Identification Of Microorganisms Research Articles

Accuracy of various matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based rapid identification methods—As a tool to augment diagnostic stewardship in blood culture laboratory, South India

BackgroundBlood stream infection is a medical emergency associated with high morbidity and mortality. Prompt identification of bloodstream infection-causing microorganisms directly from positive blood culture will significantly enhance patient care by reducing the turnaround time of pathogen recognition. MethodsA total of 256 freshly flagged positive blood culture bottles were subjected to Gram staining. Direct MALDI-TOF MS analysis was performed following sample preparation techniques such as lysis centrifugation, lysis filtration and VITEK® MS BC kit to directly identify microorganisms from positive blood cultures. Along with these short-term incubation methods of Choco spot and minute colony(8–10h) were also performed. All those positive bottles were identified by the routine (reference) laboratory method. Results177 isolates (69.14 %) were correctly identified by Lysis centrifugation, 163 isolates (63.67 %) were correctly identified by Lysis filtration, 206 isolates (80.47 %) were correctly identified by Choco spot,250 isolates (97.65 %) were correctly identified from minute colony (8–10h) of incubation. Of 162 isolates,115 isolates (70.99 %) were correctly identified by VITEK® MS Blood culture kit, (BioMérieux). VITEK® MS BC kit method revealed higher agreement with the kappa value of 0.697 than lysis centrifugation (0.672) followed by lysis filtration (0.611). ConclusionsIn house method of lysis centrifugation is found to be equivalent to VITEK® MS BC kit method and superior to lysis filtration method in correct direct identification of bacteria from positive blood cultures by MALDI-TOF MS analysis. As lysis centrifugation requires only 10 min of processing time as compared to overnight incubation, thus it offers a less expensive substitute for the VITEK® MS BC kit in the clinical laboratory. As a consequence of this study, we have implemented direct MALDI-TOF-based identification from positive BCs in our daily routine diagnostic management.

Influence of soil microplastic contamination on maize (Zea mays) development and microbial dynamics

BackgroundMicroplastics are contaminants in soil ecosystems that alter biophysical processes, affect plant growth, ecosystems and humans in the long run. This study was conducted to investigate the influence of soil microplastic contamination on the growth, development, and microbial dynamics of maize (Zea mays).Composite soil samples were collected from the farm from 0 to 10 cm depth. Physicochemical properties were assessed before maize seed sowing. Three soil weight categories were treated with 30 g, 20 g and 10 g microplastics. Soil physicochemical properties and heavy metal content in maize plants were analyzed. Morphological parameters, microbial counts, and microorganism identification were conducted. FTIR analysis was carried out in selected plant parts.ResultsThe study revealed a pH range of 5.00–6.70, electrical conductivity of 70–158 µS/cm, organic carbon and organic matter, total nitrogen, average phosphorus content, clay and silt content. The concentrations of heavy metals in plant parts were within WHO standard limits of 425.5 and 500 mg/kg for iron and manganese, but above the limits of 40, 60, and 350 mg/kg for copper, zinc and magnesium. The plant had various leaf sizes and heights, with chlorosis and necrotic activity ranging from 0.33 to 2.67, and a range of 1.33 to 6.33, respectively. Soil samples showed a total bacterial count (TBC) of 2.00 × 103 cfu/g to 7.60 × 104 cfu/g and a total fungal count (TFC) of 4.00 × 103 cfu/g to 2.20 × 104 cfu/g. Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli, Staphylococcus epidermidis, Proteus mirabilis and Bacillus cereus had percentage occurrences of 44.31, 21.17, 9.12, and 3.26%, respectively. Penicillium notatum was the most prevalent, while Fusarium spp. was the least prevalent. The FTIR analysis of maize seeds from soil samples A, B, and C revealed various organic compounds, with opium powder and streptomycin sulphate being the most abundant. The study revealed that pot B had the highest microplastic concentration at 1 WAS, while pot C had the lowest at 0.10 mg/kg, indicating a differential soil microplastic accumulation.ConclusionMicroplastics impact soil health, plant growth, and nutrient cycling, necessitating sustainable agriculture. Long-term research is needed to understand ecological and physiological impacts and explore remediation techniques.

