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Background: Onychomycosis is the most common nail pathology, involving various pathogens such as dermatophytes, moulds and yeasts. Objective: The objective of this study was to observe the prevalence of onychomycosis, analyse the most appropriate diagnostic test, and assess the distribution of pathogens based on age, sex, quarter of the year, duration of symptoms and previous treatment. Methods: Retrospectively, mycological culture and PCR data and results were collected from 121 patients. Results: Of the 121 samples, 57% (69/121) tested positive when both microbiological study techniques were combined. The prevalence of onychomycosis was higher when PCR was performed (52.1%) compared to microbiological culture (33.1%). Among the 81 samples negative by microbiological culture, 31 were positive by PCR. Similarly, of the 58 samples negative by PCR, eight were positive by microbiological culture. Diagnostic accuracy data (with 95% confidence intervals) for PCR, using microbiological culture as the gold standard, were as follows: sensitivity of 0.8, specificity of 0.62, positive predictive value of 0.51 and negative predictive value of 0.86. The most frequently identified pathogen was Trichophyton rubrum, and the hallux nail plate was the most commonly affected location. However, no statistically significant associations were found between sex, age, quarter of the year and affected area with culture and PCR results. Conclusion: Combining microbiological culture and PCR can increase the detection rate of onychomycosis and help avoid false‐negative results. [ABSTRACT FROM AUTHOR]

Introduction: The Ministry of Health in Saudi Arabia provides comprehensive antenatal care for all pregnant women with all required investigations. However, it does not include urine culture for diagnosis of asymptomatic bacteriuria (ASB). This is the first study to evaluate the prevalence of ASB among pregnant females, identify the causative organisms and determine their antibiotic susceptibility patterns in the Maternity and Children's Hospital, Arar, Saudi Arabia. Methodology: This cross-sectional study included 400 pregnant women attending an antenatal clinic. Two midstream urine samples were aseptically collected and screened using standard microbiological techniques including microscopic examination, dipstick testing, and urine culture. In order to interpret the urine culture results, ≥ 105 CFUs/mL was considered significant bacteriuria. Identification of the isolates and their antibiotic sensitivity testing was performed using the Vitek 2 system (BioMérieux, Marcy l'Etoile, France) with the available test kits. Results: The prevalence of ASB was 8.25% (35/400). Significant positive correlations (p < 0.05) were detected between positive urine culture results and random blood sugar, leucocytes, nitrites, pus cells, urine red blood cells, epithelial cells, and mucus. Escherichia coli was the most common causative organism (45.7%), followed by Staphylococcus aureus (22.9%). Klebsiella pneumoniae represented 11.4% of the isolates. Most of the isolated Gram-positive organisms were sensitive to many of the tested antibiotics; most of the detected Gram-negative isolates were resistant. Conclusions: ASB caused by antibiotic resistant organisms is alarming. Screening for ASB during pregnancy using urine culture and sensitivity testing is of vital importance to improve the maternal and neonatal outcome. [ABSTRACT FROM AUTHOR]

Background and Objectives: AmpC-producing Gram-negative bacterial (GNB) pathogens are distributed worldwide, especially in clinical settings. This study aimed to determine the antibiogram and the type of AmpC-ß-lactamase gene harboured by GNB pathogens implicated in chronic suppurative otitis media (CSOM) cases. Materials and Methods: Ear swab samples (300) collected from patients with active CSOM were analysed using standard microbiological techniques. Phenotypic and molecular detection of AmpC ß-lactamase production was done by cefoxitin/cloxacillin double-disk synergy test and PCR respectively. Antibiogram was determined by disk diffusion technique. Results: Among the GNB pathogens isolated from CSOM patients, P. aeruginosa was the most predominant (36.3%); followed by K. pneumoniae (22.3%), and E. coli (13.7%). Patients with active CSOM showed increased bacteria isolation rate from bilateral ear discharges than unilateral ear discharges. E. coli and P. aeruginosa were more prevalent among patients with duration of discharge >2 weeks; recording 9.0% and 20.3% respectively. AmpC ß-lactamase producers accounted for 14.0%; they were highly resistant (60%-100%) to cephalosporins, trimethoprim-sulfamethoxazole, ofloxacin, amoxicillin, and tetracycline, but very susceptible (70.4%-100%) to ciprofloxacin, imipenem, and amikacin. Multiple antibiotic resistance indices of isolates ranged from 0.7-0.8. FOX-AmpC-ß-lactamase gene was detected in 3.9% of the isolates. Conclusion: The detection of AmpC ß-lactamase-producing multidrug-resistant GNB pathogens harbouring FOX-AmpC-ß-lactamase gene among patients with CSOM infections in our study is a serious public health problem which needs urgent intervention. [ABSTRACT FROM AUTHOR]

Objectives Pemphigus, a group of autoimmune bullous diseases, can be fatal, resulting from overwhelming opportunistic infection of lesions secondary to cutaneous bacterial infections. This study aimed to look into the cutaneous bacterial infection profile of pemphigus patients as timely identification and appropriate treatment can play a major role in reducing mortality. Materials and Methods Pus samples/swabs received from patients with pemphigus over a 2-year period from July 2018 to June 2020 were subjected to standard microbiological culture techniques and susceptibility testing. The frequency of isolation and susceptibility profile of the different bacterial pathogens toward various antimicrobial agents were interpreted and analyzed as per the Clinical and Laboratory Standards Institute's guidelines. Results Samples from 315 patients were received during the study period comprising of 203 (64.4%) males and 112 (35.5%) females. Of 211 samples which were culture-positive, a total of 245 bacterial isolates were obtained, comprising of 158 Gram-positive cocci and 87 Gram-negative bacilli. Staphylococcus aureus (138, 56.3%) was the most common isolate followed by Pseudomonas aeruginosa (41, 16.7%) and Escherichia coli (16, 6.5%). Methicillin resistance was observed in 24.6% Staphylococcus aureus isolates and carbapenem resistance in 9.5 to 14.6% Gram-negative bacilli. Conclusions Study findings emphasize the need for continuous monitoring of cutaneous pemphigus lesions for appropriate choice of antimicrobial therapy. [ABSTRACT FROM AUTHOR]

The increasing number of surgical fracture care using osteosynthesis devices poses a challenge for orthopedics and trauma surgery. While approximately 1.23 % of all fractures result in a fracture-related infection (FRI), 10-13 % of all FRIs affect the upper extremity. Accurate diagnosis of this often-complex condition is crucial for successful therapy. Since 2018, confirmatory criteria and suggestive parameters have been established that must be considered in the diagnosis. Clinical routine diagnostics are also followed for the upper extremity. In addition to the obligatory medical history and clinical examination, radiological imaging such as X-ray, CT, MRI, and nuclear medicine procedures are included. Furthermore, 3-5 tissue samples from the fracture region should be sent for microbiological diagnosis, and a histopathological examination should be arranged for diagnostic confirmation. Treatment should be interdisciplinary. In addition to surgical and usually twelve-week antibiotic therapy, an interdisciplinary treatment approach to optimize comorbidities and surgical assessment by a plastic surgeon is advisable, especially in complex cases. Special aspects of the upper extremities, such as options for joint preservation or conversion to a joint replacement procedure, should be evaluated depending on the blood flow and osteosynthesis situation. Diaphyseal bone defects of smaller extent in the humerus can be treated better with shortening, and monorail fixator systems can be applied safely without additional intramedullary implants, unlike in the lower extremity. The commonly occurring Cutibacterium acnes can often be effectively treated with antibiotics. However, the surgeon must be aware of a three-week cultivation period by microbiological diagnostics. Separate patient-reported outcome measurements (PROMs) for assessing treatment outcomes are not available. PROMs for the upper extremity, such as the disabilities of the arm, shoulder and hand questionnaire, are applied here. [ABSTRACT FROM AUTHOR]

The environment is contaminated by an array of organic pollutants, many of which can permeate into wastewater. Municipal and industrial wastewater are the primary contributors of these pollutants in the aquatic environment and typically comprise substantial amounts of different organic compounds. Therefore, finding efficient ways to manage wastewater and other contaminated water is a crucial part of protecting the environment. Among the various methods of bioremediation, in-situ biodegradation emerges as one of the most potent and eco-conscious approaches. It harnesses naturally occurring bacteria to entirely break down or detoxify harmful pollutants, detrimental to both human health and the environment. This study aims to isolate and identify bacteria exhibiting biodegradation capabilities against a spectrum of organic pollutants, including carbohydrates, proteins, industrial dyes, and hydrocarbons from Korangi river via different morphological and biochemical methods. Municipal wastewater was collected from Korangi Creek River in Karachi and its physicochemical characteristics such as pH, temperature, total dissolved solids, conductivity, and salinity were assessed. The pour plate method was subsequently employed to isolate and assess the biodegradation potential of native bacterial strains. Isolated strains were identified by using microscopic and biochemical analysis. Three bacterial strains were selected from organiccontaminated wastewater based on their amylase activity, azoreductase activity, and kerosene-degrading activity. Different features were studied for all three biodegrading bacterial strains and were identified as Klebsiella pneumoniae subsp. Ozaenae, Group A Streptococcus pyogenes and Nonenterococcus Streptococcus bovis by microbiological culture techniques and biochemical identification tests. This study assumes significance as it utilizes the ability of nature's microorganisms to efficiently degrade and eliminate organic contaminants in a costeffective and eco-friendly way. [ABSTRACT FROM AUTHOR]

