SHV Β-lactamases Research Articles

High-level expression of chromosomally encoded SHV-1 β-lactamase reduces the susceptibility to cefiderocol of clinical isolates of Klebsiella pneumoniae.

High-level expression of chromosomally encoded SHV-1 β-lactamase reduces the susceptibility to cefiderocol of clinical isolates of Klebsiella pneumoniae.

A computational odyssey: uncovering classical β-lactamase inhibitors in dry fruits

In the antibacterial arsenal, β-lactams have held a prominent position, but increasing resistance due to unauthorized use and genetic factors requires new strategies. Combining β-lactamase inhibitors with broad-spectrum β-lactams proves effective in combating this resistance. ESBL producers demand new inhibitors, leading to the exploration of plant-derived secondary metabolites for potent β-lactam antibiotics or alternative inhibitors. Using virtual screening, molecular docking, ADMET analysis, and molecular dynamic simulation, this study actively analyzed the inhibitory activity of figs, cashews, walnuts, and peanuts against SHV-1, NDM-1, KPC-2, and OXA-48 β-lactamases. Using AutoDock Vina, the docking affinities of various compounds for target enzymes were initially screened, revealing 12 bioactive compounds with higher affinities for the target enzymes compared to Avibactam and Tazobactam. Top-scoring metabolites, including Oleanolic acid, Protocatechuic acid, and Tannin, were subjected to MD simulation studies to further analyze the stability of the docked complexes using WebGro. The simulation coordinates, in terms of RMSD, RMSF, SASA, Rg, and hydrogen bonds formed, showed that these phytocompounds are stable enough to retain in the active sites at various orientations. The PCA and FEL analysis also showed the stability of the dynamic motion of Cα residues of phytochemical-bound enzymes. The pharmacokinetic analysis of the top phytochemicals was performed to analyze their bioavailability and toxicity. This study provides new insights into the therapeutic potential of phytochemicals of selected dry fruits and contributes to future experimental studies to identify βL inhibitors from plants. Communicated by Ramaswamy H. Sarma

Molecular characterization of cefepime and aztreonam nonsusceptibility in Haemophilus influenzae.

Cefepime and aztreonam are highly efficacious against H. influenzae, and resistant strains are rare. In this study, we isolated cefepime- and aztreonam-nonsusceptible H. influenzae strains and addressed the molecular basis of their resistance to cefepime and aztreonam. Two hundred and 28 specimens containing H. influenzae were screened, of which 32 isolates were enrolled and applied to antimicrobial susceptibility testing and whole-genome sequencing. Genetic variations that were detected in all nonsusceptible isolates with statistical significance by Fisher's exact tests were identified as cefepime or aztreonam nonsusceptibility related. Functional complementation assays were conducted to assess the in vitro effects of proteins with sequence substitutions on drug susceptibility. Three H. influenzae isolates were nonsusceptible to cefepime, one of which was also nonsusceptible to aztreonam. Genes encoding TEM, SHV and CTX-M extended-spectrum β-lactamases were not detected in the cefepime- and aztreonam-nonsusceptible isolates. Five genetic variations in four genes and 10 genetic variations in five genes were associated with cefepime and aztreonam nonsusceptibility, respectively. Phylogenetic analyses revealed that changes in FtsI were correlated strongly with the MIC of cefepime and moderately with aztreonam. FtsI Thr532Ser-Tyr557His cosubstitution linked to cefepime nonsusceptibility and Asn305Lys-Ser385Asn-Glu416Asp cosubstitution to aztreonam nonsusceptibility. Functional complementation assays revealed that these cosubstitutions increased MICs of cefepime and aztreonam in susceptible H. influenzae isolates, respectively. Genetic variations relevant to resistant phenotypes of cefepime and aztreonam nonsusceptibility in H. influenzae were identified. Moreover, the effects of FtsI cosubstitutions on increasing MICs of cefepime and aztreonam in H. influenzae were demonstrated.

Cefiderocol Resistance in Klebsiella pneumoniae Is Linked to SHV Extended-Spectrum β-Lactamase Activities and Functional Loss of the Outer Membrane Porin OmpK35.

