Advances in single-cell sequencing technology in microbiome research.

With the rapid development of histological techniques and the widespread application of single-cell sequencing in eukaryotes, researchers desire to explore individual microbial genotypes and functional expression, which deepens our understanding of microorganisms. In this review, the history of the development of microbial detection technologies was revealed and the difficulties in the application of single-cell sequencing in microorganisms were dissected as well. Moreover, the characteristics of the currently emerging microbial single-cell sequencing (Microbe-seq) technology were summarized, and the prospects of the application of Microbe-seq in microorganisms were distilled based on the current development status. Despite its mature development, the Microbe-seq technology was still in the optimization stage. A retrospective study was conducted, aiming to promote the widespread application of single-cell sequencing in microorganisms and facilitate further improvement in the technology.

Quantifying Dynamic Phenotypic Heterogeneity in Resistant Escherichia coli under Translation-Inhibiting Antibiotics.

Understanding the phenotypic heterogeneity of antibiotic-resistant bacteria following treatment and the transitions between different phenotypes is crucial for developing effective infection control strategies. The study expands upon previous work by explicating chloramphenicol-induced phenotypic heterogeneities in growth rate, gene expression, and morphology of resistant Escherichia coli using time-lapse microscopy. Correlating the bacterial growth rate and cspC expression, four interchangeable phenotypic subpopulations across varying antibiotic concentrations are identified, surpassing the previously described growth rate bistability. Notably, bacterial cells exhibiting either fast or slow growth rates can concurrently harbor subpopulations characterized by high and low gene expression levels, respectively. To elucidate the mechanisms behind this enhanced heterogeneity, a concise gene expression network model is proposed and the biological significance of the four phenotypes is further explored. Additionally, by employing Hidden Markov Model fitting and integrating the non-equilibrium landscape and flux theory, the real-time data encompassing diverse bacterial traits are analyzed. This approach reveals dynamic changes and switching kinetics in different cell fates, facilitating the quantification of observable behaviors and the non-equilibrium dynamics and thermodynamics at play. The results highlight the multi-dimensional heterogeneous behaviors of antibiotic-resistant bacteria under antibiotic stress, providing new insights into the compromised antibiotic efficacy, microbial response, and associated evolution processes.

Six-year analysis of key monitoring for bacterial strain distribution and antibiotic sensitivity in a hospital.

BACKGROUND: With the widespread use of antimicrobial drugs, bacterial resistance has become a significant problem, posing a serious threat to public health. The prevalence of clinical infection strains in hospitals and their drug sensitivities are key to the appropriate use of antibiotics in clinical practice. AIM: To identify prevalent bacteria and their antibiotic resistance profiles in a hospital setting, thereby guiding effective antibiotic usage by clinicians. METHODS: Specimens from across the institution were collected by the microbiology laboratory. The VITEK 2 compact fully automatic analyzer was used for bacterial identification and antibiotic sensitivity testing, and the WHONET5.6 software was utilized for statistical analysis. RESULTS: A total of 12062 bacterial strains of key monitoring significance were detected. Staphylococcus aureus demonstrated widespread resistance to penicillin, but none of the strains were resistant to vancomycin or linezolid. Moreover, 219 strains of methicillin-resistant coagulase-negative staphylococci and 110 strains of methicillin-resistant Staphylococcus aureus were detected. Enterococcus faecalis showed moderate resistance to the third-generation quinolones ciprofloxacin and levofloxacin, but its resistance to nitrofurantoin and tetracycline was low. Enterococcus faecium displayed significantly lower resistance to third- and fourth-generation quinolones than Enterococcus faecalis. The resistance of two key monitoring strains, Escherichia coli and Klebsiella pneumoniae, to piperacillin/tazobactam was 5%-8%. However, none of the Escherichia coli and Klebsiella pneumoniae strains were resistant to meropenem. The resistance of Acinetobacter baumannii to piperacillin/sulbactam was nearly 90%. Nonetheless, the resistance to tigecycline was low, and Pseudomonas aeruginosa demonstrated minimal resistance in the antibiotic sensitivity test, maintaining a resistance of < 10% to the cephalosporin antibiotics cefotetan and cefoperazone over the last 6 years. The resistance to amikacin remained at 0.2% over the past 3 years. CONCLUSION: Our hospital's overall antibiotic resistance rate was relatively stable from 2017 to 2022. The detection rates of key monitoring strains are reported quarterly and their resistance dynamics are monitored and communicated to the entire hospital, which can guide clinical antibiotic selection.

