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Summary: The study explores the enhanced functional and bioactive properties of Konjac Glucomannan Hydrolysate (KGMH) by partially degrading Konjac Glucomannan (KGM) using β‐mannanase over 60 min. KGM concentrations (40% and 50% w/w) were treated with 200–300 U g−1 of the enzyme over 60 min. The hydrolysis of KGM was monitored by colorimetry, with DP values ranging from 4.20 to 6.16 for 40% KGM and 4.10 to 4.60 for 50% KGM. MALDI‐TOF‐MS analysis confirmed typical oligosaccharides with DP values from 2 to 9 and some acetyl substitutions. The optimal conditions of 40% KGM with 250 U g−1 enzyme and varying hydrolysis times produced KGMHs with a wide range of DPs, demonstrating in vitro antioxidant and anti‐glycation activities. The results showed significant bioactivities (P < 0.05) positively correlated with lower DP values. This study emphasises the potential of KGMH as a novel functional food ingredient, highlighting its bioactive properties and the significant impact of DPs on the biological functionality of saccharides. [ABSTRACT FROM AUTHOR]

In the present work, the production of biosurfactant was studied from the bacterial strains isolated from the marine sponge Clathria sp. sample collected from oil-contaminated coastal region of V.O. Chidambaranar Harbour, Thoothukudi, Tamil Nadu, India. Six morphologically distinct biosurfactant-producing bacterial strains (SJ1-SJ6) isolated on oil agar plates were further screened for biosurfactant production (BP). Based on the screening results, the isolate SJ2 showed a surface tension reduction value of 31.6 mN m−1, oil displacement zone of 26 mm and positive results of drop collapse test with the emulsification index percentage (EI24%) of 68.14%, and the bacterial adherence to hydrocarbon percentage of 79.2% was selected as the most potential BP strain, and it was identified as Enterobacter cloacae using 16S rRNA gene sequencing. Growth and BP optimization revealed 36 h of incubation period, 150 rpm agitation, pH 7.5, 37 °C temperature, 1% salinity, 2% glucose as carbon source and 1% yeast extract as nitrogen source were the ideal conditions with the biosurfactant yield of 2.61 g L−1. FTIR and MALDI-TOF–MS analysis of the biosurfactant showed the presence of functional groups, and it was a rhamnolipid. This rhamnolipid biosurfactant showed antibacterial activity against tested bacterial pathogens. [ABSTRACT FROM AUTHOR]

Iron plays vital roles in important biological processes in fish, but can be toxic in high concentrations. The information on metalloproteins that participate in maintenance of Fe homeostasis in an esocid fish, the northern pike, as an important freshwater bioindicator species, are rather scarce. The aim of this study was to identify main cytosolic constituents that sequester Fe in the northern pike liver. The method applied consisted of two-dimensional HPLC separation of Fe-binding biomolecules, based on anion-exchange followed by size-exclusion fractionation. Apparent molecular masses of two main Fe-metalloproteins isolated by this procedure were ~360 kDa and ~50 kDa, with the former having more acidic pI, and indicated presence of ferritin and hemoglobin, respectively. MALDI-TOF-MS provided confirmation of ferritin subunit with a m/z peak at 20.65 kDa, and hemoglobin with spectra containing main m/z peak at 16.1 kDa, and smaller peaks at 32.1, 48.2, and 7.95 kDa (single-charged Hb-monomer, dimer, and trimer, and double-charged monomer, respectively). LC-MS/MS with subsequent MASCOT database search confirmed the presence of Hb-β subunits and pointed to close relation between esocid and salmonid fishes. Further efforts should be directed towards optimization of the conditions for metalloprotein analysis by mass spectrometry, to extend the knowledge on intracellular metal-handling mechanisms. [ABSTRACT FROM AUTHOR]

