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Hard ticks (Acari: Ixodidae) have been historically identified by morphological methods which require highly specialized expertise and more recently by DNA-based molecular assays that involve high costs. Although both approaches provide complementary data for tick identification, each method has limitations which restrict their use on large-scale settings such as regional or national tick surveillance programs. To overcome those obstacles, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been introduced as a cost-efficient method for the identification of various organisms, as it balances performance, speed, and high data output. Here we describe the use of this technology to validate the distinction of two closely related Dermacentor tick species based on the development of the first nationwide MALDI-TOF MS reference database described to date. The dataset obtained from this protein-based approach confirms that tick specimens collected from United States regions west of the Rocky Mountains and identified previously as Dermacentor variabilis are the recently described species, Dermacentor similis. Therefore, we propose that this integrative taxonomic tool can facilitate vector and vector-borne pathogen surveillance programs in the United States and elsewhere.

Background: Detecting monoclonal protein (M-protein), a hallmark of plasma cell disorders, traditionally relies on methods such as protein electrophoresis, immune-electrophoresis, and immunofixation electrophoresis (IFE). Mass spectrometry (MS)-based methods, such as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray ionization-quadrupole time-of-flight (ESI-qTOF) MS, have emerged as sensitive methods. We explored the M-protein-detection efficacies of different MS techniques. Methods: To isolate immunoglobulin and light chain proteins, six types of beads (IgG, IgA, IgM, kappa, lambda, and mixed kappa and lambda) were used to prepare samples along with CaptureSelect nanobody affinity beads (NBs). After purification, both MALDI-TOF MS and liquid chromatography coupled with Synapt G2 ESI-qTOF high-resolution MS analysis were performed. We purified 25 normal and 25 abnormal IFE samples using NBs and MALDI-TOF MS (NB-MALDI-TOF). Results: Abnormal samples showed monoclonal peaks, whereas normal samples showed polyclonal peaks. The IgG and mixed kappa and lambda beads showed monoclonal peaks following the use of daratumumab (an IgG/kappa type of monoclonal antibody) with both MALDI-TOF and ESI-qTOF MS analysis. The limits of detection for MALDI-TOF MS and ESI-qTOF MS were established as 0.1 g/dL and 0.025 g/dL, respectively. NB-MALDI-TOF and IFE exhibited comparable sensitivity and specificity (92% and 92%, respectively). Conclusions: NBs for M-protein detection, particularly with mixed kappa-lambda beads, identified monoclonal peaks with both MALDI-TOF and ESI-qTOF analyses. Qualitative analysis using MALDI-TOF yielded results comparable with that of IFE. NB-MALDI-TOF might be used as an alternative method to replace conventional tests (such as IFE) to detect M-protein with high sensitivity. [ABSTRACT FROM AUTHOR]

Summary: This study explored the fermentation of defatted wheat germ (DWG) using Lactobacillus plantarum, leading to the release of bioactive peptides with improved technological and health properties. Proteomic analysis of fermented defatted wheat germ (FDWG) and DWG revealed a higher abundance of functional peptides in FDWG, with 27 peaks ranging from 503–1391 m/z compared to 22 peaks ranging from 537–1362 m/z for DWG. The peptides identified in FDWG were enriched in hydrophobic amino acids, known for their antioxidant and radical scavenging capabilities. Furthermore, FDWG‐based bread demonstrated the lowest gliadin content, making it a promising option for coeliac patients. The bread also exhibited a soft texture and higher consumer acceptance, positioning it as a viable alternative in reduced‐gluten bakery products. Overall, fermented wheat germ presents an exciting avenue for obtaining bioactive peptides with diverse potential applications in functional foods and nutraceuticals. [ABSTRACT FROM AUTHOR]

This study describes the extraction and identification by electrophoretic and spectrometric techniques of protease inhibitor from the medicinal plant Alocasia macrorrhizos as well as investigates their immunomodulatory properties and cell viability. The A. macrorrhizos tubers were subjected to protease inhibitor extractions and characterised using SDS-PAGE and MALDI-TOF. The protein extracts were assessed for activities trypsin inhibition stoichiometry, haemagglutinating, cell viability, NO and TNF-α production inhibition. Concerning the protease inhibitors analysis through SDS-PAGE, the results showed two bands with 11 and 24 kDa, and the MS analysis detected the ions more intense of m/z 4276.795 and 8563.361 in the roasted protein extract. The IC50 of trypsin inhibition was 0.119 and 0.302 mg L−1 in the roasted and crude tuber, respectively. The protease inhibitors extract from the roasted tubers showed a reduction in the production of NO and TNF-α at concentrations lower than 100 µg mL−1, without a reduction in cell viability. [ABSTRACT FROM AUTHOR]

Colibacillosis is an intestinal tract infection in poultry caused by Escherichia coli. It is one of the leading causes of mortality and results in significant economic losses for the poultry farming sector due to its high incidence rate. Recently, E. coli has been considered a predominant bacterial pathogen that is responsible for diarrhea and bloodstream infections in chickens and ducks in Vietnam. In this study, E. coli strains associated with diarrhea were isolated from chicken and duck feces and intestines collected from poultry farms in Hai Phong province using a Macconkey selective medium. The obtained isolates were initially identified by the MALDITOF MS method, screened for hemolytic activity, and finally identified by 16S rRNA gene sequencing. As a result, eight E. coli strains exhibiting hemolytic activity were identified. Among them, E. coli_5, E. coli_6, and E. coli_8 strains were clustered with E. coli O78:H51, whereas E. coli_3 was grouped with E. coli O78:H4; E. coli_4 and E. coli_7 were clusterd with E. coli O25b: H4; and E. coli_9 and E. coli_16 were grouped with E. coli O26:H11. Our results provide insights into the genetic diversity of E. coli strains associated with diarrhea in poultry farms in Hai Phong province. The isolated strains will be further characterized for the development of effective strategies to control colibacillosis in poultry. [ABSTRACT FROM AUTHOR]

Enterococcus spp. are isolated from infections of domestic animals and can present resistance to different antimicrobials. This study aimed to identify the main species of Enterococcus and determine the phenotypic resistance to antimicrobials of isolates of this bacterial genus obtained from dogs and cats admitted to a veterinary hospital in southern Brazil. A total of 57 Enterococcus spp. were isolated from different clinical samples (urine, feces, ears and skin) in domestic animals admitted to the hospital over a period of three years (2016 to 2019). MALDI-TOF results demonstrated the occurrence of Enterococcus faecium (39; 68.4%), Enterococcus faecalis (17; 29.8%) and Enterococcus avium (1; 1.8%). In an overall analysis, resistance was observed to the antimicrobials rifampicin (46; 80.7%), tetracycline and streptomycin (42; 73.7%), ampicillin and imipenem (41; 71.9%), erythromycin (39; 68.4%), gentamicin (38; 66.7%), ciprofloxacin (36; 63.2%), norfloxacin (32; 56.1%), nitrofurantoin (10; 17.5%) and chloramphenicol (9; 15.7%). None of the Enterococcus spp. showed resistance to vancomycin and linezolid. Multidrug resistance (MDR) was detected in 45 (78.9%) isolates. In conclusion, E. faecium and E. faecalis with MDR are frequent in infections of hospitalized dogs and cats from southern Brazil. [ABSTRACT FROM AUTHOR]