Klebsiella pneumoniae AR 0047 from the CDC and FDA Antibiotic Resistance Isolate Bank is resistant to cefiderocol, a siderophore-conjugated cephalosporin. Genomics analysis and genetic complementation revealed that a frameshift mutation in ompK35 contributed to cefiderocol resistance. Heterologous expression of blaSHV-5 or blaSHV-12 in Escherichia coli increased the host resistance to cefiderocol. Moreover, avibactam, a β-lactamase inhibitor, enhanced cefiderocol activity against the resistant strain. Therefore, cefiderocol resistance is linked to SHV and the loss of ompK35. IMPORTANCE Understanding cefiderocol resistance mechanisms is essential for providing solutions to treat infections and to prevent resistance development. Cefiderocol resistance in Klebsiella pneumoniae AR 0047 is linked to SHV β-lactamase activities and functional loss of outer membrane porin. The cefiderocol-avibactam combination represents an opportunity to increase potency against cefiderocol-resistant pathogens.

Extended-Spectrum Β-Lactamases in E. coli Isolates From Hospitalized Patients: A Single-Center Snapshot From Croatia

Background: A significant increasing trend in the prevalence of Escherichia coli strains that produce extended-spectrum β-lactamases (ESBLs) has been observed in recent years, both in the community setting and in the healthcare arena. We aimed to provide a snapshot of the current situation with E. coli β-lactamase–producing strains in a single general hospital by appraising their β-lactamase content and plasmid types, which will inform further clinical and research efforts. Methods: Our study population consisted of all hospitalized patients in different clinical units of the General Hospital in Slavonski Brod during a 1-year period: internal medicine, infectious disease, surgery, urology and ICU. Phenotypic tests for the detection of ESBLs and plasmid-mediated AmpC β-lactamases were initially pursued, followed by the molecular detection (polymerase chain reaction, PCR) of resistance genes using primers for blaTEM, blaCTX-M, and blaSHV. PCR-based replicon typing (PBRT) was conducted to type resistance plasmids carrying ESBL genes. Results: During the study period, 30 E. coli isolates with reduced susceptibility to third-generation cephalosporins (ie, ceftazidime, cefotaxime, and ceftriaxone) were detected in hospitalized patients. These isolates stemmed from blood culture (66.7%), wound swabs (13.3%), urine (13.3%), and drainage content (6.7%). Alongside complete resistance to β-lactam antimicrobial agents, they were also characterized by high resistance to gentamicin (93.3%) and ciprofloxacin (96.7%), whereas 23.3% of isolates were also resistant to ertapenem. Most isolates harbored both blaCTX-M and blaTEM genes concurrently (46.7%), while solitary blaTEM and blaCTX-M genes were found in 33.3% and 20% of these isolates, respectively. The presence of SHV β-lactamases was not found in any of the isolates. PBRT revealed a wide array of diverse plasmid groups, with most of the isolates harboring different combinations; however, 80% of isolates were characterized by plasmid incompatibility group B/O (IncB/O). Conclusions: We detected increased frequency of both TEM and CTX-M type β-lactamases in E. coli isolates from a single-hospital setting, with significant consequences for further treatment approaches. The high prevalence of broad- and extended-spectrum β-lactamase producers tends to prompt an increased carbapenem use (potentially resulting in increased resistance to carbapenems); thus, this type of analytical work should become a standard approach (where possible) in hospital centers in our country and worldwide.Funding: NoDisclosures: None

Molecular characterization of ESBL- producing uropathogenic Escherichia coli recovered from urine samples of patients attending a University Teaching hospital in Nigeria.

Infection of the urinary tract ranks as one of the most common infections affecting people worldwide and its treatment is made complicated by the rising incidence of antibiotic resistance. This study aimed to detect extended spectrum beta-lactamase (ESBL) genes and antibiotic resistance profile of uropathogenic Escherichia coli (E. coli) recovered from patients attending a University Teaching hospital in Nigeria. Uropathogenic E. coli isolates were obtained from the culture collection of Department of Microbiology and Parasitology of the University Teaching hospital for a period of four months (October 2019-January, 2020). Antibiotic susceptibility testing was done using the disc diffusion method while phenotypic ESBL production was detected using double disc synergy test (DDST). Detection of β-lactamase genes was done using Real-Time PCR. Forty-nine E. coli isolates were recovered from 120 urine samples, with 24 (49%) being ESBL positive. The resistance to antibiotics in the ESBL producers was: ciprofloxacin (100%), cefotaxime (100%), cefpodoxime (100%), tetracycline (95.7%), ceftazidime (56.7%), amoxicillin-clavulanate (50%), gentamicin (33.3%), and imipenem (0%). All the ESBL producers carried blaTEM, blaCTX-M-1 and blaCTX-M-9, 75% (18/24) carried blaSHV, while blaCTX-M-2, blaCTX-M-8 and blaCTX-M-25 groups were detected in 20.8% (5/24) of the isolates. There was co-occurrence of CTX-M, SHV and TEM β-lactamases in 79.2% (19/24) isolates, while five isolates (20.8%) co-harbored blaCTX-M and blaTEM. This study showed a high level of multidrug resistance and ESBL gene carriage in uropathogenic E. coli obtained in this study, suggesting a likely review of therapeutic options in the treatment of UTI to clamp down on the rising cases of antibiotic resistance.