Lectin (TfgL) was purified from the seeds of Trigonella foenum-graecum (Fenugreek) belonging to fabaceae family by ammonium sulphate precipitation, ion exchange and followed by size exclusion chromatography. SDS–PAGE analysis revealed that TfgL molecular weight is approximately 27 kDa. 2D-PAGE reveals the existence of two isolectins (pI values of 6.3 and 6.7) with acidic nature and charge heterogeneity. The MALDI-TOF–MS and peptide mass fingerprinting investigation of TfgL showed sequence similarity with a lectin. The hemagglutinating activity of TfgL was stable in broad range of temperature 37–90 °C and at varied pHs 3, 7.6 and 10. Far-UV circular dichroism measurements showed that TfgL is mostly composed of α-helix (84.5%), β-sheet (6.5%), β-turns (5%) and unordered structures (4%). TfgL showed conformational stability in wide range of temperatures (20‒90 °C) and pHs (3, 7.6 and 10) but lost its secondary structure in the presence of 6 M Gdn.HCl. Quenching titrations were carried out with acrylamide and iodide quenchers in order to investigate the exposure and accessibility of the protein tryptophan residues. Maximum quenching observed with acrylamide compared to iodide revealed that the Trp residues of TfgL are buried in the protein core, which is hydrophobic in nature. TfgL showed binding affinity towards porphyrin, the association constant (Ka), for MnTSPP and MnTMPyP was calculated to be 1.2 × 106 M‒1 and 3.45 × 106 M‒1, respectively. [ABSTRACT FROM AUTHOR]

The exploration of microbial genetic resources for the production of fFen-flavor Baijiu has not only enriched the microbial library for baijiu production but has also laid the foundation for process improvement and strain optimization in baijiu brewing. In this study, a total of 177 fungal isolates were screened, including Saccharomyces cerevisiae, non-Saccharomyces cerevisiae, molds, and some pathogenic bacteria. Among them, Saccharomyces cerevisiae was the most abundant with 119 isolates, playing a major role in the fermentation of baijiu production. A total of 148 bacterial isolates were obtained from the fermentation mash samples, showing greater diversity compared to fungi. Bacillus species were the most abundant, with 94 isolates. Bacillus licheniformis, in particular, can produce a rich enzymatic system and flavor precursors, making it an important contributor to the sensory quality of baijiu. Lactic acid bacteria were the second most abundant, with 16 isolates. Additionally, five pathogenic fungal species were identified, including Candida pelliculosa, Candida lusitaniae, Fusarium oxysporum, Fusarium solani and Talaromyces marneffei. Six pathogenic bacterial species were also isolated, namely Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus xylosus, Moraxella osloensis, Actinomyces meyeri and Stenotrophomonas maltophilia. Finally, two strains of high acetate ethyl ester-producing yeast and lactate-degrading bacteria with good tolerance to temperature, pH, and ethanol concentration were identified as Saccharomyces cerevisiae and Bacillus licheniformis, respectively. [ABSTRACT FROM AUTHOR]

The purpose of this study was to isolate, identify, and evaluate the antibacterial and probiotic potential of bacteria from honeydew honey collected in Poland. Isolates (189 colonies from 10 honey samples) were evaluated for their antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Yersinia enterocolitica, and then identified by MALDI-TOF-MS. The isolates with the greatest antimicrobial properties were screened for their probiotic potential. The total number of bacteria isolated from honey did not exceed the value of 2.5 × 102 CFU/mL. The Bacillus pumilus/altitudinis, B. licheniformis, and Bacillus cereus groups were the dominant identified bacteria. Almost 16% of the isolates expressed antibacterial potential against three pathogenic bacteria, over 20% against two, while almost 34% of the isolates did not inhibit any. The survival rate of the isolates under gastrointestinal tract conditions was higher after 4 h of exposure to bile salts (>60% survival rate for 66.66% of the isolates), while at pH 2.0, it was lower (>50% survival rate for 44% of the isolates). The most resistant isolate B. pumilus/altitudinis survived at a rate of 77% at low pH and 108% with bile salts. These results confirmed that honeydew honey is a promising reservoir of bacteria that produces metabolites with antimicrobial and probiotic potential. [ABSTRACT FROM AUTHOR]