Nowadays, mastitis is one of the most common diseases in breeding dairy cattle. Treatment with conventional antibiotics is ineffective because many pathogens are resistant. This study aims to investigate the presence of mastitis in milk samples from cows exhibiting clinical symptoms, along with assessing the sensitivity of the bacteria isolated from these samples to various antibiotics. The research involved analysing 70 milk samples obtained from cows with clinical mastitis at the Veterinary Department, Banas Dairy, Palanpur. From these samples, a total of 12 different bacterial strains were identified. Identification methods included conventional biochemical techniques and MALDI-TOF analysis. Furthermore, the study evaluated the antibacterial properties of Copper oxide nanoparticles (CuO-NPs), Triphala nanoparticles, and Chitosan nanoparticles, the nanoparticles were characterized by SEM and FTIR. The mean size of chitosan, Copper and Triphala were 380 nm, 400 nm and 400 nm with zeta potential + 44.3 mv, -42.2 mv and -27.5 mv respectively. These nanoparticles were explored as potential alternatives for mastitis treatment in cows. The research also aimed to determine the minimum inhibitory concentration (MIC) of CuO-NPs, Triphala NPs, and Chitosan NPs against the bacterial strains isolated from the milk samples. By examining the efficacy of these nanoparticles and their MIC against mastitis-causing bacteria, this study contributes to the ongoing efforts to find effective treatments for mastitis in dairy cows. [ABSTRACT FROM AUTHOR]

Abstract Hard ticks (Acari: Ixodidae) have been historically identified by morphological methods which require highly specialized expertise and more recently by DNA-based molecular assays that involve high costs. Although both approaches provide complementary data for tick identification, each method has limitations which restrict their use on large-scale settings such as regional or national tick surveillance programs. To overcome those obstacles, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been introduced as a cost-efficient method for the identification of various organisms, as it balances performance, speed, and high data output. Here we describe the use of this technology to validate the distinction of two closely related Dermacentor tick species based on the development of the first nationwide MALDI-TOF MS reference database described to date. The dataset obtained from this protein-based approach confirms that tick specimens collected from United States regions west of the Rocky Mountains and identified previously as Dermacentor variabilis are the recently described species, Dermacentor similis. Therefore, we propose that this integrative taxonomic tool can facilitate vector and vector-borne pathogen surveillance programs in the United States and elsewhere.

Enterococcus spp. are isolated from infections of domestic animals and can present resistance to different antimicrobials. This study aimed to identify the main species of Enterococcus and determine the phenotypic resistance to antimicrobials of isolates of this bacterial genus obtained from dogs and cats admitted to a veterinary hospital in southern Brazil. A total of 57 Enterococcus spp. were isolated from different clinical samples (urine, feces, ears and skin) in domestic animals admitted to the hospital over a period of three years (2016 to 2019). MALDI-TOF results demonstrated the occurrence of Enterococcus faecium (39; 68.4%), Enterococcus faecalis (17; 29.8%) and Enterococcus avium (1; 1.8%). In an overall analysis, resistance was observed to the antimicrobials rifampicin (46; 80.7%), tetracycline and streptomycin (42; 73.7%), ampicillin and imipenem (41; 71.9%), erythromycin (39; 68.4%), gentamicin (38; 66.7%), ciprofloxacin (36; 63.2%), norfloxacin (32; 56.1%), nitrofurantoin (10; 17.5%) and chloramphenicol (9; 15.7%). None of the Enterococcus spp. showed resistance to vancomycin and linezolid. Multidrug resistance (MDR) was detected in 45 (78.9%) isolates. In conclusion, E. faecium and E. faecalis with MDR are frequent in infections of hospitalized dogs and cats from southern Brazil.

Abstract Background Among gynaecological malignancies, endometrial cancer (EC) is the most prevalent type of uterine cancer affecting women. This study explored the proteomic profiles of plasma samples obtained from EC patients, those with hyperplasia (Hy), and a control group (CO). A combination of techniques, such as 2D-DIGE, mass spectrometry, and bioinformatics, including pathway analysis, was used to identify proteins with modified expression levels, biomarkers and their associated metabolic pathways in these groups. Methods Thirty-four patients, categorized into three groups—10 with EC, 12 with Hy, and 12 CO—between the ages of 46 and 75 years old were included in the study. Untargeted proteomic analysis was carried out using two-dimensional difference in gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Results In all three groups, 114 proteins that were significantly (p ≤ 0.05 and fold change ≥ 1.5) altered were successfully identified using peptide mass fingerprints (PMFs). Compared with those in the control group (CO), the EC samples had 85 differentially expressed proteins (39 upregulated and 46 downregulated), and in the Hy group, 81 proteins were dysregulated (40 upregulated and 41 downregulated) compared to those in the CO group, while 33 proteins exhibited differential regulation (12 upregulated and 21 downregulated) in the EC plasma samples compared to those in the Hy group. Vitamin D binding protein and complement C3 distinguished Hy and EC from CO with the greatest changes in expression. Among the differentially expressed proteins identified, enzymes with catalytic activity represented the largest group (42.9%). In terms of biological processes, most of the proteins were involved in cellular processes (28.8%), followed by metabolic processes (16.7%). STRING analysis for protein interactions revealed that the significantly differentially abundant proteins in the three groups are involved in three main biological processes: signalling of complement and coagulation cascades, regulation of insulin-like growth factor (IGF) transport and uptake by insulin-like growth factor binding proteins (IGFBPs), and plasma lipoprotein assembly, remodelling, and clearance. Conclusion The identified plasma protein markers have the potential to serve as biomarkers for differentiating between EC and Hy, as well as for early diagnosis and monitoring of cancer progression.

Background: Agonal bacteremia, diagnosed with postmortem positive blood culture results, is considered a possible contributing factor to death. We hypothesized that some premortem organ damage, such as kidney damage, can enhance agonal bacteremia. Methods: We performed a postmortem blood and alveolar fluid culture study in 30 cadavers and evaluated the relationship between blood culture results and clinical parameters, including organ damage (brain, heart, lung, kidney, liver and gastrointestinal tract). Results: A total of 23 cases (76.7%) were positive for blood culture; the number of cultured species was one in 12 cases, two in 7 cases, and three in 4 cases. The ratio of agonal bacteremia was significantly higher in patients with heart damage (100%, n = 13) and those with kidney damage (end-stage kidney damage, acute kidney injury, obstructive kidney failure, or metastatic kidney tumours) (100%, n = 13). The mean number of cultured species was 0.67 ± 0.98 in heart or kidney damage, 1.40 ± 0.55 in heart damage only, 1.40 ± 0.55 in kidney damage only, and 2.00 ± 0.93 in heart and kidney damage. As the number of damaged organs increased (0 organs, no heart/kidney damage; 1 organ, heart or kidney damage; and 2 organs, heart and kidney damage), the mean number of cultured species increased significantly (p for trend = 0.001964). Conclusion: Premortem kidney damage relates to agonal bacteremia. [ABSTRACT FROM AUTHOR]