Recently, bone and joint infections such as fracture-related infection (FRI) and prosthetic joint infections (PJI) have been defined for clinical and research purposes. National and international guidelines are being produced to guide clinicians towards what is considered best practice. However, much of what we do regarding the microbiological diagnosis and management of orthopaedic infection is based on expert opinion, rather than evidence-based medicine. This is largely because, prior to the publication of the clinical definitions of orthopaedic infection, studies included a heterogeneous population making comparisons between them difficult. More recently the publication of large clinical trials in the antimicrobial management of orthopaedic infection has demonstrated that such clinical research is possible, and can both challenge the existing dogma in this field and lead to universal changes in clinical practice. There is robust evidence for microbiological sampling techniques and their interpretation in diagnosis of orthopaedic infection, as well as the choice of empiric broad-spectrum antibiotics and the non-inferiority of oral versus intravenous systemic antibiotics. For other elements of the diagnosis and management of orthopaedic infection this is not the case. For example, there are limited or no data to support recommendations regarding time off antibiotics prior to microbiological sampling, systemic antibiotic duration, when to do a joint aspirate in PJI revisions or the timing of definitive joint implantation in the two-stage management of PJI. In summary, this review highlights what is currently considered best practice within the field of orthopaedic infection and discusses both the evidence behind it and where there is controversy or where further research is warranted. [ABSTRACT FROM AUTHOR]

Introduction: The Scalp Eschar and Neck Lymph Adenopathy After a Tick Bite (SENLAT) syndrome is frequently caused by Rickettsia slovaca and Rickettsia raoultii. Only six microbiologically confirmed SENLAT cases have been reported in Italy between 1996 and 2021. We report ten cases of SENLAT seen between 2015 and 2022 in a tertiary care center in Tuscany, Italy. Cases presentation: All patients were women; most common symptoms were scalp eschar on the site of tick bite (100%) and cervical lymphadenopathy (90%). No microbiological identification was obtained. Persistent alopecia, for several months to years, was observed in four patients. The known difficulty of microbiological diagnosis in SENLAT was worsened, in our cases, by factors as the absence of ticks available for identification and microbiological study, and antibiotic treatment administration previous to microbiological tests. Conclusion: The report highlights the presence of SENLAT in Italy, aiming to raise the awareness toward the emergence of this clinical entity. [ABSTRACT FROM AUTHOR]

Background: Data on the benefits of rapid microbiological testing on antimicrobial consumption (AC) and antimicrobial resistance patterns (ARPs) are scarce. We evaluated the impact of a protocol based on rapid techniques on AC and ARP in intensive care (ICU) patients. Methods: A retrospective pre- (2018) and post-intervention (2019–2021) study was conducted in ICU patients. A rapid diagnostic algorithm was applied starting in 2019 in patients with a lower respiratory tract infection. The incidence of nosocomial infections, ARPs, and AC as DDDs (defined daily doses) were monitored. Results: A total of 3635 patients were included: 987 in the pre-intervention group and 2648 in the post-intervention group. The median age was 60 years, the sample was 64% male, and the average APACHE II and SOFA scores were 19 points and 3 points. The overall ICU mortality was 17.2% without any differences between the groups. An increase in the number of infections was observed in the post-intervention group (44.5% vs. 17.9%, p < 0.01), especially due to an increase in the incidence of ventilator-associated pneumonia (44.6% vs. 25%, p < 0.001). AC decreased from 128.7 DDD in 2018 to 66.0 DDD in 2021 (rate ratio = 0.51). An increase in Pseudomonas aeruginosa susceptibility of 23% for Piperacillin/tazobactam and 31% for Meropenem was observed. Conclusion: The implementation of an algorithm based on rapid microbiological diagnostic techniques allowed for a significant reduction in AC and ARPs without affecting the prognosis of critically ill patients. [ABSTRACT FROM AUTHOR]

Composition of pulmonary microbiome of patients with severe pneumonia is poorly known. The aim of this work was to analyse the lung microbiome of patients admitted to the intensive care unit (ICU) with severe community acquired pneumonia (CAP) between 2019 and 2021 in comparison with a control group of 6 patients undergoing digestive surgery. As a second objective, the diagnostic capabilities of metagenomics was also studied in a small group of selected patients. The lung microbiome of patients with viral (5 with Influenza A and 8 with SARS-CoV-2) pneumonia at admission showed a similar diversity as the control group (p = 0.140 and p = 0.213 respectively). Contrarily, the group of 12 patients with pneumococcal pneumonia showed a significant lower Simpson´s index (p = 0.002). In the control group (n = 6) Proteobacteria (36.6%), Firmicutes (24.2%) and Actinobacteria (23.0%) were the predominant phyla. In SARS-CoV-2 patients (n = 8), there was a predominance of Proteobacteria (mean 41.6%) (Moraxella and Pelomonas at the genus level), Actinobacteria (24.6%) (Microbacterium) and Firmicutes (22.8%) mainly Streptococcus, Staphylococcus and Veillonella. In patients with Influenza A pneumonia (n = 5) there was a predominance of Firmicutes (35.1%) mainly Streptococcus followed by Proteobacteria (29.2%) (Moraxella, Acinetobacter and Pelomonas). In the group of pneumococcal pneumonia (n = 12) two phyla predominated: Firmicutes (53.1%) (Streptococcus) and Proteobacteria (36.5%) (Haemophilus). In the 7 patients with non-pneumococcal bacterial pneumonia Haemophilus influenzae (n = 2), Legionella pneumophila (n = 2), Klebsiella pneumoniae, Streptococcus pyogenes and Leptospira were detected by metagenomics, confirming the diagnosis done using conventional microbiological techniques. The diversity of the respiratory microbiome in patients with severe viral pneumonia at ICU admission was similar to that of the control group. Contrarily, patients with pneumococcal pneumonia showed a lower grade of diversity. At initial stages of SARS-CoV-2 infection, no important alterations in the pulmonary microbiome were observed. The analysis of bacterial microbiome showed promising results as a diagnostic tool. [ABSTRACT FROM AUTHOR]

Aimed to explore the relationships between infection localization, bacterial species, and procalcitonin (PCT) and C-reactive protein (CRP) levels in urinary tract infections (UTIs). A retrospective study included 314 UTI hospitalized patients divided into two groups (268 with lower UTI, 46 with upper UTI) in a tertiary care hospital. PCT and CRP were performed. Bacterial isolates were identified using standard microbiological techniques, and statistical analyses were performed to assess associations between infection localization, bacterial species, PCT, and CRP levels. Age and gender showed no significant differences between the lower and upper UTIs. Escherichia coli dominated as the leading UTI pathogen. A positive correlation (r = 0.646, P < 0.001) between PCT and CRP levels was found. The subgroup with ureteritis in the upper UTI category exhibited the highest PCT and CRP levels. PCT and CRP exhibited favorable diagnostic potential in predicting upper UTIs, with AUCs of 0.644 and 0.629, respectively. The optimal cutoff values were 0.21 ng/mL for PCT and 60.77 mg/L for CRP. Sensitivities were 69.03% and 77.99%, while specificities were 56.52% and 47.83%, respectively. E. coli emerged as the predominant bacterium in UTIs. PCT and CRP demonstrated moderate diagnostic efficacy in distinguishing between upper and lower UTIs. Notably, PCT and CRP exhibited enhanced utility in identifying ureteritis. [ABSTRACT FROM AUTHOR]

Background. Bacterial biofilm is a significant virulence factor threatening patients, leading to chronic infections and economic burdens. Therefore, it is crucial to identify biofilm production, its inhibition, and reduction. In this study, we investigated biofilm production among Gram-negative isolates and assessed the inhibitory and reduction potential of ethylene diamine tetra acetic acid (EDTA) and dimethyl sulfoxide (DMSO) towards them. In addition, we studied the antimicrobial resistance pattern of the Gram-negative isolates. Methods. Bacterial isolation and identification was done using standard microbiological techniques, following the Clinical and Laboratory Standards Institute (CLSI) guideline, 28th edition. The Kirby–Bauer disk diffusion method was used to determine the antibiotic susceptibility pattern of the isolates, and β-lactamase production was tested via the combination disk method. Biofilm formation was detected through the tissue culture plate (TCP) method. Different concentrations of EDTA and DMSO were used to determine their inhibitory and reduction properties against the biofilm. Both inhibition and reduction by the various concentrations of EDTA and DMSO were analyzed using paired t-tests. Results. Among the 110 clinical isolates, 61.8% (68) were found to be multidrug resistant (MDR). 30% (33/110) of the isolates were extended-spectrum β-lactamase (ESBL) producers, 14.5% (16/110) were metallo-β-lactamase (MBL), and 8% (9/110) were Klebsiella pneumoniae carbapenemase (KPC) producers. Biofilm formation was detected in 35.4% of the isolates. Biofilm-producing organisms showed the highest resistance to antibiotics such as cephalosporins, chloramphenicol, gentamicin, and carbapenem. The inhibition and reduction of biofilm were significantly lower (p < 0.05) for 1 mM of EDTA and 2% of DMSO. Conclusion. Isolates forming biofilm had a higher resistance rate and β-lactamase production compared to biofilm nonproducers. EDTA and DMSO were found to be potential antibiofilm agents. Hence, EDTA and DMSO might be an effective antibiofilm agent to control biofilm-associated infections. [ABSTRACT FROM AUTHOR]

