Microbiological Testing Methods Research Articles

An Optical-Electrochemical Microbiological Test Method Applied to Assessment of the Properties of Various Essential Oils

An Optical-Electrochemical Microbiological Test Method Applied to Assessment of the Properties of Various Essential Oils

A validated high-performance liquid chromatography-ultraviolet method for the determination of valproic acid derivatives in pharmaceutical formulations with a microbiological suitability evaluation

Valproic acid and its derivatives are common drugs used in the treatment of epilepsy, a result of CNS disorders. Because the molecular structure of valproic acid, as a fatty acid, does not contain any chromophore groups, its peak response demanded special conditions to be detected in HPLC, ensuring the best precision results. Still, the current approach illustrates simple, validated procedures for conduction. The conducted chromatographic system consists of the BDS Hypersil C8, 150 × 4.6 mm, 5 µm column using a mobile phase of acetonitrile: phosphate buffer (4:6) at a detection wavelength of 215 nm at room temperature. A full method validation study was conducted and approved to ensure precise, repeatable, and accurate results by implementing system suitability parameters. In microbiological terms, a suitability test is used to validate microbiological testing methods, ensuring their efficacy despite potential interference from antimicrobial properties in the tested materials. The HPLC-UV developed and validated analytical method was evaluated, and it was found to be sensitive for use in the detection of low concentrations of methylparaben, propylparaben, and valproate with an optimum run time of six minutes. limit of detection (LODs) were statistically estimated and found to be 2.27 µg/mL, 39.77 ng/mL, and 1.84 µg/mL for methylparaben, propylparaben, and valproate, respectively. Additionally, the method demonstrated a high recovery of methylparaben, propylparaben, and valproate with an excellent closed-accuracy range (98.84% - 101.34%). An excellent regression coefficient (r) of 0.99924, 0.99998, and 0.99997 for methylparaben, valproate, and propylparaben, respectively, was achieved. Assay determination of various pharmaceutical dosage forms in the local market was implemented, yielding admirable results.

Non-inferiority testing on the accuracy of two qualitative microbiological methods on paired binary data

Before implementing a new microbiological test method in pharmaceutical industry, it is required to assess whether it matches or surpasses the detection of microorganisms of the current microbiological method. The United Stated Pharmacopoeia Chapter 1223 (USP < 1223>) proposes two statistical approaches for testing non-inferiority of two qualitative microbiological methods when they can both test the same samples (paired design). The first approach compares the positive rates of the two methods. The non-inferiority test is not influenced by the performance of the methods alone, it is also affected by the spiked bacterial density. The second approach compares most probable numbers (MPNs) between the two methods, but the MPN assumes that the microbiological methods are perfect. To address the two abovementioned flaws, a generalized MPN (gMPN) approach is being proposed. Estimators for the accuracy, detection proportion, and bacterial density parameter are being derived using the method of moments. It will be shown that estimation of the accuracy and detection proportion has limited to no bias, but the bias for the bacterial density can become substantial. A test statistic for non-inferiority of accuracy, along with a sample size formula, is presented, showing nominal type I error and appropriate power. The proposed approach is demonstrated using real data.

Prevalence and Antibiotic Resistance of Klebsiella pneumoniae in Diabetic Foot Ulcer.