Because of the special culture requirements of anaerobic bacteria, their low growth-rate and the difficulties to isolate them, MALDI-TOF MS has become a reliable identification tool for these microorganisms due to the little amount of bacteria required and the accuracy of MALDI-TOF MS identifications. In this study, the performance of MALDI-TOF MS for the identification of anaerobic isolates during a 4-year period is described. Biomass from colonies grown on Brucella agar was directly smeared onto the MALDI-TOF target plate and submitted to on-plate protein extraction with 1ml of 100% formic acid. Sequencing analysis of the 16S rRNA gene was used as a reference method for the identification of isolates unreliably or not identified by MALDI-TOF MS. Overall, 95.7% of the isolates were identified to the species level using the updated V6 database vs 93.8% with previous databases lacking some anaerobic species; 68.5% of the total were reliably identified with high-confidence score values (=2.0) and 95.0% with low-confidence values (score value =1.7). Besides, no differences between Gram-positive and Gram-negative isolates were detected beyond a slight decrease of correct species assignment for gram positive cocci (94.1% vs 95.7% globally). MALDI-TOF MS has demonstrated its usefulness for the identification of anaerobes, with high correlation with phenotypic and conventional methods. Over the study period, only 2.1% of the isolates could not be reliably identified and required molecular methods for a final identification. Therefore, MALDI-TOF MS provided reliable identification of anaerobic isolates, allowing clinicians to streamline the most appropriate antibiotic therapy and manage patients accordingly. [ABSTRACT FROM AUTHOR]

Tannin was extracted from different subspecies of Acacia nilotica, Acacia nilotica nilotica (Ann), Acacia nilotica tomentosa (Ant) and Acacia nilotica adansonii (Ana). The aim was to elucidate their structure and evaluate their reactivity as bioadhesives in the wood industry. The extracts were prepared by hot water extraction (90°C temperature). Their gel time with paraformaldehyde was used at first to compare their reactivity. The tannin contents and the percentage of total polyphenolic materials in different solutions of the extracts spray dried powder were determined by the hide powder method. Concentrated solutions (47%) were tested by both MALDI ToF, CNMR. The thermomechanical analysis (TMA) was performed to evaluate their modulus of elasticity (MOE) at different pHs. The gel times of all the three tannin extracts showed that their reactivity and it was comparable to other known procyanidin/prodelphinidin tannin extract types. Ana, Ann and Ant showed highest percent of total polyphenolic materials at 70%, 64%, and 57%, respectively. The 13CNMR spectra showed that the three subspecies of condensed tannins were mainly constituted of procyanidins (PC) and prodelphinidins (PD) in slightly different ratios. Ann (56.5% PC and 43.4% PD), Ant (57%PC and 43% PD) and Ana (58% PC and 42% PD). MALDI–TOF spectra showed the presence of flavonoid monomers, and oligomers some of which linked to short carbohydrates monomers or dimers. TMA revealed that the three types of tannins had high MOE at their initial pH (5). [ABSTRACT FROM AUTHOR]

Mycobacterium ulcerans ecovar Liflandii (Mu Liflandii) was identified as the causative agent of mycobacteriosis in a research colony of Zaire dwarf clawed frogs (Hymenochirus boettgeri) at the University of Michigan. Clinical presentation included lethargy, generalized septicemia, cutaneous granulomas, coelomic effusion, and acute mortality. Identification of the mycobacterial species was based on molecular, microbiological, and histopathologic characteristics. These findings indicate that Mu Liflandii is a primary cause of morbidity and mortality in Zaire dwarf clawed frogs and should be considered in the differential diagnosis of sepsis and coelomic effusion in amphibians. Mycobacterial speciation is important given the variability in pathogenesis within the family Mycobacteriaceae and the implications for both animal and human health as potential zoonoses. The Zaire dwarf clawed frog is a species common in the pet trade, and these findings provide consideration for this pathogen as a potentially important public health concern. This is the first report of Mu Liflandii infection in the genus Hymenochirus and illustrates the diagnostic challenges of differentiating among both mycolactone-producing mycobacteria and Mycobacterium marinum. Furthermore, we demonstrate the utility of environmental sampling for this pathogen within the tank system, suggesting this mode of sampling could replace the need for direct frog surveillance. [ABSTRACT FROM AUTHOR]

Background: Among gynaecological malignancies, endometrial cancer (EC) is the most prevalent type of uterine cancer affecting women. This study explored the proteomic profiles of plasma samples obtained from EC patients, those with hyperplasia (Hy), and a control group (CO). A combination of techniques, such as 2D-DIGE, mass spectrometry, and bioinformatics, including pathway analysis, was used to identify proteins with modified expression levels, biomarkers and their associated metabolic pathways in these groups. Methods: Thirty-four patients, categorized into three groups—10 with EC, 12 with Hy, and 12 CO—between the ages of 46 and 75 years old were included in the study. Untargeted proteomic analysis was carried out using two-dimensional difference in gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Results: In all three groups, 114 proteins that were significantly (p ≤ 0.05 and fold change ≥ 1.5) altered were successfully identified using peptide mass fingerprints (PMFs). Compared with those in the control group (CO), the EC samples had 85 differentially expressed proteins (39 upregulated and 46 downregulated), and in the Hy group, 81 proteins were dysregulated (40 upregulated and 41 downregulated) compared to those in the CO group, while 33 proteins exhibited differential regulation (12 upregulated and 21 downregulated) in the EC plasma samples compared to those in the Hy group. Vitamin D binding protein and complement C3 distinguished Hy and EC from CO with the greatest changes in expression. Among the differentially expressed proteins identified, enzymes with catalytic activity represented the largest group (42.9%). In terms of biological processes, most of the proteins were involved in cellular processes (28.8%), followed by metabolic processes (16.7%). STRING analysis for protein interactions revealed that the significantly differentially abundant proteins in the three groups are involved in three main biological processes: signalling of complement and coagulation cascades, regulation of insulin-like growth factor (IGF) transport and uptake by insulin-like growth factor binding proteins (IGFBPs), and plasma lipoprotein assembly, remodelling, and clearance. Conclusion: The identified plasma protein markers have the potential to serve as biomarkers for differentiating between EC and Hy, as well as for early diagnosis and monitoring of cancer progression. [ABSTRACT FROM AUTHOR]

Simple Summary: Septic exudates in the body cavities of dogs and cats are considered a critical clinical condition. The current elective diagnostic tools for detecting septic effusion are bacterial culture and fluid cytology. Although culture is considered the gold standard, clinicians may not have access to the results for several days. This may result in a delayed diagnosis of septic effusion, which may have adverse effects on patient outcomes. This study compared the performances of cytology and bacterial culture in the identification of septic exudative body cavity effusions in dogs and cats. The results of our investigation indicated moderate agreement between cytology and microbiology. Cytology and bacterial culture results for exudative body cavity effusions in dogs and cats can be misleading when conducted individually. To improve diagnostic accuracy, these two methodologies should be integrated. The elective test for the determination of the effusions etiopathogenesis is represented by physico-chemical analysis and cytology. Nevertheless, the bacterial culture and antibiotic sensitivity tests are crucial for setting therapy and for the outcome. This study compared cytology with microbiology in the etiologic diagnosis of exudative body cavity effusions in dogs and cats collected from October 2018 to October 2022. All samples underwent aerobic and anaerobic culture and cytology examination. Bacterial identifications were confirmed using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, whereas cytological samples were blindly evaluated either in May Grunwald-Giemsa (MGG) or Gram-stained samples by two board-certified clinical pathologists. A moderate agreement (κ = 0.454) between cytology and bacterial culture was revealed. The sensitivity of the cytological evaluation in our study ranged from 38.5% to 67.9%, and the specificity ranged from 88.9% to 100%, depending on the type of the effusion, so cytology may not be representative of the etiopathogenesis, whereas bacterial culture can misidentify or fail to isolate the correct pathogen for difficult in vitro growing due to the presence of inhibitory substances or contamination. Cytology and bacterial culture results for exudative body cavity effusions in dogs and cats can be misleading if conducted separately, so these two tests should be performed together to increase diagnostic accuracy. [ABSTRACT FROM AUTHOR]

