Matrix-assisted Laser Desorption/ionization Time-of-flight Research Articles

Prevalence of Contagious Mastitis Pathogens in Bulk Tank Milk from Dairy Farms in Lower Saxony, Germany

The aim of this study was to determine the prevalence of Streptococcus (Sc.) agalactiae, Prototheca spp., Staphylococcus (S.) aureus, and especially methicillin-resistant S. aureus as well as Myco-plasmopsis (M.) spp. and M. bovis in bulk tank milk (BTM) on dairy farms in Lower Saxony, Germany. BTM samples were collected in January 2023 from 208 selected dairy farms. The samples were quantitatively culturally analyzed for S. aureus and Prototheca spp. Presumptive S. aureus colonies were further confirmed by MALDI-TOF. Presumptive Prototheca spp. colonies were confirmed by light microscopy. Sc. agalactiae and Mycoplasmopsis spp. were detected by real-time polymerase chain reaction (rtPCR). Sc. agalactiae was detected in two herds (1% (Confidence Interval 95% (CI) 0.3–3.4)). S. aureus was confirmed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) in 38 herds (18.3% (CI 13.6–24.1)), assuming a threshold of >10 cfu/mL milk. A total of 154 isolates identified as S. aureus by MALDI-TOF were transferred to agar with added oxacillin for resistance testing, of which 19 isolates (12.3% (CI 8–18.5)) showed growth. The 19 isolates came from eight different farms (3.8% (2–7.4)). Prototheca spp. were identified in 13 herds (6.3% (CI 3.7–10.4)). Mycoplasmopsis spp. were detected by PCR in 18 herds (8.7% (CI 5.5–13.3)). Of these, M. bovis was present in three herds (1.4% (0.5–4.2)). The herd prevalence of Sc. agalactiae in BTM appears to be at low levels in the sampled area. The prevalence of Mycoplasmopsis spp. in the herds was higher than expected compared to previous studies. It is interesting to note that the percentage of M. bovis in the total Mycoplasmopsis spp. was only 16.7%.

Analysis of molecular structure and topological properties of chitosan isolated from crab shell and mushroom

This investigation aimed to scrutinize the chemical and structural analogies between chitosan extracted from crab exoskeleton (High Molecular Weight Chitosan, HMWC) and chitosan obtained from mushrooms (Mushroom-derived Chitosan, MRC), and to assess their biological functionalities. The resulting hydrolysates from the hydrolysis of HMWC by chitosanase were categorized as chitosan oligosaccharides (csCOS), while those from MRC were denoted as mrCOS. The molecular weights (MW) of csCOS and mrCOS were determined using Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) mass spectrometry. Furthermore, structural resemblances of csCOS and mrCOS were assessed utilizing X-ray powder diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Intriguingly, no apparent structural disparity between csCOS and mrCOS was noted in terms of the glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) composition ratios. Consequently, the enzymatic activities of chitosanase for HMWC and MRC exhibited remarkable similarity. A topological examination was performed between the enzyme and the substrate to deduce the alteration in MW of COSs following enzymatic hydrolysis. Moreover, the evaluation of antioxidant activity for each COS revealed insignificance in the structural disparity between HMWC and MRC. In summary, grounded on the chemical structural similarity of HMWC and MRC, we propose the potential substitution of HMWC with MRC, incorporating diverse biological functionalities.

Reappraisal of p16 for Determining HPV Status of Head and Neck Carcinomas Arising in HPV Hotspots.

In an era of head and neck oncology where HPV status will soon dictate patient management, reliable HPV detection is critical. P16 immunohistochemistry (IHC) is currently recommended as the test of choice for oropharyngeal squamous cell carcinomas (OPSCCs). The purpose of this study was to determine the performance characteristics of p16 IHC based on a large clinical experience of squamous cell carcinomas (SCC) arising from HPV hot-spot regions of the head and neck. Consecutive OPSCCs, sinonasal SCCs, and metastatic SCCs of unknown primary sites were evaluated for the presence of HPV by p16 IHC and PCR-based HPV DNA testing as part of clinical care. For discrepant cases, high-risk HPV E6/E7 mRNA in situ hybridization (ISH) and, when possible, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MassArray) genotyping were performed. 746 cancers underwent HPV testing by p16 IHC and DNA PCR genotyping. There was a 95.6% concordance between the 2 assays. Of the 33 discrepant cases, 32 cases (4.3%) were p16 positive but HPV DNA negative. In these cases, 68% were positive for mRNA ISH, invariably related to a non-16 HPV genotype. P16 IHC had an overall accuracy of 98.8%, a sensitivity of 99.8%, and a specificity of 92.1%. P16 IHC is a sensitive and specific assay for determining HPV status. HPV DNA PCR appears vulnerable to HPV genotype diversity and is prone to missing rare non-16 genotypes. HPV mRNA ISH is a practical and reliable direct measure of HPV that may help eliminate the small number of false-positive p16 cases and avoid potential patient harm related to erroneous HPV classification.