Introduction A serious global threat of antimicrobial resistance has emerged due tothe improper use of antibiotics, including polypharmacy and inappropriate prescribing. This misuse has led bacteria to develop immunity against these drugs.Klebsiella pneumoniae, a concerning gram-negative bacterium, has become resistant, especially among immunocompromised diabetic patients for multiple antibiotics. To fight effectively this growing crisis and regain control of these infections, it is crucial to comprehend the resistance mechanisms utilized by the bacteria anddevelop a new therapeutic strategy to prevent antibiotic resistance. Materials and methods A five-month study from January 2023 to May 2023 was conducted at the tertiary healthcare facility of Saveetha Medical Collegeby collecting 122 clinical specimens from patients with diabetic foot ulcers (DFUs) and ulcer-related infections. The microbiological testing methods followed by the identification of bacteria using matrix-assisted laser desorption/ionizationtime-of-flight mass spectrometry (MALDI-TOF) and antimicrobial susceptibility testing (AST) by the VITEK 2 Compact system were performed. Results A stout, rod-shaped, gram-negative bacilli was observed in gram staining, and growth of mucoid γ-hemolytic colonies and lactose-fermenting mucoid colonies were seen in blood and MacConkey agar plates. MALDI-TOF analysis confirmed the presence ofKlebsiella pneumoniaealong with other bacteria such as Klebsiella oxytoca, Staphylococcus spp., Proteus spp., and​​Escherichia ​coli. VITEKshowed high resistance rates to commonly used antibiotics, including carbapenems. Notably, isolates showed sensitivity and intermediate to tigecycline and colistin. Resistance patterns varied across specimen types, emphasizing the importance of considering clinical sources when interpreting data. Conclusion Hence, this study underscores the urgent need for novel antimicrobial agents and effective infection control measures to combat multidrug-resistant Klebsiella pneumoniae infections. Understanding resistance mechanisms is essential for the incorporation of treatment strategies and preserving antibiotic efficacy.

Rhizobium Inoculant and Seed-Applied Fungicide Effects Improve the Drought Tolerance of Soybean Plants as an Effective Agroecological Solution under Climate Change Conditions.

Rhizobial inoculation in combination with fungicidal seed treatment is an effective solution for improving soybean resistance to modern climate changes due to the maximum implementation of the plant's stress-protective antioxidant properties and their nitrogen-fixing potential, which will contribute to the preservation of the environment. Model ecosystems at different stages of legume-rhizobial symbiosis formation, created by treatment before sowing soybean seeds with a fungicide (fludioxonil, 25 g/L) and inoculation with an active strain of Bradyrhizobium japonicum (titer 109 cells per mL), were subjected to microbiological, biochemical, and physiological testing methods in controlled and field conditions. Seed treatment with fungicide and rhizobia showed different patterns in the dynamics of key antioxidant enzymes in soybean nodules under drought conditions. Superoxide dismutase activity increased by 32.7% under moderate stress, while catalase increased by 90.6% under long-term stress. An increase in the antioxidant enzyme activity induced the regulation of lipoperoxidation processes during drought and after the restoration of irrigation. Regeneration after stress was evident in soybean plants with a combination of fungicide seed treatment and rhizobial inoculant, where enzyme levels and lipoperoxidation processes returned to control plant levels. Applying seed treatment with fungicide and Rhizobium led to the preservation of the symbiotic apparatus functioning in drought conditions. As proof of this, molecular nitrogen fixation by nodules has a higher efficiency of 25.6% compared to soybeans without fungicide treatment. In the field, fungicidal treatment of seeds in a complex with rhizobia inoculant induced prolongation of the symbiotic apparatus functioning in the reproductive period of soybean ontogenesis. This positively affected the nitrogen-fixing activity of soybeans during the pod formation stage by more than 71.7%, as well as increasing soybean yield by 12.7% in the field. The application of Rhizobium inoculant and fungicide to seeds contributed to the development of antioxidant protection of soybean plants during droughts due to the activation of key enzymatic complexes and regulation of lipoperoxidation processes, which have a positive effect on nitrogen fixation and productivity of soybeans. This is a necessary element in soybean agrotechnologies to improve plant adaptation and resilience in the context of modern climate change.

MassARRAY: a high-throughput solution for rapid detection of foodborne pathogens in real-world settings.

