Clinical Microbiology Laboratory Research Articles

Blood Culture Optimization: Practical Tips

Sepsis is a serious medical condition that continues to be a leading cause of hospital admission and mortality worldwide. Blood cultures play a crucial role in identifying the source of sepsis and thus directing appropriate treatment for improved patient outcomes. Blood culture data are not only utilized for patient treatment but also have utility for infection control and epidemiological purposes. An important challenge facing blood culture results and interpretation is contamination, which can have negative effects on patient care and health care costs, including in the clinical microbiology laboratory. This article reviews the process of blood culture collection, from the decision to collect blood cultures to processing, testing, and reporting. Strategies for blood culture optimization are discussed, along with data to support practices that can lead to improved patient care, diagnostic stewardship, and fiscal responsibility.

PCR-based ORF typing of Klebsiella pneumoniae for rapid identification of global clones and transmission events.

Klebsiella pneumoniae is a major cause of healthcare-associated infections. In this study, we aimed to develop a rapid and simple genotyping method that can characterize strains causing nosocomial infections. The PCR-based open reading frame (ORF) typing (POT) method consists of two multiplex PCR reactions that were designed to detect 25 ORFs specific to bacterial genetic lineages, species, antimicrobial-resistant genes (blaCTX-M group-1 , blaCTX-M group-9 , blaIMP and blaKPC ), a capsular K1-specific gene and a virulence factor gene (rmpA/A2). The electrophoresis results are then digitized. A total of 192 strains (136 clinical and 8 reference strains of K. pneumoniae, 33 clinical and 1 reference strains of K. variicola and 14 clinical strains of K. quasipneumoniae) were classified into 95, 26 and 11 POT values, respectively. The distribution patterns of ORFs among K. pneumoniae correlated well with multilocus sequence typing (MLST). Furthermore, closely related species could be distinguished and key antimicrobial resistance and hypervirulence genes were identified as part of POT. The POT method was developed and validated for K. pneumoniae. In comparison to MLST, the POT method is a rapid and easy genotyping method for monitoring transmission events by K. pneumoniae in clinical microbiology laboratories. The POT method supplies clear and informative molecular typing results for K. pneumoniae. The method would facilitate molecular epidemiological analysis in infection control and hospital epidemiology investigations.

Next Generation and Other Sequencing Technologies in Diagnostic Microbiology and Infectious Diseases.

Next-generation sequencing (NGS) technologies have become increasingly available for use in the clinical microbiology diagnostic environment. There are three main applications of these technologies in the clinical microbiology laboratory: whole genome sequencing (WGS), targeted metagenomics sequencing and shotgun metagenomics sequencing. These applications are being utilized for initial identification of pathogenic organisms, the detection of antimicrobial resistance mechanisms and for epidemiologic tracking of organisms within and outside hospital systems. In this review, we analyze these three applications and provide a comprehensive summary of how these applications are currently being used in public health, basic research, and clinical microbiology laboratory environments. In the public health arena, WGS is being used to identify and epidemiologically track food borne outbreaks and disease surveillance. In clinical hospital systems, WGS is used to identify multi-drug-resistant nosocomial infections and track the transmission of these organisms. In addition, we examine how metagenomics sequencing approaches (targeted and shotgun) are being used to circumvent the traditional and biased microbiology culture methods to identify potential pathogens directly from specimens. We also expand on the important factors to consider when implementing these technologies, and what is possible for these technologies in infectious disease diagnosis in the next 5 years.

Impact of biofilm on cefotaxime killing effect against Escherichia coli

Biofilms cause chronic infections that are difficult to treat because of the resistance of microorganisms inside them. The goal is to assess the killing potential of cefotaxime against Escherichia coli in biofilms obtained from clinical isolates kept in the clinical microbiology laboratory of Dr. Soetomo Surabaya Hospital. The study was conducted by laboratory experimental design. Two steps challenged the bacterial isolates by applying cefotaxime to the planktonic bacterial state and the biofilm state. Minimum inhibitory concentration (MIC), minimum biofilm inhibitory concentration (MBIC), minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) were used as an indicator for comparison. Nineteen isolates of E coli were used for this experiment, 7 isolates with MIC of 0.125-0.25 μg/ml (36,84%), 6 isolates with MBC value 0.25-0.5 μg/ml (31,57%). The MBIC > 128 μg/ml is 6 isolates (31,57%), MBEC > 128 μg/ml is 14 isolates (73,68%). Cefotaxime had lower killing efficacy against E. coli in biofilm than in the planktonic phase. MBIC of E. coli requires cefotaxime at minimal 5 times two-fold increase from MIC, with an average 7-8 times. MBEC of E. coli requires cefotaxime concentration minimal 5 times two-fold dilution increase from MBC, with an average 8 - 9 times.