Available evidence and potential for vaccines for reduction in antibiotic prescriptions.

Bacterial antibiotic resistance is a public health issue. It means that drugs become ineffective, infections persist and have a huge impact on the health of patients and their spreading increases. To address a complex threat such as bacterial antibiotic resistance different and integrated approaches are needed including discovery of new antibiotics, improvement of diagnostics tools and improvement of antibiotic stewardship. Absolutely relevant are prevention of infections as well as decrease in the use of antibiotics. Vaccines are an important tool in the fight against bacterial antibiotic resistance and can help prevent it in several ways. Indeed, vaccines are highly effective in preventing diseases that might otherwise require the use of antibiotics to treat symptoms and associated complications. Preventing infections through vaccination helps reduce the need for and widespread and inappropriate use of antibiotics, including for secondary bacterial infections.

The role of bacterial signaling networks in antibiotics response and resistance regulation.

Excessive use of antibiotics poses a threat to public health and the environment. In ecosystems, such as the marine environment, antibiotic contamination has led to an increase in bacterial resistance. Therefore, the study of bacterial response to antibiotics and the regulation of resistance formation have become an important research field. Traditionally, the processes related to antibiotic responses and resistance regulation have mainly included the activation of efflux pumps, mutation of antibiotic targets, production of biofilms, and production of inactivated or passivation enzymes. In recent years, studies have shown that bacterial signaling networks can affect antibiotic responses and resistance regulation. Signaling systems mostly alter resistance by regulating biofilms, efflux pumps, and mobile genetic elements. Here we provide an overview of how bacterial intraspecific and interspecific signaling networks affect the response to environmental antibiotics. In doing so, this review provides theoretical support for inhibiting bacterial antibiotic resistance and alleviating health and ecological problems caused by antibiotic contamination.

Evolution of Ceftriaxone Resistance of Penicillin-Binding Proteins 2 Revealed by Molecular Modeling.

Penicillin-binding proteins 2 (PBP2) are critically important enzymes in the formation of the bacterial cell wall. Inhibition of PBP2 is utilized in the treatment of various diseases, including gonorrhea. Ceftriaxone is the only drug used to treat gonorrhea currently, and recent growth in PBP2 resistance to this antibiotic is a serious threat to human health. Our study reveals mechanistic aspects of the inhibition reaction of PBP2 from the wild-type FA19 strain and mutant 35/02 and H041 strains of Neisseria Gonorrhoeae by ceftriaxone. QM(PBE0-D3/6-31G**)/MM MD simulations show that the reaction mechanism for the wild-type PBP2 consists of three elementary steps including nucleophilic attack, C-N bond cleavage in the ß-lactam ring and elimination of the leaving group in ceftriaxone. In PBP2 from the mutant strains, the second and third steps occur simultaneously. For all considered systems, the acylation rate is determined by the energy barrier of the first step that increases in the order of PBP2 from FA19, 35/02 and H041 strains. Dynamic behavior of ES complexes is analyzed using geometry and electron density features including Fukui electrophilicity index and Laplacian of electron density maps. It reveals that more efficient activation of the carbonyl group of the antibiotic leads to the lower energy barrier of nucleophilic attack and larger stabilization of the first reaction intermediate. Dynamical network analysis of MD trajectories explains the differences in ceftriaxone binding affinity: in PBP2 from the wild-type strain, the ß3-ß4 loop conformation facilitates substrate binding, whereas in PBP2 from the mutant strains, it exists in the conformation that is unfavorable for complex formation. Thus, we clarify that the experimentally observed decrease in the second-order rate constant of acylation (k2/KS) in PBP2 from the mutant strains is due to both a decrease in the acylation rate constant k2 and an increase in the dissociation constant KS.

Surveillance of bloodstream infections in intensive care units in England, May 2016-April 2017: epidemiology and ecology.