Cefazolin versus ceftriaxone as definitive treatment for Klebsiella pneumoniae bacteraemia: a retrospective multicentre study in Singapore.

Ceftriaxone is the preferred treatment for bacteraemia caused by non-MDR (antibiotic-susceptible) Klebsiella pneumoniae. Excessive and widespread ceftriaxone use creates selection pressure for ESBLs. Cefazolin is an alternative, although there are theoretical concerns that SHV-1 β-lactamase in K. pneumoniae may inactivate cefazolin in an inoculum-dependent manner. In this retrospective study, we investigated the outcomes in K. pneumoniae bacteraemia patients treated with IV cefazolin versus IV ceftriaxone as definitive therapy. A total of 917 patients infected with K. pneumoniae from 1 January to 31 December 2016 in three public acute care hospitals in Singapore were screened for study eligibility. Consecutive unique episodes of monomicrobial bacteraemia caused by cefazolin- and/or ceftriaxone-susceptible K. pneumoniae were analysed (n = 284). There were 143 patients (50.4%) in the cefazolin group and 141 patients (49.6%) in the ceftriaxone group. Demographics, baseline illness severity and risk factors for healthcare-associated bacteraemia were comparable in the two treatment groups. The primary outcome of 28 day all-cause mortality was not significantly different between the cefazolin and ceftriaxone groups (10.5% versus 7.1%, P = 0.403). Both in the crude analysis and using a multivariable logistic regression model with inverse probability weighting based on propensity score, cefazolin treatment was not associated with increased risk of 28 day mortality (OR 1.51 with ceftriaxone as the reference group, 95% CI 0.67-3.53; adjusted OR 1.55, 95% CI 0.33-7.40). Cefazolin may be a ceftriaxone-sparing alternative treatment for antibiotic-susceptible K. pneumoniae bacteraemia. This observation may provide sufficient clinical equipoise for a randomized controlled trial.

Occurrence of CTX-M, SHV and TEM β-lactamase genes in Extended Spectrum Beta-Lactamase (ESBL)- producing bacteria recovered from wastewater of a privately-owned hospital in Nigeria and a hand-dug well within its vicinity

BackgroundThe possibility of the dissemination of resistant bacteria and antimicrobials from healthcare facilities into shared environment is a threat to human health. This study investigated the occurrence of ESBL and AmpC β-lactamase genes in ESBL- producing bacteria isolated from wastewater of a privately-owned hospital in South-western Nigeria and underground water sources within its vicinity. MethodsHospital wastewater and water samples from borehole and hand-dug well, were collected over three months for the isolation of gram-negative bacteria. Phenotypic detection of ESBL production was done using the double disc synergy test (DDST), while characterization of positive isolates was done using API 20E identification kit. Susceptibility to antibiotics and detection of resistance genes were done using the disc diffusion method and Real-Time PCR respectively. Pulse field gel electrophoresis was used to determine the relationship between the isolates. ResultsSixteen ESBL- producing bacteria identified as Serratia fonticola (14), Enterobacter cloacae (1) and Pantoea agglomerans (1) were recovered. Resistance to ampicillin was 100%, ceftazidime (93.75%), amoxicillin-clavulanate (75.0%), trimethoprim-sulfamethoxazole (68.75%), gentamicin (50%), cefotaxime (43.75%), cefpodoxime (31.25%), ceftriaxone (18.75%) and 56.25% each to tetracycline and ciprofloxacin. No resistance was observed to cefoxitin. All the ESBL positive bacteria carried blaSHV, six carried blaSHV + blaTEM, while blaCTX-M-2, blaCTX-M-8 and blaCTX-M-25 were detected in three isolates. No isolate carried blaCTX-M-1, blaCTX-M-9, blaKPC, blaNDM-1, blaCMY-1 and ampC. All the ESBL- producing Serratia fonticola showed a high degree of genetic relatedness. ConclusionHospital wastewater and hand-dug wells in proximity to hospitals could be important reservoirs of ESBL- producing bacteria.

