Beta-lactamase Expression Research Articles

FTIR analysis and antimicrobial activity of sodium tungstate and calcium tungstate

In recent years, many antibacterial agents have been produced with the aim of eradicating infectious diseases, but many of these agents are ineffective against the resistance presented by bacteria. It is currently estimated that more than 60 % of current antibiotics are ineffective, so the discovery of new drugs is vital. Among the compounds studied in recent years are polyoxotungstates, inorganic compounds targeted for their pharmacological properties. The aim of this study was therefore to chemically characterize two tungstates: calcium and sodium, and to evaluate their microbiological properties, both in combination with antibiotics and due to their ability to reverse the resistance process represented by the expression of the enzyme betalactamase. The microbiological tests were carried out using the microdilution technique, with colorimetric disclosure, using resazurin, and the chemical characterization and vibrational modes of the compounds were evaluated using Fourier transform infrared spectroscopy with attenuated total reflectance. Calcium tungstate showed four spectroscopic bands, located between 84 and 1915 cm−1, while sodium tungstate showed two bands at 335 and 935 cm−1. Calcium tungstate intensified the effect of gentamicin against the bacterium Escherichia coli 06, as well as reversing the mechanism of enzymatic resistance presented by the bacteria Staphylococcus aureus K-4100 and K-4414. Given the current scenario of resistance, these results represent new alternatives for the treatment of bacterial infections, allowing a better understanding of the properties of polyoxometalates. These results are unprecedented as far as the literature is concerned.

Efficacy of an inulin-based treatment on intestinal colonization by multidrug-resistant E. coli: insight into the mechanism of action

ABSTRACT Inulin, an increasingly studied dietary fiber, alters intestinal microbiota. The aim of this study was to assess whether inulin decreases intestinal colonization by multidrug resistant E. coli and to investigate its potential mechanisms of action. Mice with amoxicillin-induced intestinal dysbiosis mice were inoculated with extended spectrum beta-lactamase producing E. coli (ESBL-E. coli). The combination of inulin and pantoprazole (IP) significantly reduced ESBL-E. coli fecal titers, whereas pantoprazole alone did not and inulin had a delayed and limited effect. Fecal microbiome was assessed using shotgun metagenomic sequencing and qPCR. The efficacy of IP was predicted by increased abundance of 74 taxa, including two species of Adlercreutzia. Preventive treatments with A. caecimuris or A. muris also reduced ESBL-E. coli fecal titers. Fecal microbiota of mice effectively treated by IP was enriched in genes involved in inulin catabolism, production of propionate and expression of beta-lactamases. They also had increased beta-lactamase activity and decreased amoxicillin concentration. These results suggest that IP act through production of propionate and degradation of amoxicillin by the microbiota. The combination of pantoprazole and inulin is a potential treatment of intestinal colonization by multidrug-resistant E. coli. The ability of prebiotics to promote propionate and/or beta-lactamase producing bacteria may be used as a screening tool to identify potential treatments of intestinal colonization by multidrug resistant Enterobacterales.

Spread of Pseudomonas aeruginosa ST274 Clone in Different Niches: Resistome, Virulome, and Phylogenetic Relationship.

Pseudomonas aeruginosa ST274 is an international epidemic high-risk clone, mostly associated with hospital settings and appears to colonize cystic fibrosis (CF) patients worldwide. To understand the relevant mechanisms for its success, the biological and genomic characteristics of 11 ST274-P. aeruginosa strains from clinical and non-clinical origins were analyzed. The extensively drug-resistant (XDR/DTR), the non-susceptible to at least one agent (modR), and the lasR-truncated (by ISPsp7) strains showed a chronic infection phenotype characterized by loss of serotype-specific antigenicity and low motility. Furthermore, the XDR/DTR and modR strains presented low pigment production and biofilm formation, which were very high in the lasR-truncated strain. Their whole genome sequences were compared with other 14 ST274-P. aeruginosa genomes available in the NCBI database, and certain associations have been primarily detected: blaOXA-486 and blaPDC-24 genes, serotype O:3, exoS+/exoU- genotype, group V of type IV pili, and pyoverdine locus class II. Other general molecular markers highlight the absence of vqsM and pldA/tleS genes and the presence of the same mutational pattern in genes involving two-component sensor-regulator systems PmrAB and CreBD, exotoxin A, quorum-sensing RhlI, beta-lactamase expression regulator AmpD, PBP1A, or FusA2 elongation factor G. The proportionated ST274-P. aeruginosa results could serve as the basis for more specific studies focused on better antibiotic stewardship and new therapeutic developments.