Constructing Structural Isomers to Reveal and Enhance Lithium Storage in a Conducting Polymer

AbstractOrganic electrodes have gained great interest for batteries. Conducting polymers have the merits of high conductivity, and hence are attempted as electrode materials in batteries as soon as their discoveries. However, conducting polymers showe poor electrochemical performance and are baring vague charge storage mechanisms. Here, a strategy is reported to reveal the charge storage mechanisms of conducting polymers by constructing isomers and using polypyridines as a proof‐of‐concept. Polypyridines are rarely studied and its redox activity is still not clear. The results indicate that every pyridine ring can accept one electron and the dimer unit with adjacent C═N groups can gain one more electron through chelation effects with Li ions. The electron transferred number and the chelation effects can be quantified by the matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) mass spectroscopy and verified by density functional theory (DFT) calculations. As a result, one of the polypyridine isomers delivers the highest capacity of 504 mAh g−1 after deducting the contribution of conductive additives. This work provides a universal way to reveal the charge storage mechanism and highlights the significance of adjacent active centers to form chelation with Li ions and hence enhance the performance of organic batteries.

Precise diagnosis and risk stratification of prostate cancer by comprehensive serum metabolic fingerprints: a prediction model study.

Prostate cancer (PCa) is one of the most common malignancies in men worldwide and has caused increasing clinical morbidity and mortality, making timely diagnosis and accurate staging crucial. The authors introduced a novel approach based on mass spectrometry for precise diagnosis and stratification of PCa to facilitate clinical decision-making. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of trace blood samples was combined with machine learning algorithms to construct diagnostic and stratification models. A total of 367 subjects, comprising 181 with PCa and 186 with non-PCa were enrolled. Additional 60 subjects, comprising 30 with PCa and 30 with non-PCa were enrolled as an external cohort for validation. Subsequent metabolomic analysis was carried out using Autoflex MALDI-TOF, and the mass spectra were introduced into various algorithms to construct different models. Serum metabolic fingerprints were successfully obtained from 181 patients with PCa and 186 patients with non-PCa. The diagnostic model based on the eight signals demonstrated a remarkable area under curve of 100% and was validated in the external cohort with the area under curve of 87.3%. Fifteen signals were selected for enrichment analysis, revealing the potential metabolic pathways that facilitate tumorigenesis. Furthermore, the stage prediction model with an overall accuracy of 85.9% precisely classified subjects with localized disease and those with metastasis. The risk stratification model, with an overall accuracy of 89.6%, precisely classified the subjects as low-risk and high-risk. Our study facilitated the timely diagnosis and risk stratification of PCa and provided new insights into the underlying mechanisms of metabolic alterations in PCa.

Clinical and microbiological profile of Viridans group streptococcal bacteraemia; experience from South India.