Prevalence of non-extended spectrum β-lactamases SHV-1 and TEM-1 or -2 types in multidrug-resistant Enterobacteriaceae in northern Iran.

The present study aimed to evaluate TEM-1 or -2 and SHV-1 β-lactamases frequency in multidrug-resistant (MDR) Enterobacteriaceae isolated from patients' urine in northern Iran. The resistance pattern to 20 antibiotics and ESBL production in 200 MDR Enterobacteriaceae was detected using the disk diffusion test and double-disk synergy test (DDST), respectively. Multiplex PCR was applied to detect blaTEM-1 or -2 and blaSHV genes in isolates. DDST findings were inconsistent with multiplex PCR results. The distribution of each of blaTEM-1 or -2 and blaSHV genes, either alone or in combination, in the ESBL-producing isolates was higher than the non-ESBL-producing isolates. There was a significant effect of the presence of blaTEM-1 or -2 gene on resistance to cephalotin at the p < 0.01 level and cefepime, tetracycline, and streptomycin at the P < 0.05 level, and the presence of blaSHV-1 gene on resistance to fosfomycin at the P < 0.05 level as well as the presence both blaTEM-1 or -2 and blaSHV-1 genes on resistance to cephalotin and fosfomycin at the P < 0.01 level. In all isolates, ESBL production, except for cephalotin resistance, did not improve resistance to other antibiotics used and even non-ESBL-producing isolates showed higher resistance to antibiotics compared to ESBL-producing isolates. It seems that mechanisms other than production of ESBL to be involved as part of the resistance mechanisms of the studied isolates against the used antibiotics. For epidemiological studies, both phenotypic and molecular tests must be included to identify the blaTEM-1 or -2 and blaSHV-1 genotypes to ensure infection prevention and control.

Isolation of Serratia fonticola Producing FONA, a Minor Extended-Spectrum β-Lactamase (ESBL), from Imported Chicken Meat in Japan.

Five novel strains of Serratia fonticola that produce FONA, a minor extended-spectrum beta-lactamase (ESBL), were isolated during routine surveillance of ESBL-producing Enterobacteriaceae in imported chicken meat in Japan in 2017 and 2018. These strains exhibited a clear ESBL phenotype in susceptibility tests carried out in the presence of clavulanic acid; however, all strains tested negative in a multiplex polymerase chain reaction assay used to detect TEM, SHV, and CTX-M β-lactamase genes. After identification of the bacterial species as S. fonticola, full length blaFONA genes were amplified and the DNA sequences were determined. The blaFONA genes from all 5 strains were different from those previously reported (blaFONA-1 to blaFONA-6); they clustered close to one another but were distinct from previously reported blaFONA genes in a phylogenic analysis based on amino acid sequences.

A Genotype-Phenotype Correlation Study of SHV β-Lactamases Offers New Insight into SHV Resistance Profiles.

The SHV β-lactamases (BLs) have undergone strong allele diversification that has changed their substrate specificities. Based on 147 NCBI entries for SHV alleles, in silico mathematical models predicted 5 positions as relevant for the β-lactamase inhibitor (BLI)-resistant (2br) phenotype, 12 positions as relevant for the extended-spectrum BL (ESBL) (2be) phenotype, and 2 positions as related solely to the narrow-spectrum (2b) phenotype. These positions and six additional positions described in other studies (including one promoter mutation) were systematically substituted and investigated for their substrate specificities in a BL-free Escherichia coli background, representing, to our knowledge, the most comprehensive substrate and substitution analysis for SHV alleles to date. An in vitro analysis confirmed the essentiality of positions 238 and 179 for the 2be phenotype and of position 69 for the 2br phenotype. The E240K and E240R substitutions, which do not occur alone in known 2br SHV variants, led to a 2br phenotype, indicating a latent BLI resistance potential of these substitutions. The M129V, A234G, S271I, and R292Q substitutions conferred latent resistance to cefotaxime. In addition, seven positions that were found not always to be associated with the ESBL phenotype resulted in increased resistance to ceftaroline. We also observed that coupling of a strong promoter (IS26) to an A146V mutant with the 2b phenotype resulted in highly increased resistance to BLIs, cefepime, and ceftaroline but not to third-generation cephalosporins, indicating that SHV enzymes represent an underestimated risk for empirical therapies that use piperacillin-tazobactam or cefepime to treat different infectious diseases caused by Gram-negative bacteria.