Magnetic nanoparticles attached to hydrophilic polymer brushes containing immobilized Fe3+ ions were synthesized and used as a new strategy for phosphopeptide enrichment from digested tryptic proteins. To test the performance of nanoparticles in phosphopeptide enrichment, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used. Magnetic nanoparticles with the following features such as high adsorption capacity (about 360 mg/g), easy control and transfer by external magnetic field (high magnetic saturation, about 50 emu/g), excellent phosphopeptide recovery (93.4%) and no shadow effect (due to their non-porous structure), were synthesized. It has been shown that the presence of other proteins in high concentrations does not affect on the performance of nanoparticles. Furthermore, the lowest concentration of digested β-casein phosphopeptides detectable by nanoparticles was determined to be approximately 0.5 fmol µL− 1. The function of synthesized nanoparticles to enrich phosphopeptides in complex biological samples was also demonstrated by isolating four endogenous phosphopeptides from human serum. [ABSTRACT FROM AUTHOR]

With the ongoing mutation of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) leading to various variants, there is an urgent need for new diagnostic methods for SARS‐CoV‐2 infection. The existing nucleic acid test and antigen test suffer from long assay time and low sensitivity, respectively. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS)‐based nasal swabs analysis have been demonstrated as a promising technique in SARS‐CoV‐2 infection screening. However, the applicability of the technique in the different variants of SARS‐CoV‐2 is uncertain. Given the prevalence of the Omicron variant since 2022, we developed a MALDI‐TOF‐based diagnosis method with nasal swab samples to detect the infection by this variant. We collected 325 SARS‐CoV‐2‐positive and 221 SARS‐CoV‐2‐negative nasal swab samples, and the molecular mass fingerprints were acquired from the samples by MALDI‐TOF MS. Using a random forest machine learning classification model to analyze the molecular mass fingerprints MALDI‐TOF mass spectra, the accuracy of 97%, false negative rate of 0%, and false positive rate of 7.6% were achieved for the diagnosis of SARS‐CoV‐2 infection. Combining the MALDI‐TOF analysis with top‐down proteomics, we identified four potential protein biomarkers, that is, humanin‐like 4, thymosin beta‐10, thymosin beta‐4 and statherin, in the nasal swab for the diagnosis of coronavirus disease 2019. It was further found that the four protein biomarkers can also differentiate the SARS‐CoV‐2 original strains infection and Omicron strains infection. These results suggest that the MALDI‐TOF MS‐based nasal swab analysis holds effective diagnostic capabilities of SARS‐CoV‐2 infection, and shows promising potential for global application and extension to other infectious diseases. [ABSTRACT FROM AUTHOR]

The purpose of the research was microbiological screening using MALDI-TOF technology starting from bulk raw milk to the finished dairy product and analyzing microorganisms that were being detected during the technological process of production of Ukrainskyi hard rennet cheese and which were clinically significant for human and animal health. Methods. Microbial detection was performed by accumulation and inoculation using the sector inoculation method on differential media for aerobic and anaerobic microorganisms with further MALDI-TOF identification. Sampling was carried out at 7 stages of cheese production: starting from bulk raw milk to bactofugation, after bactofugation to a mixture normalized in fat content, a pasteurized mixture, a mixture prepared for coagulation, cheese after pressing, and cheese after maturation. Microflora studies were repeated three times, with 405 samples examined. Microbiological studies of Ukrainskyi hard rennet cheese using Maldi TOF technology starting from raw materials to finished dairy products showed the presence of microorganisms at all stages of production – from bulk milk to the finished product. During the entire period of experiments, 43 species of various microorganisms have been isolated and identified. However, the number and individual types of microorganisms differed at different stages of production. Some microorganisms that have been isolated in raw milk are also found in the final product, such as Acinetobacter baumannii and Escherichia coli. In total, 18 types of microorganisms have been isolated and identified in the final product – hard rennet cheese, including Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli, which are of particular concern in the context of safe consumption of this cheese.

To address the fungal wilt of pigeon pea caused by Fusarium oxysporium f. sp. udum, farmers currently rely on chemical fungicides, despite their harmful effects. However, there is a growing need for safer alternatives like green pesticides. Bacterial biocontrol agents and their derivatives serve as potential green pesticides in the management of plant pathogens. In the present study, we aimed to identify indigenous Bacillus subtilis strains effective against F. oxysporium f. sp. udum. We used PCR and MALDI-TOF analysis to identify the active components responsible for the efficiency of efficient strain. Biochemical studies of cell-free extracts extracted from B. subtilis strains demonstrated the highest biosurfactant activity in NBAIR BSWG1, with an oil displacement of 2 cm and an emulsification index of 60 %. Molecular characterization confirmed the presence of surfactin, fengycin, and iturin coding genes in the B. subtilis strains, among them, NBAIR BSWG1 showed the highest number of lipopeptide-producing genes. Meanwhile, NBAIR BSWG1 showed inhibition of 79.84 % against F. oxysporium f. sp. udum using cell-free extract. Further metabolite profiling of NBAIR BSWG1 using MALDI-TOF analysis further confirmed surfactin, fengycin, and iturin in the purified cell-free extract of NBAIR BSWG1. Two peaks with m/z of 923.77 and 1149.92 were identified as novel lipopeptide compounds which need further characterization. The present study identified NBAIR BSWG1 as an efficient bacterial strain for the inhibition of F. oxysporium f. sp. udum and its antifungal properties are mainly due to the production of cyclic lipopeptides.

Newborn screening (NBS) for sickle cell disease (SCD) requires a robust, high-throughput method to detect hemoglobin S (HbS). Screening for SCD is performed by qualitative methods, such as isoelectric focusing (IEF), and both qualitative and quantitative methods such as high performance liquid chromatography (HPLC), capillary electrophoresis (CE), and tandem mass spectrometry (MS/MS). All these methods detect HbS, as well as low-level or absent HbA, and also other variants of hemoglobin. HPLC is considered as a reference method for NBS, because of its high sensitivity and specificity in detecting HbS. NeoSickle®, a fully automated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platform, combined with automated sample processing, a laboratory information management system and NeoSickle® software for automatic data interpretation, has increased the throughput of SCD testing. The purpose of this study was to compare the performances of NeoSickle® and HPLC. A prospective study was conducted including 9,571 samples from the NBS program to compare MALDI-MS using NeoSickle® with an HPLC method. Correlation between the two methods was studied. For the MALDI-MS method, sensitivity, specificity, NPV, and PPV were calculated. We found over 99.4 % correlation between the HPLC and MALDI-MS results. NeoSickle® showed 100 % of sensitivity and specificity in detecting SCD syndrome, leading to positive and negative predictive values of 100 %. NeoSickle® is adapted to NBS for SCD, and can be used in first-line high-throughput screening to detect HbS, and beta-thalassemia major warning. When HbS is detected, second-line use of another specific method as HPLC is necessary. [ABSTRACT FROM AUTHOR]