Effective technology of disinfecting ozonation of hatching chicken eggs

Relevance. The positive aspects of disinfecting ozonation of hatching eggs allow us to expect successful application of the method not only in large but also in small-scale farms, as well as for experimental and industrial purposes in laboratories and bio-enterprises related to the technological process based on the incubation of a small number of eggs. This determines the importance of expanding the range of ozonizers due to numerous portable devices. There are no clear recommendations for these devices and this causes the need to search for the most effective and harmless modes for the embryo, and schemes of disinfecting ozonation.Methods. The study used fertilized chicken eggs “Hysex Brown” and a portable ozonizer “OZON-OviV”. Ozone concentration 2.0 mg / l. Eggs were treated in a specially made chamber. Technology-1: twice for 30 minutes before incubation and on the 3rd day of incubation. Technology-2: three times for 30 minutes before incubation, on days 3 and 5. The range of studies included: assessment of total microbial contamination (densitometry); identification of microorganisms (MALDI-TOF-spectrometry); biological control of incubation (fertility, hatchability, mortality, developmental abnormalities); assessment of the adequacy of the internal organs (MicroCT); embryo morphometry (weight, length, chest circumference) and calculation of development proportionality indices; histological assessment of the liver.Results. The total ozone concentration during treatment by two methods was 240 mg/l and 360 mg/l, respectively. The disinfecting efficiency of ozonation has been proven, providing a decrease in the level of total microbial contamination by 30% and 40% with double and triple treatment. A tendency to maintain a low total microbial contamination, compared to intact eggs, up to 14 days of incubation has been revealed. The dynamics of the microbial landscape indicate the bacteriostatic effect of ozone in the concentrations used on a wide range of microorganisms. Microtomographic and histological methods confirmed the harmlessness of the technologies used. Along with the more pronounced antibacterial effect of technology-2, the presence of a stimulating effect on the body of the developing embryo was revealed, which determines the preference for its choice.

Can bone SPECT/CT determine optimal sites for microbiological identification in post-traumatic or chronic osteomyelitis of extremities?

IntroductionAccurate microbiological identification is crucial when managing chronic osteomyelitis (COM) and post-traumatic osteomyelitis (PTO). Although bone single photon emission computed tomography/computed tomography (SPECT/CT) has helped in localizing osteomyelitis lesions, its effectiveness in guiding microbiological sampling remains unclear. This study aimed to determine whether bone SPECT/CT can improve microbiological identification rates in COM or PTO of the extremities. Patients and methodsFrom February 2020 to August 2024, 53 patients with suspected COM or PTO in the extremities were retrospectively analyzed. All patients underwent bone SPECT/CT, followed by microbiological sampling during surgery. Tissue samples were taken from the areas of high SPECT/CT uptake or based on intraoperative findings where no uptake was observed. Microorganism identification rates were analyzed, including a sub-group analysis based on antibiotic discontinuation. ResultsOf the 53 patients, 42 had positive bone SPECT/CT scan findings, with pathogen identification in 30 patients (71.4 %). In contrast, pathogen identification occurred in one out of twelve patients (9.1 %) with negative findings (odds ratio 25, p< 0.001). Bone SPECT/CT demonstrated a sensitivity of 96.8 % and an overall accuracy of 75.5 %. When antibiotics had been discontinued for ≥2 weeks, the pathogen identification rate increased to 90 %, compared with 50 % for <2 weeks of discontinuation (odds ratio 10.0, p= 0.006). In a sub-group of 30 patients with adequate antibiotic discontinuation duration, a positive bone SPECT/CT scan yielded a pathogen identification rate of 90.1 % (odds ratio 60.0, p= 0.001). ConclusionBone SPECT/CT effectively identifies optimal sites for microbiological sampling in COM and PTO of the extremities, particularly when antibiotics have been discontinued for ≥2 weeks, enhancing pathogen detection rates.

Impact of bacterial co-isolation on treatment initiation and long-term prognosis of patients with nontuberculous mycobacterial pulmonary disease: analysis of a prospective cohort study.