Background: Neonatal sepsis is the major cause of neonatal mortality and morbidity, especially in low and middle-income countries. Continuous monitoring of pathogens and their antibiotic resistance pattern is crucial for managing neonatal sepsis. This study aimed to determine neonatal sepsis due to bacteria, antibiotic resistance patterns, associated risk factors and patient outcomes at St. Paul's Hospital Millennium Medical College. Method: An institutional-based cross-sectional study was conducted on 400 neonates suspected of sepsis at St. Paul's Hospital Millennium Medical College from March 2020 to July 2020. A questionnaire was used to collect socio-demographic information, clinical parameters and potential risk factors from study participants. About 2ml of blood was drawn aseptically and inoculated into Tryptone Soya Broth at the patient's bedside. Bacterial identification was performed by using standard microbiological techniques. The disk diffusion method was used to determine the antibiotic susceptibility patterns of each isolated bacteria. Data entry and analysis were done using Statistical Package for Social Sciences (SPSS) version 20 software. Bivariate and multivariable logistic regressions were used to assess associated risk factors of neonatal sepsis. A p-value less than 0.05 was considered statically significant with a 95% confidence interval. Results: The overall prevalence of neonatal septicemia was 21% (84/400). Of these, 67 (79.8%) and 17 (20.2%) were gram-negative and gram-positive bacteria, respectively. Klebsiella spp, 37 (44%), E. coli 19 (21.6%) and Coagulase negative Staphylococci 13 (15.47%) were the leading cause of neonatal sepsis. Ciprofloxacin and amikacin were the most effective antibiotics for gram-negative and gram-positive bacteria. Multidrug resistance was observed in 84% of the bacterial isolates. Low birth weight and preterm were associated with neonatal septicemia (AOR = 49.90, 95% CI = 15.14-123.081, P = 0.002) and (AOR = 18.20, 95% CI = 6.835–27.541, P = 0.004) respectively. Conclusion: Klebsiella spp and E. coli were frequently isolated bacteria in our study. The proportion of multidrug-resistance was significantly high. Most isolated bacteria were resistant to ampicillin, ceftazidime, cefotaxime and gentamycin, which indicates the necessity of continuous evaluation of antibiotic resistance rate. [ABSTRACT FROM AUTHOR]

Introduction: Proteus mirabilis is a key pathobiont in catheter-associated urinary tract infections (CA-UTIs), which is well known to form crystalline biofilms that occlude catheters. Urease activity alkylates urine through the release of ammonia, consequentially resulting in higher levels of Mg2+ and Ca2+ and formation of crystals. In this study, we showed that N-acetyl cysteine (NAC), a thiol antioxidant, is a potent urease inhibitor that prevents crystalline biofilm formation. Methods: To quantify urease activity, Berthelot's method was done on bacterial extracts treated with NAC. We also used an in vitro catheterised glass bladder model to study the effect of NAC treatment on catheter occlusion and biofilm encrustation in P. mirabilis infections. Inductively-coupled plasma mass spectrometry (ICP-MS) was performed on catheter samples to decipher elemental profiles. Results: NAC inhibits urease activity of clinical P. mirabilis isolates at concentrations as low as 1 mM, independent of bacterial killing. The study also showed that NAC is bacteriostatic on P. mirabilis, and inhibited biofilm formation and catheter occlusion in an in vitro. A significant 4-8log10 reduction in viable bacteria was observed in catheters infected in this model. Additionally, biofilms in NAC treated catheters displayed a depletion of calcium, magnesium, or phosphates (>10 fold reduction), thus confirming the absence of any urease activity in the presence of NAC. Interestingly, we also showed that not only is NAC anti-inflammatory in bladder epithelial cells (BECs), but that it mutes its inflammatory response to urease and P. mirabilis infection by reducing the production of IL-6, IL-8 and IL-1b. Discussion: Using biochemical, microbiological and immunological techniques, this study displays the functionality of NAC in preventing catheter occlusion by inhibiting urease activity. The study also highlights NAC as a strong antiinflammatory antibiofilm agent that can target both bacterial and host factors in the treatment of CA-UTIs. [ABSTRACT FROM AUTHOR]

Advancements in DNA sequencing technologies within the last decade have stimulated an unprecedented interest in the human microbiome, largely due the broad diversity of human diseases found to correlate with microbiome dysbiosis. As a direct consequence of these studies, a vast number of understudied and uncharacterized microbes have been identified as potential drivers of mucosal health and disease. The looming challenge in the field is to transition these observations into defined molecular mechanistic studies of symbiosis and dysbiosis. In order to meet this challenge, many of these newly identified microbes will need to be adapted for use in experimental models. Consequently, this review presents a comprehensive overview of the molecular microbiology tools and techniques that have played crucial roles in genetic studies of the bacteria found within the human oral microbiota. Here, we will use specific examples from the oral microbiome literature to illustrate the biology supporting these techniques, why they are needed in the field, and how such technologies have been implemented. It is hoped that this information can serve as a useful reference guide to help catalyze molecular microbiology studies of the many new understudied and uncharacterized species identified at different mucosal sites in the body. [ABSTRACT FROM AUTHOR]

Using phages as salvage therapy for nonhealing infections is gaining recognition as a viable solution for patients with such infections. The escalating issue of antibiotic resistance further emphasizes the significance of using phages in treating bacterial infections, encompassing compassionate-use scenarios and clinical trials. Given the high specificity of phages, selecting the suitable phage(s) targeting the causative bacteria becomes critical for achieving treatment success. However, in contrast to conventional antibiotics, where susceptibility-testing procedures were well established for phage therapy, there is a lack of standard frameworks for matching phages from a panel to target bacterial strains and assessing their interactions with antibiotics or other agents. This review discusses and compares published methods for clinical phage microbiology, also known as phage susceptibility testing, and proposes guidelines for establishing a standard pipeline based on our findings over the past 5 years of phage therapy at the Israeli Phage Therapy Center. [ABSTRACT FROM AUTHOR]

Background: Dental caries is a preventable multifactorial disease, with Streptococcus mutans being suggested to be its primary pathogen. Our study aim was to compare the effects of three different low-cost and easy-to-use regimens with that of the gold standard (chlorhexidine [CHX] mouthwash) on the count of salivary S. mutans in dental students over 30 days. Materials and Methods: In this single-blinded parallel randomized controlled clinical trial, a total of 120 dental students were included and randomly allocated into four intervention groups: (1) CHX mouthwash (control(, (2) probiotic yogurt, (3) casein phosphopeptide-amorphous calcium phosphate chewing gum, and (4) xylitol chewing gum. Salivary S. mutans counts were evaluated at baseline, 15 days, and 30 days after initiation of the study and compared at different times and among different groups using the repeated measures analysis of variance design analysis and least significant difference test with SPSS software version 20. The level of significance was determined to be 0.05. Results: The microorganism count variable at baseline, first, and second follow-ups was significantly different for all groups except the probiotic yogurt group (P = 0.340). S. mutans count was significantly different when comparing the first follow-up and baseline values in the CHX and xylitol gum groups (P = 0.027, P = 0.037). When comparing the second follow-up with baseline values, a significant difference was observed in the xylitol gum group (P = 0.003). Conclusion: Xylitol chewing gum seems to be a viable alternative to the gold standard (CHX mouthwash) in reducing the salivary count of S. mutans. [ABSTRACT FROM AUTHOR]

Ultraviolet (UV) light is an effective disinfection technology, able to inactivate a wide range of microorganisms, including bacteria and fungi. A safer UV wavelength of 222 nm, also known as far-UVC, has been proposed to minimize these harmful effects while retaining the light's disinfection capability. This study is aimed at exploring the antimicrobial activity of filtered far-UVC (222 nm) on a panel of pathogens commonly found in nosocomial installations. A panel of Gram-positive and Gram-negative bacteria and yeast pathogens was tested. Microorganisms were deposited on a plastic surface, allowing them to dry before exposure to the far-UVC light at a distance of 50 cm. Results showed that far-UVC light successfully inhibits the growth of the tested pathogens, although at different exposure times. In conclusion, the results of this study provide fundamental information to achieve reliable disinfection performance with far-UVC lamps with potential applications in healthcare facilities like hospitals and long-term care homes. [ABSTRACT FROM AUTHOR]