Microorganisms isolated in patients with post-cardiac surgery mediastinitis at a cardiology hospital of Mexico City.

Mediastinitis after cardiovascular surgery gives rise to prolonged hospital stay and increased medical care costs, and is associated with high in-hospital mortality. To describe the clinical characteristics of patients with post-surgical mediastinitis, including the isolated microorganisms, resistance profile, and in-hospital survival. Cross-sectional study of patients with bacteriologically-confirmed post-surgical mediastinitis cared for at a cardiology hospital in Mexico City between January 2017 and March 2019. Fifty-eight cases of mediastinitis were included. Median age was 67 years. Most subjects were males who underwent myocardial revascularization. During in-hospital follow-up, all-cause and mediastinitis-related mortality were 27.6% and 20.7%, respectively. Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli were the most commonly isolated microorganisms. High resistance to methicillin was found in coagulase-negative staphylococci, as well as high expression of extended-spectrum beta-lactamases in Escherichia coli and Klebsiella pneumoniae strains. High mortality and significant antimicrobial resistance were found in patients with post-cardiac surgery mediastinitis.

Cinnamaldehyde Restores Ceftriaxone Susceptibility against Multidrug-Resistant Salmonella.

In recent years, infections caused by multidrug-resistant (MDR) bacteria have greatly threatened human health and imposed a burden on global public health. To overcome this crisis, there is an urgent need to seek effective alternatives to single antibiotic therapy to circumvent drug resistance and prevent MDR bacteria. According to previous reports, cinnamaldehyde exerts antibacterial activity against drug-resistant Salmonella spp. This study was conducted to investigate whether cinnamaldehyde has a synergistic effect on antibiotics when used in combination, we found that cinnamaldehyde enhanced the antibacterial activity of ceftriaxone sodium against MDR Salmonella in vitro by significantly reduced the expression of extended-spectrum beta-lactamase, inhibiting the development of drug resistance under ceftriaxone selective pressure in vitro, damaging the cell membrane, and affecting its basic metabolism. In addition, it restored the activity of ceftriaxone sodium against MDR Salmonella in vivo and inhibited peritonitis caused by ceftriaxone resistant strain of Salmonella in mice. Collectively, these results revealed that cinnamaldehyde can be used as a novel ceftriaxone adjuvant to prevent and treat infections caused by MDR Salmonella, mitigating the possibility of producing further mutant strains.

Comparative genomic analyses of the clinically-derived Winkia strain NY0527: the reassignment of W. neuii subsp. neuii and W. neuii subsp. antitratus into two separate species and insights into their virulence characteristics.

Winkia neuii, previously known as Actinomyces neuii, is increasingly recognized as a causative agent of various human infections, while its taxonomy and genomic insights are still understudied. A Winkia strain NY0527 was isolated from the hip abscess of a patient, and its antibiotic susceptibility was assessed. The genome was hybrid assembled from long-reads and short-reads sequencing. Whole-genome-based analyses on taxa assignment, strain diversity, and pathogenesis were conducted. The strain was found to be highly susceptible to beta-lactam antibiotics, but resistant to erythromycin, clindamycin, and amikacin. The complete genome sequences of this strain were assembled and found to consist of a circular chromosome and a circular plasmid. Sequence alignment to the NCBI-nt database revealed that the plasmid had high sequence identity (>90%) to four Corynebacterium plasmids, with 40-50% query sequence coverage. Furthermore, the plasmid was discovered to possibly originate from the sequence recombination events of two Corynebacterium plasmid families. Phylogenomic tree and genomic average nucleotide identity analyses indicated that many Winkia sp. strains were still erroneously assigned as Actinomyces sp. strains, and the documented subspecies within W. neuii should be reclassified as two separate species (i.e., W. neuii and W. anitratus). The core genome of each species carried a chromosome-coded beta-lactamase expression repressor gene, which may account for their broadly observed susceptibility to beta-lactam antibiotics in clinical settings. Additionally, an ermX gene that expresses fluoroquinolone resistance was shared by some W. neuii and W. anitratus strains, possibly acquired by IS6 transposase-directed gene transfer events. In contrast, tetracycline resistance genes were exclusively carried by W. neuii strains. In particular, W. neuii was found to be more pathogenic than W. anitratus by encoding more virulence factors (i.e., 35-38 in W. neuii vs 27-31 in W. anitratus). Moreover, both species encoded two core pathogenic virulence factors, namely hemolysin and sialidase, which may facilitate their infections by expressing poreformation, adhesion, and immunoglobulin deglycosylation activities. This study highlights the underappreciated taxonomic diversity of Winkia spp. and provides populational genomic insights into their antibiotic susceptibility and pathogenesis for the first time, which could be helpful in the clinical diagnosis and treatment of Winkia spp. infections.

