agar with 0.25 μg/mL in 15 of 38 isolates and 0.5 μg/mL in 10 of 38 isolates. In conclusion, although ERT and TIG were effective against B. melitensis isolates in vitro, further studies are needed in order to determine the use of these novel drugs in treatment of brucellosis.     

Klebsiella pneumoniae: development of a mixed population of carbapenem and tigecycline resistance during antimicrobial therapy in a kidney transplant patient

Nine isolates of Klebsiella pneumoniae were isolated from a renal transplant patient suffering from recurrent urosepsis over a period of 4 months. Imipenem resistance was detected after imipenem-ertapenem therapy. When treatment was switched to tigecycline the K. pneumoniae developed resistance to tigecycline (MIC = 8 mg/L). The nine isolates were tested by determination of agar dilution MICs, phenotypic carbapenemase, extended-spectrum beta-lactamases and metallo-beta-lactamase (MBL) testing and pulsed-field gel electrophoresis. Polymerase chain reaction and sequencing analysis were employed for identification of bla genes and mapping of the integron carrying the MBL gene. The nine isolates were clonally related and all produced the SHV-12 enzyme. Five MBL-producing isolates showed imipenem MICs ranging from 2 to 64 mg/L and all were detected by testing with imipenem and EDTA. The five isolates harboured the bla_VIM-1 gene. Three isolates showed increased tigecycline MICs (4–8 mg/L). Serial blood cultures obtained on the same day resulted in a VIM-positive/tigecycline-susceptible and a VIM-negative/tigecycline-resistant K. pneumoniae isolate. No isolate developed concurrent imipenem and tigecycline resistance. The patient had a persistent urinary tract infection and recurrent bacteraemia caused by a mixed population of Klebesiella pneumoniae isolates adapting to the selective pressure of antimicrobial therapy at the time. The present study is a worrisome example of what could happen when an immunocompromised host is subjected to the pressures of antimicrobial therapy. In addition, we report the first treatment-emergent MIC increase of tigecycline from 0.5 to 8 mg/L in K. pneumoniae.     

In vitro activity of tigecycline against clinical isolates of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae, Serratia marcescens and Enterobacter cloacae

BACKGROUND AND PURPOSE: Strains of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have spread widely in Taiwan hospitals. In this study, we evaluated the in vitro antimicrobial activity of tigecycline against ESBL-producing Enterobacteriaceae, including Klebsiella pneumoniae, Serratia marcescens and Enterobacter cloacae. METHODS: 104 confirmed ESBL-producing bacteria were isolated from 4 hospitals in mid- and southern Taiwan between 2000 and 2006. The in vitro activity of tigecycline against these ESBL producers was tested by use of Etest strips. RESULTS: The minimal tigecycline concentration at which 50% of isolates were inhibited and minimal concentration at which 90% of isolates were inhibited for ESBL-producing isolates ranged from 0.38 to 0.75 mug/mL and 0.5 to 1.5 mug/mL, respectively. CONCLUSIONS: Tigecycline, a new semisynthetic glycylcycline, may be considered an alternative drug of choice for patients infected with ESBL-producing bacteria.     

Imported Klebsiella pneumoniae carbapenemase-producing K. pneumoniae clones in a Greek hospital: impact of infection control measures for restraining their dissemination

The recent emergence of carbapenemase-producing Enterobacteriaceae strains represents a major threat for hospitalized patients. We document the dissemination and control of carbapenemase-producing Klebsiella pneumoniae clones in a Greek hospital. During a 3-year study period (January 2009 to December 2011), carbapenemase-producing K. pneumoniae strains were isolated from clinical samples from 73 individual patients. Phenotyping and molecular testing confirmed that 52 patients were infected with K. pneumoniae carbapenemase 2 (KPC-2) producers, 12 were infected with VIM-1 producers, and the remaining 9 were infected with isolates producing both KPC-2 and VIM-1 enzymes. Twenty-eight of these clinical cases were characterized as imported health care associated, and 23 of these were attributed to KPC producers and 5 were attributed to KPC and VIM producers. The remaining 45 cases were deemed hospital acquired. In the second year of the study, intensified infection control intervention was implemented, followed by active surveillance and carrier isolation in the third year. The incidence of carbapenemase-producing K. pneumoniae patient cases decreased from 0.52/1,000 patient days in 2009 to 0.32/1,000 patient days in 2010 (P = 0.075). Following these additional infection control measures, the incidence fell to 0.21/1,000 patient days in 2011 and differed significantly from that in 2009 (P = 0.0028). Despite the fact that the imported cases of carbapenemase-producing K. pneumoniae were equally distributed over this 3-year period, the incidence of hospital-acquired cases decreased from 0.36/1,000 patient days in 2009 to 0.19/1,000 patient days in 2010 (P = 0.058) and to 0.1/1,000 patient days in 2011 (P = 0.0012). Our findings suggest that rigorous infection control measures and active surveillance can effectively reduce the incidence of secondary transmission due to KPC-producing pathogens.     