Development of Online Clinical Microbiology Laboratory Unknowns: Active Learning for Online, Hybrid, and In-Person Courses.

Microbiology laboratory classes allow students to make connections between theoretical concepts and clinical practice through experiential learning. In the past, most laboratory sessions were taught in-person, on-site. However, advances in multimedia and the ability of learning management systems to provide online course delivery allow laboratory assignments to be taught through online simulations. An online unknown identification activity promotes active learning while allowing instructors to assess student knowledge. This article describes the creation of online clinical microbiology unknown assignments which are suitable for complementing a variety of microbiology courses in an online, hybrid, or in-person format.

Automated antimicrobial susceptibility testing and antimicrobial resistance genotyping using Illumina and Oxford Nanopore Technologies sequencing data among Enterobacteriaceae.

Whole-genome sequencing (WGS) enables the molecular characterization of bacterial pathogens. We compared the accuracy of the Illumina and Oxford Nanopore Technologies (ONT) sequencing platforms for the determination of AMR classes and antimicrobial susceptibility testing (AST) among 181 clinical Enterobacteriaceae isolates. Sequencing reads for each isolate were uploaded to AREScloud (Ares Genetics) to determine the presence of AMR markers and the predicted WGS-AST profile. The profiles of both sequencing platforms were compared to broth microdilution (BMD) AST. Isolates were delineated by resistance to third-generation cephalosporins and carbapenems as well as the presence of AMR markers to determine clinically relevant AMR classes. The overall categorical agreement (CA) was 90% (Illumina) and 88% (ONT) across all antimicrobials, 96% for the prediction of resistance to third-generation cephalosporins for both platforms, and 94% (Illumina) and 91% (ONT) for the prediction of resistance to carbapenems. Carbapenem resistance was overestimated on ONT with a major error of 16%. Sensitivity for the detection of carbapenemases, extended-spectrum β-lactamases, and plasmid-mediated ampC genes was 98, 95, and 70% by ONT compared to the Illumina dataset as the reference. Our results highlight the potential of the ONT platform’s use in clinical microbiology laboratories. When combined with robust bioinformatics methods, WGS-AST predictions may be a future approach to guide effective antimicrobial decision-making.

Bacterial Profile and Antibiotic Resistance Pattern among Children with Urinary Tract Infections in Dr. Soetomo Hospital, Surabaya, Indonesia

Urinary tract infections (UTIs) are the most common infections in pediatric patients characterized by the growth of bacteria in the urine in significant numbers. Antibiotics remain the primary treatment of UTI in children. However, there has been an increase in antibiotic resistance to uropathogens worldwide due to their inappropriate and extensive uses. There is considerable geographical variation in the distribution of bacteria and antibiotic resistance pattern. Thus, to prevent further resistance and provide empirical antibiotic options, this study aims to determine the profile of bacteria and antibiotics resistance pattern among UTI pediatric patients in Dr. Soetomo Hospital. This study was performed by collecting data from the urine culture logbook at the Clinical Microbiology Laboratory of Dr. Soetomo Hospital in July-October 2019. The sample was UTI patients aged one day – 18 years due to bacterial infection with a colony count of ≥100,000 CFU/ml. In this study, 131 patients showed significant bacterial growth dominated by males and ages one month – 2 years. UTI were caused by gram-negative bacteria (74%) and gram-positive bacteria (26%), with the most bacteria found in each group were Escherichia coli and Enterococcus faecalis. E. coli showed ≥70% resistance to ampicillin, cefazoline, piperacillin, tetracycline, and trimethoprim-sulfamethoxazole. Comorbidities were dominated by hydronephrosis (10.98%), chronic kidney disease (9.79%) and hydrocephalus (8.09%). In conclusion, gram-negative bacteria were the leading cause of UTI in children with E. coli as the most common uropathogen, highly resistant to ampicillin and cefazolin. Gram-positive bacteria were less frequent with varied resistance patterns. Most common comorbidity was hydronephrosis.