The emergence of multidrug-resistant (MDR) Klebsiella pneumoniae remains a global health threat due to its alarming rates of becoming resistant to antibiotics. Therefore, identifying plant-based treatment options to target this pathogen's virulence factors is a priority. This study examined the antivirulence activities of twelve plant extracts obtained from three South African medicinal plants (Lippia javanica, Carpobrotus dimidiatus, and Helichrysum populifolium) against carbapenem-resistant (CBR) and extended-spectrum beta-lactamase (ESBL) positive K. pneumoniae strains. The plant extracts (ethyl acetate, dichloromethane, methanol, and water) were validated for their inhibitory activities against bacterial growth and virulence factors such as biofilm formation, exopolysaccharide (EPS) production, curli expression, and hypermucoviscosity. The potent extract on K. pneumoniae biofilm was observed with a scanning electron microscope (SEM), while exopolysaccharide topography and surface parameters were observed using atomic force microscopy (AFM). Chemical profiling of the potent extract in vitro was analysed using liquid chromatography-mass spectrometry (LC-MS). Results revealed a noteworthy minimum inhibitory concentration (MIC) value for the C. dimidiatus dichloromethane extract at 0.78 mg/mL on CBR- K. pneumoniae. L. javanica (ethyl acetate) showed the highest cell attachment inhibition (67.25%) for CBR- K. pneumoniae. SEM correlated the in-vitro findings, evidenced by a significant alteration of the biofilm architecture. The highest EPS reduction of 34.18% was also noted for L. javanica (ethyl acetate) and correlated by noticeable changes observed using AFM. L. javanica (ethyl acetate) further reduced hypermucoviscosity to the least length mucoid string (1 mm-2 mm) at 1.00 mg/mL on both strains. C. dimidiatus (aqueous) showed biofilm inhibition of 45.91% for the ESBL-positive K. pneumoniae and inhibited curli expression at 0.50 mg/mL in both K. pneumoniae strains as observed for H. populifolium (aqueous) extract. Chemical profiling of L. javanica (ethyl acetate), C. dimidiatus (aqueous), and H. populifolium (aqueous) identified diterpene (10.29%), hydroxy-dimethoxyflavone (10.24%), and 4,5-dicaffeoylquinic acid (13.41%), respectively, as dominant compounds. Overall, the ethyl acetate extract of L. javanica revealed potent antivirulence properties against the studied MDR K. pneumoniae strains. Hence, it is a promising medicinal plant that can be investigated further to develop alternative therapy for managing K. pneumoniae-associated infections. [ABSTRACT FROM AUTHOR]

Introduction: The emergence of resistance and interference mechanisms to phage infection can hinder the success of bacteriophage-based applications, but the significance of these mechanisms in phage therapy has not been determined. This work studies the emergence of Salmonella isolates with reduced susceptibility to a cocktail of three phages under three scenarios: i) Salmonella cultures (LAB), ii) biocontrol of cooked ham slices as a model of food safety (FOOD), and iii) oral phage therapy in broilers (PT). Methods: S. Typhimurium ATCC 14028 RifR variants with reduced phage susceptibility were isolated from the three scenarios and conventional and molecular microbiology techniques were applied to study them. Results and discussion: In LAB, 92% of Salmonella isolates lost susceptibility to all three phages 24 h after phage infection. This percentage was lower in FOOD, with 4.3% of isolates not susceptible to at least two of the three phages after seven days at 4°C following phage treatment. In PT, 9.7% and 3.3 % of isolates from untreated and treated broilers, respectively, displayed some mechanism of interference with the life cycle of some of the phages. In LAB and FOOD scenarios, resistant variants carrying mutations in rfc and rfaJ genes involved in lipopolysaccharide synthesis (phage receptor) were identified. However, in PT, the significant decrease of EOP, ECOI, and burst size observed in isolates was prompted by lateral gene transfer of large IncI1 plasmids, which may encode phage defense mechanisms. These data indicate that the acquisition of specific conjugative plasmids has a stronger impact than mutagenesis on the emergence of reduced phage-susceptibility bacteria in certain environments. In spite of this, neither mechanism seems to significantly impair the success of Salmonella biocontrol and oral phage therapy. [ABSTRACT FROM AUTHOR]

Background: Iodophors are known to be a treatment for biofilm-related periprosthetic joint infection. However, the efficacy and mechanism of eradicating biofilms from different artificial joint materials after iodophor treatment are unknown. This study was conducted to understand the effect and mechanism of iodophors with respect to the adhesion and virulence of Staphylococcus aureus biofilms attached to artificial joint materials. Methods: Biofilms of Staphylococcus aureus strains were grown on titanium alloy, cobalt chromium molybdenum and polyethylene coupons, which are commonly used materials for artificial joints, for 24 h. Afterward, all coupons were divided into experimental and control groups: (1) exposed to a 0.5 ± 0.05% iodophor for 5 min and (2) exposed to phosphate-buffered saline for 5 min. To gauge the level of biofilm, colony forming units (CFU), live/dead staining confocal microscopy and crystal violet staining were used. Meanwhile, the expression of icaACDR and clfA, which are related to virulence and adhesion, was examined in both the experimental and control groups. Results: A roughly three-log decrease in CFU/cm2 was seen in the viable plate count compared to the control group. Confocal imaging and crystal violet staining verified the CFU data. Moreover, the expression of icaACDR was reduced on three different orthopaedic implant materials, and the expression of clfA was also inhibited on titanium alloy coupons exposed to the iodophor. Conclusions: Our results indicated that exposure to an iodophor for 5 min could significantly eliminate biofilms. When Staphylococcus aureus that had adhered to these three materials, which were used for artificial joints, was treated with an iodophor for 5 min, the expression of icaACDR was significantly reduced. This provides strong evidence for clinically clearing periprosthetic joint infections without removing the artificial joints. [ABSTRACT FROM AUTHOR]

The catalytic activity of monoamine oxidase A (MAO-A) has been linked to tumorigenesis due to the production of reactive oxygen species (ROS) and the resulting oxidative stress. MAO-A inhibition revealed a beneficial role in prostate and lung cancer treatment. This study is aimed at evaluating the effect of different monoamine oxidase A inhibitors (MAO-AIs) on the proliferation and progression of breast cancer cell lines. The cell viability assay was used to evaluate the antiproliferative and combined effects of MAO-AIs. Cell migration was evaluated using wound healing, invasion, and colony formation assays. The underlying mechanism of cell death was studied using flow cytometry. The real-time polymerase chain reaction was used to determine the relative gene expression. Finally, MAO-A activity in breast cancer cells was evaluated using an MAO-A activity assay. According to the results, the examined MAO-AIs significantly inhibited the proliferation of breast cancer cells in a dose-dependent manner. In breast cancer cells, the combination of anticancer drugs (doxorubicin or raloxifene) with MAO-AIs resulted in a synergistic effect. MAO-AIs significantly reduced wound closure and invasion ability in breast cancer cells. Also, MAO-AIs reduced the colony count and size of breast cancer cells. MAO-AIs resulted in significant proapoptotic activity in breast cancer cells. Finally, the MAO-AIs suppressed MAO-A, Bcl-2, and VEGF gene expressions in breast cancer cells relative to untreated cells. This study provides solid evidence supporting the anticancer effect of MAO-A inhibitors in breast cancer cells. [ABSTRACT FROM AUTHOR]

Aerosols produced by dental handpieces represent a permanent risk of disease transmission in the dental environment. The current study evaluated the effects of natural ventilation (open windows) on Streptococcus mutans airborne contamination by dental handpieces in simulated clinical conditions. A dental phantom was placed on a dental chair at a standard university dental clinic operatory. An S. mutans suspension was infused into the phantom's mouth while an operator performed standardized dental procedures using low (contra‐angle) and high speed (turbine) dental handpieces or an ultrasonic scaler, with windows open or closed. Selective medium Petri dishes were placed in 18 sites of the operatory environment to evaluate contamination topographically. Sites were clustered as: wall, floor, ceiling, dental chair, and cabinet. Contamination was expressed as log10CFU/cm2. A linear mixed model analysis was used, nesting the sites within each ventilation and handpiece combination. Open windows significantly reduced contamination. The high‐speed handpiece provided the highest contamination, followed by the ultrasonic scaler and the low‐speed handpiece. Contamination values were much smaller at the ceiling, and much larger at the chair. Opening windows produced more homogeneous contamination of the operatory compared to closed windows. Natural ventilation during dental procedures decreases contamination and affects its topographical distribution. [ABSTRACT FROM AUTHOR]

The study of the microbiota associated to clams is important not only to know their sanitary status but also to prevent pathobiology events. The use of different microbiological techniques can help to obtain a better picture of the bacterial diversity of clams as well as to isolate new bacterial taxa. In this study, two clam species, Ruditapes decussatus and R. philippinarum, were analyzed in two locations of Galicia (northwest of Spain) in April and October, by combining classic culturing, dilution-to-extinction approach, and 16S rRNA gene target sequencing. 16S rRNA gene target sequencing revealed a great diversity within the clam samples, shedding light into the vast microbial communities associated to these bivalves. All samples were dominated by the same bacterial genera in the different periods, namely Mycoplasma, Vibrio, and Cutibacterium. The α-diversity in the samples obtained during the month of October was lower and showed the dominance of rare bacterial taxa, such as Methylobacterium or Psychrobacter. Dilution-to-extinction technique demonstrated its usefulness to culture rare bacterial taxa that were not found in clams under the classic culturing techniques, including Rahnella, Brachybacterium, Micrococcus, Jantinobacter, and Lelliottia. Altogether, our study provides valuable information on the microbiota associated to R. decussatus and R. philippinarum, demonstrating the high complexity and dynamics of these microbial populations. [ABSTRACT FROM AUTHOR]

Objective. To determine the diagnostic performance of blood culture positivity times for distinguishing true bacteremia from contaminants in the automated "BACT/ALERT®" system. Materials and methods. A cross-sectional, diagnostic test-type study was conducted from a database of blood culture samples processed between January 2016 and August 2021. All blood culture samples from patients with suspected bacteremia were included; blood culture samples were entered into the "BACT/ALERT®" system to differentiate true bacteremia from contaminants. Results. We obtained 33,951 blood cultures samples, of which 3875 were positive. Of the total number of positive blood cultures, 75.2% (n=2913) were true bacteremia and 24.8% (n=962) were contaminants. The median time to positivity in blood cultures with true bacteremia was significantly shorter (16.3 hours; IQR: 11.2 - 24.9) than the median time to positivity of blood cultures with contaminants (22.5 hours; IQR: 18.4 - 31.8; p<0.001). The positivity time showed the capacity to differentiate true bacteremia from contaminants, with an AUC-ROC of 0.73 (95%CI: 0.71 - 0.75), with 85% and 63% sensitivity and specificity respectively for the diagnosis of contaminants when the positivity time exceeds 16.5 hours. The use of antibiotics prior to sampling delayed the time to positivity, while having fever before sampling shortened the time to positivity. Conclusions. Our results show good diagnostic performance of blood culture positivity times to differentiate true bacteremia from contaminants using the "BACT/ ALERT®" system when the positivity time was longer than 16.5 hours. [ABSTRACT FROM AUTHOR]