Viridans Group Streptococci (VGS) are a group of distinct species that can cause bacteraemia and other invasive infections. They are also among the common organisms causing infective endocarditis. Data on the epidemiology and clinical profile of VGS is limited, especially from India. We conducted an electronic medical record-based retrospective analysis of patients with VGS bacteraemia admitted to our hospital between January 2012 to December 2021. Blood cultures were incubated by BacT/ALERT system and bacterial identification and susceptibility testing were done by using the VITEK 2 microbial identification system. Susceptibility test reporting was as per Clinical and Laboratory Standards Institute (CLSI) guidelines. The incidence, clinical profile, source of bacteraemia, co-morbidities and antimicrobial resistance among VGS bacteraemia were analyzed. VGS were isolated in 219 patients, accounting for 3.2% of positive blood cultures during the period studied. The median age of the patients was 58 years and 69% were males. Diabetes mellitus was the most common co-morbidity (55%) followed by chronic kidney disease and chronic liver disease. Patients with haematological malignancy and neutropenia were few. Intra-abdominal infections were the most common source of infection and was noted in 26%. Infective endocarditis was diagnosed in only 10% of the cases. Streptococcus mitis was the most common species isolated followed by S. gallolyticus and S. sanguinis. 9.58% of the isolates could not be identified up to the species level. Overall penicillin susceptibility was 71% and ceftriaxone susceptibility was 92%, with individual species variation. In-hospital mortality was 19%. VGS are an important cause of bacteraemia and was associated with 19% mortality in our study. High rates of penicillin and ceftriaxone resistance are a reason of concern. Molecular diagnostics like matrix assisted laser desorption ionization-time of flight (MALDI-TOF) identification must be increasingly applied for species identification considering that a substantial number of isolates were not identified to species level.

Construction of Biosensing System for Glycated Albumin Using an Electron Transfer Peptide-Modified Protein Probe.

Glycated albumin (GA) is one of the proteins that replaces several sugar moieties and can be used as an indicator of diabetes mellitus. We developed a sensing system that uses GA in the early detection of diabetes mellitus. In this study, H6Y4C acetylated (Ac-) at the N-terminals of the peptide was combined with wheat germ agglutinin (WGA) to recognize glucose moieties. The Ac-H6Y4C-WGA was constructed as a GA-sensing probe. The tyrosine residues of Y4C exhibited an oxidation peak, and His-tag moieties were introduced to separate Ac-H6Y4C-WGA in the synthesis of the probe. The Ac-H6Y4C-WGA probe binds with the 1-2 molecules of Ac-H6Y4C per WGA using matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-MS. Next, the functions of Ac-H6Y4C-WGA were evaluated using voltammetry. The number of electron-transfers was calculated based on the relationship between the peak potential and logarithm of scan rate and was 3.03. In the electrochemical measurements with mannose and bovine serum albumin, the peak currents were similar to that of GA alone. By contrast, a decrease in the peak current was suppressed when glucose was added to the solution containing the probe. As a result, Ac-H6Y4C-WGA was selectively bound to the glucose moieties of GA. The calibration curve via differential pulse voltammetry was proportional to the concentrations of GA and ranged from 1.0 × 10-12 to 2.0 × 10-11 M with a detection limit of 3.3 × 10-13 M.

MALDI-TOF imaging analysis of benzalkonium chloride penetration in ex vivo human skin.

Benzalkonium chloride (BZK), alkyldimethylbenzlamonium chloride, is a cationic surfactant that is used as an antiseptic. BZK is classified as a quaternary ammonium compound composed of molecules of several alkyl chains of differing lengths, that dictate its effectiveness towards different microbes. As a result, BZK has become one of the most used preservatives in antibacterial solutions. Despite its widespread use, it is not clear whether BZK penetrates human skin. To answer this question, BZK treated skin was analyzed using matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry imaging. Solutions containing BZK and differing excipients, including citric acid, caprylyl glycol, and vitamin E, were applied ex vivo to excised human skin using Franz diffusion cells. Treated skin was embedded in gelatin and sectioned prior to MALDI-TOF imaging. BZK penetrates through the epidermis and into the dermis, and the penetration depth was significantly altered by pH and additives in tested solutions.

Deep Impact: Shifts of Native Cultivable Microbial Communities on Fresh Lettuce after Treatment with Plasma-Treated Water.