Bacterial foodborne pathogens pose a substantial global public health concern, prompting government agencies and public health organizations to establish food safety guidelines and regulations aimed at mitigating the risk of foodborne illness. The advent of DNA-based amplification coupled with mass spectrometry, known as MassARRAY analysis, has proven to be a highly precise, sensitive, high-throughput, and cost-effective method for bacterial detection. This study aimed to develop, validate, and evaluate a MassARRAY-based assay for the detection and identification of significant enteropathogenic bacteria. The MassARRAY-based assay was developed for the detection of 10 crucial bacterial foodborne pathogens, including Campylobacter coli, Campylobacter jejuni, Clostridium perfringens, Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Listeria monocytogenes, Salmonella spp., Shigella spp., and Staphylococcus aureus. The assay was optimized using the reference gDNA (n = 19), followed by validation using gDNA (n = 85) of reference and laboratory isolates. Additionally, the evaluation of the assay's reaction using a mixture of gDNA from all nine targeted species was performed. The limit of detection of the developed MassARRAY-based assay was determined using bacterial cells. Moreover, the validation method for field samples was evaluated by comparing it with standard microbiological testing methods routinely analyzed. The developed MassARRAY-based assay demonstrated 100% concordance with known bacterial pure cultures. The assay's reaction using a mixture of gDNA from all nine targeted species revealed the MassARRAY's capability to detect all targeted species in a single assay with the lowest concentration of 1 ng/μL of gDNA. The limits of detection of the assay range from 357 ± 101 to 282,000 ± 79,196 cells. Moreover, the validation of the assay in field samples revealed a 100% correlation between the data obtained from the standard microbiological method and the MassARRAY-based assay. These findings suggested that the developed MassARRAY-based assay exhibited the excellence in high-throughput detection of foodborne bacterial pathogens with high accuracy, reliability, and potential applicability within real-world field samples.

Molecular Assessment of Resistance and Virulence Potential of Vibrio Species Isolated From Dumpsites in Port Harcourt, Nigeria

Background: Dumpsites have the potential to serve as reservoirs for various medically important bacteria and their virulence and resistance gene markers. For Vibrio spp., numerous genes associated with virulence have been identified in environmental strains. Due to the specific growth requirements of Vibrio spp., such strains can often be overlooked. This study aimed to assess the potential of Vibrio spp. isolated from two dumpsites in Port Harcourt, Nigeria, to serve as reservoirs of virulence and resistance genes. Methods: The soil samples were evaluated for the presence of Vibrio spp. following enrichment, using standard microbiological and biochemical test methods. DNA from Vibrio spp. was extracted using the boiling method, and isolates were tested for the presence of four resistance (sxt, strB, BlaTEM, and dfrA1) and four virulence (ctxA, hlyA, tcpA, and toxR) genes. Results: The study found a 40% occurrence of resistance genes and a 10% occurrence of virulence genes, with the strB streptomycin resistance gene being the most commonly detected (42%). Two of the virulence genes (ctxA and tcpA) were not detected. Seven of the test isolates exhibited multiple gene markers, with four gene markers present in each of two isolates. Conclusion: Overall, the study revealed a generally low potential for Vibrio sp. isolated from the dumpsites in Port Harcourt, Nigeria, to act as reservoirs of virulence and resistance genes. Additionally, the study reported an absence of major virulence markers associated with V. cholerae. A concerning finding was the high occurrence (42%) of the strB gene among these environmental isolates.

Development of an expert system for assessing failures in oil and gas pipelines due to microbiologically influenced corrosion (MIC)

This study highlights the modeling of an expert system for the classification of internal microbiologically influenced corrosion (MIC) failures related to pipelines in the upstream oil and gas industry. The model is based on artificial neural networks (ANNs) and involves the participation of 15 MIC experts. Each expert evaluated a number of model case studies ranging from MIC- to non-MIC-driven upstream pipeline failures. The model accounts for variations in microbiological testing methods, microbiological sample types, and degradation morphology, among other variables. It also accounts for missing datasets, which is commonly the case in actual failure assessments. The outcome is an expert system model whose outputs are classes (classification ANN) which comprises MIC potential and data confidence. The performances of two approaches are contrasted in this study. One classifies the output in 5 classes, a 5-output classification (5OC) model; the other in 3 classes, a 3-output classification (3OC) model. The 5OC model had an accuracy of 62.0% while the simpler 3OC model had a better accuracy of 74.8%. This modelling exercise has demonstrated that knowledge from experts can be captured in a reasonably effective model to screen for possible MIC failures. It is hoped that this study not only may contribute to a better understanding of the prevalence of MIC in the oil and gas sector, but also may highlight the key areas necessary to improve the diagnosis of MIC failures in the future.