Antibacterial effect of vitamin C against uropathogenic E. coli in vitro and in vivo

BackgroundResistance to antibiotics has increased steadily over time, thus there is a pressing need for safer alternatives to antibiotics. Current study aims to evaluate the influence of vitamin C as an antibacterial and anti-biofilm agent against uropathogenic E. coli (UPEC) strains. The expression of beta-lactamases and biofilm encoding genes among E. coli isolates before and after treating the isolates with sub MIC of vitamin C was analyzed by Real-time PCR. The in vivo assessment of the antibacterial and anti-biofilm effects of vitamin C against uropathogenic E. coli strains was done using a urinary tract infection (UTI) rat model.ResultsThe effective concentration of vitamin C that could inhibit the growth of most study isolates (70%) was 1.25 mg/ml. Vitamin C showed a synergistic effect with most of the studied antibiotics; no antagonistic effect was detected at all. Vitamin C showed an excellent anti-biofilm effect against studied isolates, where 43 biofilm-producing isolates were converted to non-biofilm at a concentration of 0.312 mg/ml. The expression levels of most studied genes were down-regulated after treatment of E. coli isolates with vitamin C. In vivo assessment of vitamin C in treating UTIs showed that vitamin C has a rapid curative effect as the comparable antibiotic. Administration of both vitamin C and nitrofurantoin at a lower dose for treatment of UTI in rats had a better effect.ConclusionVitamin C as an antibacterial and anti-biofilm agent either alone or in combination with antibiotics could markedly improve UTI in experimental rats.

Incidence of multidrug resistance and extended spectrum beta-lactamase expression in community-acquired urinary tract infection among different age groups of patients

Background and Aim: Escherichia coli was the most prevalent cause for urinary tract infections (UTIs). These bacteria also generate extended spectrum b-lactamases (ESBL), which inactivate penicillins and cephalosporins. The purpose of the current study is to determine the incidence of multidrug resistance and extended beta-lactamase expression in community-acquired urinary tract infection (UTIs) among patients of different age groups. Patients and Methods: This prospective cross-sectional study was conducted on 186 uropathogens investigated in the diagnostic center of Tertiary Care Hospital Peshawar from January 2022 to December 2022. Patients of either gender or any age with suspected UTIs associated with K. pneumoniae and E. coli were derived from urine specimen were enrolled. Identification and analysis of antimicrobial susceptibility was done using Disc diffusion method and Double Disc synergy test. SPSS version 27 was used for data analysis. Results: Of the total 186 Gram-negative uropathogens, there were 57 (30.6%) male and 129 (59.4%) female. The incidence of E. coli and K. pneumoniae was 138 (74.2%) and 48 (25.8%) respectively. The prevalence of uropathogens identified was in 71% females and 29% males.