Foods consumed raw, such as lettuce, can host food-borne human-pathogenic bacteria. In the worst-case, these diseases cause to death. To limit illness and industrial losses, one innovative sanitation method is non-thermal plasma, which offers an extremely efficient reduction of living microbial biomass. Unfortunately, the total viable count (TVC), one of the most common methods for quantifying antimicrobial effects, provides no detailed insights into the composition of the surviving microbial community after treatment. To address this information gap, different special agars were used to investigate the reduction efficiency of plasma-treated water (PTW) on different native cultivable microorganisms. All tested cultivable microbial groups were reduced using PTW. Gram-negative bacteria showed a reduction of 3.81 log10, and Gram-positive bacteria showed a reduction of 3.49 log10. Fungi were reduced by 3.89 log10. These results were further validated using a live/dead assay. MALDI-ToF (matrix-assisted laser-desorption-ionization time-of-flight)-based determination was used for a diversified overview. The results demonstrated that Gram-negative bacteria were strongly reduced. Interestingly, Gram-positive bacteria and fungi were reduced by nearly equal amounts, but could still recover from PTW treatment. MALDI-ToF mainly identified Pseudomonas spp. and groups of Bacillus on the tested lettuce. These results indicate that the PTW treatment could efficiently achieve a ubiquitous, spectrum-wide reduction of microbial life.

Isolation and detection of bacterial species on mitis salivarius bacitracin agar from the plaque samples of caries active children.

In cariology studies, mitis-salivarius-bacitracin (MSB) agar has been commonly considered as the selective medium for Streptococcusmutans growth. The present study was the part of a funded project (a noninferiority randomized controlled trial) which compared the efficacy of a plant extract-based mouth rinse with that of a fluoride mouth rinse on the S.mutans counts of the children. This study aimed to identify the frequency of detection of S.mutans and nonstreptococcal bacterial species from the dental plaque of caries active children using a combined technique of anaerobic culture and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Caries-active children (8-12 years old) were enrolled from a pediatric dental outpatient department at a tertiary care hospital. From each participant, dental plaque samples were collected from carious surfaces under sterilized conditions and then subjected to anaerobic culture. After 48 h of incubation, the bacterial colonies were isolated by sub-culture and identified by the MALDI-TOF. In all, 13 different bacterial species were isolated from the MSB agar medium. Other than S.mutans species, colonies of bacterial species such as Veillonelladispar,Streptococcusanginosus, Veillonellaparvula, and Streptococcusgordonii were also frequently observed from the medium. The study concluded that several bacterial strains, both streptococcal and nonstreptococcal, could be isolated from the MSB agar medium; hence, this medium should no longer be considered selective medium for the culture of S.mutans in clinical and epidemiological studies.

A Case of Successfully Treated Roseomonas mucosa-induced Peritonitis Diagnosed by Mass Spectrometry.

Roseomonas mucosa is difficult to identify using routine analytical techniques. We herein report a case of peritoneal dialysis (PD)-related peritonitis caused by R. mucosa identified using matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS). A 70-year-old woman was admitted to our hospital with PD-related peritonitis. Blood agar medium of dialysate culture derived colony pale pink in color, and the organism was identified as R. mucosa using MALDI-TOF MS. She was successfully treated with ciprofloxacin and meropenem without catheter removal. To our knowledge, this is the first case of R. mucosa peritonitis in which technique failure has been avoided.

Nosocomial transmission of Aspergillus flavus in a neonatal intensive care unit: Long-term persistence in environment and interest of MALDI-ToF mass-spectrometry coupled with convolutional neural network for rapid clone recognition.

Aspergillosis of the newborn remains a rare but severe disease. We report four cases of primary cutaneous Aspergillus flavus infections in premature newborns linked to incubators contamination by putative clonal strains. Our objective was to evaluate the ability of matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) coupled to convolutional neural network (CNN) for clone recognition in a context where only a very small number of strains are available for machine learning. Clinical and environmental A. flavus isolates (n=64) were studied, 15 were epidemiologically related to the four cases. All strains were typed using microsatellite length polymorphism. We found a common genotype for 9/15 related strains. The isolates of this common genotype were selected to obtain a training dataset (6 clonal isolates/25 non-clonal) and a test dataset (3 clonal isolates/31 non-clonal), and spectra were analysed with a simple CNN model. On the test dataset using CNN model, all 31 non-clonal isolates were correctly classified, 2/3 clonal isolates were unambiguously correctly classified, whereas the third strain was undetermined (i.e., the CNN model was unable to discriminate between GT8 and non-GT8). Clonal strains of A. flavus have persisted in the neonatal intensive care unit for several years. Indeed, two strains of A. flavus isolated from incubators in September 2007 are identical to the strain responsible for the second case that occurred 3 years later. MALDI-TOF is a promising tool for detecting clonal isolates of A. flavus using CNN even with a limited training set for limited cost and handling time.