A metagenomics-based workflow for the detection and genomic characterization of GBS in raw freshwater fish.

The need for a rapid and accurate food microbiological testing method is apparent for a timely and effective foodborne outbreak response. This is particularly relevant for emerging foodborne pathogens such as Group B Streptococcus (GBS) whose associated food safety risk might be undercharacterized. By using GBS in raw freshwater fish as a case example, this study describes the development of a metagenomics-based workflow for rapid food microbiological safety testing and surveillance. This study can inform as a working model for various foodborne pathogens in other complex food matrices, paving the way for future methodological development of metagenomics for food microbiological safety testing.

Investigation of Pseudomonas aeruginosa bacteria in a number of Baghdad schools and extent of their resistance to disinfectants and sterilizers

PurposeThis study was design to investigate of P. aeruginosa, an example of Gram-negative bacteria, in seven primary and secondary schools of Baghdad city, and the effects of Ethanol and Dettol of P. aeruginosa biofilm.Design/methodology/approachSeventy swabs were collected from seven primary and secondary schools of Baghdad city, Iraq, during November -December 2022. Swabs were collected from classes desk, doors handles, students hands and water taps. Standard microbiological testing methods were used on the samples for isolation and identification. The ability of bacteria to form biofilm and the effects of Ethanol and Dettol on"preformed” biofilms was examined by microtiter plate with the use of an ELISA reader.FindingsIn 70 swabs from seven primary and secondary schools, growth was observed in 33 swabs as P. aeruginosa. Primary schools were higher contaminated than secondary and water taps and door handles represented the main source of this contamination. The ability of bacteria to produce biofilm was observed in 19 (57.6%) isolates and 14 (42.4%) nonbiofilm producers. As well as, Ethanol (70%) treatment of preformed biofilms led to enhance biofilm formation and revealed significantly greater staining after 4 and 24h than Dettol (3%) compared to an untreated control (tryptic soy broth (TSB) incubation).Originality/valueStudies on P. aeruginosa in Iraqi schools are quite rare. This work is considered distinctive because it drew attention to the presence of pathogenic bacteria within primary and secondary schools, which are not considered their natural environment.

Analysis of the Effectiveness of Microbiological Testing Methods in Gynecological Inflammatory Infections

Analysis of the Effectiveness of Microbiological Testing Methods in Gynecological Inflammatory Infections

Validation studies of the spiral plating method to determine the potency of lactobacillus-containing probiotics

Scientific relevance. Viable cell counting is an important microbiological test for the quality assessment of medicinal products containing live microorganisms. To optimise labour and material costs and enhance testing precision and reproducibility, it is practical to use partially automated instrumental methods, such as the spiral plating method.Aim. The aim was to conduct a validation study of the spiral plating method for assessing the potency of biologicals containing live bacterial cells, using a lactobacillus-containing probiotic medicinal product as a case study.Materials and methods. This study used a culture of Lactiplantibacillus plantarum isolated from a probiotic medicinal product. Spiral plating on agar-based biological culture media was performed using an automatic Eddy Jet 2 plating system. The study used an IUL Flash &amp; Go colony counter for automatic reporting of the results. The validation study was conducted according to the general chapter on the validation of microbiological testing methods (OFS.1.1.0021.18) of the State Pharmacopoeia of the Russian Federation.Results. According to the validation results for the main parameters, the spiral plating method had the range of 104–105 CFU/mL; the limit of quantification of 102 CFU/mL; and the coefficient of linear determination, R2, of 0.99. The accuracy of the spiral plating method in determining the potency of the test sample was 93%; the repeatability was 4.9%.Conclusions. The study results confirm the similarity of the spiral plating method with an automatic colony counter to Koch’s plating method, which is currently used in accordance with the State Pharmacopoeia of the Russian Federation, in terms of the validated parameters. Therefore, spiral plating can be used to evaluate the potency of lactobacillus-containing probiotics. Spiral plating can help improve the cost-effectiveness and accuracy of testing.