Huge Diversity of TEM and SHV β-Lactamases Types Among CTX-M-15-Producing Enterobacteriaceae Species in Tunisia.

The aim of our study was to characterize third-generation cephalosporin (3GC)-resistant Enterobacteriaceae isolated over two different periods from patients hospitalized in the Military Hospital of Tunis with special focus to class A β-lactamases. This study included 180 Enterobacteriaceae resistant to 3GC isolated from samples of patients hospitalized in various services of the hospital. Enterobacteriaceae species detected by the Vitek 2 Compact® (BioMérieux®) automated system showed the dominance of Klebsiella pneumoniae followed by Escherichia coli during both periods. These strains were mainly isolated from urine samples and rectal swabs of patients hospitalized mostly in neonatology service and intensive care unit. The molecular research of genes encoding CTX-M, TEM, and SHV β-lactamase types showed a high rate of strains producing CTX-M β-lactamases, all of them harbored the blaCTX-M-15 gene. However, a huge diversity of SHV and TEM β-lactamases types was discovered in our study. In fact, nine various subvariants of blaSHV gene (blaSHV-1, blaSHV-11, blaSHV-12, blaSHV-27, blaSHV-28, blaSHV-31, blaSHV-38, blaSHV-79, and blaSHV-81) and eight subvariants of blaTEM gene (blaTEM-70, blaTEM-71, blaTEM-76, blaTEM-77, blaTEM-79, blaTEM-105, blaTEM-148, and blaTEM-186) were identified among our Enterobacteriaceae species during both periods. All subvariants of blaTEM gene and some subvariants of blaSHV gene (blaSHV-31, blaSHV-38, blaSHV-79, and blaSHV-81) have not been previously detected in our country.

Diversity of CTX-M-positive Escherichia coli recovered from animals in Canada

Historically, extended-spectrum cephalosporin resistance in bacteria from animals in Canada has been attributed to the SHV and CMY β-lactamase families. This pattern is beginning to change with the emergence of the blaCTX-M gene family among Escherichia coli recovered from various animal species. Here we analyze and compare whole genome sequences of blaCTX-M-positive E. coli isolates (n = 173) from dogs, chicken, swine, horses and beef cattle in Canada. Ten blaCTX-M variants were identified with blaCTX-M-1,-14, -15, -27 and blaCTX-M-55 being identified in most animal species. These variants occurred across many sequence types, suggesting that mobile genetic elements mediate the spread of blaCTX-M. The variants blaCTX-M-14, -15, -27 and blaCTX-M-55 are associated with the global spread of blaCTX-M in human clinical isolates and their presence could be indicative of transfer between humans and animals. These variants were also the principal variants identified among sequence type 131 isolates, which were not associated with any other species than dogs. These isolates carried the same blaCTX-M variants as E. coli isolates found in humans. Close contact may promote the transmission of these isolates between humans and companion animals.

Determination of Antibiotic Resistance Pattern and frequency of CTX-M, TEM, and SHV Β-Lactamase Encoding Genes among Shigella Isolates from Inpatients in Tehran, Iran

Determination of Antibiotic Resistance Pattern and frequency of CTX-M, TEM, and SHV Β-Lactamase Encoding Genes among Shigella Isolates from Inpatients in Tehran, Iran

Molecular Detection of New SHV β-lactamase Variants in Clinical Escherichia coli and Klebsiella pneumoniae Isolates from Egypt.