Chromosome Mediated Fluoroquinolone and Extended Spectrum Beta-lactamase Resistant Genes in E. coli of Poultry Origin in Ekiti State

Background: One health approach aimed at solving global health crisis links human, animal, and environmental health. This inclusive strategy has contributed to antibiotic classification in both human and animal medicine.
 Aims: The aims of this research work are to determine the phylogenetic relationship of E. coli isolated from poultry and waste sources. The presence of chromosome mediated fluoroquinolone and extended spectrum beta-lactamase resistant genes will also be detected in the isolates.
 Study Design: Experimental design.
 Methodology: Data on farming attitudes of poultry farmers were collected using a questionnaire. E. coli was isolated from fresh poultry droppings and waste disposal sites using eosine methylene blue agar. The antibiotic sensitivity profile of the isolates was determined using the modified Kirby Bauer disc diffusion method. Phenotypic expression of fluoroquinolone (qnrS) and beta-lactamase (blaCMY) resistant traits were further detected using Polymerase Chain Reaction. The 16S rRNA gene sequencing was carried out followed by sequence alignment of E. coli genes with those from GenBank sources to determine the molecular identity of the isolates. Spearman’s correlation coefficient (rs) was run to determine the relationship between antibiotic treatment and resistant profile of the isolates. The phylogenetic relationship of the isolates was determined using Bio edit and Mega 6 software.
 Results: Organic poultry farming was practiced by small-scaled, peasant farmers who raised free range birds while antibiotics were widely used on farms that adopted intensive mode of farming. The percentage occurrence of E. coli from waste disposal sources was lesser than that from fresh poultry droppings. Highest percentage of antibiotic resistance to the fluoroquinolones was found while the carbapenemase recorded the lowest. Statistical analysis shows that antibiotic treatment in poultry and resistant profile of isolates to antibiotics are directly related. The percentage similarity of gene sequence with those from Gene Data Bank (≥99.29%) validates the identity of the isolates as E. coli. About, 60% of the sampled population had the qnrS gene with a band size of approximately 322 base pair. Besides, 40% of the sampled isolates possessed the blaCMY gene with a band size of approximately 460 base pair. Both genes co-existed in the chromosome of 15% of the sampled isolates sourced from poultry droppings and waste sources. Phylogenetic classification links the origin of isolates from waste disposal sources to poultry production sites. Besides, variant strains of multiple antibiotic resistant E. coli from poultry with antibiotic treatment were more diverse compared to those obtained from birds raised without antibiotics.
 Conclusion: The qnrS and blaCMY genes found in multiple antibiotic resistant E. coli mediated resistance to critically important antibiotics. The co-existence of these genes in variants strains of E. coli occupying different phylogenetic clusters suggests that antibiotics were widely used on the birds. Antibiotic treatment regimen in poultry may be responsible for the expression of antibiotic resistant genes found in the chromosome of the variant strains of E. coli.

Veterinary clinic surfaces as reservoirs of multi-drug- and biocide-resistant Gram-negative bacteria.

This cross-sectional study was carried out to determine the common Gram-negative bacteria (GNB) contaminating veterinary clinic environments, and to evaluate the susceptibility of the isolates to commonly used antibiotics and biocides. A total of 62 swab samples were collected from different frequently touched surfaces in the 4 veterinary clinics visited. The samples were processed for isolation and identification of GNB using standard microbiological procedures. The susceptibility of the isolates to disinfectants and antibiotics was determined using agar dilution and disc diffusion techniques, respectively. A total of 114 GNB were isolated from the 4 clinics with isolation rates of 21.9, 22.8, 23.7 and 31.6% in clinics A, B, C and D, respectively. The surfaces of treatment tables were more contaminated (16.7 %) than receptionist/clinician desks (15.8%), weighing balances (10.5 %), door handles (7.9 %), drip stands (7.9 %), handwashing basins (7.0 %) and client chairs (7.0%). The surface-contaminating isolates were distributed into 20 genera, with members of Enterobacteriaceae predominating (n=97). Fifty-nine per cent of the isolates were resistant to the disinfectant Septol, while 5.3 and 0.9% were resistant to Purit and Dettol disinfectants, respectively. Multiple drug resistance was observed among 99% of the isolates with approximately 100% resistance to beta-lactams. Phenotypic expression of extended-spectrum (3.5 %) and AmpC beta-lactamase (38.6 %) production was detected. These findings highlight the role of clinic environments in serving as reservoirs for potential pathogens and sources for the spread of multi-drug resistant GNB.

Mass Spectrometry-Based Detection of Beta Lactam Hydrolysis Enables Rapid Detection of Beta Lactamase Mediated Antibiotic Resistance.