Oxygen Sensor-Based Respirometry and the Landscape of Microbial Testing Methods as Applicable to Food and Beverage Matrices.

The current status of microbiological testing methods for the determination of viable bacteria in complex sample matrices, such as food samples, is the focus of this review. Established methods for the enumeration of microorganisms, particularly, the 'gold standard' agar plating method for the determination of total aerobic viable counts (TVC), bioluminescent detection of total ATP, selective molecular methods (immunoassays, DNA/RNA amplification, sequencing) and instrumental methods (flow cytometry, Raman spectroscopy, mass spectrometry, calorimetry), are analyzed and compared with emerging oxygen sensor-based respirometry techniques. The basic principles of optical O2 sensing and respirometry and the primary materials, detection modes and assay formats employed are described. The existing platforms for bacterial cell respirometry are then described, and examples of particular assays are provided, including the use of rapid TVC tests of food samples and swabs, the toxicological screening and profiling of cells and antimicrobial sterility testing. Overall, O2 sensor-based respirometry and TVC assays have high application potential in the food industry and related areas. They detect viable bacteria via their growth and respiration; the assay is fast (time to result is 2-8 h and dependent on TVC load), operates with complex samples (crude homogenates of food samples) in a simple mix-and-measure format, has low set-up and instrumentation costs and is inexpensive and portable.

430. Oritavancin Activity Against Gram-positive Pathogens Causing Bloodstream Infections in Hematology/Oncology and Transplant Units in US Medical Centers (2010-2019)

Abstract Background Patients undergoing hematologic, oncologic, and/or transplant treatment are at risk for infections. Appropriate antimicrobial management is crucial for patients with suspected or confirmed bloodstream infections (BSI). We evaluated the in vitro activity of oritavancin (ORI) and comparators against Gram-positive (GP) isolates causing BSI in patients from hematology/oncology and transplant units (HTU) in US medical centers. Methods 1,217 isolates (1/patient) were consecutively collected from HTU patients in 33 US hospitals during 2010-2019. Isolates were identified by MALDI-TOF and/or standard microbiological testing methods, and susceptibility (S) tested by CLSI reference broth microdilution in a central laboratory. CLSI breakpoints were applied. Results Enterococcus spp. (overall, 36.1%; E. faecium [EFM], 20.2%; E. faecalis [EF], 14.3%) and S. aureus (SA, 35.7%; 36.4% methicillin-resistant [MRSA]) were the most common organism groups, followed by coagulase-negative Staphylococcus (CoNS; 11.8%; 76.4% MR), Viridans group streptococci (VHS; 9.6%), and beta-haemolytic streptococci (BHS; 3.5%). ORI inhibited 96.7%/98.9% of EFM/EF at ≤0.12mg/L (S breakpoint for vancomycin [VAN]-S EF). Ampicillin (100.0%S), linezolid (LZD; 100.0%S), daptomycin (DAP; 98.3%S), and VAN (97.1%S) were also active against EF. Only ORI (96.7%S) and LZD (100.0%S) remained active against EFM. A VRE phenotype and elevated daptomycin MIC (2-4 mg/L) were noted in 72.8% and 55.3% of EFM, with ORI inhibiting 95.5% and 95.6% of these isolates at ≤0.12mg/L, respectively. ORI displayed equivalent MIC50/90 (0.03/0.06 mg/L) values against MSSA (99.3%S) and MRSA (99.4%S). ORI inhibited 95.5% of MRCoNS at ≤0.12mg/L (SA breakpoint). VAN, DAP, and LZD remained active against MRSA and MRCoNS. ORI was also active against BHS (MIC50/90, 0.03/0.25 mg/L; 95.2%S) and VGS (MIC50/90, 0.015/0.25 mg/L; 96.6%S) as was VAN (100.0%), DAP (100.0%), and LZD (100.0%/99.1%S, respectively). Conclusion VRE EFM, MRCoNS, and MRSA rates were generally high in BSI GP isolates recovered from patients in HTU. ORI was highly active and inhibited &amp;gt; 95% of isolates at MIC of ≤ 0.12mg/L from this unique collection, including R subsets such as VAN-R EFM, EFM displaying elevated DAP MIC (2-4 mg/L), MRSA, and MRCoNS. Disclosures Cecilia G. Carvalhaes, MD, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|Melinta: Grant/Research Support|Pfizer: Grant/Research Support Dee Shortridge, PhD, AbbVie: Grant/Research Support|JMI Laboratory: Employee|Melinta: Grant/Research Support|Menarini: Grant/Research Support|Shionogi: Grant/Research Support Helio S. Sader, MD, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|Melinta: Grant/Research Support|Nabriva Therapeutics: Grant/Research Support|Pfizer: Grant/Research Support Rodrigo E. Mendes, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|GSK: Grant/Research Support|Melinta: Grant/Research Support|Nabriva Therapeutics: Grant/Research Support|Office for Assistant Secretary of Defense for Health Affairs: Grant/Research Support|Pfizer: Grant/Research Support|Shionogi: Grant/Research Support|Spero Therapeutics: Grant/Research Support.