Background: With the increase in immunosuppressed patients and antimicrobial misuse, Non-diphtheriae Corynebacterium species (NDC spp.) have risen as opportunistic pathogens in hospitalized patients. The current work aimed at comparing Vitek 2C automated identification cards versus MALDI-TOF (Matrix-associated laser-induced desorption ionization-time of flight) as the gold standard method for the identification of different NDC spp. and determining the antimicrobial susceptibility of NDC spp. by the disc diffusion method for proper management of patients. Methods: Thirty NDC spp. isolates were subjected to identification by MALDI-TOF MS and Vitek 2C and antibiotic susceptibility testing by the disc diffusion method. Results: Our study showed that Corynebacterium (C.) striatum was the most commonly isolated species (83.3%), followed by C. amycolatum (10%). 36.7 % of the isolates were recovered from blood culture while 20 % were recovered from deep wounds. As regard isolate identification by Vitek-2C ANC ID CARD compared to MALDI-TOF as the gold standard method, the agreement was 86.7%. Regarding the antimicrobial susceptibility by disc diffusion method, all the isolates showed increased resistance against penicillin (100%), ciprofloxacin (70%), gentamicin (93.3%), tetracycline (76.6%), and clindamycin (70%). On the other hand, vancomycin and linezolid showed promising results where 100 % of the isolates were susceptible. Conclusion: We spotlight that NDC spp. is not viewed as a contaminant organism anymore, particularly in immunosuppressed patients. Timely identification, proper clinical correlation, and appropriate therapeutic intervention can lead to favorable outcomes. [ABSTRACT FROM AUTHOR]

Introduction: Antimicrobial resistance (AMR) is a global health problem that requires early and effective treatments to prevent the indiscriminate use of antimicrobial drugs and the outcome of infections. Mass Spectrometry (MS), and more particularly MALDI-TOF, have been widely adopted by routine clinical microbiology laboratories to identify bacterial species and detect AMR. The analysis of AMR with deep learning is still recent, and most models depend on filters and preprocessing techniques manually applied on spectra. Methods: This study propose a deep neural network, MSDeepAMR, to learn from raw mass spectra to predict AMR. MSDeepAMR model was implemented for Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus under different antibiotic resistance profiles. Additionally, a transfer learning test was performed to study the benefits of adapting the previously trained models to external data. Results: MSDeepAMR models showed a good classification performance to detect antibiotic resistance. The AUROC of the model was above 0.83 in most cases studied, improving the results of previous investigations by over 10%. The adapted models improved the AUROC by up to 20% when compared to a model trained only with external data. Discussion: This study demonstrate the potential of the MSDeepAMR model to predict antibiotic resistance and their use on external MS data. This allow the extrapolation of the MSDeepAMR model to de used in different laboratories that need to study AMR and do not have the capacity for an extensive sample collection. [ABSTRACT FROM AUTHOR]

Featured Application: This research developed three cost-effective testing techniques to quickly and efficiently detect suspected samples to determine the authenticity of cosmetics utilizing MALDI-TOF, GC-MS and mid-IR instruments. (1) Background: The adulteration of cosmetics has become increasingly common, which seriously harms ordinary consumers. The counterfeit cosmetics pointed out in this study mainly refer to imitating genuine products in terms of ingredients and packaging. Ordinary consumers cannot distinguish their authenticity solely based on appearance and daily use. If there is a convenient and low-cost detection method that can expose this phenomenon of adulteration, it will be able to expose adulteration and protect the interests of consumers quickly and conveniently. (2) Methods: MALDI-TOF, GC-MS, and mid-IR were used to develop low-cost and fast methods for identifying the authenticity of cosmetics. Five types of liquid and five types of emulsion cosmetics purchased from container and wholesale markets were analyzed using the three instruments mentioned above, and their spectra and acquired data were carefully compared to determine their authenticity. MALDI-TOF and GC-MS directly tested cosmetic samples, and mid-IR spectroscopy tested the ink on the outer packaging of cosmetics. (3) Results: The data procured by MALDI-TOF can provide a representation of its product attributes; two liquid samples and one paste sample demonstrated inconsistent test outcomes with the corresponding reference samples, suggesting contamination. The results of GC-MS can illustrate the substance count within cosmetic samples; the comparison outcomes of the total ion chromatogram indicate that one paste sample was a counterfeit. The results attained from mid-IR were consonant with those acquired from the MALDI-TOF analysis and GC-MS. (4) Conclusions: These three newly developed techniques can all be effectively utilized for the task of detecting cosmetic adulteration and quality control in the manufacturing process. With regard to user-friendliness and rapidity, both MALDI-TOF and mid-IR outperform GC-MS, demonstrating consistently superior levels of detection. Conversely, GC-MS has unique advantages in identifying emulsion cosmetics containing a high amount of weak polarity and volatile substances. Consequently, these corresponding methods could serve as efficient and cost-effective ways to detect authenticity issues in real-world cosmetic products. [ABSTRACT FROM AUTHOR]

Objectives Matrix-assisted laser desorption ionization time of flight (MALDI-TOF) has revolutionized clinical microbiology laboratories, enabling the identification of microorganisms in minutes. Although this advancement has significantly enhanced patient care by providing early diagnosis, the use of MALDI-TOF for routine detection of antimicrobial resistance (AMR) remains an ongoing area of research. The purpose of this scoping review was to identify and summarize the existing literature on MALDI-TOF–based methodologies for detecting AMR. Methods The PubMed, Embase, and Cumulative Index to Nursing and Allied Health Literature databases were searched for articles published up to July 2023. Publications were carefully screened, and inclusion was based on their relevance to the topic and detailed explanations of the methodologies employed. Results We reviewed 49 articles and identified various MALDI-TOF–based approaches for detecting AMR that we grouped in 2 main categories: (1) bacterial growth detection following a brief incubation with antibiotics and (2) identification of specific mass spectral profiles associated with drug resistance. Recently, the use of machine learning algorithms has been explored to detect characteristic patterns associated with AMR. Thorough evaluation of each study's procedures was conducted to determine their potential for implementation in clinical laboratories. Conclusions MALDI-TOF shows promise as a tool for AMR detection, but its readiness for wider clinical adoption relies on ongoing research, development of algorithms and software for automated interpretation of mass spectra, and validation efforts. Its role in routine clinical practice may evolve as technology and knowledge progress. [ABSTRACT FROM AUTHOR]

This study is the first to evaluate the phytochemical content and biological properties of Tunisian T. platyphyllos Scop. A total of 23 compounds of essential oils were identified by gas chromatography-mass spectrometry (GC-MS) analysis of bracts and fruit extracts. The results show that oxygenated monoterpenes were the dominant class of essential oils. The phenolic composition was investigated by matrix-assisted laser desorption/ionizationtime of flight (MALDI-TOF). The analysis showed that the chemical profiles of the ethanolic extracts of bracts and fruits are substantially similar. The highest polyphenol content was found in the ethanolic extracts of the fruits (7.65 mg gallic acid equivalents (GAE).g−1 on dry weight (DW). As for the antioxidant capacity, it was recorded in the essential oils of the fruits (IC50 = 0.91 mg.mL−1). The results obtained showed that the antioxidant activity of the fruit essential oil was higher than that of the ethanol extract. The fruit essential oil was also found to have the highest reducing power (IC50 = 0.67 mg.mL−1). In terms of iron-chelating power, fruit essential oil has the highest chelating power (IC50 = 2.03 mg.mL−1). Compared to the ethanolic extract, the essential oil had the maximum antioxidant capacity. The enzymatic activity of acetylcholinesterase (AChE) inhibition of the essential oil from T. platyphyllos bracts had the most potent inhibitory effect (IC50 = 0.77 mg.mL−1), followed by the essential oil from the fruits (IC50 = 0.95 mg.mL−1). The results suggest that T. platyphyllos can be used as a potential source of naturally occurring bioactive compounds and antioxidants. [ABSTRACT FROM AUTHOR]