Enhanced exopolysaccharide production in gamma irradiated Bacillus subtilis: A biofilm-mediated strategy for ZnO nanoparticles removal

Biofilm-mediated strategy was studied for ZnO nanoparticle removal from aqueous media. Bacillus subtilis isolated from the soil rhizosphere was selected based on its high viscosity (133 Pa/s) of the cultivated culture and biofilm formation. The bacterium was exposed to gamma-irradiation to enhance EPS production along with its cultivation in EPS-producing media. The results show an increase in viscosity that reached 160 Pa/s at 2 kGy. EPS production increased from 4.45 to 7.95 mg/mL and the protein/carbohydrate ratio increased from 3 to 4.4 which reflects the stickiness of EPS. Thermal Gravimetric Analysis (TGA) showed 2 phase weight loss for gamma irradiated EPS and defined protein peaks when characterized using Matrix Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF). Native and gamma-irradiated Bacillus subtilis cells with their enhanced EPS were grown as a biofilm on sterile waste gauze fabric, Scanning Electron Microscopy (SEM) showed an increased biofilm attachment in gamma-irradiated samples. The latter was used for the removal of ZnO NP from aqueous media. Energy dispersive X-ray (EDX) mapping confirms that ZnO NPs were entrapped within the carbon and oxygen elements forming the biofilm with net intensities of 14.04, 1713, and 1190, respectively. The results confirm that biofilm-mediated strategy is effective in nanoparticles removal.

Fatal Lodderomyces elongisporus Fungaemia in COVID-19 Patient

Lodderomyces elongisporus, an ascomycetous yeast, is often misidentified as Candida parapsilosis due to its physiologic similarity. Compared to Candida parapsilosis complex, L. elongisporus is still inferior in regard to incidence and virulence. We report a case of L. elongisporus fungaemia in a patient with diabetic ketoacidosis (DKA) precipitated by COVID-19 category 5 infection. A 52-year-old diabetic and hypertensive lady was brought in for fever, cough and lethargy for four days, with sudden onset of dyspnoea. Upon arrival, she was febrile, and in metabolic ketoacidosis. Her lung auscultation was clear. SARS-CoV-2 RNA was detected via real-time polymerase chain reaction (RT-PCR). Her blood culture grew L. elongisporus, identified via matrix-assisted laser desorption/ionization time of flight (MALDI-TOF). Although treated with intravenous (IV) amphotericin B for four days, she succumbed on the sixth day of admission. Accurate identification of this yeast, especially in differentiating it with Candida parapsilosis complex, remains a diagnostic challenge for routine diagnostic laboratories. MALDI-TOF offers a reliable alternative for accurate and prompt diagnosis. Co-infections with COVID-19 have never been recorded worldwide. We highlight the first case of L. elongisporus isolation co-existing with SARS-CoV-2 infection during this pandemic.

Base‐assisted anionic polymerization of alkyl sorbates and subsequent ring‐closure without extreme dilution for synthesis of cyclic stereoregular poly(alkyl sorbates)

AbstractA typical base, lithium diisopropylamide (LDA) or lithium bis(trimethylsilyl)amide (LiHMDS), was found to initiate the anionic chain‐growth polymerization of ethyl sorbate (ES) in the presence of an aluminum Lewis acid, that is, methylaluminum bis(2,6‐di‐tert‐butyl‐4‐methylphenoxide) (MAD) and, after ES consumption, ring‐closure occurred without highly dilute conditions to give the cyclic poly(ES). The finding of ring closure was supported by matrix‐assisted laser desorption/ionization‐time of flight (MALDI‐TOF) mass spectroscopy and direct transmittance electron microscope (TEM) observations, in which a series of spectral patterns separated by the mass of ES and cyclic objects with 7.4–25.9 nm rings were observed, respectively. The results indicated that instead of the use of an expensive N‐heterocyclic carbene (NHC), employing these commodity bases to initiate chain polymerization enabled us to fabricate the intended cyclic vinyl polymers without highly dilute conditions. Also, this polymerization proceeded even at −80°C to give threo (98%)‐disyndiotactic (91%) stereoregular poly(ES) in an excellent yield with narrow molecular weight distributions (Mn = 5.9 × 103–8.4 × 103, Mw/Mn = 1.14).