BACKGROUND: Bloodstream infections (BSIs) in patients in intensive care units (ICUs) are associated with increased morbidity, mortality and economic costs. Many BSIs are associated with central venous catheters (CVCs). The Infection in Critical Care Quality Improvement Programme (ICCQIP) was established to initiate surveillance of BSIs in English ICUs. METHODS: A web-based data capture system was launched on 1st May 2016 to collect all positive blood cultures (PBCs), patient-days and CVC-days. National Health Service (NHS) trusts in England were invited to participate in the surveillance programme. Data were linked to the antimicrobial resistance dataset maintained by Public Health England and to mortality data. FINDINGS: Between 1st May 2016 and 30th April 2017, 84 ICUs (72 adult ICUs, seven paediatric ICUs and five neonatal ICUs) based in 57 of 147 NHS trusts provided data. In total, 1474 PBCs were reported, with coagulase-negative staphylococci, Escherichia coli, Staphylococcus aureus and Enterococcus faecium being the most commonly reported organisms. The rates of BSI and ICU-associated CVC-BSI were 5.7, 1.5 and 1.3 per 1000 bed-days and 2.3, 1.0 and 1.5 per 1000 ICU-CVC-days in adult, paediatric and neonatal ICUs, respectively. There was wide variation in BSI and CVC-BSI rates within ICU types, particularly in adult ICUs (0-44.0 per 1000 bed-days and 0-18.3 per 1000 ICU-CVC-days). CONCLUSIONS: While the overall rates of ICU-associated CVC-BSIs were lower than 2.5 per 1000 ICU-CVC-days across all age ranges, large differences were observed between ICUs, highlighting the importance of a national standardized surveillance system to identify opportunities for improvement. Data linkage provided clinically important information on resistance patterns and patient outcomes at no extra cost to participating trusts.

Co-selection and stability of bacterial antibiotic resistance by arsenic pollution accidents in source water.

Frequent heavy-metal pollution accidents severely deteriorated the source water quality of drinking water treatment plants (DWTP). Limited data have explicitly addressed the impact of these incidents on bacterial antibiotic resistance (BAR). In present study, we investigated the shift of antibiotic resistome caused by heavy metal pollution incidents via simulating an arsenic shock loading [As (III)], along with the associated risks imposed on drinking water systems. The results indicated that a quick co-selection of antibiotic resistant bacteria (ARB) was achieved after exposure to 0.2-1 mg/L As (III) for only 6 h, meanwhile, there was an increase of relative abundance of antibiotic resistance genes (ARGs) and mobile genetic elements. Most of the co-selected BAR could be maintained for at least 4 days in the absence of As (III) and antibiotics, implying that the pollution in source water possibly contributed to the preservation and proliferation of antibiotic resistance determinants in the subsequent DWTP. Bacterial community structure analysis showed a strong correlation between bacterial community shift and BAR promotion, and enrichment of opportunistic bacteria (e.g. Escherichia-Shigella, Empedobacter sp. and Elizabethkingia sp.). The results indicated a potential epidemiological threat to the public due to accident-level arsenic contamination in the source water. This study gave insight into understanding the source water pollution accidents from the perspective of bio-hazard and biological risks, and highlighted a neglected important source of BAR in drinking water systems.

Tryptoline-based benzothiazoles re-sensitize MRSA to ß-lactam antibiotics.

Resistance-modifying agents (RMAs) offer a promising solution to combat bacterial antibiotic resistance. Here we report the discovery and structure-activity relationships of a new class of RMAs with a novel tryptoline-based benzothiazole scaffold. Our most potent compound in this series (4ad) re-sensitizes multiple MRSA strains to cephalosporins at low concentrations (2 µg/mL) and has low mammalian cytotoxicity with a half growth inhibitory concentration (GI50) > 100 µg/mL in human cervical carcinoma (HeLa) cells. In addition, the same core scaffold with different substitutions also gives good antibacterial activity against MRSA.

Effect of hydraulic conditions on the prevalence of antibiotic resistance in water supply systems.