Antibiotic resistance by beta lactamase expression is a serious and growing threat. We aimed to determine whether beta-lactamase activity is detectable in urine specimens to enable faster identification of resistance. Urine specimens from patients with extended spectrum beta lactamase (ESBL)-expressing urinary infections were incubated with beta lactam antibiotics. Beta lactam hydrolysis was determined by mass spectrometry methods. Ceftriaxone hydrolysis was observed in 45 of 45 ESBL-containing specimens from patients not treated with a beta lactamase inhibitor before specimen collection. Ceftriaxone hydrolysis was not observed in 108 of 108 non-ESBL-containing specimens. Spiking studies show that beta lactam hydrolysis can be observed within 30 minutes. Beta lactam hydrolysis is evidenced by mass spectrometry preceded by either liquid chromatography or matrix-assisted laser desorption ionization specimen processing methods. Clinically significant beta lactamase activity is detectable directly from urine specimens. The described methods would enable the detection of beta lactam resistance 24 to 48 hours sooner than culture based methods.

Chlorine disinfectants promote microbial resistance in Pseudomonas sp.

The substantial use of disinfectants has increased antibiotic resistance, thereby mediating serious ecological safety issues worldwide. Accumulating studies have reported the role of chlorine disinfectants in promoting disinfectant resistance. The present study sought to investigate the role of chlorine disinfectants in developing multiple resistance in Pseudomonas sp. isolated from the river through antioxidant enzyme measurement, global transcriptional analyses, Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The results demonstrated that 100 mg/L sodium hypochlorite could increase disinfectant resistance and antibiotic resistance. The SOS response (a conserved response to DNA damage) triggered by oxidative stress makes bacteria resistant to chlorine. An increase in antibiotic resistance could be attributed to a decreased membrane permeability, increased expression of MuxABC-OpmB efflux pump, beta-lactamase, and antioxidant enzymes. Additionally, KEGG enrichment analysis suggested that the differentially expressed genes were highly enriched in the metabolic pathways. In summary, the study results revealed the impact of chlorine disinfectants in promoting microbial disinfectant resistance and antibiotic resistance. This study will provide insight into disinfectant resistance mechanisms.

Clinical and microbiological characterization of sepsis and evaluation of sepsis scores.

BackgroundDespite the necessity of early recognition for an optimal outcome, sepsis often remains unrecognized. Available tools for early recognition are rarely evaluated in low- and middle-income countries. In this study, we analyzed the spectrum, treatment and outcome of sepsis at an Ethiopian tertiary hospital and evaluated recommended sepsis scores.MethodsPatients with an infection and ≥2 SIRS criteria were screened for sepsis by SOFA scoring. From septic patients, socioeconomic and clinical data as well as blood cultures were collected and they were followed until discharge or death; 28-day mortality was determined.ResultsIn 170 patients with sepsis, the overall mortality rate was 29.4%. The recognition rate by treating physicians after initial clinical assessment was low (12.4%). Increased risk of mortality was significantly associated with level of SOFA and qSOFA score, Gram-negative bacteremia (in comparison to Gram-positive bacteremia; 42.9 versus 16.7%), and antimicrobial regimen including ceftriaxone (35.7% versus 19.2%) or metronidazole (43.8% versus 25.0%), but not with an increased respiratory rate (≥22/min) or decreased systolic blood pressure (≤100mmHg). In Gram-negative isolates, extended antimicrobial resistance with expression of extended-spectrum beta-lactamase and carbapenemase genes was common. Among adult patients, sensitivity and specificity of qSOFA score for detection of sepsis were 54.3% and 66.7%, respectively.ConclusionSepsis is commonly unrecognized and associated with high mortality, showing the need for reliable and easy-applicable tools to support early recognition. The established sepsis scores were either of limited applicability (SOFA) or, as in the case of qSOFA, were significantly impaired in their sensitivity and specificity, demonstrating the need for further evaluation and adaptation to local settings. Regional factors like malaria endemicity and HIV prevalence might influence the performance of different scores. Ineffective empirical treatment due to antimicrobial resistance is common and associated with mortality. Local antimicrobial resistance statistics are needed for guidance of calculated antimicrobial therapy to support reduction of sepsis mortality.