Nontuberculous mycobacterial pulmonary disease (NTM-PD), a chronic respiratory condition, presents a growing challenge globally. Uncertainties exist regarding the impact of concurrent bacterial co-isolation on treatment initiation and long-term prognosis. This study analysed data from participants enrolled in an ongoing prospective observational cohort study on NTM-PD (NCT01616745) between 1 July 2011, and 31 December 2022, who provided sputum samples for bacterial culture at enrolment. Identification of potential pathogenic microorganisms (PPMs) was defined as a positive bacterial culture. Clinical characteristics were compared between NTM-PD patients with Pseudomonas, non-pseudomonal PPMs, and those without PPM co-isolation. Cox proportional hazard regression models were employed to assess the association of bacterial co-isolation with rates of NTM-PD treatment initiation and all-cause mortality. Overall, 453 patients (median age, 62 years; 30% male) were included in the analysis. PPMs were co-isolated in 77 patients (17%), including 13 with Pseudomonas species. Co-isolation of Pseudomonas was associated with a significantly higher erythrocyte sedimentation rate (P = 0.02) and St. George's Respiratory Questionnaire score (P = 0.01). Non-pseudomonal PPM co-isolation was significantly associated with a higher likelihood of NTM-PD treatment initiation (adjusted hazards ratio [aHR], 1.56, 95% confidence interval [CI], 1.03-2.36, P = 0.036), whereas co-isolation of Pseudomonas was independently correlated with increased all-cause mortality (aHR, 3.25, 95% CI, 1.08-9.84, P = 0.037). Our findings emphasize the importance of microbial surveillance, as bacterial co-isolation affects treatment initiation and prognosis in patients with NTM-PD.

The Antimicrobial Efficacy and Quality Changes in Pineapple Juice Treated with Natural Preservatives

The study investigated effects of natural preservatives on the quality of fresh pineapple fruit juice. The natural preservatives used are ginger, garlic, moringa seed and cloves and were processed into powder. The treatment of pineapple with the natural preservatives was at a concentration of 20% with 3 minutes dipping time. The enumeration and identification of microorganisms on the fresh pineapple fruit, shows that the total bacteria count was 1.8×105 colony forming unit per gram (CFU/g) and 4.6×103 CFU/g respectively. The identified bacteria are staphylococcus aureus and streptococcus spp and identified fungi were Aspergillus Niger, penicillin spp and Aspergillus perfrigens respectively. Moringa seed extract exhibited moderate antibacterial activity, with zones of inhibitions (ZOIs) of 16 mm against Staphylococcus aureus and 15 mm against Streptococcus spp. Garlic (GL) showed the least antibacterial activity among the natural preservative samples, with 9mm zones of inhibition against Staphylococcus aureus and 4 mm against Streptococcus spp. Clove extract displayed significant antibacterial activity, with 14 mm zones of inhibition against Staphylococcus aureus and 19 mm against Streptococcus spp. Ginger extract exhibited slight antibacterial activity, with zones of inhibition of 4 mm against both bacterial strains. Chemical composition of the treated pineapple juice result ranges from 0.77-0.865% TTA, 64.12-63.10 Vit C, 26.50-25.06 Brix Content,3.20-3.39 pH and 13.23-13.17 TSS respectively. The control (100% pineapple Juice) displayed balanced properties serving as an effective baseline3. Storage studies, ambient stored samples were analyzed at 0 day and 21 days intervals. All samples were subjected to standard analytical methods. Results showed increase in number of bacteria and fungi with increase in the number of days. The use of cloves, garlic, ginger, and moringa seeds as natural antimicrobial agents in pineapple juice is effective in controlling microbial growth, thus enhancing the safety and shelf life of the juice. These treatments also influence the chemical and sensory properties of the juice, contributing to consumer acceptability.

Biofilm infections of endobronchial valves in COPD patients after endoscopic lung volume reduction: a pilot study with FISHseq

Endoscopic lung volume reduction (ELVR) using endobronchial valves (EBV) is a treatment option for a subset of patients with severe chronic obstructive pulmonary disease (COPD), suffering from emphysema and hyperinflation. In this pilot study, we aimed to determine the presence of bacterial biofilm infections on EBV and investigate their involvement in lack of clinical benefits, worsening symptomatology, and increased exacerbations that lead to the decision to remove EBVs. We analyzed ten COPD patients with ELVR who underwent EBV removal. Clinical data were compared to the microbiological findings from conventional EBV culture. In addition, EBV were analyzed by FISHseq, a combination of Fluorescence in situ hybridization (FISH) with PCR and sequencing, for visualization and identification of microorganisms and biofilms. All ten patients presented with clinical symptoms, including pneumonia and recurrent exacerbations. Microbiological cultures from EBV detected several microorganisms in all ten patients. FISHseq showed either mixed or monospecies colonization on the EBV, including oropharyngeal bacterial flora, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus spp., and Fusobacterium sp. On 5/10 EBV, FISHseq visualized biofilms, on 1/10 microbial microcolonies, on 3/10 single microorganisms, and on 1/10 no microorganisms. The results of the study demonstrate the presence of biofilms on EBV for the first time and its potential involvement in increased exacerbations and clinical worsening in patients with ELVR. However, further prospective studies are needed to evaluate the clinical relevance of biofilm formation on EBV and appropriate treatment options to avoid infections in patients with ELVR.