First isolation of Mycobacterium saskatchewanense from medical devices

Mycobacterium saskatchewanense is a species of pigmented slow-growing Non-Tuberculous Mycobacteria (NTM), positive for Mycobacterium avium complex (MAC) by AccuProbe system. MAC organisms have frequently been isolated from different medical devices. This is the first study reporting isolation of M. saskatchewanense from medical devices and highlights the importance of correctly identifying the NTMs that often colonize sanitary water. GenoType Mycobacterium CM CE-IVD kit (CM) was used as the first step of NTM strain identification, and all positive cultures were found to be components of MAC. Then, GenoType NTM-DR CE-IVD kit (NTM-DR) was used to differentiate the different species. Sub-culture on solid media were used for: (i) phenotypical confirmation by colony morphology and Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometry; (ii) molecular confirmation by Next Generation Sequencing. All positive cultures were identified as M. intracellulare by CM and NTM-DR assays, whereas colony morphology showed bright yellow scotochromogenic growth. MALDI-TOF analyses identified the strains as M. saskatchewanense with a high score, and identification was confirmed by NGS analysis based on the hsp-65 region. This paper suggests that it is important to actively monitor NTM contamination in medical devices that use sanitary water, to prevent the possibility of patients becoming infected.

Multilocus Sequence Typing of Emerging Azole-Resistant Candida auris Isolates from Critically Ill Patients in a New York City Medical Center

Abstract Candida auris has developed into a slow-moving global pandemic since its emergence in 2009, particularly as a source of nosocomial infection amongst immunocompromised and critically-ill patients. According to CDC data, C. auris emergence has accelerated in the US during the COVID-19 pandemic and has garnered particular attention for its high level of resistance to multiple classes of antifungals. C. auris has been categorized into five genetically diverse clades, each with unique antifungal drug resistance patterns. We have observed a sharp increase in cases identified at our tertiary medical center in New York City over the past three years. In this study, we designed a set of C. auris-specific oligonucleotides to determine the clonality of isolates through Multilocus Sequence Typing (MLST) of four genetic loci known to have a high level of divergence amongst Candida species. From 2020 through 2022 we identified 22 patients with C. auris bloodstream infection at our institution by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and performed MLST on isolates that were available from 19 patients. We assessed four genetic loci: ITS1/ITS2, D1/D2, RPB1 and RPB2; ERG11 was included to assess for mutations associated with fluconazole resistance. We found that the number of blood stream infections increased from 2 cases in 2020 to 12 in 2022. Our MLST results showed that all C. auris isolates from blood and body fluids belong to CDC-defined Clade 1c, which carries the ERG11 gene mutation resulting the amino acid substitution K143R in 14-α-Demethylase associated with fluconazole resistance. These isolates also demonstrated fluconazole resistance by in vitro testing (19/19), amphotericin resistance was present in 44% of isolates with available data (7/16), and all isolates were susceptible to micafungin (19/19). Of 19 patients with blood cultures positive for C. auris, 13 (68%) were in a critical care setting at time of collection, 17 (89%) had been in an ICU during the same admission, and 8 (42%) expired within 3 months of C. auris positivity. MLST is considered a relatively fast and inexpensive method of sequence-based strain typing and has recently been applied to assessing the clonality of C. auris isolates. We show that the use of C. auris-specific oligonucleotides for sequencing, rather than those derived from other Candida species, produces high quality data for MLST, as well as the detection of specific mutations involved in antifungal resistance, such as ERG11. Further assessment is needed by the gold standard in sequence typing, Whole Genome Sequencing (WGS), to confirm our analysis of clonality here. MLST may therefore represent a useful tool in understanding the interrelatedness of invasive C. auris infections in hospitalized patients as well as inform clinical decision-making around infection control and empiric antifungal therapy.

Presence of the blaTEM Gene in Commensal Neisseria spp.: A Possible Cause for the Acquired Drug Resistance Among Pathogenic Respiratory Bacteria.