High Prevalence and Mechanism Associated With Extended Spectrum Beta-Lactamase-Positive Phenotype in Laribacter hongkongensis.

In this study, we reported the prevalence and mechanism associated with the extended-spectrum beta-lactamase (ESBL)-positive phenotype in Laribacter hongkongensis isolated from patients and fish. Using the inhibition zone enhancement test, 20 (95.2%) of the 21 patient strains and 8 (57.1%) of the 14 fish strains were tested ESBL-positive. However, ESBL genes, including SHV, TEM, CTX-M, GES, and PER, were not detected in all of these 28 L. hongkongensis isolates. No ESBL gene could be detected in either the complete genome of L. hongkongensis HLHK9 or the draft genome of PW3643. PCR and DNA sequencing revealed that all the 35 L. hongkongensis isolates (showing both ESBL-positive and ESBL-negative phenotypes) were positive for the ampC gene. When the AmpC deletion mutant, HLHK9ΔampC, was subject to the zone enhancement test, the difference of zone size between ceftazidime/clavulanate and ceftazidime was less than 5 mm. When boronic acid was added to the antibiotic disks, none of the 28 “ESBL-positive” isolates showed a ≥ 5 mm enhancement of inhibition zone size diameter between ceftazidime/clavulanate and ceftazidime and between cefotaxime/clavulanate and cefotaxime. A high prevalence (80%) of ESBL-positive phenotype is present in L. hongkongensis. Overall, our results suggested that the ESBL-positive phenotype in L. hongkongensis results from the expression of the intrinsic AmpC beta-lactamase. Confirmatory tests should be performed before issuing laboratory reports for L. hongkongensis isolates that are tested ESBL-positive by disk diffusion clavulanate inhibition test.

Austria-wide survey on resistant, potentially pathogenic bacteria at Austrian bathing sites, 2017

Summary There is growing concern about human-induced antibiotic resistance and on the occurrence of antibiotic-resistant, potentially pathogenic bacteria in the environment. The aim of this study was to investigate the incidence of resistant, clinically relevant bacteria at bathing sites. In total, 27 of 263 bathing sites authorized under the EU Bathing Water Directive (3 per Austrian state) were sampled during the summer of 2017. Samples were tested for antibiotic-resistant bacteria by enrichment in thioglycollate broth and cultivation on chromogenic media. The screening for potentially pathogenic antibiotic-resistant bacteria was negative in 23 of the 27 samples. Antibiotic-resistant bacteria were detected from 4 of the 27 bathing sites: one Pseudomonas aeruginosa and three resistant Enterobacteriaceae (piperacillin/tazobactam-resistant Enterobacter cloacae with high-level expression of AmpC beta-lactamase, carbapenem-resistant Enterobacter mori, extended-spectrum beta-lactamase-producing Escherichia coli). Despite the occurrence of resistant bacteria, we consider the public health risk at Austrian bathing sites as low.

Expression of virulence and antimicrobial resistance genes among Escherichia coli clinical isolates from blood and stool samples

Objective: As major nosocomial pathogens, Escherichia coli isolates exhibit antibiotic resistance and also express adhesive structures and antibiotic resistance genes. The objective of this study was the comparison of virulence gene expression of extended-spectrum beta-lactamase (ESBL)-producing E. coli between blood and stool samples. Methods: In this study, 20 E. coli clinical isolates (10 ESBL-producers including 5 from blood, 5 from stool samples and 10 non-ESBL-producer strains) were included. The existence of fimA, kpsMII and cdt (adhesives and toxin), acr-ab (efflux-encoding) and bla(CTX-M1) genes were confirmed by PCR. The quantitative real-time PCR was performed for evaluation of gene expression. Results: ESBL-producing E. coli isolates from stool samples could express fimA, kpsMII and cdt genes significantly higher than blood samples, whereas those isolates from blood samples significantly expressed the acr-ab (efflux-encoding) genes. In addition, the bla(CTXM1) gene was expressed among isolates from stool samples significantly higher (P = 0.022) than those from blood samples according to the analysis of variance (ANOVA) test. In addition, among non-ESBL-producers, the expression of fimA, kpsMII and cdt genes was significantly lower than ESBL-producing isolates from blood samples, but not significantly different than those from stool samples. Moreover, the expression of acr-ab genes was significantly lower than those from stool samples. Conclusion: The results exhibited that the expression of virulence genes among clinical isolates of E. coli is not the same or similar in various conditions or from various clinical origins. Thus determining the profile of gene expression in each of clinical situations can be helpful in tracking the infectious pathogens. ESBL-producing strains possibly have regulatory factors for inducing higher virulence gene expression. Copyright (C) 2019 Wolters Kluwer Health, Inc. All rights reserved.