Background: The lungs are enveloped by a serous membrane called visceral pleura, while the chest wall and pericardium are covered by parietal pleura. Lung hila connect these layers, and a small amount of fluid in the pleural space lubricates movement. Pleural effusion, excess fluid in the pleural cavity, indicates underlying pulmonary, pleural, or extra-pulmonary diseases. Objective: To study the various causes of unilateral pleural effusion in patients presenting to a District General Hospital in the United Kingdom. Methodology: This was a prospective study conducted at Worsecter Royal Infirmary, UK, during July 2021 to June 2022. All (58) adult patients (age above 18 years) admitted to chest ward from various sources with unilateral pleural effusion were included. Case notes were reviewed and data collected. Diagnostic pleural tap was performed in every case and aspirate was sent for biochemical, microbiological and cytological analysis. Pleural biopsies were taken where diagnosis could not be established with the help of other investigations. Results: Among 58 patients with unilateral pleural effusion (34 men, 24 women), most were aged 50-89 (mean age 69). Among study cases, 16% were referred for surgery (VATS or thoracotomies), with an average 11-day waiting period. Effusions were transudate (15) or exudate (43), and malignancy was confirmed in 31 cases. Complications included intercostal drain dislodgement (19%) and subcutaneous emphysema (12%). Malignancy was the most common cause of unilateral pleural effusion, followed by parapneumonic effusion, left ventricular failure, TB, and asbestos-related benign effusion. Conclusion: In conclusion, this study highlights malignancy as the primary cause of unilateral pleural effusion, confirmed through cytology and biopsies. Managing these patients is complex, involving prolonged hospitalization, multiple thoracocentesis attempts, and often prompt surgical intervention. Imaging, especially contrast CT chest, is crucial for lesion identification. Complications like drain dislodgement and subcutaneous emphysema underscore the challenges in pleural effusion management. [ABSTRACT FROM AUTHOR]

We aimed to investigate the association between the presence of cutaneous urease‐producing bacteria and the development of incontinence‐associated dermatitis (IAD) using an original urea agar medium as a step toward developing advanced preventive measures. In previous clinical assessments, we developed an original urea agar medium to detect urease‐producing bacteria via the medium's colour changes. In a cross‐sectional study, specimens were collected via the swabbing technique at genital skin sites in 52 stroke patients hospitalised in a university hospital. The primary objective was to compare the presence of urease‐producing bacteria between the IAD and no‐IAD groups. Determining the bacterial count was the secondary objective. The prevalence of IAD was 48%. A significantly higher detection rate of urease‐producing bacteria was observed in the IAD group than in the no‐IAD group (P =.002) despite the total number of bacteria being equivalent between them. In conclusion, we discovered that there was a significant association between the presence of urease‐producing bacteria and IAD development in hospitalised stroke patients. [ABSTRACT FROM AUTHOR]

Interdisciplinary studies on cultural heritage artworks provide efficient solutions to control fungal growth and the negative effects of biodeterioration. In this study, we aimed to identify the population of filamentous fungi colonizing an engraving by the Dutch painter Rembrandt, whose conservation status was compromised and showed visible stains of biodeterioration. Microbiological techniques, such as cultivation‐dependent approaches and molecular biology, have been used to identify fungal populations. In addition, the anaerobic atmosphere technique and eco‐friendly antifungal agents, such as essential oils (EOs) of Curcuma longa, Thymus vulgaris, and Melaleuca alternifolia, were tested against the metabolically active fungal population Cladoposporium spinulosum. Furthermore, in vitro assays revealed that the interaction between the fungal strains and EO was positive, inhibiting the growth of these fungi, and the EOs from T. vulgaris and M. alternifolia showed low minimum inhibitory concentration values. Exposure to anaerobic conditions for 35 days was effective in the total elimination of isolated fungal strains. In conclusion, this study demonstrated the effectiveness of a nondestructive technique for artwork on engraving colonized by fungal strains and using EO as an alternative to toxic antifungals used in conventional treatments in artworks. Thus, this interdisciplinary study involving applied microbiology and botanical and preventive conservation presents a tool to control microbial growth while maintaining artwork integrity. [ABSTRACT FROM AUTHOR]

Manufacturers' desire to sell "healthy" food in response to the consumers' desire to lead a healthy lifestyle has increased the use of probiotics during the past few decades. Probiotics are used in dairy products, as well as non-dairy items as a starter culture, encompassing a wide range of goods. Numerous phenotyping, physical characterisation, and genotyping techniques have been developed to identify probiotic lactobacilli to ensure quality management. These techniques are frequently precise enough to categorise probiotic strains by genus and species. Traditional microbiological methods were initially employed for genus and species identification. However, due to their numerous shortcomings as the probiotic ability is often strain-dependent, and that there is no way to differentiate between strains using simple microbiological techniques, new methods that are mostly based on the examination of nucleic acids have been developed. Therefore, the objective of the present review was to provide critical assessment on existing methods for identifying members of the genus Lactobacillus, together with newly discovered approaches. The present review aimed to give the most recent information on the scientific techniques used to measure and describe the possible probiotic properties of microorganisms. It will also emphasise molecular and non-molecular tools. Most of these tools are based on 16S ribosomal DNA sequencing, and employ PCR techniques. [ABSTRACT FROM AUTHOR]

Multidrug-resistance (MDR) strains of Pseudomonas aeruginosa has resulted in the medical therapy complicated. Antimicrobial resistance and biofilm production are virulence factors responsible for the persistence of prolonged infections. In this study, antimicrobial susceptibility pattern and phenotypic characteristics of biofilm of P. aeruginosa were investigated. The aim and objective of this study is to evaluate the antibacterial susceptibility testing and the presence of Biofilm Formation Potential among clinical isolates of Pseudomonas aeruginosa at a Tertiary care centre. This was a cross sectional study carried out in the Department of Microbiology, Rama Medical College Hospital and Research Centre, Mandhana, Kanpur for a period of 1 year i.e, between March 2022 to March 2023. A Total of 355 clinical isolates from different clinical samples including urine, sputum, blood, Et secreation, pus, throat swab were identified by Standard microbiological techniques and the isolates were further tested for the detection of biofilm along with the MBL by Imipenem-EDTA combined disc test and MBL Etest (Imipenem) according to the CLSI guidelines 2021. A total of 355 clinical samples were studied out of which 132 P. aeruginosa isolates were obtained with the prevalence rate of 37.1% with the males 75(56.8%) being affected the most as compared to the females 57 (43.1%). The age group of 21-30 years of age followed by 31-40 was affected the most. In the age group of 0-10 years and above 71 years was the least affected with the Pseudomonas aeruginosa infection. It was also observed that the maximum number of isolates observed were from the Urine sample and least was from the blood and throat swab. The Sensitivities of Colistin was (96.9%), Piperacillin tazobactam (73.4%), Amikacin (77.2 %) and cefepime (73.4%) were found to be the most effective Antibiotics. The resistance to ciprofloxacin was (42.5%), Levofloxacin (50%), Gentamicin(63.6%), Imipenem (63.6%), Tobramycin( 69.6%), Ceftazidime (69.6%). Of which 20(15.1%%) were MBL positive by Imipenem-EDTA combined disc test, and 19(20.45%) by MBL E- test. It was observed that 50.75% of the Pseudomonas aeruginosa isolates were Biofilm producers with 1.5% strong and 3.7% were moderate biofilm producers. In conclusion, the present study showed combination therapy including an anti-pseudomonal beta-lactamand an aminoglycoside along with anti-biofilm agents can also be selected to eradicate biofilm-associated Pseudomonal infections, with strict infection control practices. MBL E-Test was found to be the best phenotypic method for detection of MBL is recommended into the daily routine of clinical laboratory. [ABSTRACT FROM AUTHOR]

The article discusses the challenges in diagnosing pneumonia in immunocompromised patients and the potential of plasma cell-free DNA metagenomic sequencing (mcfDNA) to enhance diagnostic capabilities. Topics include the limitations of traditional microbiologic methods, the use of mcfDNA sequencing for detecting pneumonia pathogens, and the results and implications of the PICKUP study, which evaluated the diagnostic value of mcfDNA sequencing in patients with hematologic cancer.