Identification of micro-organism from positive blood cultures: comparison of three different short culturing methods to the Rapid Sepsityper workflow.

Sepsis is one of the leading causes of death worldwide. The rapid identification (ID) of the causative micro-organisms is crucial for the patients' clinical outcome. MALDI-TOF MS has been widely investigated to speed up the time-to-report for ID from positive blood cultures, and many different procedures and protocols were developed, all of them attributable either to the direct separation of microbial cells from the blood cells, or to a short subculture approach. In this study, the Rapid Sepsityper workflow (MBT Sepsityper IVD Kit, Bruker Daltonics GmbH and Co. KG, Bremen, Germany) was compared to three different short subculturing methods, established into the routine practice of three different clinical microbiology laboratories. A total of N=503 routine samples were included in this study and tested in parallel with the two approaches. Results of the rapid procedures were finally compared to routine proceedings with Gram-staining and overnight subculture. Among monomicrobial samples, the Rapid Sepsityper workflow enabled overall the correct identification of 388/443 (87.6 %) micro-organisms, while the short subculturing methods of 267/435 (61.8 %). Except for the performance with Streptococcus pneumoniae, in each one of the three sites the Rapid Sepsityper workflow proved to be superior to the short subculture method, regardless of the protocol applied, and it delivered a result from 1 to 5 h earlier.

Molecular detection of fluoroquinolone-resistant Neisseria meningitidis by using mismatched PCR-restriction fragment length polymorphism technique.

Ciprofloxacin (CIP) is a commonly used antibiotic for meningococcal chemoprophylaxis, and the mutations in the quinolone resistance-determining region of gyrA are associated with CIP-resistant Neisseria meningitidis. Here, we established a mismatched PCR-restriction fragment length polymorphism (RFLP) assay to detect a mutation at codon 91 of gyrA, followed by high-level CIP-resistant meningococci. We designed PCR-RFLP primers to detect the T91I mutation in gyrA by introducing an artificial AciI cleavage site. This assay was performed using 26 N. meningitidis strains whose gyrA sequences have been characterized. The amplified 160 bp PCR product from gyrA was digested into three fragments (80, 66, and 14 bp) when there was no mutation, or two fragments (146 and 14 bp) when there was a mutation at codon 91. A correlation was observed between the mismatched PCR-RFLP assay and gyrA sequencing. This rapid, simple, and accurate assay has the potential to detect CIP-resistant N. meningitidis in clinical microbiology laboratories, contributing to the appropriate antibiotic selection for meningococcal chemoprophylaxis, will help maintain an effective treatment for close contacts of IMD patients, and prevent the spread of CIP-resistant N. meningitidis.

Discrimination of Escherichia coli, Shigella flexneri, and Shigella sonnei using lipid profiling by MALDI-TOF mass spectrometry paired with machine learning.

Matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry (MALDI‐TOF MS) has become a staple in clinical microbiology laboratories. Protein‐profiling of bacteria using this technique has accelerated the identification of pathogens in diagnostic workflows. Recently, lipid profiling has emerged as a way to complement bacterial identification where protein‐based methods fail to provide accurate results. This study aimed to address the challenge of rapid discrimination between Escherichia coli and Shigella spp. using MALDI‐TOF MS in the negative ion mode for lipid profiling coupled with machine learning. Both E. coli and Shigella species are closely related; they share high sequence homology, reported for 16S rRNA gene sequence similarities between E. coli and Shigella spp. exceeding 99%, and a similar protein expression pattern but are epidemiologically distinct. A bacterial collection of 45 E. coli, 48 Shigella flexneri, and 62 Shigella sonnei clinical isolates were submitted to lipid profiling in negative ion mode using the MALDI Biotyper Sirius® system after treatment with mild‐acid hydrolysis (acetic acid 1% v/v for 15 min at 98°C). Spectra were then analyzed using our in‐house machine learning algorithm and top‐ranked features used for the discrimination of the bacterial species. Here, as a proof‐of‐concept, we showed that lipid profiling might have the potential to differentiate E. coli from Shigella species using the analysis of the top five ranked features obtained by MALDI‐TOF MS in the negative ion mode of the MALDI Biotyper Sirius® system. Based on this new approach, MALDI‐TOF MS analysis of lipids might help pave the way toward these goals.