Comparative analysis of biocontrol agent Trichoderma asperellum ACCC30536 transcriptome during its interaction with Populus davidiana × P. alba var. pyramidalis

After exposure to with Populus davidiana × P. alba var. pyramidalis, the expression of genes in Trichoderma asperellum were compared in four transcriptomes. The top 20 high expression genes included six heat shock proteins and three hydrophobins, indicating that Trichoderma can rapidly adapt to environment stresses and elicit a plant defense response. The genes, involved in the interaction between Trichoderma and plant, showed an increasing expression level, for example sugar transporters, EPL1s, endoxylanases, pectin lyases, and nitrilases. Interestingly, sugar transporters also showed high expression when T. asperellum was cultured on medium lacking a carbon substrate, which would contribute to T. asperellum’s survival and domination in ecological niche competition. And the genes related to mycoparasitism were expressed abundantly following T. asperellum’s interaction with PdPap, indicating the PdPap induction could enhance the mycoparasitic ability of T. asperellum. Twelve chitinases and five glucanases showed higher expression in transcriptome Cs, indicating that T. asperellum secretes both types of enzyme before interacting with pathogens, allowing T. asperellum to implement mycoparasitism and obtain more energy. Many novel transcripts were obtained in each transcriptome, which may play important roles in the biocontrol process of T. asperellum. Interestingly, T. asperellum undergo constitutive alternative splicing in the biocontrol process: Seven biocontrol genes were alternative spliced via intron retention. qRT-PCR analysis proved that intron retention is negatively associated with the expression of chitinase, oligopeptide transporters, and beta-lactamase. However, the percentage of MAPK intron retention was quite low, suggesting that intron retention has little effect on the function of MAPK.

Association of systemic antimicrobials with the expression of beta-lactamases in bacteria cultured from urological patients

BackgroundPatients with abnormalities of the genitourinary tract are at high risk for infections with antimicrobial-resistant pathogens. MethodsAll urine cultures ordered by members of the Division of Urology from four quarterly one-week periods were included. All gram-negative bacilli isolated were analyzed using the Check-Points Check-MDR CT103XL assay to identify the presence of genes associated with resistance to beta-lactam antibiotics. Association between the days of antibiotics and the presence of an ESBL-producing organism was determined. ResultsOne hundred eleven positive cultures were included in this analysis, of which 5 (4.5%) contained ESBL-producing species. Days of systemic antibiotics within 30 days of urine culture was associated with an increased risk of isolating an ESBL-producing pathogen. ConclusionThe overall prevalence of ESBL-producing organisms is low in this cohort. The number of days of systemic antibiotics within 30 days of a urine culture was significantly associated with increased risk of isolating an ESBL-producing organism.

Multilocus sequence analysis, biofilm production, antibiotic susceptibility and synergy tests of Burkholderia species in patients with and without cystic fibrosis