Quinoa is an Andean crop that stands out as a high-quality protein-rich and gluten-free food. However, its increasing popularity exposes quinoa products to the potential risk of adulteration with cheaper cereals. Consequently, there is a need for novel methodologies to accurately characterize the composition of quinoa, which is influenced not only by the variety type but also by the farming and processing conditions. In this study, we present a rapid and straightforward method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to generate global fingerprints of quinoa proteins from white quinoa varieties, which were cultivated under conventional and organic farming and processed through boiling and extrusion. The mass spectra of the different protein extracts were processed using the MALDIquant software (version 1.19.3), detecting 49 proteins (with 31 tentatively identified). Intensity values from these proteins were then considered protein fingerprints for multivariate data analysis. Our results revealed reliable partial least squares-discriminant analysis (PLS-DA) classification models for distinguishing between farming and processing conditions, and the detected proteins that were critical for differentiation. They confirm the effectiveness of tracing the agricultural origins and technological treatments of quinoa grains through protein fingerprinting by MALDI-TOF-MS and chemometrics. This untargeted approach offers promising applications in food control and the food-processing industry. [ABSTRACT FROM AUTHOR]

Objective: To understand the virulence genes and molecular epidemiological characteristics of human-infected strains of Streptococcus suis in Rui'an, Zhejiang Province, from 2021 to 2022, and to provide a scientific basis for diagnosis, treatment, prevention, and control. Methods: Three blood-borne strains of Streptococcus suis were analysed by morphological observation, identification, and drug sensitivity tests. We performed polymerase chain reaction (PCR) amplification of their main seven virulence factors and housekeeping genes. This was followed by virulence analysis and multilocus sequence typing. We analysed their relationships with local pathogens from previous years. Results: Three Streptococcus suis strains were isolated from the blood samples of three patients. From these, the virulence genotypes demonstrated that the two strains were orf2+ and ef+/orf2+/sly+, respectively. The Multilocus sequence typing (MLST) typing results demonstrated that the two strains were ST25 and ST7, respectively. Conclusion: The first isolation of ST25 Streptococcus suis in Rui'an was presumed to have a close affinity with the endemic strain in North America. The other strain was an ST7 clone, consistent with the endemic strain in Sichuan, and which may have originated from Sichuan. Virulence genotype analysis demonstrated that different virulence genes of the pathogens resulted in different clinical manifestations.

A facile and mild method based on self-assembled lysozyme (LYZ) to fabricate bifunctional MNPs@UIO-66-Arg core-shell-satellite nanocomposites (CSSNCs) is reported for the high-efficiency enrichment of phosphopeptides. Under physiological conditions, LYZ rapidly self-assembled into a robust coating on Fe3O4@SiO2 magnetic nanoparticles (MNPs) with abundant surface functional groups, which effectively mediate heterogeneous nucleation and growth of UIO-66 nanocrystals. Well-defined MNPs@UIO-66 CSSNCs with stacked pores, showing high specific surface area (333.65 m2 g− 1) and low mass transfer resistance, were successfully fabricated by fine-tuning of the reaction conditions including reaction time and acetic acid content. Furthermore, the UIO-66 shells were further modified with arginine to obtain bifunctional MNPs@UIO-66-Arg CSSNCs. Thanks to the unique morphology and synergistic effect of Zr-O clusters and guanidine groups, the bifunctional MNPs@UIO-66-Arg CSSNCs exhibited outstanding enrichment performance for phosphopeptides, delivering a low limit of detection (0.1 fmol), high selectivity (β-casein/BSA, mass ratio 1:2000), and good capture capacity (120 mg g− 1). The mechanism for phosphopeptides capture may attribute to the hydrogen bonds, electrostatic interactions, and Zr–O–P bonds between phosphate groups in peptides and guanidyl/Zr-O clusters on bifunctional MNPs@UIO-66-Arg CSSNCs. In addition, the small stacking pores on the core-shell-satellite architecture may selectively capture phosphopeptides with low molecular weight, eliminating interference of other large molecular proteins in complex biological samples. [ABSTRACT FROM AUTHOR]