Changes in the incidence of Candida-related central line-associated bloodstream infections in pediatric intensive care unit: Could central line bundle have a role?

Candida species are among the most prevalent microorganisms in pediatric critical care units that cause central line-associated bloodstream infections . The goal of this study was to assess the therapeutic benefit of central line bundle for the prevention of Candida species-related bloodstream infections in pediatric intensive care units. The study covered the period from January 1, 2009, to December 31, 2019. Pre-bundle and bundle phases were included in the research. The Clinical Microbiology Laboratory's records revealed episodes of Candida-related central line-associated bloodstream infections. The study was conducted in the Dr. Behçet Uz Child Disease and Pediatric Surgery Training and Research Hospital's PICU (which has 24 beds and admits 350 patients per year). This study included pediatric patients in the pediatric intensive care unit with non-tunneled central venous catheters . In the pediatric intensive care unit, a central line bundle was started. A total of 236 Candida-related central line-associated bloodstream infections were discovered during the study period. Non-albicans Candida accounted for 83.5% (197) of the total, whereas C.albicans accounted for 16.5%(39). During the pre-bundle period, 137 Candida species were isolated from the patients, while 99 Candida species were isolated during the bundle period. Candida-related central line-associated bloodstream infections dropped from 13.68 to 5.93 per 1000 CL-days after the central line bundle was used ( p < 0.001). According to our findings, the central line bundle greatly reduced central line-associated Candida species bloodstream infections. Central line bundles are an effective scientific solution for preventing Candida-related central line-associated bloodstream infections in hospitals with high Candida prevalence.

Ampicillinase C Beta-lactamase Producers among Isolates of Enterobacteriaceae in a Tertiary Care Centre: A Descriptive Cross-sectional Study.

Ampicillinase C beta-lactamase-producing organisms are often resistant to multiple antimicrobial agents, and therapeutic options against these pathogens are limited. Limited information is available regarding Ampicillinase C beta-lactamase producers. The aim of this study was to find out the prevalence of Ampicillinase C beta-lactamase producers among isolates of Enterobacteriaceae in a tertiary care centre. A descriptive cross-sectional study was carried out in the Clinical Microbiology Laboratory of a tertiary care centre from May 2021 to October 2021. Ethical approval was received from the Institutional Review Committee (Reference number: 044-077/078). Isolates of Enterobacteriaceae from various clinical samples were collected by convenience sampling. Ampicillinase C screening for beta-lactamase producers among the Enterobacteriaceae isolates was done using cefoxitin (30 μg) disc. Detection of Ampicillinase C beta-lactamase producers among the screen-positive isolates was done by cefoxitin-cloxacillin double-disc synergy test. An increase in the zone size of ≥4 mm was considered as Ampicillinase C beta-lactamase producers. Point estimate and 95% Confidence Interval were calculated. Among the total 481 isolates of Enterobacteriaceae, 49 (10.19%) (7.50-12.90, 95 % Confidence Interval) were detected as Ampicillinase C beta-lactamase producers among isolates of Enterobacteriaceae. The prevalence of Ampicillinase C beta-lactamase producers was lower than in other studies done in similar settings. Meropenem could be a drug of choice for the treatment of infections due to Ampicillinase C beta-lactamase-producing gram-negative bacteria. antibiotic; beta-lactamase; Enterobacteriaceae; gram-negative bacteria.

Bioactivity against Staphylococcus aureus of New Nanoparticles Derived from Iron Oxide and Beta vulgaris Extracts and Synthesis Characterization

In this study an opportunity was obtained that combines the VITEK system and the detection system of clinical microbiology laboratories to reach the rapid microbial identification and sensitivity test for Staph aureus bacteria, where the results of the susceptibility test were compared to 3925 combinations of antibiotics. The total rate was for the normal antimicrobials as well as the category approval for direct sensitivity test. 92% with less than 3% significant errors and the average time required to identify the identity with a sensitivity test is 7.5 hours (3.0 SD hours), compared to an average of 32.3 hours 9-14 hours for the normal method. The study focuses on the effectiveness of anti-bacterial iron nanoparticles from Beta vulgaris extract in various concentrations (1 mg/ml. 2 mg/ml. 3 mg/ml.4 mg/ml. against Stphylococcus aureus. Where in an inhibition zone was obtained for the highest concentration of iron nanoparticles for Beta vulgaris when compared to other samples.