The occurrence of Listeria monocytogenes in marine fish and fish market areas was investigated. Two hundred and eighty-eight samples (123 environmental samples—siphons, knives, cutting boards, floor, sinks, water, and ice—and 165 marine fish samples) were examined. Twenty-four isolates were characterized as Listeria monocytogenes (five from environmental samples (4.0%) and 19 from fish samples (11.5%)). The strains were further characterized according to their antibiotic resistance, pathogenicity, and biofilm formation ability. They were molecularly serotyped as IIc (n = 22) and IVb (n = 2) and possessed all the virulence genes tested (inlA, inlB, inlC, inlJ, actA, hlyA, iap, plcA, and prfA), except for two strains lacking the hlyA and iap genes, respectively. All strains showed strong (41.7%) or moderate biofilm-producing ability (58.3%) and almost all showed resistance to at least one antibiotic, with the highest rates being observed against clindamycin and vancomycin. The proteomic analysis by MALDI-TOF revealed two distinct clusters that involved strains from fish only and those from both fish and the environment. The presence of Listeria monocytogenes in the fish-market environment and marine fish, along with the pathogenicity and persistence characteristics of the seafood-related strains, emphasize the need for vigilance concerning the spread of this notorious foodborne pathogen. [ABSTRACT FROM AUTHOR]

In this study, an antimicrobial protein (AMP) from the epidermis of catfish (Clarias batrachus) was purified and characterized for its molecular and antimicrobial properties. The crude epidermal extract was subjected to precipitation of proteins using 70% ammonium sulphate saturation followed by dialysis and protein separation by reverse phase ultra fast liquid chromotography (UFLC) using the C18 column. The active peptides obtained from fractions 1, 2, and 3 were screened for antimicrobial activity against different bacterial pathogens with a minimum 5 mg/ml inhibitory concentration. An active fraction with a retention time (RT) of 27.069 from the UFLC chromatogram exhibited significant antimicrobial activity against broad-spectrum bacterial and fungal organisms, including multidrug-resistant clinical isolates. The RT 27.069, identified as cationic AMP of fraction-3, has a molecular weight of 25 kDa as determined by MALDI-TOF mass spectrometry. Further studies by Mascot search and BLAST analysis using the partial sequence of AMP showed that the protein has high homologs to pleurocidin-like peptide (Plp) from a fish Pleuronectus americanus. Moreover, the identified AMP is a cationic peptide with a good stability index, having a score of 31.50 predicted by the ProtParam. Therefore, the identified antimicrobial peptide (Plp) indicates that this cost-effective natural substance obtained from fish could potentially be used as a treatment for several bacterial and fungal infections in humans. [ABSTRACT FROM AUTHOR]

Background. Honey is a nutritious food made by bees from nectar and sweet deposits of flowering plants and has been used for centuries as a natural remedy for wound healing and other bacterial infections due to its antibacterial properties. Honey contains a diverse community of bacteria, especially probiotic bacteria, that greatly affect the health of bees and their consumers. Therefore, understanding the microorganisms in honey can help to ensure the quality of honey and lead to the identification of potential probiotic bacteria. Methods. Herein, the bacteria community in honey produced by Apis cerana was investigated by applying the next-generation sequencing (NGS) method for the V3≥V4 hypervariable regions of the bacterial 16S rRNA gene. In addition, lactic acid bacteria (LAB) in the honey sample were also isolated and screened for in vitro antimicrobial activity. Results. The results showed that the microbiota of A. cerana honey consisted of two major bacterial phyla, Firmicutes (50%; Clostridia, 48.2%) and Proteobacteria (49%; Gammaproteobacteria, 47.7%). Among the 67 identified bacterial genera, the three most predominant genera were beneficial obligate anaerobic bacteria, Lachnospiraceae (48.14%), followed by Gilliamella (26.80%), and Enterobacter (10.16%). Remarkably, among the identified LAB, Lactobacillus kunkeei was found to be the most abundant species. Interestingly, the isolated L. kunkeei strains exhibited antimicrobial activity against some pathogenic bacteria in honeybees, including Klebsiella spp., Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus. This underscores the potential candidacy of L. kunkeei for developing probiotics for medical use. Taken together, our results provided new insights into the microbiota community in the A. cerana honey in Hanoi, Vietnam, highlighting evidence that honey can be an unexplored source for isolating bacterial strains with potential probiotic applications in honeybees and humans. [ABSTRACT FROM AUTHOR]

Background and Objectives: Nucleotide delivery has emerged as a noteworthy research trend in recent years because of its potential utility in addressing a range of genetic defects resulting in the presence of incorrect nucleotides. The primary goals of this research were to create and to characterize polyurethane microstructures, with the aim of utilizing them for nucleotide transport. Materials and Methods: Two samples were prepared using an aliphatic diisocyanate in reaction with a mixture of polyethylene glycol and polycaprolactone diol, where 2′-deoxycytidinic acid was used as the active agent and glycerol 1,2-diacetate was used as an enhancer of the aqueous solubility. The solubility, pH, size distribution, and surface charge of the samples were measured, and encapsulation efficacy and release, cell proliferation, and irritation tests on mouse skin were conducted. Results: The results showed almost neutral acidic–basic structures with a high heterogeneity, and a medium tendency to form clusters with non-cytotoxic and non-irritative potentials. Conclusions: Future research could explore the efficacy of this carrier in delivering other nucleotides, as well as investigating the long-term effects and safety of these microstructures in vivo. [ABSTRACT FROM AUTHOR]

Introduction: In hospitalized patients, candidemia accounts for considerable morbidity and mortality. Knowledge of local species epidemiology and antifungal susceptibility is important when considering therapy. This study was performed to speciate Candida isolates from inpatients admitted with blood stream infections in our hospital and to assess their risk factors. Materials and Methods: This present study of 2 years' duration was performed from April 2022 to March 2024. BacT/Alert aerobic culture bottle (bioMérieux, France) was used for all routine blood cultures and incubated for 5 days at 37°C. Any blood culture growing yeast was identified by Gram stain and MALDI-TOF MS (Bruker Daltonics). Disk diffusion method was used for antifungal susceptibility testing and test was performed according to clinical laboratory standard institute (CLSI) guidelines. Results: About 132 Candida isolates from blood were isolated in 2 years. Predominance of males (59%) was noted with the median age of 52 years. Majority were non- albicans Candida (81.06 %) with C. tropicalis (31.81%) and C. auris (13.63%). Susceptibility to fluconazole and voriconazole was 100% for both C. albicans and C. tropicalis isolates. Fluconazole resistance of 11.77% and 100% was observed in C. parapsilosis and C. auris isolates respectively. Conclusion: The predominance of non-albicans Candida with increasing fluconazole resistance and isolation of C. auris as the third most common non-albicans Candida species in this study is of concern. Accurate identification of non-albicans Candida specially C. auris and strict infection control policies should be in place to combat the spread of C. auris in the hospital.

Dichlorodiphenyltrichloroethane is an organo-chlorine insecticide used for malaria and agricultural pest control, but it is the most persistent pollutant, endangering both human and environmental health. The primary aim of the research is to screen, characterize, and assess putative fungi that degrade DDT for mycoremediation. Samples of soil and wastewater were gathered from Addis Ababa, Koka, and Ziway. Fungi were isolated and purified using potato dextrose media. Matrix-Assisted Laser Desorption, Ionization, and Flight Duration The technique of mass spectrometry was employed to identify fungi. It was found that the finally selected isolate, AS1, was Aspergillus niger. Based on growth factor optimization at DDT concentrations (0, 3500, and 7000 ppm), temperatures (25, 30, and 35 °C), and pH levels (4, 7, and 10), the potential DDT-tolerant fungal isolates were investigated. A Box-Behnken experimental design was used to analyze and optimize fungal biomass and sporulation. The highest biomass (0.981 ± 0.22 g) and spore count (5.60 ± 0.32 log/mL) of A. niger were found through optimization assessment, and this fungus was chosen as a potential DDT-degrader. For DDT degradation investigations by A. niger in DDT-amended liquid media, gas chromatograph-electron capture detector technology was employed. DDT and its main metabolites, DDE and DDD, were eliminated from both media to the tune of 96–99 % at initial DDT concentrations of 1750, 3500, 5250, and 7000 ppm. In conclusion, it is a promising candidate for detoxifying and/or removing DDT and its breakdown products from contaminated environments.