New Insights on the Glyphosate-Degrading Enzymes C-P Lyase and Glyphosate Oxidoreductase Based on Bioinformatics

Bioremediation, the degradation of environmental pollutants by living organisms, has immense potential to lead to a greener planet. Bioinformatics analysis can contribute to the identification of novel microorganisms, which biodegrade contaminants, or of participating proteins and enzymes, and the elucidation of the complex metabolic pathways involved. In this study, we focus on C-P lyase and glyphosate oxidoreductase (Gox), two enzymes which degrade glyphosate, a widely used pesticide. Amino acid sequences of the two enzymes were collected from a broad range of microorganisms using the KEGG database and BLAST. Based on this, we identified additional lineages, with putative glyphosate-degrading activity, for which no glyphosate-degrading species have been reported yet. The conserved residues in each enzyme were identified via multiple alignments and mapped onto the 3D structures of the enzymes, using PyMOL, leading to novel insights into their function. As the experimental structure of Gox is still unknown, we created structural models based on three different programs and compared the results. This approach can be used to yield insights into the characteristics of potential glyphosate-degrading enzymes. Given the limited information available, such a step is important to gain further knowledge about them, which can contribute to their application in bioremediation in the future.

TCT-938 Identification and Characterization of Microparticles and Microorganisms in Blood Clots of STEMI Patients: A Comparative Study of Rescue and Primary Angioplasty

TCT-938 Identification and Characterization of Microparticles and Microorganisms in Blood Clots of STEMI Patients: A Comparative Study of Rescue and Primary Angioplasty

Identification of Microorganisms (Fungi) in Fermented Ganjang from Korea, China, and Japan Using Next-Generation Sequencing Analysis

Identification of Microorganisms (Fungi) in Fermented Ganjang from Korea, China, and Japan Using Next-Generation Sequencing Analysis

Molecular detection and phylogenetic tree assay of Pseudomonus aeruginosa isolated from otitis cases of cats and humans, Iraq

The aim of this study was toward isolation and identification of P. aeruginosa from cats with otitis externa in Basra city Using culture media, Biochemical tests and Polymerase chain reaction. Pseudomonas aeruginosa was isolated from 46 isolates that involved 32 cases out of 75 of feline otitis externa (21.3%) and in 14 (9.3%) from 75 human samples .the clinical signs of otitis externa were head shaking, scratching, excessive ear wax, malodor, pain during palpation, alopecia and pus and/or blood. Culture was done on Pseudomonas chrome agar revealed positive samples with production of greenish-blue pigmentation by the bacteria .polymerase chain reaction revealed that isolates were Pseudomonas aeruginosa using specific primers for P. aeruginosa identification 16S rRNA and the result was positive for the 15 isolates. a phylogenetic tree analysis of P. aeruginosa was performed using the 16S rRNA gene sequence analysis which illustrated the phylogenetic relationship between P. aeruginosa strains isolated from cats and other related genera. In conclusion, this study confirmed the higher prevalence of P. aeruginosa in otitis externa infection of cats when compared to human. The using of molecular assay and 16S rRNA gene contributed effectively in detection of P. aeruginosa. Therefore, this study suggests the application of molecular techniques in identification of different microorganisms. Additionally, moreover studies are necessary to detect the prevalence of P. aeruginosa in different infections in animals as well as human.

Application of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry for Identification of Foodborne Pathogens: Current Developments and Future Trends.

Foodborne pathogens have gained sustained public attention, exerted significant pressure on food manufacturers, and posed serious health risks to human. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been employed for quick and accurate identification of microorganisms in the prevention of foodborne epidemics in recent years. Herein, we first summarize the principle of MALDI and its workflow for foodborne pathogens. Subsequently, we review the recent progress and applications of MALDI-TOF MS in foodborne pathogen determination. Additionally, we outline the expanded utilization of MALDI-based techniques for the identification of closely related species. We also assess the current gaps and propose possible solutions to address the existing challenges. MALDI-TOF MS is a promising biotool for rapid and accurate identification of foodborne microbes at the species and genus level in food samples. Database expansion and direct quantification of spoilage microbes are two promising areas for future progress in MALDI-TOF MS applications.