Background The oral microbiome consists of various bacterial genera, withNeisseriaspp. being a prominent part of this niche. WhileNeisseria gonorrhoeaeandNeisseriameningitidisare human-restricted pathogens, non-pathogenicNeisseriaspecies likeNeisseria sicca,Neisseria perflava, etc.,are primarily commensals that can also behave as opportunistic pathogens. With increasing penicillin resistance in commensalNeisseria, there is a concern that these bacteria might harbor resistance genes that can be transferred to other pathogens. This study aimed to characterize the blaTEM gene (encodes for the plasmid-mediated β-lactamase enzyme that hydrolyzes the β-lactam ring) of commensalNeisseriaspp. isolated from respiratory samples. Methodology The research was conducted in the Department of Clinical Microbiology at Sri Ramachandra University, Chennai. The specimens used were sputum and throat swabs, which were subjected to a series of phenotypic methods and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) for speciation. The antibiogram was determined using the Kirby-Bauer disk diffusion method, and a PCR assay was utilized to identify theblaTEMgene responsible for β-lactamase production. Results Out of 274 processed samples, 65 unique commensalNeisseriaspp. were identified. The study highlighted the presence of theblaTEMgene in 93.9% (61) of the isolates, which is responsible for β-lactamase production. All isolates exhibitedresistance to penicillin. MostblaTEM-positive commensalNeisseria spp. were susceptible to cefuroxime (83.6%), ceftriaxone (85.2%), and cefotaxime (85.2%). The high prevalence of theblaTEMgene in commensalNeisseriais alarming. The gene, found on plasmids, could potentially transfer to other related species likeNeisseria gonorrhoeaeandNeisseriameningitidis, as well as other Gram-negative bacilli. Conclusion The presence of resistance genes in commensal bacteria is of concern, as they might be reservoirs for resistance transfer to pathogenic strains. The study emphasizes the importance of continuous monitoring and deeper investigations into commensal bacteria, emphasizing the need for a broader community screening approach to understand resistance mechanisms in the normal microbiome.

Laccase-Catalyzed Synthesis of Added-Value Polymers from Cork and Grape Extracts.

The development of products from natural plant sources, including agriculture and food wastes, contributes significantly to the circular economy and global sustainability. Cork and grape wastes were employed as the primary sources in this study to obtain compounds of interest under mild extraction conditions. Laccase was applied to oxidize the cork and grape extracts, with the aim of producing value-added molecules with improved properties. Ultraviolet-visible (UV-vis) spectroscopy was assessed to monitor the oxidation process, and characterization of the end products was performed by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) spectroscopy. The antioxidant and antiaging properties were evaluated by means of ABTS, DPPH, FRAP, and SPF testing. Overall, as compared to their monomeric counterparts, the polymeric compounds displayed remarkable antioxidant and antiaging characteristics after laccase oxidation, showing tremendous potential for applications in the food, pharmaceutical, cosmetic, and textile industries.

Antibacterial Susceptibility Pattern of S. maltophilia Isolates at A Tertiary Care Hospital, India

A sudden emergence of Stenotrophomonas maltophilia as a primary pathogen both in immunocompromised and immunocompetent individuals has raised a serious concern, as it is associated with significant case fatality ratio. We intended to study the clinico-microbiological profile of S. maltophilia isolates from various samples and outcome of the infections in a tertiary healthcare center, Pune, India. This is an observational cross-sectional study was conducted from January 2021 to June 2022 at Department of Microbiology of a tertiary care Centre in Pune, India. Of the 12049 samples received for culture, S. maltophilia was isolated in 57 samples. Only 42 samples with pure growth of S. maltophilia were included in the study with 15 excluded due to mixed growth. All isolates were confirmed by VITEK-MS (bioMerieux, SA, France) which uses Matrix Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) technology. Of the 42 isolates, majority were isolated from pus(28.6%) and most of patients (61.9%) were from acute health care settings. The isolates had high susceptibility to Cotrimoxazole (85.7%) and Minocycline (85.7%) and low susceptibility to Ceftazidime (45.2%). A case fatality rate of 7.1% (3/42 cases) was noted and 39 cases were discharged after complete treatment. All the three fatal cases were susceptible to levofloxacin, ciprofloxacin, cotrimoxazole and minocycline and all three fatal cases were resistant to ceftazidime. S. maltophilia has recently shown an increase in nosocomial infections especially in acute healthcare settings like ICU and other critical care wards. The isolates of the present study had high susceptibility to trimethoprim-sulfamethoxazole (TMP-SXT) and Minocycline and low susceptible to Ceftazidime.