Abstract With the ongoing mutation of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) leading to various variants, there is an urgent need for new diagnostic methods for SARS‐CoV‐2 infection. The existing nucleic acid test and antigen test suffer from long assay time and low sensitivity, respectively. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS)‐based nasal swabs analysis have been demonstrated as a promising technique in SARS‐CoV‐2 infection screening. However, the applicability of the technique in the different variants of SARS‐CoV‐2 is uncertain. Given the prevalence of the Omicron variant since 2022, we developed a MALDI‐TOF‐based diagnosis method with nasal swab samples to detect the infection by this variant. We collected 325 SARS‐CoV‐2‐positive and 221 SARS‐CoV‐2‐negative nasal swab samples, and the molecular mass fingerprints were acquired from the samples by MALDI‐TOF MS. Using a random forest machine learning classification model to analyze the molecular mass fingerprints MALDI‐TOF mass spectra, the accuracy of 97%, false negative rate of 0%, and false positive rate of 7.6% were achieved for the diagnosis of SARS‐CoV‐2 infection. Combining the MALDI‐TOF analysis with top‐down proteomics, we identified four potential protein biomarkers, that is, humanin‐like 4, thymosin beta‐10, thymosin beta‐4 and statherin, in the nasal swab for the diagnosis of coronavirus disease 2019. It was further found that the four protein biomarkers can also differentiate the SARS‐CoV‐2 original strains infection and Omicron strains infection. These results suggest that the MALDI‐TOF MS‐based nasal swab analysis holds effective diagnostic capabilities of SARS‐CoV‐2 infection, and shows promising potential for global application and extension to other infectious diseases.

Turquoise blue dye is frequently used for industrial dyeing applications. But the release of untreated colored wastewater became an environmental and public health hazard. Microbial remediation of Azodye is environmentally safe and an alternative to a physicochemical approach. The aim of this research is to isolate and characterize turquoise blue dye degrading microbes from polluted environment. Microbial isolation and purification from soil and effluent sample was done on PDA and NA. Turquoise blue dye degrading test was investigated under optimized conditions using -the definitive screening design method. UV–Vis spectrophotometer used to measure the degradation percentage at 620 nm and 25 °C. The results revealed that 24 fungi and 6 bacterial species were identified from the contaminated site using Biolog Microstation and MALDI-TOF. Among all identified microbial species Pencilium citrinum Thom BCA & Penicillium heriquei show the highest percentage decolorization of turquoise blue dye up to 300 ppm with 90 % removal at pH4 and 87 % at pH 7 up to 400 ppm respectively. The azodye degradation ability of these fungi species used in the development of mycoremediation technologies provide an alternative option for Azodye removal after HPLC analysis, molecular characterization, and toxic analysis.

Bryndza cheese is a traditional Slovakian product. In this research, we have investigated whether it would be possible to freeze bryndza, store it at a temperature of -18°C, and then thaw and place it on the market during the off-production season. The current legislation in Slovakia does not allow this procedure. The freezing process was chosen based on the request of several small food business operators who would like to replace the process of preserving the primary raw material, matured salted ewe’s lump cheese, in barrels due to acrid-sour taste. Bryndza cheese is preferred by consumers due to its unique microbial composition, which is beneficial for their health. Many microorganisms present in bryndza are probiotics. For this reason, we wanted to determine how the microflora in the bryndza cheese changes after freezing. These findings have practical implications for the food industry, particularly for small food business operators, who can potentially adopt freezing to preserve bryndza, thereby extending its shelf life and availability to consumers. Additionally, in many households, people store bryndza in their freezers after purchasing and use it to prepare dishes. Understanding the role of microorganisms in the ripening process and during storage can provide valuable data on Brynzda quality and safety. The present study aimed to analyse the representation of microorganisms in “Bryndza” samples at the beginning of storage and after 6 months of storage at a temperature of -18 °C. A total of 10 samples of “Bryndza” cheese made from pasteurised milk were analysed. Analysis of total viable counts of viable bacteria (TVC), coliform bacteria (CB), lactic acid bacteria (LAB), and microscopic filamentous fungi (MFF) was performed using the plate dilution method. Isolated strains of microorganisms were identified with mass spectrometry MALDI-TOF MS Biotyper. A total of 295 isolates from Bryndza cheese were identified at the start of storage and 220 isolates at the end of storage of samples. The dominant species of microorganisms found in Bryndza cheese were lactic acid bacteria, especially Lactococcus lactis, with 68 isolates and Lactobacillus fermentum, with 41 isolates at the start of storage. The most frequently isolated species were Lactococcus lactis, with 62 isolates, and Limosilactobacillus fermentum, with 33 isolates. Our results show that important lactic acid bacteria were present in the bryndza even after 6 months of freezing, but coliform bacteria were absent. Experimental outputs: TVC: showed no significant decrease (p-value = 0.0137); LAB: No significant decrease in lactic acid bacteria counts post-storage; MFF: Significant decrease in microscopic filamentous fungi post-storage; CB: Qualitative analysis indicates a significant reduction to undetectable levels after storage. Long-term storage of bryndza at -18°C is safe from a microbiological point of view.

Nocardiosis is a rare infection of the lung, brain and skin and can be a localized or disseminated infection. Here, we report a case of a 56-year-old male patient who presented with a right testicular torsion for which right orchiectomy was done in the hospital. In the post-operative period, patient developed increased respiratory insufficiency and was intubated. Chest X-ray revealed bilateral fluffy opacities most likely due to infective etiology. Endotracheal tube aspirate sample was sent for Gram staining and culture, organism present showed characteristics like Nocardia. MALDI-TOF identified organism as Nocardia otitidiscaviarum. The patient was treated with cotrimoxazole and ceftriaxone. His clinical condition improved. We report this case because Nocardia otitidiscaviarum is a rare cause of pulmonary nocardiosis and although there is high mortality reported in nocardiosis, early diagnosis because of the availability of advanced diagnostic technology and prompt treatment can help save the patient.