Introducing First-Year Undergraduate Students the Fundamentals of Antibiotic Sensitivity Testing through a Combined Computer Simulation and Face-to-Face Laboratory Session.

ABSTRACTDetermining the antibiotic sensitivity of disease-causing microorganisms is a fundamental process in a clinical microbiology laboratory. With the continued use of antibiotics, the emergence of antibiotic resistance has become a significant health issue. However, the principles and laboratory testing to determine antibiotic sensitivity are generally not taught to first-year undergraduate students. This is partly due to the limited time to cover the fundamental biology of microorganisms and the mechanism of action of antibiotics in an introductory course. We overcame these limitations by teaching first-year students the fundamental principles of antibiotic sensitivity using an online data generator/simulation. Using the Kirby-Bauer disk diffusion test, students replicated the effects of antibiotic dose on bacterial growth and determined the antimicrobial susceptibility testing of their allocated bacterium. After 2–3 weeks, the antimicrobial sensitivity testing was replicated in an authentic face-to-face laboratory setting over 2 days. The impact of the intervention on student learning was assessed using a written laboratory report and a short questionnaire containing Likert and free-text questions. Student self-reported understanding of the content rose significantly, with nearly all students passing the written assessment. The approach was found to be enjoyable and interactive and facilitated authentic learning in first-year students. This cohort of students will continue to use more advanced versions of this simulation in future years, allowing for the long-term benefits of this approach to be assessed.

Evaluation of agents grown from urine culture and antibiotic susceptibility in patients followed up with the diagnosis of urinary system infection

Aim: In this study, it was aimed to determine the reproducing agents and the antibiotic susceptibility of the agents in a total of 89 urine samples cultured in Ankara Training and Research Hospital, Infectious Diseases and Clinical Microbiology laboratory during the 6-month period between 2021-2022. Material and Method: While growth was detected in urine culture in 68 (76%) of 89 urine samples taken in the study, no growth was found in 21 (24%) of them. Antibiotic susceptibility was studied on Mueller-Hinton agar by the disc diffusion method in isolates with growth in urine culture. Results: Reproducing factors are in order of frequency; 25 E. coli, 17 extended-spectrum beta-lactamase (ESBL) positive E. coli, 10 Klebsiella pneumoniae, 3 Pseudomonas aeruginosa, 3 Proteus mirabilis, 3 Enterococcus faecalis, 1 Enterobacter cloacae, 1 Candida krusei, 1 Staphylococcus aureus, 1 patient E. coli and Proteus species were determined. Resistance rates in E. coli strains isolated from urine culture; 20% (4/20) for ciprofloxacin, 5% (1/19) for nitrofurantoin, 9.5% (2/21) for trimethoprim/ sulfamethoxazole (TMP/SMZ). No resistance was found to fosfomycin 0% (0/19) and to carbapenems (imipenem, meropenem, and ertapenem) (0/21). The resistance rates in ESBL positive E. coli strains are; 100% (13/13) for ciprofloxacin, 7% (1/14) for nitrofurantoin, 12.5% (1/8) for fosfomycin, 26.6% (4/15) for TMP-SMZ. No resistance was found to carbapenems (imipenem, meropenem and ertapenem). Conclusion: In our study, it was determined that the most common infectious agent causing urinary tract infection was E. coli. In our study, it was determined that the resistance rates for ciprofloxacin and TMP-SMZ, which are used in empirical treatment, are high, and the resistance rates for fosfomycin and nitrofurantoin are low in E. coli strains that cause urinary system infection. Due to the low rates of resistance in the empirical treatment of urinary system infections, it would be a suitable approach to prefer oral treatment options such as fosfomycin and nitrofurantoin in outpatients.