Combined disc methods for the detection of KPC- and/or VIM-positive Klebsiella pneumoniae: improving reliability for the double carbapenemase producers

Klebsiella pneumoniae strains co-producing klebsiella pneumoniae carbapenemase (KPC) and verona integron-encoded metallo-beta-lactamase (VIM) are frequently isolated in Greece and have also occurred in other European countries. Conventional combined disc tests exhibit low sensitivity against these emerging pathogens. We have evaluated modifications of the KPC/Metallo-β-Lactamase Confirmation kit (ROSCO) exhibiting high diagnostic value against KPC, VIM and KPC + VIM producers. The key changes were the inclusion of additional combined tablets containing meropenem plus two inhibitors (dipicolinic acid (1000 μg per tablet) for metallo-β-lactamases and a boronic acid derivative for KPCs) and the replacement of aminophenylboronic acid by phenylboronic acid (400 μg per tablet).     

Murine monoclonal antibodies to Klebsiella pneumoniae protect against lethal endotoxemia and experimental infection with capsulated K. pneumoniae.

To prepare monoclonal antibodies (MAbs) directed against the core-lipid A fractions of smooth lipopoly-saccharide (LPS) from Klebsiella pneumoniae O1:K2, we immunized BALB/c mice with the LPS-associated proteins plus LPS. This preparation exposed the core-lipid A moiety, which is normally hidden in the micellar structure of classical LPS preparations. Among 10 MAbs selected for their reactivity with LPS-associated proteins plus LPS from K. pneumoniae O1:K2, 6 (3A3, 3C2, 3C4, 7D2, 11C3, and 12B6) were directed against the core fraction and 2 (6C5 and 10A5) were directed against the lipid A fraction. Only one (2A4) recognized the O antigen, and one (6D5) had an undefined specificity. When injected before challenge with K. pneumoniae O1:K2 LPS in galactosamine-sensitized mice, five of the MAbs (3C4, 6D5, 7D2, 11C3, and 12B6) provided protection in this model of lethal endotoxemia. MAb 7D2 was also protective in an experimental infection with capsulated K. pneumoniae O1:K2.     

Clearance of carbapenemase-producing Enterobacteriaceae (CPE) carriage: a comparative study of NDM-1 and KPC CPE

ObjectivesThe aim of this study was to compare clearance rates and related characteristics of patients carrying KPC-producing carbapenemase-producing Enterobacteriaceae (CPE) with those of patients carrying NDM-1-producing CPE.MethodsFrom November 2010 to October 2016, consecutive patients whose clinical or surveillance cultures yielded CPE were prospectively identified and followed in a 2700-bed tertiary referral hospital. CPE control protocols included strict single-room isolation, contact precautions and weekly surveillance cultures. CPE clearance was defined as three or more consecutive CPE-negative cultures without relapse. We compared patients carrying NDM-1 CPE and KPC and those with and without clearance. The time to CPE clearance or discharge was assessed using the Kaplan–Meier method and NDM-1 CPE and KPC CPE groups were compared.ResultsA total of 147 patients carrying CPE, 106 with NDM-1 and 41 with KPC, were included in the study. At the time of hospital discharge, 12 of the 106 patients carrying NDM-1 CPE were clear of CPE, whereas none of the KPC CPE patients were (NDM-1, 11.3% (12/106) versus KPC, 0% (0/41), p 0.02). There was no significant association between CPE clearance and factors such as an immunocompromised condition, antibiotic usage, or species of colonizing organism. Among 40 patients who were readmitted, CPE non-clearance was significantly higher in patients carrying KPC CPE (NDM-1, 36.7% (11/30) versus KPC, 80.0% (8/10), p 0.03).ConclusionsCompared with NDM-1 CPE patients, patients carrying KPC CPE had a significantly lower probability of clearance during hospitalization. Furthermore, KPC CPE carriage persisted for a substantial period of time following patient discharge.     