Burkholderia spp. emerged as important pathogens in the airways of immunocompromised humans, especially those with cystic fibrosis (CF). Failure of identification with conventional techniques, high intrinsic resistance to most antibiotics and biofilm formation can cause difficulties in the treatment of these infections. The aim of this study was to identify Burkholderia spp. strains isolated from CF and non-CF patients with with routine microbiological methods, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and multilocus sequence analysis (MLSA), to determine of the antibiotic susceptibility and synergies, and to evaluate biofilm formation of these isolates. A total of 38 Burkholderia spp. (25 CF, 13 non-CF) from 26 patients were identified by biochemical, phenotypical and matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and sequence types were revealed by multilocus sequence analysis (MLSA). Sequence types of isolates were identified using the PubMLST database. Characteristics of biofilm formation of clinical isolates were evaluated by microplate method. Antibiotic susceptibilities of ceftazidime, meropenem, trimethoprim-sulfamethoxazole (TMP-SXT) and levofloxacin were determined by broth microdilution method according to CLSI (2017) guidelines. Synergy tests were performed by checkerboard method. Clinical isolates were identified as Burkholderia cenocepacia (n= 16), Burkholderia contaminans (n= 11), Burkholderia gladioli (n= 4), Burkholderia dolosa (n= 4), Burkholderia multivorans (n= 2) and Burkholderia seminalis (n= 1). Sequence types of these isolates were determined as ST19, ST72, ST102, ST180, ST482, ST602, ST629, ST740, ST839 and ST1392. The correct identification at the species-level with MALDI-TOF MS was 94-100% for all isolates except B.contaminans. Biofilm formation among the identified species in the study was determined as 53% (n= 20). There was no statistical difference when the biofilm production was evaluated separately among Burkholderia species and biofilm production rates between CF (56%, 14/25) and non-CF (46%, 6/13) Burkholderia isolates (p> 0.05). Overall rates of resistance to ceftazidime, meropenem, TMP-SXT, and levofloxacin of the isolates were 35%, 66%, 50% and 40%, respectively. The antibiotic resistance against Burkholderia spp., isolates obtained from CF patients were more susceptible to ceftazidime, but no significant difference was found for other antibiotics. Synergy was determined between meropenem and TMP-SXT in two isolates. Antagonism was detected in 15 isolates, 12 of them were between meropenem and ceftazidime, three of them were between ceftazidime and TMP-SXT. Numerous resistance mechanisms may lead to higher resistance in this bacteria, whereas the antagonism between meropenem and ceftazidime in this study might be attributed to the expression of beta-lactamases. In this study, the distinctness of sequence types between Burkholderia spp. isolated from CF and non-CF patient, provided a better understanding about the importance of biofilm formation for the infections with these bacteria and emphasized that the management of therapy should be driven by the antibiotic test results.

RESISTENCIA BACTERIANA EN UROCULTIVOS DE UNA POBLACIÓN DE EMBARAZADAS DE CONTROL PRENATAL EN BOGOTÁ JUNIO 2013 – JUNIO 2015

La infección urinaria baja, específicamente la bacteriuria asintomática afecta el 2 al 7% de los embarazos; sin tratamiento, evoluciona a pielonefritis, aumenta la probabilidad de parto pretérmino, bajo peso al nacer y preeclampsia. La identificación y tratamiento de la infección urinaria baja es prioritaria. Conocer la sensibilidad antibiótica de los diferentes patógenos permite revisar las estrategias de tratamiento y disminuir los desenlaces adversos perinatales. Objetivo: Determinar el perfil de resistencia de los principales patógenos aislados en los urocultivos de las pacientes embarazadas que acudieron a control prenatal en Clínica Colsanitas. Materiales y Métodos: Se realizó un estudio observacional con pacientes gestantes que asistieron a control prenatal. Se tomaron todos los resultados de urocultivos de las pacientes gestantes y se excluyeron urocultivos sugestivos de contaminación. Se analizaron los patógenos aislados y la sensibilidad a los diferentes antibióticos. Resultados: De 14054 muestras para urocultivo 1177 resultaron positivas. El principal patógeno aislado fue Escherichia coli con una prevalencia del 71,4%, seguido en frecuencia por Enterococcus faecalis, Proteus mirabilis y Klebsiella pneumoniae. La resistencia de Escherichia coli a ampicilina fue de 37,3%, a trimetoprim sulfametoxazol 23,3%, cefalotina 11,1%. La sensibilidad a fosfomicina fue de 98%. En general los patógenos más frecuentemente aislados presentaron bajas tasas de expresión de betalactamasas. Conclusiones: E. coli fue el patógeno más frecuente. La alta resistencia a la ampicilina contraindica su uso empírico. El uso de otros antibióticos como cefalexina o nitrofurantoína es adecuado. Fosfomicina puede ser una opción cuando no sea posible usar los anteriormente descritos.