The incidence of antibiotic resistance genes (ARGs) in tap water leads to potential risks to human health and draws more and more attention from the public. However, ARGs harbored in drinking water remain largely unexplored. In this study, a simulated water supply system was designed to study the effects of different pipe flow rates on the transmission of antibiotic resistance in water supply systems. We observed that the biofilm in low flow rate pipeline (0.1 m/s, 0.3 m/s) had higher concentration of both antibiotic resistant bacteria (ARB) and ARGs, while high flow rate (0.5 m/s and 0.7 m/s) resulted in low relative abundance of ARB and high relative abundance of ARGs in biofilms. The results showed that the high flow rate led to an abundance in non-culturable bacteria and a scarcity of nutrients in the biofilm, giving rise to its antibiotic resistance. High-throughput sequencing pointed out that the high content of Caulobacteraceae and Paenibacillus were determined in biofilms of high flow rate pipelines. Similarity analysis of microbial community composition of inlet water (IW), biofilms and outlet water (OW) showed that the composition of microbial community in OW was more similar to that in biofilms than in IW. Genera of bacteria in biofilms and OW (Brevundimonas, Brevibacillus and Pseudomonas) which had relationship with sulⅠ, sulⅡ in biofilms (P < 0.05) had higher relative abundance than that in IW. Different flow rate conditions had an impact on the biomass, microbial community, ARB and ARGs composition of biofilms. Thus, the detachment of biofilms can increased the antibiotic resistance of the water.

Impact of disinfectant on bacterial antibiotic resistance transfer between biofilm and tap water in a simulated distribution network.

Bacterial antibiotic resistance (BAR) is profoundly important to human health, but the environmental reservoirs of resistance determinants are poorly understood. BAR of biofilm and tap water were analyzed by using a water distribution simulator where different doses of chlorine and chloramine were used in this study. The results revealed that the disinfectants (≥2 mg/L) suppressed antibiotic resistant bacteria (ARB) in tap water and biofilms, while disinfected water and biofilms had a high relative abundance of ARB. The difference of ARB concentration and ARB percentage between the samples obtained from a disinfected pipeline and a non-disinfected pipeline became smaller over time. Because the water supply system is a unidirectional process, it is unclear how planktonic bacteria in water transfer BAR over time, although biofilm is suspected to play a role in this process. Compared with the biofilm samples without disinfectant, the disinfected biofilm had lower ICC and HPC/ICC percentage, lower AOC and AOC/TOC percentage, indicating that the disinfectant inhibited the bacteria growth in biofilm, and the disinfected biofilm had high proportion of non-culturable bacteria and low biodegradability, which affected BAR in biofilms. High throughput sequencing showed that in biofilms, the relative abundance of genera (uncultured_f_Rhodocyclaceae, Brevundimonas, and Brevibacillus in chlorinated systems, and Brevundimonas, Brevibacillus in chloraminated systems) with multiple antibiotic resistance and high abundance (up to 78.5%), were positively associated with disinfectant concentration and ARB percentage. The major prevalent genera in biofilms were also detected in tap water, suggesting that biofilm growth or biofilm detachment caused by external environmental factors will allow the movement of biofilm clusters with higher ARB concentration and percentage into bulk water, thereby increasing the antibiotic resistance of bacteria in tap water.

First acetyl-proteome profiling of Salmonella Typhimurium revealed involvement of lysine acetylation in drug resistance.

Salmonella are becoming increasingly resistant to fluoroquinolones (FQs), therefore determining the resistance mechanism is very important. Recent studies have shown that protein post-translational modifications (PTM) play a role in bacterial antibiotic resistance. One such type of PTM, lysine acetylation, is a reversible and highly regulated PTM which has been found to be associated with antibiotic resistance in Mycobacterium and Acinetobacter species. Salmonella Typhimurium are major zoonotic pathogens, which are becoming increasingly resistant to FQs, the antibiotics of choice where therapy is indicated. To date, however, there have been no studies on the relationship between PTM and drug resistance in Salmonella. Therefore, in the present study, ciprofloxacin-resistant and susceptible strains of Salmonella were used as the research objects, and tandem mass tag labeling and acetylation enrichment techniques were used to screen for the different expression of actylated proteins between the two strains, and for quantitative and bioinformatics analysis. We identified a total of 631 acetylated proteins involving 1259 lysine acetylation sites. Among the quantified sites, compared with the susceptible strain, the expression of lysine acetylation was upregulated for 112 sites and downregulated for 149 sites in the resistant strain. Bioinformatic analyses showed that the main enrichment pathways for these differentially acetylated proteins are microbial metabolic process, biosynthesis of antibiotics, and bacterial chemotaxis. Among the differentially acetylated proteins, 14 proteins related to bacterial antibiotic resistance were identified (excluding metabolic and virulence-related proteins), and the lysine acetylation expression of these proteins was significantly different between the resistant and susceptible strains. These results indicated that protein lysine acetylation is not only related to metabolism and virulence, but also to antibiotic resistance. The results provide an important basis for in-depth studies of the relationship between protein lysine acetylation and bacterial antibiotic resistance.