The augmentation mammoplasty with prosthesis implantation is the most frequent aesthetic surgical intervention. One of the most frequent complications associated to this surgery is the bacteriological contamination of the implants. In order to avoid it, different protocols like the 14-points have been proposed. One of the measures included is the avoidance of the aerial exposure of the prosthesis before implantation. However, the risk of contamination without direct contact has not been well-established. Our aim has been to evaluate the risk of aerial contamination breast prostheses during the surgical procedure. During the course of the surgery of augmentation mammoplasty with prosthesis implantation, we rubbed a hyssop over the prosthesis before the full unwrapping; these samples were used as the control group. Afterwards, the prostheses were irrigated with either saline serum or povidone-iodine and exposed to air in the operating room. New samples were taken with a hyssop after 2 and 5 min. All these samples were cultured in the microbiology lab. In total, 36 prostheses, microtexturized and smooth ones, were analyzed. All microbiological cultures were negative at 0′, 2′, and 5′, regardless of the implant texture and the irrigation fluid used. There is no microbial contamination of the breast implants during the course of the surgical intervention preimplantation due to aerial exposure, and there might be therefore other different sources of prosthesis contamination. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the effects of 0.2% sodium hypochlorite, Efferdent (Prestige Consumer Healthcare), and 6.25% Ricinus communis on biofilm removal and antimicrobial action on dentures and brushes using nonimmersion or immersion protocols for the brushes.Materials and Methods: A total of 45 denture wearers were randomly assigned to a denture immersion protocol for 7 days: 0.85% saline solution for 20 minutes (control); 0.2% sodium hypochlorite for 20 minutes (SH); Efferdent for 3 minutes; or 6.25% Ricinus communis for 20 minutes (RC). The participants were also randomized to immersion (n = 23) or no immersion (n = 22) of their brushes with their dentures in the same solutions. For biofilm evaluation, the dentures were stained and photographed, and the area of the biofilm was measured using Image Tool 3.0 (University of Texas Health Science Center). To evaluate microbial load on dentures and brushes, the biofilm was collected, and the Candida spp and Streptococcus mutans colonies were counted.Results: The SH, Efferdent, and RC groups showed reduced biofilm and Candida spp on dentures regardless of the immersion protocol for the brushes. However, no difference was found in the Candida spp counts collected from the brushes immersed compared to the brushes not immersed in the solutions. The SH and Efferdent groups showed reduced S mutans on both dentures and brushes, except for in the nonimmersion subgroups.Conclusion: All solutions reduced denture biofilm and microbial load. However, immersion of brushes in the solutions did not contribute to reducing the microbial load. [ABSTRACT FROM AUTHOR]

Anaerobic gut fungi (Neocallimastigomycetes) live in the digestive tract of large herbivores, where they are vastly outnumbered by bacteria. It has been suggested that anaerobic fungi challenge growth of bacteria owing to the wealth of biosynthetic genes in fungal genomes, although this relationship has not been experimentally tested. Here, we cocultivated the rumen bacteria Fibrobacter succinogenes strain UWB7 with the anaerobic gut fungi Anaeromyces robustus or Caecomyces churrovis on a range of carbon substrates and quantified the bacterial and fungal transcriptomic response. Synthetic cocultures were established for at least 24 h, as verified by active fungal and bacterial transcription. A. robustus upregulated components of its secondary metabolism in the presence of Fibrobacter succinogenes strain UWB7, including six nonribosomal peptide synthetases, one polyketide synthase-like enzyme, and five polyketide synthesis O-type methyltransferases. Both A. robustus and C. churrovis cocultures upregulated S-adenosyl-l-methionine (SAM)-dependent methyltransferases, histone methyltransferases, and an acetyltransferase. Fungal histone 3 lysine 27 trimethylation marks were more abundant in coculture, and heterochromatin protein-1 was downregulated. Together, these findings suggest that fungal chromatin remodeling occurs when bacteria are present. F. succinogenes strain UWB7 upregulated four genes in coculture encoding drug efflux pumps, which likely protect the cell against toxins. Furthermore, untargeted nonpolar metabolomics data revealed at least one novel fungal metabolite enriched in coculture, which may be a defense compound. Taken together, these data suggest that A. robustus and C. churrovis produce antimicrobials when exposed to rumen bacteria and, more broadly, that anaerobic gut fungi are a source of novel antibiotics. IMPORTANCE Anaerobic fungi are outnumbered by bacteria by 4 orders of magnitude in the herbivore rumen. Despite their numerical disadvantage, they are resilient members of the rumen microbiome. Previous studies mining the genomes of anaerobic fungi identified genes encoding enzymes to produce natural products, which are small molecules that are often antimicrobials. In this work, we cocultured the anaerobic fungus Anaeromyces robustus or Caecomyes churrovis with rumen bacteria Fibrobacter succinogenes strain UWB7 and sequenced fungal and bacterial active genes via transcriptome sequencing (RNA-seq). Consistent with production of a fungal defense compound, bacteria upregulated genes encoding drug efflux pumps, which often export toxic molecules, and fungi upregulated genes encoding biosynthetic enzymes of natural products. Furthermore, tandem mass spectrometry detected an unknown fungal metabolite enriched in the coculture. Together, these findings point to an antagonistic relationship between anaerobic fungi and rumen bacteria resulting in the production of a fungal compound with potential antimicrobial activity.

Objective: To evaluate the effect of different finishing and polishing procedures on surface roughness and microbial adhesion and viability of Streptococcus mutans on novel highly‐filled composites for injectable mold technique. Materials and Methods: One hundred sixty specimens were divided into four material groups: FSF (Filtek Supreme Flowable Restorative), TE (Tetric EvoFlow), GUI (G‐aenial Universal Injectable) and GUF (G‐aenial Universal Flo). Within each group, specimens were split regarding finishing and polishing (F/P) procedures into: SLD (Sof‐Lex Discs), SLS (Sof‐Lex Spirals), OG (OneGloss) and PG (PoGo). Surface roughness was analyzed using profilometer (Ra and Rz) and scanning electron microscopy (SEM) and Strep. mutans biofilm formation was analyzed using colony forming unit (CFU) and cell viability assay. Two‐way analysis of variance (ANOVA) followed by Tukey's post hoc test were used for comparison among groups, Pearson's coefficient was applied for the correlation between Ra and CFU/ml and all data were presented as mean ± SD. Results: Both materials and F/P procedures affect Ra, Rz and Strep. mutans CFU/ml values (p ≤ 0.05). Considering the Ra and Rz, GUI and GUF revealed lower values, compared to FSF and TE and SLD and SLS revealed lower values, compared to OG and PG. Considering the Strep. mutans CFU/ml and viability, GUI and TE presented lower values, compared to GUF and FSF and SLD and SLS presented lower values, compared to OG and PG. Moderate positive correlation was found between Ra and CFU/ml (r = 0.552). Conclusions: The smoothest surfaces possess GUI and GUF, among materials and SLD and SLS, among F/P procedures. GUI adhered the lowest amount of Strep. mutans, due to the smoothest surfaces. FSF and GUF revealed the highest amount of Strep. mutans, due to bis‐GMA, bis‐MEPP and TEGDMA in their composition. Clinical Significance: The findings of the present study may be beneficial for the proper selection of highly‐filled composites and an adequate finishing and polishing procedure when performing the injectable mold composite resin veneer technique. [ABSTRACT FROM AUTHOR]

Objectives: Air pollutants in dentistry offices may cause problems for the health of staff and patients. Accordingly, the present study was performed to the assessment of bacterial bioaerosols and particulate matter (PM) characteristics in the indoor air of dentistry clinics. Methods: The sampling points were restoration, endodontics, and prosthesis wards. The PM specimens were taken using Grimm Dust Monitor and the microbial specimens were taken using QuickTake 30. Results: The maximum and minimum PM10 concentration across the different measure wards occurred as 70.31 and 30.32 μg/m3 in the Ketabchi Clinic restoration and its endodontics ward, respectively. The total number of bioaerosols was 2079. Gram‑positive Staphylococcus spp.(872), Gram‑positive Micrococcus spp.(706), and Gram‑negative Bacillus spp.(501) were present in the indoor air. The results showed a significant relationship for PM1 and PM2.5 in each sampling site. Conclusion: The low PM and bacterial bioaerosols contamination can be due to the coincidence with coronavirus pandemic, as during this period, health and hygienic protocols have been strictly observed. [ABSTRACT FROM AUTHOR]