Concordance between epidemiological evaluation of probability of transmission and whole genome sequence relatedness among hospitalized patients acquiring Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae

ObjectivesWe aimed to evaluate the concordance between epidemiologically determined transmission and genetic linkage of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-Kp).MethodsWe included consecutive KPC-Kp carriers between December 2016 and April 2017 in a hospital endemic for KPC-Kp. We assessed epidemiological relatedness between patients by prospective investigations by the infection control team. The probability of epidemiological relatedness was classified into four groups: no suspected transmission, low, moderate and high probability of transmission. Whole-genome sequencing of isolates was performed. Genetic linkage between KPC-Kp isolates was expressed by distance between isolates in single nucleotide polymorphisms (SNPs). We established an SNP cut-off defining a different strain based on the reconstructed phylogenetic tree. We compared the epidemiological and genetic linkage of all isolates from all patients.ResultsThe study included 25 KPC-Kp carriers with 49 isolates. SNP variance was available for 1129 crossed patient-isolate pairs. Genomic linkage, based on a cut-off of 80 SNPs to define related isolates, was found in 115/708 (16.2%) of isolates with no transmission suspected epidemiologically, 27/319 (8.5%) of low, 11/26 (42.3%) of moderate and 64/76 (84.2%) of high epidemiological transmission risk determination (p < 0.001 for trend). Similar results and significant trends were shown on sensitivity analyses using a lower SNP cut-off (six SNPs) and patient-isolate unique pairs, analysing the first isolate from each patient.ConclusionsWhile significant concordance between epidemiological and genomic transmission patterns was found, epidemiological investigations of transmission are limited by the possibility of unidentified transmissions or over-estimation of associations. Genetic linkage analysis is an important aid to epidemiological transmission assessment.     

Evaluation of polymyxin B in combination with 13 other antibiotics against carbapenemase-producing Klebsiella pneumoniae in time-lapse microscopy and time-kill experiments

ObjectivesThis study aimed to explore the interactions of polymyxin B in combination with 13 other antibiotics against carbapenemase-producing Klebsiella pneumoniae.MethodsFive clinical isolates of multidrug-resistant K. pneumoniae producing KPC-2, KPC-3, NDM-1, OXA-48 and VIM-1 carbapenemases were used. Polymyxin B was tested alone and in combination with amikacin, aztreonam, cefepime, chloramphenicol, ciprofloxacin, fosfomycin, linezolid, meropenem, minocycline, rifampicin, temocillin, thiamphenicol and trimethoprim. Inhibition of growth during antibiotic exposure was evaluated in 24-hr automated time-lapse microscopy experiments. Combinations that showed positive interactions were subsequently evaluated in static time-kill experiments.ResultsAll strains carried multiple (≥9) resistance genes as determined by whole-genome sequencing. In the initial screening the combination of polymyxin B and minocycline was most active with enhanced activity compared with the single antibiotics detected against all strains. Positive interactions were also observed with polymyxin B in combination with rifampicin and fosfomycin against four of five strains and less frequently with other antibiotics. Time-kill experiments demonstrated an additive or synergistic activity (1–2 log₁₀ or ≥2 log₁₀ CFU/mL reduction, respectively, compared with the most potent single antibiotic) with 21 of 23 tested combinations. However, because of regrowth, only 13 combinations were synergistic at 24 hr. Combinations with minocycline or rifampicin were most active, each showing synergy and bacteriostatic or bactericidal effects resulting in 1.93–3.97 and 2.55–5.91 log₁₀ CFU/mL reductions, respectively, after 24 hr against four strains.DiscussionPolymyxin B in combination with minocycline, rifampicin or fosfomycin could be of potential clinical interest. Time-lapse microscopy showed some discrepancy in results compared with the time-kill data but was useful for screening purposes.     

Infections caused by carbapenem-resistant Klebsiella pneumoniae among patients in intensive care units in Greece: a multi-centre study on clinical outcome and therapeutic options

Infections due to carbapenem-resistant Klebsiella pneumoniae (CR-KP) have emerged as a public health problem worldwide given their spread dynamics and the limited therapeutic options. Our aim was to study the clinical outcome of patients with CR-KP infections in relation to antimicrobial treatment. CR-KP infections that occurred in a 10-month period (September 2009 to June 2010) in patients admitted to 19 intensive care units all over Greece were studied. A total of 127 CR-KP infections were reported. Central venous catheter bacteraemia was the most frequent infection, followed by ventilator-associated pneumonia (39 (30.7%) and 35 (27.6%) cases, respectively). Resistance to colistin, tigecycline, gentamicin and amikacin was detected in 20%, 33%, 21% and 64% of isolates, respectively. Regarding treatment, 107 cases received active treatment, including 1 or ≥2 active antibiotics in 65 (60.7%) and 42 (39.3%) cases, respectively. The most frequent combination was colistin plus aminoglycoside and tigecycline plus aminoglycoside (17 and 11 cases, respectively). Forty-eight (45.2%) of the cases that received active treatment were considered clinical failures, with 23.5% mortality at 14 days. Logistic regression analysis revealed that age ≤55 years, non-immunocompromised patients and patients who received colistin had higher successful response rates, while patients ≤55 years old had lower mortality rates at 14 days after the introduction of active treatment. CR-KP infections are associated with a significant clinical failure rate. Colistin remains a valuable antimicrobial agent for treating these infections, while the rise of resistance to the last available antibiotics further limits treatment options.     