Impact of biofilm formation and detachment on the transmission of bacterial antibiotic resistance in drinking water distribution systems.

There is growing awareness of the antibiotic-resistance crisis and its implications for public health among clinicians, researchers, politicians, and the public. We studied bacterial antibiotic resistance transition and the role of biofilms in a drinking water distribution system (DWDS). We tracked several different antibiotic resistant bacteria (ARB) with resistance to tetracycline, sulfamethoxazole, clindamycin, and norfloxacin for one year in a DWDS. The results indicated that the amount of ARB increased in tap water, presumably due to biofilm detachment. The effect of biofilm detachment on the transmission of antibiotic resistance from biofilms to tap water was explored by using a bacterial annular reactor. The percentage of ARB of inlet water, outlet water, and biofilms ranged from 0.26% to 9.85%, 1.08%-16.29%, and 0.52%-29.97%, respectively in a chlorinated system, and from 0.23% to 9.89%, 0.84%-16.84%, and 0.35%-17.77%, respectively, in a chloraminated system. The relative abundances of antibiotic resistance Acinetobacter, Sphingomonas, and Bradyrhizobium were higher in outlet water than in inlet water, as determined by high throughout sequencing. The amount of ARB percentage varied with the concentration of viable but non-culturable (VBNC) cells (r = 0.21, n = 160, P < 0.05) in biofilm, suggesting a higher antibiotic resistance mutation rate in VBNC cells. Our results suggest that biofilm detachment was promoted by disinfectant and affected the overall bacterial antibiotic resistance of microbes in tap water.

Resistencia antimicrobiana de Streptococcus agalactiae de origen humano y bovino / Antimicrobial resistance of Streptococcus agalactiae of human and bovine origin / Resistência antimicrobiana de Streptococcus agalactiae de origem humana e bovina

Resumen Streptococcus agalactiae (SAG) es un agente etiológico importante en un amplio espectro de infecciones humanas y bovinas. En humanos, este patógeno es el principal responsable de septicemias severas y muertes neonatales, debido a la enfermedad conocida como "sepsis neonatal", la cual ha sido reportada en diferentes países, incluyendo a Colombia. Cerca del 36% de las mujeres embarazadas son colonizadas por esta bacteria y de ellas el 45% de los neonatos adquiere la infección por SAG. En adultos, la colonización de SAG asintomática ocurre frecuentemente en el tracto gastrointestinal y genitourinario. Sin embargo, puede llegar a causar enfermedades tales como meningitis, septicemia, abscesos, infecciones del tracto urinario y artritis especialmente en adultos inmunocomprometidos. Adicionalmente, SAG es considerado un patógeno de alta importancia en la producción lechera, por ser responsable de cuadros generalmente subclínicos y crónicos de mastitis en vacas, afectando la sanidad del hato, así como la calidad y cantidad de leche producida. La principal herramienta para el control de SAG es el uso de antimicrobianos tipo betalactámicos o tipo macrólidos en casos de pacientes alérgicos a las penicilinas. Sin embargo, se ha reportado aislamientos de SAG resistentes o con susceptibilidad disminuida a los antimicrobianos utilizados para su control en ambas especies: humanos y bovinos. El hallazgo de resistencia antimicrobiana en SAG está recibiendo atención entre la comunidad científica en todo el mundo debido a su impacto negativo en la salud pública. El presente trabajo es una revisión de literatura científica, no sistemática, que tiene como objetivo analizar los mecanismos y la prevalencia de la resistencia antimicrobiana de SAG, así como los genes asociados a esta condición en aislamientos de origen humano y bovino.