Context: Infective endocarditis (IE) is a dynamic disease with high morbidity and mortality despite advancements in diagnostics, antimicrobial therapy, and surgery. India faces a complex situation due to changing factors such as host characteristics, diagnostic techniques, antibiotics, and aggressive surgical approaches. Monitoring local epidemiology is crucial for prompt recognition and treatment. However, studies on IE trends in India are scarce and outdated. Aims: To investigate the clinical, microbiological, predisposing conditions, echocardiographic characteristics, and treatment outcomes of IE patients admitted to a Tertiary Care Academic Institute in Western India. Settings and Design: This retrospective observational study was done in a 2011-bedded tertiary care referral hospital in Western Maharashtra, between June 2019 and May 2023. The records of all adult patients (age >14 years) admitted during the study period with a definite diagnosis of IE, both native valve endocarditis and prosthetic valve endocarditis according to the modified Duke criteria, were included in the study. Baseline demographic data on age, sex, underlying heart disease, and predisposing conditions for bacteremia (infections, central venous catheters, surgeries, and invasive procedures) were collected. Subjects and Methods: The echocardiographic and microbiological findings, complications, treatment, and outcomes were noted. All blood cultures done for the diagnosis of IE were processed using standard methods to identify bacterial and fungal species and antimicrobial susceptibility profiles. For all the patients in this study, blood (5-8 mL per bottle) was collected in adult blood culture bottles using standard precautions and processed by a semiautomated blood culture system (BioMérieux, Marcyľ Etoile). Identification of causative organisms was performed by standard microbiological methods. Transthoracic and transesophageal echocardiography were performed to determine the location of vegetation, type of valve infected, and cardiac complications. Sample Size Calculation: Considering the Indian prevalence of IE to be 14.5% as observed by Gupta et al., margin of error as 9%, and 95% of confidence level, the minimum required sample size calculated using WinPepi software (version: 11.65) for conducting the study was 59 patients. Statistical Analysis: Data was collected and entered into a database using Microsoft Excel. Data was represented as the frequency with percentage and Chi-square test (two-sample proportionality test) were applied for comparison of categorical variables. Quantitative data was summarized using mean and standard deviation and a two-sample t-test was applied to compare differences. P < 0.05 was considered statistically significant. Results: The mean age in the study was 44.1 ± 10.6 years with a female preponderance (male:female - 0.85:1). Rheumatic heart disease (RHD) (44.3%) was the most common underlying heart disease etiology, whereas 33% of patients did not have any cardiac disease. Blood culture was positive in 34.9% of episodes, with the most common organisms being Gram-positive Staphylococci (38%). Medical complications such as congestive heart failure (CHF) and acute kidney injury were seen in 41.2% of patients, followed by cerebrovascular accident and stroke in 20.6%. The majority of patients, 61.9%, had received antibiotics before referral to our center. Thirty-seven (58.7%) patients underwent definitive surgery for IE. The total in-hospital mortality rate was 12 (19.1%). In the univariate analysis, younger patients (<50 years) with CHF, diabetes mellitus, and Staphylococcus culture-positive IE were independent predictors of mortality. Conclusions: Over the past four decades, the landscape of IE in India has experienced noteworthy transformations. The disease profile of patients has shifted from predominantly older individuals with RHD to a rising number of younger patients with prosthetic valves or those without identified valvular conditions. Moreover, the yield of blood cultures has declined due to the increased utilization of antibiotics over the last two decades, although Staphylococcus species continue to be the most common organisms detected. Despite significant advancements in medical technology, the mortality rate remains unchanged. Our study provides compelling evidence to advocate for early surgical intervention, which holds the potential for improved outcomes. [ABSTRACT FROM AUTHOR]

An editorial is presented on the transformative potential of metagenomics in microbiology, discussing advancements and implications. Topics discussed include the historical evolution of microbiological diagnostics, the role of metagenomic next-generation sequencing (mNGS) in clinical diagnostics, opportunities, and challenges associated with mNGS, and the potential for implementing mNGS on a national scale in Australia.

Candida albicans is the most common organism responsible for both mucosal and systemic infections, accounting for approximately 70% of fungal infections worldwide. Miana, scientifically known as Coleus scuttellarioides (L.) Benth., is recognized for its use in traditional medicinal practices. Miana plants contain endophytic bacteria that possess the ability to produce secondary metabolites with potential antifungal agents. The objective of this study was to assess the antifungal activity of nine endophytic bacteria isolates derived from Coleus scuttellarioides against Candida albicans. This study was laboratory-based qualitative experimental research that applied the Kirby-Bauer diffusion method and several modifications. The Candida albicans specimens were spread throughout the entire potato dextrose agar medium. Afterwards, paper discs that had been soaked in a liquid culture of endophytic bacterial isolates were carefully placed on the surface of the medium. The complete setup was then incubated for 1-2 days. The potential antifungal activity of endophytic bacteria was assessed by observing the emergence of a clear zone surrounding their growth, which would indicate inhibition. An additional observation was performed in the follow-up test, involving the use of Sabouraud dextrose agar medium to confirm the initial test result. The results from the inhibitory test revealed that none of the bacterial isolates exhibited any inhibition zone. Conversely, ketoconazole as the positive control showed an inhibition zone with an average diameter of 28.5 mm. In conclusion, endophytic bacterial isolates obtained from Coleus scuttellarioides have no discernible antifungal properties against Candida albicans. This study implies that ketoconazole remains effective in treating infections caused by Candida albicans. [ABSTRACT FROM AUTHOR]

Introduction: beta-lactamase-producing bacteria, especially extended-spectrum beta-lactamase (ESBL) producers have strong clinical relevance and have been implicated in chronic suppurative otitis media (CSOM) treatment failures. This study aimed to determine the frequency, antibiogram, and molecular characteristics of ESBL-producing gramnegative bacterial (GNB) pathogens isolated from patients with CSOM. Methods: three hundred (300) ear swab samples collected from patients with active CSOM were analysed using standard microbiological techniques. Antibiogram of pathogens was determined by Kirby-Bauer disk diffusion technique. Phenotypic detection and molecular characterization of ESBL-producing GNB pathogens were performed by double disk synergy test (DDST) and polymerase chain reaction (PCR). Results: Escherichia coli and P. aeruginosa were more prevalent among CSOM patients with a duration of discharge >2 weeks. The frequency of ESBL producers among the GNB pathogens was 18.3%. Isolates were generally multidrug-resistant but very susceptible (100% - 70.4%) to ciprofloxacin, imipenem, and amikacin. Multiple antibiotic resistance values of the isolates ranged from 0.7-0.8. Polymerase chain reaction showed that blaSHV (47.6%) was the most predominant ESBL genotype. This was followed by blaTEM (25.2%) and blaCTX-M (10.7%) as the least predominant ESBL gene. Concomitant expression of ESBL gene was observed in 13.6% of the isolates. Conclusion: this study reported the occurrence and spread of β-lactamase-producing bacteria in patients with CSOM infections. It is therefore very crucial to screen for antibiotic-resistant pathogens at early stages of CSOM infections, for proper antimicrobial therapy and to curb the increasing spread of antimicrobial resistance. [ABSTRACT FROM AUTHOR]

This study seeks to determine the feacal carriage of extended spectrum beta-lactamase and fluoroquinolone resistant non-typhoidal Salmonella enterica isolates from food-producing animals and humans. A total of three hundred (300) fecal samples were collected using sterile universal containers from food-producing animals namely (Chicken [100], Pig [100] and humans (100) from Onicha Local Government Area of Ebonyi State and analyzed for the presence of non-typhoidal Salmonella enterica using standard microbiological techniques. Phenotypic detection of extendedspectrum beta-lactamase (ESBL) were done by disc diffusion and Double Disk Synergy Test. Molecular characterization for ESBL and fluoroquinolone-resistant genes were done by PCR with specific primers. The result shows that non-typhoidal Salmonella species (NTS) accounted for 25 % and 17 % in poultry and pig fecal sample respectively while 60 % and 40% were phenotypic ESBL producers respectively. When compared statistically there is significant difference among isolates confirmed ESBL-positive (P< 0.05). Also, none of the 16 (58 %) NTS isolated from humans harbored ESBL phenotype. PCR analysis with β-lactam specific primer detected the presence of blaOXA 50 % and 50 %, blaSHV 36 %, and 64 %, blaTEM 43 % and 57 %, blaCTX-M 36 % and 64 % in poultry and pig respectively. Fluoroquinolone resistant gene QnrA was present in 0 and 100 % of poultry and pig respectively. QnrB was 40 % and 60 % in poultry and pig isolates respectively. QnrS was present in 64 % isolates of poultry and 13 % isolates in pig. The high prevalence of genes encoding beta-lactamases and fluoroquinolone resistance (TEM, SHV, CTX-M and OXA, (qnrA, qnrB and qnrS) were present more in poultry and pig than in humans and demonstrate a significant public health threat from consumption of food-producing animal harboring such pathogenic resistant genotype if not properly controlled. [ABSTRACT FROM AUTHOR]

The current study determines the density of Vibrio spp. and isolates V. cholerae and Vibrio mimicus from fish-anatomical-sites, prawn, crab and mussel samples recovered from fish markets, freshwater and brackish water. Virulence and antibiotic resistance profiling of isolates were carried out using standard molecular and microbiology techniques. Vibrio spp. was detected in more than 90% of samples [134/144] and its density was significantly more in fish than in other samples. Vibrio. cholerae and V. mimicus were isolated in at least one sample of each sample type with higher isolation frequency in fish samples. All the V. cholerae isolates belong to non-O1/non-O139 serogroup. One or more V. cholerae isolates exhibited intermediate or resistance against each of the eighteen panels of antibiotics used but 100% of the V. mimicus were susceptible to amikacin, gentamycin and chloramphenicol. Vibrio cholerae exhibited relatively high resistance against polymyxin, ampicillin and amoxicillin/clavulanate while V. mimicus isolates exhibited relatively high resistance against nitrofurantoin, ampicillin and polymixin. The multiple-antibiotic-resistance-index [MARI] for isolates ranges between 0 and 0.67 and 48% of the isolates have MARI that is >0.2 while 55% of the isolates exhibit MultiDrug Resistance Phenotypes. The percentage detection of acc, ant, drf18, sul1, mcr-1, blasvh, blaoxa, blatem, blaoxa48, gyrA, gyrB and parC resistance-associated genes were 2%, 9%, 14%, 7%, 2%, 25%, 7%, 2%, 2%, 32%, 25% and 27% respectively while that for virulence-associated genes in increasing other was ace [2%], tcp [11%], vpi [16%], ompU [34%], toxR [43%], rtxC [70%], rtxA [73%] and hyla [77%]. The study confirmed the potential of environmental non-O1/non-O139 V. cholerae and V. mimicus to cause cholera-like infection and other vibriosis which could be difficult to manage with commonly recommended antibiotics. Thus, regular monitoring of the environment to create necessary awareness for this kind of pathogens is important in the interest of public health. [ABSTRACT FROM AUTHOR]