The emergence of multidrug-resistant (MDR) pathogens was reported worldwide. Herein, SHV extended-spectrum β-lactamase (SHV-ESBL) variants detection was investigated in MDR E. coli and K. pneumoniae isolates recovered from human subjects (n = 144), one day-old chicks (n = 36) and broiler clinical samples (n = 90). All examined samples were positive for E. coli (n = 246/270; 91.11%) and Klebsiella pneumoniae (n = 24/270; 8.89%). Antimicrobial susceptibility testing was performed on E. coli and K. pneumoniae. SHV-ESBL producing isolates were defined followed by SHV-ESBL amino acids sequence and proteins structure-function analyses. Phylogenetic analysis of 11 MDR isolates resistant to at least 6 β-lactams was designed to determine their genetic relationship with those previously identified in Egypt. SHV-ESBL variants were detected in 28% and 16% of E. coli and K. pneumoniae isolates, respectively. Among the 11 SHV-ESBL producing isolates, one isolate displayed 100% blaSHV-12 similarity with three point mutations, while the other 10 isolates displayed amino acid substitutions at previously non-reported sites. Amino acid sequence analyses of these 10 isolates displayed 96-100% identity to blaSHV-10 (2 isolates with 3-6 point mutations), blaSHV-18 (one isolate with 4 point mutations), blaSHV-58 (4 isolates with 4-5 point mutations), and blaSHV-91 (3 isolates with 3-7 point mutations). These mutations altered SHV-enzyme pocket dimensions and its binding sites chargeability. The blaSHV phylogeny analysis revealed occurrence of variants in closely related lineages with blaSHV-5 and blaSHV-12 with possibility of blaSHV gene transfer between human and birds. The occurrence of these variants in Egypt could help in epidemiological studies and could explain the emergent resistance to β-lactams.

Epidemiology and molecular characterization of multidrug-resistant Escherichia coli isolates harboring blaCTX-M group 1 extended-spectrum β-lactamases causing bacteremia and urinary tract infection in Manhiça, Mozambique.

BackgroundThe emergence and spread of extended-spectrum β-lactamases (ESBLs), especially CTX-M, is an important public health problem with serious implications for low-income countries where second-line treatment is often unavailable. Knowledge of the local prevalence of ESBL is critical to define appropriate empirical therapeutic strategies for multidrug-resistant (MDR) organisms. This study aimed to assess and characterize the presence of ESBL and especially CTX-M-producing Escherichia coli MDR isolates from patients with urinary tract infections (UTIs) and bacteremia in a rural hospital in Mozambique.Materials and methodsOne hundred and fifty-one E. coli isolates from bacteremia and UTI in children were screened for CTX-M, TEM, SHV and OXA β-lactamases by polymerase chain reaction and sequencing. Isolates carrying CTX-M group 1 β-lactamases were further studied. The resistance to other antibiotic families was determined by phenotypic and genotypic methods, the location of the blaCTX-M gene and the epidemiology of the isolates were studied, and extensive plasmid characterization was performed.ResultsApproximately 11% (17/151) of E. coli isolates causing bacteremia and UTI were ESBL producers. CTX-M-15 was the most frequently detected ESBL, accounting for 75% of the total isolates characterized. The blaCTX-M gene is located in different plasmids belonging to different incompatibility groups and can be found in non-epidemiologically related isolates, indicating the high capacity of this resistance determinant to spread widely.ConclusionOur data suggest the presence of a co-selection of third-generation cephalosporin-resistant determinants in the study area despite limited access to these antibiotics. This highlights the importance of continuous surveillance of antimicrobial resistance of both genetic elements of resistance and resistant isolates in order to monitor the emergence and trends of ESBL-producing isolates to promote adequate therapeutic strategies for the management of MDR bacterial infections.

Typhoid Fever due to Extended Spectrum β-Lactamase-Producing Salmonella enterica Serovar Typhi: A Case Report and Literature Review

Emergence of cephalosporin-resistant strains of Salmonella enterica serovar Typhi is a cause of concern in the management of enteric fever. Cephalosporin resistance in Salmonella species is mainly due to the production of extended-spectrum β-lactamases (ESBLs). The majority of ESBLs in Salmonella enterica serovar Typhi are derivatives of the TEM, SHV, and CTX-M β-lactamase families. Of these, CTX-M appears to be predominant. This paper discusses the detection and molecular characterization of an ESBL-producing Salmonella enterica serovar Typhi strain isolated from a patient who was admitted to a private hospital in Sri Lanka. The three main types of β-lactamases such as TEM, SHV, and CTX-M were identified in this isolate. This case report from Sri Lanka contributes to the knowledge of the increasingly reported cases of typhoid fever due to Salmonella enterica serovar Typhi resistant to β-lactamase by ESBL production.

Induction of β-Lactamase Activity and Decreased β-Lactam Susceptibility by CO2 in Clinical Bacterial Isolates.