Development of novel nanostructured biosensors for rapid detection of pathogens in clinical diagnostics

&lt;p class="MsoNormal" style="text-align: justify; text-justify: inter-ideograph; line-height: 120%; mso-pagination: none; layout-grid-mode: char; mso-layout-grid-align: none; margin: 12.0pt 0cm 6.0pt 0cm;"&gt;&lt;span lang="EN-US" style="font-size: 10.0pt; line-height: 120%; font-family: 'Times New Roman',serif;"&gt;The prompt and precise identification of microorganisms is crucial for successful clinical diagnostics and the prevention of infectious disease outbreaks. Traditional diagnostic methods often suffer from limitations such as extended processing durations, elevated expenses, and the necessity for specialized laboratory equipment. In this research, we propose the development of novel nanostructured biosensors that utilize the distinct characteristics of nanomaterials to improve the accuracy, specificity, and efficiency of identifying pathogens. These biosensors are created with the intention of offering point-of-care testing functionality, thus rendering them appropriate for utilization in a range of clinical settings. The integration of advanced nanotechnology with bioanalytical methods aims to create a reliable system for the real-time identification of bacterial, viral, and fungal pathogens. This review encompasses the design, fabrication, and testing of the biosensors, along with a comprehensive analysis of their performance in comparison to conventional diagnostic techniques. The results demonstrate the potential of nanostructured biosensors to revolutionize pathogen detection, offering significant improvements in efficiency and accuracy, which are essential for timely medical intervention and public health management.&lt;/span&gt;&lt;/p&gt;

Expression of CD14+CD282+, CD14+CD284+, CD14+HLA-DR+, CD14+CD11b+ receptors and sIgA level in premature infants with K. pneumoniae

K. pneumoniae is one of the leading microorganisms causing nosocomial infections among premature newborns. The ineffectiveness of immune defense, morphofunctional immaturity, length of hospitalization and invasive procedures create the prerequisites for the implementation of the infectious process in a hospital setting. The question of the reasons for the development of infection, the etiological agent of which colonizes the intestines, remains open. Purpose of the study: to evaluate the expression of CD14+CD282+, CD14+CD284+, CD14+HLA-DR+, and CD14+CD11b+ receptors on blood monocytes and the level of sIgA in coprofiltrates in premature infants with intestinal colonization by K. pneumonia with different genetic profiles. We examined 11 children with the uge gene (group 1), 20 newborns with the uge + fim genes (group 2), and 12 children with the kfu + uge + fim genes. Microbiological examination of feces included identification and antibiotic sensitivity of microorganisms. Detection of the uge, fim and kfu genes in K. pneumoniae strains was carried out by PCR. The expression level of monocyte activation markers was determined by flow cytometry. Gestational age and anthropometric parameters did not differ between newborns. Children identified with the fim gene in combination with other genes were more often discharged home with K. pneumoniae than with the uge gene. In these children, a decrease in the level of expression of CD14+CD282+, CD14+CD284+, CD14+CD11b+, CD14+HLA-DR+ receptors at birth and upon reaching postconceptional age, and a low sIgA content in coprofiltrates during 24 days of life were recorded. Thus, a decrease in the expression of CD14+CD282+, CD14+CD284+, CD14+CD282+ and CD14+HLA-DR+ receptors by blood monocytes and insufficiency of sIgA production in the large intestine determine a long period of colonization of K. pneumoniae strains with the presence of the fim gene in combination with other genes (from 15 to 180 days), as well as the possibility of Klebsiella infection occurring in subsequent periods of the child’s life. Much more often, children with the combination of genes uge + fim and kfu + uge + fim were discharged from the hospital with a diagnosis of anemia; only in these groups of children was the development of bronchopulmonary dysplasia recorded.