Bio-Mapping Indicators and Pathogen Loads in a Commercial Broiler Processing Facility Operating with High and Low Antimicrobial Intervention Levels.

The poultry industry in the United States has traditionally implemented non-chemical and chemical interventions against Salmonella spp. and Campylobacter spp. on the basis of experience and word-of-mouth information shared among poultry processors. The effects of individual interventions have been assessed with microbiological testing methods for Salmonella spp. and Campylobacter spp. prevalence as well as quantification of indicator organisms, such as aerobic plate counts (APC), to demonstrate efficacy. The current study evaluated the loads of both indicators and pathogens in a commercial chicken processing facility, comparing the “normal chemical”, with all chemical interventions turned-on, at typical chemical concentrations set by the processing plant versus low-chemical process (“reduced chemical”), where all interventions were turned off or reduced to the minimum concentrations considered in the facility’s HACCP system. Enumeration and prevalence of Salmonella spp. and Campylobacter spp. as well as indicator organisms (APC and Enterobacteriaceae—EB) enumeration were evaluated to compare both treatments throughout a 25-month sampling period. Ten locations were selected in the current bio-mapping study, including live receiving, rehanger, post eviscerator, post cropper, post neck breaker, post IOBW #1, post IOBW #2, prechilling, post chilling, and parts (wings). Statistical process control parameters for each location and processing schemes were developed for each pathogen and indicator evaluated. Despite demonstrating significant statistical differences between the normal and naked processes in Salmonella spp. counts (“normal” significantly lower counts than the “reduced” at each location except for post-eviscerator and post-cropper locations), the prevalence of Salmonella spp. after chilling is comparable on both treatments (~10%), whereas for Campylobacter spp. counts, only at the parts’ location was there significant statistical difference between the “normal chemical” and the “reduced chemical”. Therefore, not all chemical intervention locations show an overall impact on Salmonella spp. or Campylobacter spp., and certain interventions can be turned off to achieve the same or better microbial performance if strategic intervention locations are enhanced.

Antimicrobial Activity Comparison of Multipurpose Solutions

Purpose: This study compares the antimicrobial activity of seven multipurpose contact lens multipurpose solutions (MPS) available in European markets by microbiological test methods ISO 14729 and ISO 18259.