Background. In recent times, cervical dysbiosis which mostly causes and aggravates infections is highlighted for its role in immune modulation in cervical dysplasia, which promotes the shifting of Th1 phenotype immunity to Th2 phenotype immunity. This study therefore estimated and compared the levels of circulatory IL-4, IL-6, IL-10, TNF-α, and IFN-γ cytokines among adult women identified to have different grades of cervical intraepithelial neoplasia (CIN) and with cervicovaginal infection. Methods. A total of 157 participants were recruited from the Akyemansa District of Ghana, and cervical swabs and blood samples were taken. The Pap smear test, microbiological culture, and ELISA were employed for cytology analysis, bacteria isolation, and identification and estimation of IL-4, IL-6, IL-10, TNF-α, and IFN-γ cytokines, respectively. Results. Overall, 14/157 (8.9%) had CIN with 7.6% having CIN 1 and 1.3% having CIN 2. The main predictor for CIN was age above 46 years (OR 11.16, 95% CI: 2.4-51.8). Bacterial vaginosis (p = 0.003) and Candida infection (p = 0.012) were significantly higher in CIN. Again, Staphylococcus aureus (60% vs. 17.6%, p = 0.005), Citrobacter sp. (40.0% vs. 13.2%, p = 0.017), and Morganella morganii (40.0% vs. 4.4%, p = 0.002) isolates were significantly higher in CIN-positive participants. IL-10 and TNF-α concentrations were elevated in participants with CIN 1+ (TNF-α NIL vs. CIN 1+ only, p < 0.05) while IL-6 was decreased among participants with CIN 1+. In the presence of vaginal infection, TNF-α decreased among CIN 1+ participants while IL-10 remained elevated. Conclusion. The findings of this study suggest that cervical dysbiosis causes immune suppression, which creates a suitable microenvironment for the development of CIN. [ABSTRACT FROM AUTHOR]

Many grape endophytic microorganisms exhibit high potential for suppressing the development of grape diseases and stimulating grapevine growth and fitness, as well as beneficial properties of the crop. The microbiome of wild grapevines is a promising source of biocontrol agents, which can be beneficial for domesticated grapevines. Using next-generation sequencing (NGS) and classical microbiology techniques, we performed an analysis of bacterial and fungal endophytic communities of wild grapevines Vitis amurensis Rupr. and Vitis coignetiae Pulliat growing in the Russian Far East. According to the NGS analysis, 24 and 18 bacterial taxa from the class level were present in V. amurensis and V. coignetiae grapevines, respectively. Gammaproteobacteria (35%) was the predominant class of endophytic bacteria in V. amurensis and Alphaproteobacteria (46%) in V. coignetiae. Three taxa, namely Sphingomonas, Methylobacterium, and Hymenobacter, were the most common bacterial genera for V. amurensis and V. coignetiae. Metagenomic analysis showed the presence of 23 and 22 fungi and fungus-like taxa of class level in V. amurensis and V. coignetiae, respectively. The predominant fungal classes were Dothideomycetes (61–65%) and Tremellomycetes (10–11%), while Cladosporium and Aureobasidium were the most common fungal genera in V. amurensis and V. coignetiae, respectively. A comparative analysis of the endophytic communities of V. amurensis and V. coignetiae with the previously reported endophytic communities of V. vinifera revealed that the bacterial biodiversity of V. amurensis and V. coignetiae was similar in alpha diversity to V. vinifera's bacterial biodiversity. The fungal alpha diversity of V. amurensis and V. coignetiae was statistically different from that of V. vinifera. The beta diversity analysis of bacterial and fungal endophytes showed that samples of V. vinifera formed separate clusters, while V. amurensis samples formed a separate cluster including V. coignetiae samples. The data revealed that the endophytic community of bacteria and fungi from wild V. amurensis was richer than that from V. coignetiae grapes and cultivated V. vinifera grapes. Therefore, the data obtained in this work could be of high value in the search for potentially useful microorganisms for viticulture. [ABSTRACT FROM AUTHOR]

Background: Laminar airflow (LAF) technologies minimize infectious microorganisms to enhance air quality and surgical site infections (SSIs). LAF lowers SSIs in some clinical studies but not others. This study analyzes laminar airflow ventilation's capacity to reduce orthopaedic surgery-related SSIs. Methods: The PRISMA-compliant keywords were utilized to conduct a search for pertinent articles in various databases including PubMed, MEDLINE, CENTRAL, Web of Sciences, and the Cochrane databases. Observational studies, including retrospective, prospective, and cohort designs, satisfy the PICOS criteria for research methodology. The assessment of quality was conducted utilizing the Robvis software, while the meta-analysis was performed using the RevMan application. The study's results were assessed based on effect sizes of odds ratio (OR) and risk ratio (RR). Results: From 2000 to 2022, 10 randomized controlled clinical trials with 10,06,587 orthopaedic surgery patients met the inclusion criteria. The primary outcomes were: (1) Risk of SSI, (2) Bacterial count in sampled air and (3) Reduction in SSIs. The overall pooled OR of all included studies was 1.70 (95% CI 1.10–2.64), and the overall pooled RR was 1.27 (95% CI 1.02–1.59) with p < 0.05. LAF is ineffective at preventing SSIs in orthopaedic procedures due to its high-risk ratio and odds ratio. Conclusions: The present meta-analysis has determined that the implementation of LAF systems does not result in a significant reduction in the incidence of surgical site infections (SSIs), bacterial count in the air, or SSIs occurrence in orthopaedic operating rooms. Consequently, the installation of said equipment in operating rooms has been found to be both expensive and inefficient. [ABSTRACT FROM AUTHOR]

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Background: Thorough disinfection of root canals in primary molars may be complicated by the complex root canal morphology. This in-vitro study aimed to compare direct and residual antimicrobial effect of 2% chlorhexidine (CHX) gel, 500 mg/ml double antibiotic paste (DAP) and chitosan-chlorhexidine nanoparticles (CS-CHX NPs) as intracanal medicaments against Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) in primary molars. Methods: Mesial roots of 63 mandibular second primary molars were infected with E. faecalis and C. albicans. Teeth were divided into 9 groups: Ia: (CS-CHX NPs), IIa: (CHX), IIIa: (DAP), IVa: chitosan nanoparticles (CSNPs) in which medicaments were placed for 3 days, groups Ib: (CS-CHX NPs), IIb: CHX, IIIb: (DAP), IVb: (CSNPs) in which medicaments were placed for 7 days, and Group V (control): teeth were infected, irrigated with saline, and sampled 3- and 7-days post-infection. Microbiological samples were obtained after infection, 3, and 7 days after medicament placement and 7 days after medicament removal for both time points. One-way ANOVA, Tukey's post hoc test and paired t-test were used at p < 0.05. Results: CS-CHX NPs had the highest anticandidal effect which was comparable to CHX and significantly higher than other medicaments (p < 0.001) at both time points. CS-CHX NPs had the highest effect against E. faecalis which was comparable to DAP and significantly higher than other medicaments (p < 0.001) at 3 days. All medicaments showed similar effect against E. faecalis after 7 days. The 7-days placement significantly increased the antimicrobial effect against both micro-organisms in all groups, except CS-CHX NPs which showed an insignificant increase. CS-CHX NPs showed the highest residual effect against both micro-organisms that increased with 7-days placement. Conclusion: CSNPs and CHX combination showed a synergistic effect against both micro-organisms. CS-CHX NPs displayed a higher effect at a shorter period compared to other medicaments, yet its residual effect was higher with 7-days placement. [ABSTRACT FROM AUTHOR]

Simple Summary: Mastitis is one of the most common diseases in the dairy industry in the world, and its treatment cost is also the most expensive of all diseases in the dairy industry. The research on the rapid diagnosis of cow mastitis is of great significance to the development of the dairy cow industry all over the world. In this study, we use microbiology and metabolomics techniques to explore the differential microbiota and metabolites in cow milk at different somatic levels, to find new biomarkers for the diagnosis of cow mastitis and provide a new plan for the diagnosis, prevention, and treatment of mastitis in the future. Dairy cow mastitis is one of the common diseases of dairy cows, which will not only endanger the health of dairy cows but also affect the quality of milk. Dairy cow mastitis is an inflammatory reaction caused by pathogenic microorganisms and physical and chemical factors in dairy cow mammary glands. The number of SCC in the milk of dairy cows with different degrees of mastitis will increase in varying degrees. The rapid diagnosis of dairy cow mastitis is of great significance for dairy cow health and farm economy. Based on the results of many studies on the relationship between mastitis and somatic cell count in dairy cows, microflora, and metabolites in the milk of Holstein cows with low somatic cell level (SCC less than 200,000), medium somatic cell level (SCC up to 200,000 but less than 500,000) and high somatic cell level (SCC up to 5000,00) were analyzed by microbiome and metabolic group techniques. The results showed that there were significant differences in milk microbiota and metabolites among the three groups (p < 0.05), and there was a significant correlation between microbiota and metabolites. Meanwhile, in this experiment, 75 differential metabolites were identified in the H group and L group, 40 differential metabolites were identified in the M group and L group, and six differential microorganisms with LDA scores more than four were found in the H group and L group. These differential metabolites and differential microorganisms may become new biomarkers for the diagnosis, prevention, and treatment of cow mastitis in the future. [ABSTRACT FROM AUTHOR]