Evolving strategies in microbe identification—a comprehensive review of biochemical, MALDI-TOF MS and molecular testing methods

Abstract Detection and identification of microorganisms are the first steps to guide susceptibility testing and enable clinicians to confirm diseases and guide therapy. The faster the pathogen identification is determined, the quicker the appropriate treatment can be started. In the clinical microbiology laboratory, multiple methodologies can be used to identify organisms, such as traditional biochemical testing or more recent methods like MALDI TOF MS and nucleic acid detection/identification assays. Each of these techniques has advantages and limitations, and clinical laboratories need to determine which methodology is best suited to their particular setting in terms of clinical needs, availability of technical expertise and cost. This article presents a concise review of the history, utilization, advantages and limitations of the main methods used for identifying microorganisms in microbiology laboratories.

Metagenomic analysis of atheroma plaques for identification of microorganisms indicates presence of Toxoplasma gondii as a possible etiological agent

BackgroundCardiovascular diseases (CVDs) are the leading cause of death worldwide. Vital organs like the heart are affected by the occlusion of blood vessels due to atherosclerotic plaque formation. However, the role of infectious agents has always been an essential subject of investigation. This study investigated the presence of microorganisms, including nanobacteria, in atherosclerotic plaques removed from human carotid arteries by microbiological and metagenomic examination. MethodsAtheroma plaque samples were obtained from 20 patients with carotid artery stenosis who had atherectomy by surgery or percutaneous intervention. Nanobacteria were grown by culturing homogenates of the atheroma plaques. Whole genome sequencing was done for samples. Because of the high percentage of Toxoplasma gondii (T. gondii) DNA, PCR investigation was applied to detect T. gondii DNA in the samples. ResultsA molecular analysis of nanobacteria revealed them to be made of human proteins, supporting the theory that they are not living organisms. According to sequencing results, samples showed that more than 50 % of the metagenomic sequences belonged to Toxoplasma gondii. PCR investigation indicated that T. gondii DNA was positive in 8 (40 %) of 20 plaques. ConclusionsFurther evidence regarding the role of T. gondii in the etiology of plaque formation may help determine the strategy for prevention and treatment of infections in preventing atheroma plaque formation in the future.

The Complex and Changing Genus Bacillus: A Diverse Bacterial Powerhouse for Many Applications

For years, the Bacillus genus has encompassed a heterogeneous group of individuals whose main common trait was their ability to sporulate in the presence of oxygen. This criterion has been revised, resulting in the reclassification of several species into new genera and to a redefinition of the characteristics of the members of this taxon. Currently, the species of the genus are grouped into the Subtilis clade and the Cereus clade. The former, called Bacillus sensu stricto, initially composed of B. subtilis, B. licheniformis, B. pumilus, and B. amyloliquefaciens, has subsequently incorporated new species related to these. The Cereus clade, Bacillus cereus sensu lato, consists of pathogenic species (B. anthracis, B. cereus, and B. thuringiensis) as well as others of significance in agriculture and industry. Furthermore, identifying these individuals remains complex, requiring alternatives to 16S rRNA sequencing. The ability to form spores resistant to stressful conditions provides a significant advantage over other genera, with observable differences in sporulation rates and spore structure among different species. Additionally, Bacillus spp. are known for their capacity to produce antimicrobial substances, lytic enzymes, and volatile organic compounds, each with diverse applications. Some species are even used as probiotics. This review delves into aspects related to the taxonomy and identification of microorganisms belonging to the genus Bacillus, which often present challenges. The aim is to provide a comprehensive overview of the topic. In addition, it highlights the characteristics and applications of the genus, emphasizing its importance in biotechnology and microbiology.

Investigating the Relationship between Awareness, Beliefs, and Performance of Operating Room Students in Hand Hygiene with Analysis of Microbial Flora on Hands during the COVID-19 Pandemic