Evaluation of agents grown from urine culture and antibiotic susceptibility in patients followed up with the diagnosis of urinary system infection

Aim: In this study, it was aimed to determine the reproducing agents and the antibiotic susceptibility of the agents in a total of 89 urine samples cultured in Ankara Training and Research Hospital, Infectious Diseases and Clinical Microbiology laboratory during the 6-month period between 2021-2022. Material and Method: While growth was detected in urine culture in 68 (76%) of 89 urine samples taken in the study, no growth was found in 21 (24%) of them. Antibiotic susceptibility was studied on Mueller-Hinton agar by the disc diffusion method in isolates with growth in urine culture. Results: Reproducing factors are in order of frequency; 25 E. coli, 17 extended-spectrum beta-lactamase (ESBL) positive E. coli, 10 Klebsiella pneumoniae, 3 Pseudomonas aeruginosa, 3 Proteus mirabilis, 3 Enterococcus faecalis, 1 Enterobacter cloacae, 1 Candida krusei, 1 Staphylococcus aureus, 1 patient E. coli and Proteus species were determined. Resistance rates in E. coli strains isolated from urine culture; 20% (4/20) for ciprofloxacin, 5% (1/19) for nitrofurantoin, 9.5% (2/21) for trimethoprim/ sulfamethoxazole (TMP/SMZ). No resistance was found to fosfomycin 0% (0/19) and to carbapenems (imipenem, meropenem, and ertapenem) (0/21). The resistance rates in ESBL positive E. coli strains are; 100% (13/13) for ciprofloxacin, 7% (1/14) for nitrofurantoin, 12.5% (1/8) for fosfomycin, 26.6% (4/15) for TMP-SMZ. No resistance was found to carbapenems (imipenem, meropenem and ertapenem). Conclusion: In our study, it was determined that the most common infectious agent causing urinary tract infection was E. coli. In our study, it was determined that the resistance rates for ciprofloxacin and TMP-SMZ, which are used in empirical treatment, are high, and the resistance rates for fosfomycin and nitrofurantoin are low in E. coli strains that cause urinary system infection. Due to the low rates of resistance in the empirical treatment of urinary system infections, it would be a suitable approach to prefer oral treatment options such as fosfomycin and nitrofurantoin in outpatients.

Correlation Between B Group Streptococcus Infection in Vagina and Maternal Serum Procalcitonin Level with Preterm Delivery

Preterm delivery is one of the biggest contributors to neonatal death because the organ systems in premature babies are not yet fully developed. The cause of preterm labor is multifactorial, one of which is complex inflammation due to Group B Streptococcus (GBS) bacterial infection. Maternal serum procalcitonin is a prohormone involved in the inflammatory process, and specifically as a marker of bacterial infection. This study time to determine the relationship between vaginal GBS bacterial infection and high maternal serum procalcitonin levels in preterm labor. This study used a cross-sectional method. Samples were 31 pregnant women who came to the Obstetrics ER at Sanglah Hospital Denpasar with a diagnosis of preterm delivery that met the inclusion criteria. Vaginal mucus samples were examined at the Clinical Microbiology Laboratory of Sanglah Hospital and procalcitonin samples were examined at the Clinical Pathology Laboratory of Sanglah Hospital. Descriptive and analytical analysis was performed on the data obtained. There were 7 positive GBS results (22.5%) and 24 negative GBS results (77.5%). The results of high procalcitonin obtained 12 (38.7%) while the normal 19 (61.3%). In the Chi-Square test, the PR value was 3.4 (95% CI: 1.61-7.30) with p value = 0.004. Therefore, it can be concluded that there is a relationship between GBS infection and serum procalcitonin levels, where positive GBS infection is a risk factor that increases the occurrence of high serum procalcitonin levels in preterm labor.

Drug sensitivity Profile of Pseudomonas spp. isolated from a tertiary care hospital against commonly used antibiotics

Background: Antimicrobials agents are being abundantly used to control bacterial growth, like Pseudomonas, leading to resistance. Objective: To find out the drug sensitivity patterns of the Pseudomonas spp. against various antibiotics in the tertiary care hospital (Ghurki Trust &amp; Teaching Hospital). Methodology: This cross-sectional study was designed on the retrospective collection of the records of 1000 sample reports, of the patients admitted in Ghurki Trust Teaching Hospital from the period of January 2018 to December 2019, who presented with multiple infections or developed pseudomonas infections during their stay in the hospital. Tests were carried out in the clinical microbiology Laboratory of Lahore Medical &amp; Dental College for bacterial culture. Specimen included were breast tissues, pus, urine and stool culture, blood culture, wound discharge, ear discharge, abscess from the wound, various infected implants, and dead necrotic tissues from bone. Data were analyzed using SPSS version 26. Results: Out of the 1000 samples, a total of 150 (15%) were positive for bacterial growth. Among these 150, 70 were positive for gram-negative. The bacterial profiles of these 150 samples have shown that frequently (42) isolated gram-negative bacteria was Pseudomonas. On further tests, these bacteria were more sensitive to Imipenem and Cilastatin (69%) followed by levofloxacin ( 31%). No sensitivity was found against vancomycin and amoxicillin. Conclusion: In this study, the Pseudomonas were isolated and showed sensitivity to Imipenem and Cilastatin followed by levofloxacin. These studies should be conducted frequently to keep a track of the sensitivity patterns of bacteria.