Occurrence of carbapenem-resistant organisms in gastrointestinal postoperative infections: A therapeutic challenge

Dissemination of multidrug resistant organisms including Carbapenem-Resistant Organisms (CRO) in Hospitals is of global concern. Such nosocomial infections are more common during surgical procedures involving prolonged post-operative care and Hospital stay. Treatment options include administration of prophylactic antibiotics, which are broad-spectrum antibiotics. However, long-term administration of these antibiotics leads to an increase in the incidence of multidrug resistant organisms in Hospital sectors. To evaluate early detection of carbapenemase producing organisms from the clinical isolates of postoperative patients by carba NP test. The study was conducted at the diagnostic laboratory in clinical samples obtained from hospitalized patients. A total of 716 clinical samples were tested by employing basic microbiological and biochemical testing methods and the isolates were screened for antimicrobial susceptibility. Carbapenem-resistant isolates were then confirmed by E-test (imipenem, meropenem) and also via carba NP test.In a total of 716 samples, 257 tested positive for various microorganisms, of which 230 gram-negative bacilli were identified. Amongst them, 93 isolates were identified as resistant to carbapenem by disc diffusion method of which 50 isolates were tested for carbapenemase production. Within the 50 isolates, 47 isolates were resistant to E-test meropenem and 40 isolates were resistant to imipenem. Of note, 35 out of the 50 CROs were identified as carbapenemase producers. Our results show that Carba NP test is a simple method that can be employed routinely for early detection of carbapenemase mediated CROs thus reducing the spread of resistant strains in Hospitals.

105. Impact of a Multiplex Polymerase Chain Reaction Panel on Duration of Empiric Antibiotic Therapy in Suspected Bacterial Meningitis

BackgroundMultiplex polymerase chain reaction (PCR) panels allow for rapid detection or exclusion of pathogens causing community-acquired meningitis and encephalitis (ME). However, the clinical impact of rapid multiplex PCR ME panel results on the duration of empiric antibiotic therapy is not well characterized.MethodsWe performed a retrospective pre-post study to evaluate the implementation of the FilmArray ME panel (BioFire Diagnostics, LLC) for diagnosis of bacterial meningitis at our institution. We included adults who presented with suspected bacterial meningitis, received empiric antibiotic therapy, and underwent cerebrospinal fluid microbiological testing in the emergency department. The primary outcome was duration of empiric antibiotic therapy. A bivariable analysis that compared baseline demographics, clinical characteristics, and study outcomes between the pre-ME panel and post-ME panel periods was performed using Mann-Whitney tests, chi-squared tests, or Fisher’s exact tests. Time-to-event analysis used the Kaplan-Meier method and log-rank statistics.ResultsIn the pre-ME panel period, the positive detection rate of bacterial pathogens was 2.2% (3/137) by cerebrospinal fluid culture and 4.3% (3/69) in the post-ME panel period. Table 1 shows baseline characteristics of patients. Compared to the pre-ME panel period, there were significant reductions in the post-ME panel period for the duration of empiric antibiotic therapy (median 34.7 h, IQR 8.5–61.7, vs. 12.3 h, IQR 3.3–40.0, P=0.01), time to targeted therapy (59.3 h, IQR 36.5–74.6, vs 7.02 h, IQR 0.9–12.4, P< 0.001), and hospital length of stay (4 d, IQR 2–7, vs. 3 d, IQR 1–5, P=0.03), as shown in Table 2. There was also significant reduction in time to discontinuation or de-escalation of empiric antibiotic therapy (P=0.049) as shown in Figure 1.Table 1. Baseline characteristics for patients with suspected bacterial meningitis Table 2. Antimicrobial use and hospitalization outcomes Compared to the pre-ME panel period, there were significant reductions in the post-ME panel period for the duration of empiric antibiotic therapy (P=0.01), time to targeted therapy (P<0.001), and hospital length of stay (P=0.03).Figure 1. Probability of Empiric Antibiotic Therapy Between Pre- and Post-ME Panel Periods Kaplan-Meier analysis of the time from initiation of empiric antibiotic therapy to discontinuation or de-escalation of empiric antibiotic therapy between the pre- and post-ME panel periods. P value from log-rank test=0.049 (n=206). There was a significant difference in the time to discontinuation or de-escalation of empiric antibiotic therapy between the groups (sex- and immunosuppressant use-adjusted hazard ratio, 1.46 [95% confidence interval, 1.08–1.97]; P=0.01).ConclusionThe implementation of the FilmArray ME panel for suspected bacterial meningitis appears to reduce the duration of empiric antibiotic therapy, time to targeted therapy, and hospital length of stay compared to traditional culture-based microbiological testing methods.Disclosures Justin J. Choi, MD, Allergan (Consultant, Grant/Research Support)Roche (Consultant, Grant/Research Support) Lars Westblade, PhD, Accelerate Diagnostics Inc (Grant/Research Support)BioFire Diagnostics (Grant/Research Support)Hardy Diagnostics (Grant/Research Support)Roche (Consultant, Advisor or Review Panel member)Shionogi Inc (Advisor or Review Panel member)Talis Biomedical (Advisor or Review Panel member) Marshall J. Glesby, MD, Enzychem (Consultant)Gilead (Grant/Research Support)ReAlta Life Sciences (Consultant)Regeneron (Consultant, Grant/Research Support)Sobi (Consultant)Springer (Other Financial or Material Support, Royalties)UpToDate (Other Financial or Material Support, Royalties)