Abstract Streptococcus agalactiae (SAG) is an important etiologic agent in a wide spectrum of human and bovine infections. In humans, this pathogen is the main responsible of severe septicemia and neonatal dead, due to the disease known as "neonatal sepsis", which has been reported in different countries, including Colombia. About 36% of pregnant women are colonized by this bacterium and of them, the 45% of the newborns acquire the SAG infection. In adults, asymptomatic SAG colonization occurs frequently in gastrointestinal and genitourinary tract. However, it can cause diseases such as meningitis, septicemia, abscesses, infections in urinary tract and arthritis particularly in immunocompromised adults. Additionally, SAG is considered a highly important pathogen in dairy production for being responsible of mastitis cases generally subclinical and chronic in cows, affecting the herd health, as well as the quality and the quantity of milk produced. The main tool for SAG control is the use of beta-lactams antimicrobials or macrolides in cases of penicillin-allergic patients. Some of the studies reported resistant SAG isolates or with decreased susceptibility to the antimicrobials used for its control in both species: humans and bovines. The finding of antimicrobial resistance in SAG is getting attention from the scientific community around the world because its negative impact in public health. The present work is a non-systematic review of scientific literature, with the objective of analyzing the mechanism and prevalence of SAG antimicrobial resistance, as well as, the genes associated to this condition in human and bovine isolates.

Resumo Streptococcus agalactiae (SAG) é um agente etiológico importante em um amplo espectro de infecções humanas e bovinas. Nos seres humanos, esse patógeno é a principal causa de septicemia grave e mortes neonatais, devido à doença conhecida como "sepse neonatal", que tem sido relatada em diferentes países, incluindo a Colômbia. Cerca de 36% das mulheres grávidas são colonizadas por esta bactéria e destes 45% dos recém-nascidos adquirem a infecção pelo SAG. Em adultos, a colonização do SAG assintomático ocorre com freqüência nos tratos gastrintestinal e genitourinário. No entanto, pode levar a doenças como meningite, septicemia, abscessos, infecções do trato urinário e artrite, especialmente em adultos imunocomprometidos. Além disso, a SAG é considerado um patógeno de grande importância na produção de leite, sendo responsável mastite geralmente subclínica e crônica em caixas de vacas, afetando a saúde do rebanho, ea qualidade e quantidade de leite produzida. A principal ferramenta para o controle do SAG é o uso de antimicrobianos beta-lactâmicos ou do tipo macrolídeo em casos de pacientes alérgicos a penicilinas. No entanto, isolados de SAG resistentes ou com reduzida susceptibilidade aos antimicrobianos usados para controlá-los foram relatados em ambas as espécies: humanos e bovinos. O achado de resistência antimicrobiana no SAG está recebendo atenção entre a comunidade científica em todo o mundo devido ao seu impacto negativo na saúde pública. Este papel é uma revisão da literatura científica, não sistemática, ou seja para analisar os mecanismos e a prevalência de resistência antimicrobiana SAG, bem como os genes associados com esta condição em isolados humanos e bovinos.

Payload hardware and experimental protocol development to enable future testing of the effect of space microgravity on the resistance to gentamicin of uropathogenic Escherichia coli and its σs-deficient mutant.

Human immune response is compromised and bacteria can become more antibiotic resistant in space microgravity (MG). We report that under low-shear modeled microgravity (LSMMG), stationary-phase uropathogenic Escherichia coli (UPEC) become more resistant to gentamicin (Gm), and that this increase is dependent on the presence of σs (a transcription regulator encoded by the rpoS gene). UPEC causes urinary tract infections (UTIs), reported to afflict astronauts; Gm is a standard treatment, so these findings could impact astronaut health. Because LSMMG findings can differ from MG, we report preparations to examine UPEC's Gm sensitivity during spaceflight using the E. coli Anti-Microbial Satellite (EcAMSat) as a free-flying "nanosatellite" in low Earth orbit. Within EcAMSat's payload, a 48-microwell fluidic card contains and supports study of bacterial cultures at constant temperature; optical absorbance changes in cell suspensions are made at three wavelengths for each microwell and a fluid-delivery system provides growth medium and predefined Gm concentrations. Performance characterization is reported here for spaceflight prototypes of this payload system. Using conventional microtiter plates, we show that Alamar Blue (AB) absorbance changes can assess the Gm effect on E. coli viability, permitting telemetric transfer of the spaceflight data to Earth. Laboratory results using payload prototypes are consistent with wellplate and flask findings of differential sensitivity of UPEC and its ∆rpoS strain to Gm. if σs plays the same role in space MG as in LSMMG and Earth gravity, countermeasures discovered in recent Earth studies (aimed at weakening the UPEC antioxidant defense) to control UPEC infections would prove useful also in space flights. Further, EcAMSat results should clarify inconsistencies from previous space experiments on bacterial antibiotic sensitivity and other issues.