Secrets of the Microbiology Lab: How What They Do Directly Affects What You Report

<h3>Background</h3> Infection preventionists rely upon the information provided by the clinical microbiology laboratory to determine whether or not an infection should be reported. However, few infection preventionists understand or are aware of what procedures, policies, and behavior occur within the microbiology laboratory that directly influences the patient data that they use for their surveillance reporting as well as their infection prevention response. This presentation will teach infection preventionists the often unknown processes performed by the microbiology laboratory and its direct impact on their reporting to the National Healthcare Safety Network (NHSN) and their institution's internal data. <h3>Methods</h3> This study will systematically address the common policies and procedures performed related to the collection, transport, and diagnostic reporting of patient specimens that influence surveillance reporting criteria of multidrug-resistant organisms and Clostridioides difficile Infection (MDRO/CDI), urinary tract infection (catheter-associated and non-catheter-associated), central line associated bloodstream infection (CLABSI), and surgical site infection (SSI) to NHSN. <h3>Results</h3> Accepted policies, procedures, and processes vary widely across clinical microbiology laboratories. Infection preventionists need to be aware of how these differences can lead to potential errors in their reporting and the true burden of infection within their institution. Without understanding the basics of how the clinical laboratory information is resulted, it is impossible to guarantee accurate reporting to NHSN and appropriate infection prevention response. <h3>Conclusions</h3> It is the goal of this presentation to equip infection preventionists with the knowledge of what may occur within their own institution's microbiology laboratory and provide guidance on how to collaborate with their laboratory colleagues to ensure that what they report to NHSN is transparent and accurate. The information provided in this presentation should ultimately lead to a more impactful infection prevention program.

A Microcurrent Dressing Reduces Cutibacterium Acnes Colonization in Patients Undergoing Shoulder Arthroplasty or Arthroscopy: A Prospective Case Series.

Background: Cutibacterium acnes infections in the shoulder remain a significant concern in the setting of shoulder arthroplasty. Purpose: We sought to evaluate the efficacy of a microcurrent dressing in reducing C. acnes skin colonization and thereby reducing the risk of periprosthetic joint infection of the shoulder. Methods: This study was designed as a prospective case series. From October 2017 to February 2019, patients undergoing elective shoulder arthroplasty or arthroscopic shoulder surgery at a major academic medical center were offered enrollment; they signed an informed consent to participate. Patients under the age of 18, scheduled for revision shoulder arthroplasty, or with sensitivity or allergy to silver, zinc, or latex were excluded. Subjects underwent skin culture swab of the shoulder in the mid-point of the planned deltopectoral incision. The JumpStart (Arthrex; Naples, FL) microcurrent dressing was then placed over the area of the planned incision, and a full-thickness skin biopsy was harvested from the incision at the initiation of the surgical procedure. All specimens were cultured for C. acnes by the hospital's clinical microbiology laboratory with standard anaerobic technique. Results: Thirty-one subjects were enrolled in the study. Those who demonstrated no growth at baseline for the control specimen were excluded from further analysis (N = 11), given the absence of preoperative C. acnes colonization. Culture results from the 20 remaining subjects revealed significantly diminished C. acnes skin growth at the time of surgery compared to baseline. Sixty percent (12 of 20) of the subjects with positive skin swabs at baseline demonstrated no growth in the skin biopsy specimens at the time of surgery. There were no adverse events associated with the application of the microcurrent dressing. Conclusion: This prospective case series found that preoperative application of a microcurrent dressing resulted in significantly diminished C. acnes skin burden at the time of surgery in patients undergoing elective shoulder arthroplasty or arthroscopic shoulder surgery. Further study is warranted to investigate whether this preoperative intervention may contribute to a reduction in perioperative infections, including prosthetic joint infection.