BackgroundHand hygiene compliance among hospital staff has always been recognized as an effective method in controlling and reducing hospital-acquired infections. Since the outbreak of the COVID-19 pandemic, hand hygiene has emerged as one of the effective preventive measures against this disease, leading to increased awareness and concerns. This study aims to examine the awareness, beliefs, and practices of operating room students regarding hand hygiene, by analyzing the microbial flora on their hands during the COVID-19 pandemic. The objective is to assess the level of hand hygiene compliance among students, who will be future hospital employees. AimThis study aimed to investigate the correlation between the level of awareness, belief, and performance of operating room students regarding hand hygiene with the analysis of microbial hand flora during the COVID-19 pandemic. The purpose was to examine the compliance with hand hygiene among students, who will be future hospital employees, during this pandemic. MethodologyThe present study included a sample of 92 students majoring in Operating Room who were enrolled in the research through a census. The data collection tool consisted of a questionnaire to assess the students' awareness, beliefs, and performance regarding hand hygiene and analysis of microbial flora on hands. Standard culture media including Blood Agar, Brilliant Green Agar, and Eosin Methylene Blue were used for isolating microorganisms. Microorganism identification was performed using biochemical tests. ResultsThe results of this study indicate that out of the participants, 65 were girls and 27 were boys with an average age of 21.9±2.3 years. The mean scores for awareness, belief, and performance were 8.9, 72.3, and 64.2, respectively. Analysis of microbiological flora also revealed that the highest growth microorganisms were epidermidis bacteria (26.6%), coagulase-negative staphylococci bacteria (22.9%), and streptococcus bacteria (21.2%). Discussion and ConclusionBased on the results of this study, operating room students have shown a high level of awareness, belief, and performance during the COVID-19 pandemic. Furthermore, the minimal growth of bacteria in the conducted culture samples confirms this finding. Due to the limited availability of studies, it is recommended that further research be conducted in this area.

Assessing Diagnostic Performance of Molecular Culture for Neonatal Sepsis: Protocol of the CHAMPIONS Study.

Managing neonatal sepsis is challenging due to nonspecific clinical signs, hematological markers with poor accuracy, and a lengthy turnaround time for the identification of microorganisms. Delaying the initiation of antibiotics in truly infected infants can lead to severe morbidity and mortality. Therefore, decisions regarding empiric antibiotic treatment are risk stratified, which exposes many uninfected infants to antibiotics. This causes gut microbiota perturbation, unnecessary hospital admissions, and the generation of multi-resistant organisms. High-speed diagnostic assays could expedite discontinuation or avert the initiation of antibiotics in uninfected infants. This study will evaluate the diagnostic performance of molecular culture (MC), a rapid broad-range PCR-based bacterial profiling technique, for diagnosing neonatal sepsis in infants below 90 days old. A multi-center prospective observational cohort study will include infants evaluated for early and late-onset sepsis. Routine evaluation for suspected sepsis includes microbiological cultures of blood. Additionally, blood for MC will be collected. For early-onset sepsis, umbilical cord blood may be used alternatively. Primary outcome is the agreement between MC and conventional blood culture results. Secondary outcome is the agreement of both assays with clinical sepsis using four different, commonly used definitions. Faster diagnostic pathways for sepsis may reduce antibiotic exposure time. Broad-range molecular assays may identify pathogens undetectable by conventional methods. Employment of umbilical cord blood samples for early-onset sepsis diagnosis can resolve challenges in collecting adequate blood volume and could further expedite treatment decisions.

Early organism identification by Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) decreases the time to appropriate antibiotic modifications for common bacterial infections.

MALDI-TOF-MS facilitates the identification of microorganisms from positive cultures in a timely and accurate manner. It eliminates the necessity for the application of biochemicals and operates on the principle of proteomics. It decreases the time required to report culture results. Prompt detection and notification of the pathogen, prior to the disclosure of antimicrobial susceptibilities, could potentially shorten the duration until the initial antibiotic adjustment is necessary, thereby influencing patients' clinical prognoses. Fifty patients in the conventional arm and one hundred patients in the interventional arm were compared in a pre and post quasi-experimental study conducted at a tertiary care centre in North India. Patients with positive cultures from medical wards and intensive care units were included. Comparing the time to first antibiotic modification after culture positivity, MALDI-TOF-MS-based identification, and clinical outcomes in both arms was the primary objective. Antibiotic modifications, escalation, and de-escalation were all recorded. The intervention arm exhibited a substantially shorter median time to first antibiotic modification (2010 mins vs 2905 mins, p=0.002) than the conventional arm. In the interventional group, a total of 44 out of 100 antibiotic modifications were implemented. Of these, 19 (43.3%) were determined solely by the MALDI report, without the anticipation of susceptibility assessments. De-escalation of antibiotics constituted the pre-dominant form of modification (47.4%). The difference between the 27% and 32% mortality rates in the intervention arm and the conventional arm was not statistically significant (p=0.52). MALDI-TOF-MS facilitates the modification of antibiotics early on. The primary benefit lies in the reduction of superfluous antibiotic usage. Early organism identification and reporting prior to the availability of susceptibility results did not result in any mortality benefit. This strategy, when combined with a strong antimicrobial stewardship programme, can aid in the reduction of antibiotic use.