Different metabolic profiles of K1 serotype and non-serotype K1 and K2 Klebsiella pneumoniae isolates in oral infection mice model

K1 or K2 serotype Klebsiella pneumoniae isolate caused clinical pyogenic liver abscess (KLA) infection is prevalent in many areas. It has been identified that K1 or K2 serotype K. pneumoniae isolates caused KLA infection in mice by oral inoculation. In our study, K1 serotype K. pneumoniae isolate Kp1002 with hypermucoviscosity (HV)-positive phenotype caused KLA infection in C57BL/6 mice by oral inoculation. Simultaneously, non-serotype K1 and K2 isolate Kp1014 with HV-negative phenotype failed to cause KLA infection in the same manner. It seems that gastrointestinal tract translocation is the pathway by which K1 or K2 serotype K. pneumoniae caused KLA infection. Liquid chromatography-tandem mass spectrometry was used to further analyze metabolic profile changes in mice with KLA infection. Data showed that after Kp1002 or Kp1014 oral inoculation, serum Phosphatidylcholine (PC) and Lysophosphatidylcholine (LPC) levels significantly changed in mice. Some PC and LPC molecules showed changes both in the Kp1002 KLA group and the Kp1014 no-KLA group compared with the control group. The level of 18:1/18:2-PC significantly changed in the Kp1002 KLA group compared with the control group, but showed no change between the Kp1014 no-KLA group and the control group. The level of 18:1/18:2-PC might have been particularly affected by KLA infection caused by K1 serotype K. pneumoniae Kp1002. It may be a potential biomarker for KLA infection.     

Comparative pathogenicity of bacteraemic isolates of Aeromonas hydrophila and Klebsiella pneumoniae

Two bacteraemic isolates of Aeromonas hydrophila and Klebsiella pneumoniae, Ah-2743 and Kp-129, respectively, were studied for their relative pathogenicity in comparison with Escherichia coli ATCC 25922. Ah-2743 caused significantly higher serum levels of tumour necrosis factor-alpha, interleukin (IL)-1beta or IL-6 in human whole blood, and higher serum IL-1beta and IL-6 levels in infected mice, than did Kp-129 and E. coli ATCC 25922. In addition, in BALB/c mice infected by intraperitoneal inoculation, Ah-2743 caused a higher fatality rate (80%) than did Kp-129 (0%). With intramuscular inoculation, Ah-2743 caused more rapid and intense local accumulation of inflammatory cells than did Kp-129, and myonecrosis was present only in Ah-2743-infected mice. These data indicate that Ah-2743 is more pathogenic than Kp-129 or E. coli ATCC 25922 for BALB/c mice. The cellular components or extracellular factors associated with increased cytokine induction and pathogenicity of A. hydrophila in mice merit further investigation.     

Detection of extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli.

Forty clinical isolates of Escherichia coli and 141 isolates of Klebsiella pneumoniae that either transferred ceftazidime resistance or showed sulbactam enhancement of oxyimino-beta-lactam susceptibility were tested by disk diffusion methodology for susceptibility to aztreonam, cefotaxime, ceftazidime, and cefoxitin. With standard 30 micrograms antibiotic disks, the fraction of these extended-spectrum beta-lactamase (ESBL)-producing isolates testing resistant by National Committee for Clinical Laboratory Standards criteria was lowest (24%) with cefotaxime disks. Forty percent of the E. coli and 29% of the K. pneumoniae isolates appeared susceptible with at least one oxyimino-beta-lactam disk. Ceftazidime and aztreonam disks were equivalent in differentiating ESBL production, and both were superior to cefotaxime disks. Over half the E. Coli and 29% of the K. pneumoniae isolates tested cefoxitin resistant. In 30 isolates, cefoxitin resistance was transmissible and due to a plasmid-mediated AmpC-type beta-lactamase. With a 5-micrograms ceftazidime disk, a breakpoint could be chosen with high sensitivity and specificity for ESBL-producing organisms. Present disk diffusion criteria underestimate the prevalence of ESBL-producing strains.     