Antimicrobial-resistant foodborne pathogens in the Middle East: a systematic review.

Foodborne pathogens are known as significant public health hazards worldwide, particularly in the Middle East region. Antimicrobial resistance (AMR) among foodborne pathogens becomes one of the top challenges for the environment, public health, and food safety sectors. However, less is known about antimicrobial-resistant foodborne pathogens in the Middle East region. Possibly because of the lack of surveillance, documentation, and reporting. This review focuses on the current status of antimicrobial resistance profiling among foodborne pathogens in the Middle East. Therefore, PubMed and other relevant databases were searched following PRISMA guidelines. Subject heading and texts were searched for "antimicrobial resistances," "foodborne," and "Middle East" to identify observational studies on AMR foodborne pathogens published during the last 10years (2011 to 2020). Article retrieval and screening were done using a structured search string and strict inclusion/exclusion criteria. Median and interquartile ranges of percent resistance were calculated for each antibiotic-bacterium combination. A total of 249 articles were included in the final analysis from ten countries, where only five countries had more than 85% of the included articles. The most commonly reported pathogens were Escherichia coli, Salmonella spp. Staphylococcus aureus, and Listeria spp. An apparent rise in drug resistance among foodborne pathogens was recorded particularly against amoxicillin-clavulanic acid, ampicillin, nalidixic acid, streptomycin, and tetracycline that are commonly prescribed in most countries in the Middle East. Besides, there is a lack of standardization and quality control for microbiological identification and susceptibility testing methods in many of the Middle East countries.

Impact of a Multiplex Polymerase Chain Reaction Panel on Duration of Empiric Antibiotic Therapy in Suspected Bacterial Meningitis.

BackgroundMultiplex polymerase chain reaction (PCR) panels allow for rapid detection or exclusion of pathogens causing meningitis and encephalitis (ME). The clinical impact of rapid multiplex PCR ME panel results on the duration of empiric antibiotic therapy is not well characterized.MethodsWe performed a retrospective prepost study at our institution that evaluated the clinical impact of a multiplex PCR ME panel among adults with suspected bacterial meningitis who received empiric antibiotic therapy and underwent lumbar puncture in the emergency department. The primary outcome was the duration of empiric antibiotic therapy.ResultsThe positive pathogen detection rates were similar between pre- and post-multiplex PCR ME panel periods (17.5%, 24 of 137 vs 20.3%, 14 of 69, respectively). The median duration of empiric antibiotic therapy was significantly reduced in the post-multiplex PCR ME panel period compared with the pre-multiplex PCR ME panel period (34.7 vs 12.3 hours, P = .01). At any point in time, 46% more patients in the post-multiplex PCR ME panel period had empiric antibiotic therapy discontinued or de-escalated compared with the pre-multiplex PCR ME panel period (sex- and immunosuppressant use-adjusted hazard ratio 1.46, P = .01). The median hospital length of stay was shorter in the post-multiplex PCR ME panel period (3 vs 4 days, P = .03).ConclusionsThe implementation of the multiplex PCR ME panel for bacterial meningitis reduced the duration of empiric antibiotic therapy and possibly hospital length of stay compared with traditional microbiological testing methods.