Antimicrobial susceptibility testing of clinical isolates is a crucial step toward appropriate treatment of infectious diseases. The clinical isolate Francisella philomiragia 14IUHPL001, recently isolated from a 63-year-old woman with atypical pneumonia, featured decreased susceptibility to β-lactam antibiotics when cultivated in 5% CO2. Quantitative β-lactamase assays demonstrated a significant (P < 0.0001) increase in enzymatic activity between bacteria cultivated in 5% CO2 over those incubated in ambient air. The presence of β-lactamase genes blaTEM and blaSHV was detected in the clinical isolate F.philomiragia 14IUHPL001 by PCR, and the genes were positively identified by nucleotide sequencing. Expression of blaTEM and blaSHV was detected by reverse transcription-PCR during growth at 5% CO2 but not during growth in ambient air. A statistically significant alkaline shift was observed following cultivation of F.philomiragia 14IUHPL001 in both ambient air and 5% CO2, allowing desegregation of the previously reported effects of acidic pH from the currently reported effect of 5% CO2 on blaTEM and blaSHV β-lactamases. To ensure that the observed phenomenon was not unique to F.philomiragia, we evaluated a clinical isolate of blaTEM-carrying Haemophilus influenzae and found parallel induction of blaTEM gene expression and β-lactamase activity at 5% CO2 relative to ambient air. IMPORTANCE β-Lactamase induction and concurrent β-lactam resistance in respiratory tract pathogens as a consequence of growth in a physiologically relevant level of CO2 are of clinical significance, particularly given the ubiquity of TEM and SHV β-lactamase genes in diverse bacterial pathogens. This is the first report of β-lactamase induction by 5% CO2.

Treating complicated carbapenem-resistant enterobacteriaceae infections with ceftazidime/avibactam: a retrospective study with molecular strain characterisation.

Ceftazidime/avibactam (CAZ/AVI) is the first antimicrobial agent with activity against carbapenem-resistant Enterobacteriaceae (CRE) approved by the US Food and Drug Administration (FDA). Notably, human clinical outcome data for this indication are limited. Therefore, a retrospective study was performed to evaluate the clinical outcomes and bacterial genomic characteristics of patients hospitalised at a tertiary medical centre with CRE infections treated for the first time with CAZ/AVI. From a total of 44 patients with CRE infections, 6 patients were treated with CAZ/AVI. The duration of CAZ/AVI treatment ranged from 7 days to 28 days. Five patients achieved clinical cure, however two relapsed with the same carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strain within 3 weeks of completion of CAZ/AVI treatment. In addition, one patient with CR-Kp pneumonia experienced clinical failure despite having a documented CAZ/AVI-susceptible CR-Kp strain [minimum inhibitory concentration (MIC) = 2 mg/L]. Consequently, the overall rate of unsuccessful outcome in this small cohort of patients was 50%. All strains carried KPC-3, OXA-9 and different TEM and SHV β-lactamases, but none carried the intrinsically avibactam-resistant class B metallo-β-lactamases. No obvious differences in antibiotic resistance genes were observed. This study provides an early glimpse of the clinical outcomes of patients with CR-Kp infections treated with CAZ/AVI. Findings of clinical failure and relapse in patients with no prior exposure to CAZ/AVI and with documented susceptibility to CAZ/AVI highlight the urgent need for well-designed clinical studies evaluating the effectiveness of CAZ/AVI in the treatment of CRE infections.

A Review of SHV Extended-Spectrum β-Lactamases: Neglected Yet Ubiquitous.

β-lactamases are the primary cause of resistance to β-lactams among members of the family Enterobacteriaceae. SHV enzymes have emerged in Enterobacteriaceae causing infections in health care in the last decades of the Twentieth century, and they are now observed in isolates in different epidemiological settings both in human, animal and the environment. Likely originated from a chromosomal penicillinase of Klebsiella pneumoniae, SHV β-lactamases currently encompass a large number of allelic variants including extended-spectrum β-lactamases (ESBL), non-ESBL and several not classified variants. SHV enzymes have evolved from a narrow- to an extended-spectrum of hydrolyzing activity, including monobactams and carbapenems, as a result of amino acid changes that altered the configuration around the active site of the β -lactamases. SHV-ESBLs are usually encoded by self-transmissible plasmids that frequently carry resistance genes to other drug classes and have become widespread throughout the world in several Enterobacteriaceae, emphasizing their clinical significance.