In-vitro synergistic activity of colistin and meropenem against clinical isolates of carbapenem resistant E.coli and Klebsiella pneumoniae by checkerboard method

ContextThe emergence of drug resistant pathogens pose major threat to hospitalized patients as well as to the community associated with increased mortality and morbidity. The treatment of carbapenem resistant enterobacteriaceae, one of the top WHO priority pathogen remains a global issue. Combination therapy with different classes of antibiotics have been tried with the aim to reduce toxicity, to increase the efficacy of the drugs and to reduce resistance. The in-vitro synergy methods have to be carried out to determine whether the combination of those antibiotics are synergistic, antagonistic or additive.AimsWe have performed in-vitro synergy testing by checkerboard method for colistin -meropenem combination to determine whether the combination of the two antibiotics were synergistic or antagonistic.Methods and materialAll the consecutive twenty five blood isolates of Escherichia coli and twenty five blood isolates of Klebsiella pneumoniae which were showing resistance to carbapenems by either disc diffusion or vitek 2 were collected over a period of 6 months and checkerboard method was performed.Statistical analysis usedThe reduction of MIC of colisin on combination with meropenem compared to MIC of colistin alone is analyzed by McNemar's chisquare test with the help of software Stata version 14 and p value < 0.05 is considered as significant.Results56% of K. pneumoniae showed synergy and 44% showed additive/indifference results. For E. coli 40% showed synergy and 60% showed additive/indifference. None of the isolates of E. coli and K. pneumoniae showed antagonism. There was more than two fold reduction in MIC of colistin (significant) on combining withmeropenem.ConclusionsThe study results support the combination therapy to treat infections by multi-drug-resistant Klebsiela pneumoniae and Escherichia coli by in-vitro checkerboard testing method which inturn will be helpful for clinicians for judicious use of antimicrobials.     

Monoclonal antibody against Klebsiella capsular polysaccharide reduces severity and hematogenic spread of experimental Klebsiella pneumoniae pneumonia.

Klebsiella pneumoniae is an important nosocomial pathogen causing severe pulmonary infections. The majority of clinical Klebsiella isolates produce a high-molecular-weight capsular polysaccharide (CPS) which is one of the dominant virulence factors. In the present study, we examined the potency of a murine immunoglobulin M monoclonal antibody (MAb) with specificity to Klebsiella type 2 CPS to protect rats against experimental Klebsiella pneumonia. The MAb did not prevent the invasion of virulent bacteria into the interalveolar space. However, the resolution of infection was accelerated in MAb-treated animals. This was demonstrated by (i) less severe weight loss and (ii) markedly reduced inflammatory reactions in the lung. The elimination of bacteria was significantly increased not only in the lungs but also in the livers of antibody-treated rats. This was reflected by reduced levels of circulating, soluble CPS and MAb-bound CPS. A mixture of human MAbs with specificity to CPS of clinically important Klebsiella serotypes may prove to be a useful tool for the prevention or supportive treatment of Klebsiella pneumonia.     

In vitro activity of tigecycline and occurrence of tetracycline resistance determinants in isolates from patients enrolled in phase 3 clinical trials for community-acquired pneumonia

The in vitro activity of tigecycline was evaluated against baseline pathogens isolated from patients enrolled in phase 3 clinical trials for community-acquired pneumonia conducted in 29 countries worldwide. Tigecycline was active against the most prevalent pathogens, including Streptococcus pneumoniae (MIC₉₀ 0.06 mg/L), Staphylococcus aureus (MIC₉₀ 0.25 mg/L), Haemophilus influenzae (MIC₉₀ 0.5 mg/L) and Klebsiella pneumoniae (MIC₉₀ 1 mg/L). Twelve isolates of S. pneumoniae expressing tet (M) and two isolates of K. pneumoniae producing extended-spectrum β-lactamases isolated during the study were susceptible to tigecycline. The excellent in vitro activity of tigecycline against these clinical isolates confirmed its potential utility against pathogens associated with community-acquired pneumonia.     

Influence of ethanol concentration in the phagocytic function of neutrophils against Klebsiella pneumoniae isolates in an experimental model

Background/Purpose

Although the prevalence of pneumonia or other extrapulmonary infections is higher in people with alcoholism or acute alcohol intoxication, the possible relationship of acute alcohol intoxication to phagocytic function has not been investigated. Our aim was to determine whether acute alcohol intoxication suppresses phagocytic function in human neutrophils.