Modified Carbapenem Inactivation Method for Phenotypic Detection of Carbapenemase Production among Enterobacteriaceae.

The ability of clinical microbiology laboratories to reliably detect carbapenemase-producing carbapenem-resistant Enterobacteriaceae (CP-CRE) is an important element of the effort to prevent and contain the spread of these pathogens and an integral part of antimicrobial stewardship. All existing methods have limitations. A new, straightforward, inexpensive, and specific phenotypic method for the detection of carbapenemase production, the carbapenem inactivation method (CIM), was recently described. Here we describe a two-stage evaluation of a modified carbapenem inactivation method (mCIM), in which tryptic soy broth was substituted for water during the inactivation step and the length of this incubation was extended. A validation study was performed in a single clinical laboratory to determine the accuracy of the mCIM, followed by a nine-laboratory study to verify the reproducibility of these results and define the zone size cutoff that best discriminated between CP-CRE and members of the family Enterobacteriaceae that do not produce carbapenemases. Bacterial isolates previously characterized through whole-genome sequencing or targeted PCR as to the presence or absence of carbapenemase genes were tested for carbapenemase production using the mCIM; isolates with Ambler class A, B, and D carbapenemases, non-CP-CRE isolates, and carbapenem-susceptible isolates were included. The sensitivity of the mCIM observed in the validation study was 99% (95% confidence interval [95% CI], 93% to 100%), and the specificity was 100% (95% CI, 82% to 100%). In the second stage of the study, the range of sensitivities observed across nine laboratories was 93% to 100%, with a mean of 97%; the range of specificities was 97% to 100%, with a mean of 99%. The mCIM was easy to perform and interpret for Enterobacteriaceae, with results in less than 24 h and excellent reproducibility across laboratories.

Broad- versus Narrow-Spectrum Oral Antibiotic Transition and Outcomes in Health Care-associated Pneumonia.

RATIONALE: Guidelines recommend a switch from intravenous to oral antibiotics once patients who are hospitalized with pneumonia achieve clinical stability. However, little evidence guides the selection of an oral antibiotic for patients with health care-associated pneumonia, especially where no microbiological diagnosis is made. OBJECTIVES: To compare outcomes between patients who were transitioned to broad- versus narrow-spectrum oral antibiotics after initially receiving broad-spectrum intravenous antibiotic coverage. METHODS: We performed a secondary analysis of an existing database of adults with community-onset pneumonia admitted to seven Utah hospitals. We identified 220 inpatients with microbiology-negative health care-associated pneumonia from 2010 to 2012. After excluding inpatient deaths and treatment failures, 173 patients remained in which broad-spectrum intravenous antibiotics were transitioned to an oral regimen. We classified oral regimens as broad-spectrum (fluoroquinolone) versus narrow-spectrum (usually a ß-lactam). We compared demographic and clinical characteristics between groups. Using a multivariable regression model, we adjusted outcomes by severity (electronically calculated CURB-65), comorbidity (Charlson Index), time to clinical stability, and length of intravenous therapy. MEASUREMENTS AND MAIN RESULTS: Age, severity, comorbidity, length of intravenous therapy, and clinical response were similar between the two groups. Observed 30-day readmission (11.9 vs. 21.4%; P = 0.26) and 30-day all-cause mortality (2.3 vs. 5.3%; P = 0.68) were also similar between the narrow and broad oral antibiotic groups. In multivariable analysis, we found no statistically significant differences for adjusted odds of 30-day readmission (adjusted odds ratio, 0.56; 95% confidence interval, 0.06-5.2; P = 0.61) or 30-day all-cause mortality (adjusted odds ratio, 0.55; 95% confidence interval, 0.19-1.6; P = 0.26) between narrow and broad oral antibiotic groups. CONCLUSIONS: On the basis of analysis of a limited number of patients observed retrospectively, our findings suggest that it may be safe to switch from broad-spectrum intravenous antibiotic coverage to a narrow-spectrum oral antibiotic once clinical stability is achieved for hospitalized patients with health care-associated pneumonia when no microbiological diagnosis is made. A larger retrospective study with propensity matching or regression-adjusted test of equivalence or ideally a prospective comparative effectiveness study will be necessary to confirm our observations.