Background. Melioidosis cases are increasing in Southeast Asia, posing a significant challenge owing to the rising number of diabetic and immunecompromised patients. Pneumonia is the most common presentation of melioidosis, while cutaneous melioidosis is rare. Objective. We report a case of primary cutaneous melioidosis (PCM) that eventually required intensive care unit (ICU) management. Method. We describe the case of a 33-year-old male with uncontrolled diabetes mellitus who initially presented with a skin lesion, which was followed by gradual multiorgan involvement diagnosed as melioidosis based on culture-positive results from blood and synovial fluid, and needed ICU admission. Results. He was successfully treated with intravenous antibiotics and invasive mechanical ventilation after ICU admission. Conclusion. Cutaneous melioidosis can lead to ICU admission and, if untreated, has a high fatality rate. Faster diagnostic methods like VITEK 2 Compact and MALDI TOF substantially reduce delays in initiating required treatment. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The composition of the vaginal microbiota is known to be influenced by various factors and to be associated with several disorders affecting women’s health. Although metagenomics is currently a widely used method for studying the human microbiota, it has certain limitations, such as a lack of information on bacterial viability. It is therefore important to use culture-based methods such as culturomics. Here, we used 35 different culture conditions to comprehensively characterize the vaginal bacterial diversity of a single woman's flora. A total of 206 bacterial species, belonging to six phyla (for a little more than half to Firmicutes, followed mainly by Actinobacteria, Bacteroidetes, and Proteobacteria) and 45 families, and 2 fungal species were cultivated. While several species of lactobacilli have been isolated, a wide variety of other bacteria were also separated, including 65 never reported before in vaginal flora, including a new bacterial species, Porphyromonas vaginalis sp. nov. Extensive culture-based methods are essential to establish a comprehensive, evidence-based repertoire of bacterial viability. If combined with molecular methods, they can provide a much more thorough understanding of the vaginal microbiota and fulfil the unknown part of metagenomic studies. [ABSTRACT FROM AUTHOR]

An understanding of the normal bacterial microbiota of any organ is essential to provide the background to conditions and interventions that might cause the microbiota to change. In the vagina of the mare, a change of bacterial microbiota could be induced by introduction of semen, treatment with antibiotics, discharge from an unhealthy uterus etc. Previous studies on equine vaginal bacteria are not all conducted in the same way and results are not altogether consistent. Therefore, this study was designed to provide a deeper understanding of the bacterial microbiota of the mare vagina, and possible changes throughout the estrous cycle. The cranial portion of the vagina was sampled on day 0 (ovulation), day 3, day 7, and day 14 of the estrous cycle. The vaginal sampling was conducted with double-guarded occluded swabs from the cranial floor of the vagina. Ovulation was determined by rectal palpation and ultrasonic examination, and the day 0 samples were taken within ±24 h of ovulation. Swabs were brought to the laboratory in Amies medium within 2–3 h and were plated out immediately on both selective and non-selective agars. Results were registered as amount of growth (qualitatively), bacterial species and number of isolates. Bacterial growth was highest on day 3 and 7, representing the beginning and middle of diestrus. The dominant bacteria were Escherichia coli and Streptococcus zooepidemicus. Escherichia coli was especially dominant in maiden mares, compared to the mares that had foaled. An increase in bacterial diversity throughout the estrous cycle was observed, being highest on day 14. These results suggest that there are changes in the bacterial microbiota of the mare vagina throughout the normal estrous cycle. • Equine vaginal bacteria were sampled on days 0, 3, 7 and 14 of the estrous cycle. • Bacterial growth was highest on day 3 and 7, during diestrus. • Bacterial diversity increased throughout the cycle, being highest on day 14. • Differences in microbiota were seen in maiden mares compared to foaled mares. • Knowledge of cyclical changes in normal flora is needed. [ABSTRACT FROM AUTHOR]

Background: Mastitis is the major disease affecting milk production of dairy cattle, and milk is an obvious substrate for the detection of both the inflammation and its causative infectious agents at quarter, cow, or herd levels. In this review, we examine the use of milk to detect inflammation based on somatic cell count (SCC) and other biomarkers, and for the detection of mastitis pathogens through culture‐based and culture‐free methods. Findings: The use of SCC at a cow or bulk milk level to guide udder health management in lactation is well‐established, and SCC is increasingly used to guide selective dry cow treatment. Other markers of inflammation include electrical conductivity, which is used commercially, and markers of disease severity such as acute phase proteins but are not pathogen‐specific. Some pathogen‐specific markers based on humoral immune responses are available, but their value in udder health management is largely untested. Commercial pathogen detection is based on culture or polymerase chain reaction, with other tests, for example, loop‐mediated isothermal amplification or 16S microbiome analysis still at the research or development stage. Matrix‐assisted laser desorption ionisation time of flight (MALDI‐ToF) is increasingly used for the identification of cultured organisms whilst application directly to milk needs further development. Details of test sensitivity, specificity, and use of the various technologies may differ between quarter, cow, and bulk milk applications. Conclusions: There is a growing array of diagnostic assays that can be used to detect markers of inflammation or infection in milk. The value of some of these methods in on‐farm udder health improvement programs is yet to be demonstrated whilst methods with proven value may be underutilised. [ABSTRACT FROM AUTHOR]

Nocardiosis is a rare infection of the lung, brain and skin and can be a localized or disseminated infection. Here, we report a case of a 56-year-old male patient who presented with a right testicular torsion for which right orchiectomy was done in the hospital. In the post-operative period, patient developed increased respiratory insufficiency and was intubated. Chest X-ray revealed bilateral fluffy opacities most likely due to infective etiology. Endotracheal tube aspirate sample was sent for Gram staining and culture, organism present showed characteristics like Nocardia. MALDI-TOF identified organism as Nocardia otitidiscaviarum. The patient was treated with cotrimoxazole and ceftriaxone. His clinical condition improved. We report this case because Nocardia otitidiscaviarum is a rare cause of pulmonary nocardiosis and although there is high mortality reported in nocardiosis, early diagnosis because of the availability of advanced diagnostic technology and prompt treatment can help save the patient. [ABSTRACT FROM AUTHOR]

Bacterial vaginosis (BV) is a common dysbiosis of unclear etiology but with potential consequences representing a public health problem. The diagnostic strategies vary widely. The Amsel criteria and Nugent score have obvious limitations, while molecular biology techniques are expensive and not yet widespread. We set out to evaluate different diagnostic strategies from vaginal samples using (1) a combination of abnormal vaginal discharge and vaginal pH > 4.5; (2) the Amsel-like criteria (replacing the "whiff test" with "malodorous discharge"); (3) the Nugent score; (4) the molecular quantification of Fannyhessea vaginae and Gardnerella vaginalis (qPCR); (5) and MALDI-TOF mass spectrometry (we also refer to it as "VAGI-TOF"). Overall, 54/129 patients (42%) were diagnosed with BV using the combination of vaginal discharge and pH, 46/118 (39%) using the Amsel-like criteria, 31/130 (24%) using qPCR, 32/130 (25%) using "VAGI-TOF", and 23/84 (27%) using the Nugent score (not including the 26 (31%) with intermediate flora). Of the 84 women for whom the five diagnostic strategies were performed, the diagnosis of BV was considered for 38% using the combination of vaginal discharge and pH, 34.5% using the Amsel-like criteria, 27% using the Nugent score, 25% using qPCR, and 25% using "VAGI-TOF". When qPCR was considered as the reference, the sensitivity rate for BV was 76.2% for the combination of vaginal discharge and pH, 90.5% for the Amsel-like criteria, 95.2% for the Nugent score, and 90.5% for "VAGI-TOF", while the specificity rates were 74.6%, 84.1%, 95.3%, and 95.3%, respectively. When the Nugent score was considered as the reference, the sensitivity for BV was 69.6% for the combination of vaginal discharge and pH, 82.6% for the Amsel-like criteria, 87% for qPCR, and 78.7% for "VAGI-TOF", while the specificity rates were 80%, 94.3%, 100%, and 97.1%, respectively. Overall, the use of qPCR and "VAGI-TOF" provided a consistent diagnosis of BV, followed by the Nugent score. If qPCR seems tedious and for some costly, "VAGI-TOF" could be an inexpensive, practical, and less time-consuming alternative. [ABSTRACT FROM AUTHOR]