Suscetibilidade à azitromicina de isolados bacterianos de processos infecciosos em cães e gatos / Susceptibility to azithromycin of bacteria isolated from infectious processes in dogs and cats

O presente estudo avaliou o perfil de suscetibilidade à azitromicina de patógenos bacterianos prevalentes em diferentes sítios infecciosos de animais de companhia. Adicionalmente, foram estudados o perfil de atividade in vitro de azitromicina contra esses patógenos e sua concentração inibitória mínima (CIM). Testes como a difusão em disco e a microdiluição em caldo detectaram resistência respectivamente em 48,6 por cento e 55 por cento dos isolados de Staphylococcus spp. e em 55,3 por cento e 72,7 por cento dos bastonetes Gram-negativos. A CIM50 para S. aureus foi 4,0mg/mL, para S. intermedius foi de 1,0mg/mL, para Staphylococcus spp. coagulase-negativas foi de e"512mg/mL e para bastonetes Gram-negativos foi de 256mg/mL. Quinze por cento (9/60) dos isolados oxacilina-resistente e multidroga-resistentes, mecA-positivos, de Staphylococcus spp. apresentaram também resistência à azitromicina. A disseminação de bactérias multidroga-resistentes aponta para a necessidade da avaliação da atividade antimicrobiana para selecionar o fármaco mais indicado e, assim, minimizar falhas terapêuticas na conduta clínica veterinária.

The susceptibility pattern to azithromycin of bacterial pathogens from various infectious sites, and the in vitro activity and minimum inhibitory concentration (MIC) of azithromycin were studied. Tests such as disc diffusion and broth microdilution detected respectively 48.6 percent and 55 percent of resistant Staphylococcus spp., and 55.3 percent and 72.7 percent resistant gram-negative rods. MIC50 for S. aureus was 4.0mg/mL, that for S. intermedius was 1.0mg/mL, for coagulase-negative Staphylococcus e"512mg/mL, and for gram-negative rods 256mg/mL. Fifteen percent (9/60) of oxacilin-resistant, multidrug-resistant and mecA-positive Staphylococcus spp. isolates were also azithromycin resistant. The dissemination of multidrug resistant bacteria points out to the need of antimicrobial evaluation activity in order to select the best indicated drug and thus minimizing therapeutic failures in veterinary practice.

Bacterial Antibiotic Resistance in Inpatients of Chronic Pulmonary Heart Disease(CPHD):Clinical Analysis / 中华医院感染学杂志

OBJECTIVE To investigate the pathogenic change and bacterial antibiotic resistance in lower respiratory tract infection in inpatients of chronic pulmonary heavt disease(CPHD) for recently 3 years and to provide reference for clinical treatment.METHODS Retrospective analysis was conducted on sputum cultivation from CPHD patients who were hospitalized from Jan 2004 to Dec 2006.RESULTS A total of 772 pathogenic strains were isolated,60.1 % of which were Gram-negative bacilli.Pseudomonas aeruginosa,Klebsiella Pneumoniae,Escherichia coli and Acinetobacter baumannii were the main Gram-negative pathogens.Gram-positive bacilli accounted for 13.7%,most of which were Streptococcus pneumoniae,Staphylococcus aureus and so on.Fungi accounted for 26.2%.Imipenem/cilastatin sodium was the most sensitive drug for P.aeruginosa,Acinebacter and Enterobacteriaceae.And vancomycin hydrochloride was the most for S.aureus.Their multiple drug-resistance to anti-microbial agents was serious.CONCLUSIONS Gram-negative bacteria are the majority of the pathogens from CPHD of lower respiratory tract infection in hospital.The pathogens show multiple drug-resistance in drug sensitive test.It is suggested that there be urgent need for surveillance of bacterial resistance and rational use of anti-microbial agents during the clinical therapy.