Predictive biomarkers of response to chemotherapy in patients with metastatic colorectal cancer (mCRC) are needed to better characterize tumors and enable more tailored therapies. Here we used serum proteomics to screen for chemotherapy predictive markers. We found that higher baseline serum inter-α-trypsin inhibitor Heavy Chain 4 (ITIH4) expression in newly diagnosed mCRC patients was associated with poorer response to standard first-line chemotherapy. In addition, the higher expression of ITIH4 in CRC tissue also suggested poorer prognosis mCRC patients. Moreover, the overexpression of ITIH4 could promote the proliferation of CRC cells and reduce the sensitivity of CRC cells to 5-fluorouracil (5-FU) by inhibiting apoptosis in vivo and vitro. Through RNA-seq combined with bioinformatics analysis, we speculated that ITIH4 may activate phosphatidyl 3-kinase-protein kinase B (PI3K-AKT) pathway to inhibit apoptosis, thereby reducing the sensitivity of CRC cells to 5-FU. In conclusion, our findings unveil that ITIH4 is associated with CRC resistance to 5-FU, and may serve as a potential predictive biomarker for the sensitivity of advanced CRC patients to standard first-line chemotherapy regimens, and also provide a potential therapeutic target to render 5-FU resistance in CRC patients.

ABSTRACT The objective was to investigate the effect of wild lactic acid bacteria (LAB) strains on the fungal and chemical composition of whole-plant corn silage. Three treatments were evaluated: CON-without inoculation, LB-inoculated with Lentilactobacillus buchneri (CCMA 1366), and LF-inoculated with L. farraginis (CCMA 1362). The silages were stored for 29, 103, and 193 d. The highest acetic acid (19.7 g/kg DM) and 1,2-propanediol (6.3 g/kg DM) concentrations were found in the LF, the highest aerobic stability was observed in this silage after 29 d (158.3 h). The yeast population was below the detection level at 29 d in the LF and 103 and 193 d in all silages. Seven genera of filamentous fungi (FF) were identified before ensiling; none of these were found after ensiling. At 29 d, Monascus pilosus was identified in the LB after aerobic exposure but did not produce citrinin. At 103 and 193 d, there was Aspergillus fumigatus predominance. A. flavus was identified at 103 d in the CON and LB; none produced B1, B2, G1, or G2 toxins. LAB strains modified the silages’ chemical and FF composition, and LF increased aerobic stability and reduced FF’s yeast population and diversity.

Background: Candidemia is associated with a high mortality rate. This study aimed to evaluate the clinical impact of a diagnostic intervention and antifungal stewardship in adults with candidemia, including effectiveness in facilitating appropriate antifungals and improving patient outcomes. Methods: A pre-post quasi-experimental study was conducted to analyze the impact of the integrated workflow of rapid species identification and antifungal stewardship intervention provided by infectious disease specialists for adults with candidemia at a medical center in southern Taiwan from March 1st, 2014 to February 29th, 2016. The primary endpoint was 30-day crude mortality, and secondary outcomes included the time to species identification, time to initial antifungal modification, and length of hospital stay. Results: Total 303 patients with candidemia were included, including 152 adults in the pre-intervention period (Mar. 1st, 2014–Feb. 28th, 2015; control group) and 151 in the intervention period (Mar. 1st, 2015–Feb. 29th, 2016; case group). Demographic and clinical characteristics of patients in two groups were similar. The case group had a shorter time to species identification (72 vs. 96 h, P

The purpose of this study was to isolate, identify, and evaluate the antibacterial and probiotic potential of bacteria from honeydew honey collected in Poland. Isolates (189 colonies from 10 honey samples) were evaluated for their antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Yersinia enterocolitica, and then identified by MALDI-TOF-MS. The isolates with the greatest antimicrobial properties were screened for their probiotic potential. The total number of bacteria isolated from honey did not exceed the value of 2.5 × 102 CFU/mL. The Bacillus pumilus/altitudinis, B. licheniformis, and Bacillus cereus groups were the dominant identified bacteria. Almost 16% of the isolates expressed antibacterial potential against three pathogenic bacteria, over 20% against two, while almost 34% of the isolates did not inhibit any. The survival rate of the isolates under gastrointestinal tract conditions was higher after 4 h of exposure to bile salts (>60% survival rate for 66.66% of the isolates), while at pH 2.0, it was lower (>50% survival rate for 44% of the isolates). The most resistant isolate B. pumilus/altitudinis survived at a rate of 77% at low pH and 108% with bile salts. These results confirmed that honeydew honey is a promising reservoir of bacteria that produces metabolites with antimicrobial and probiotic potential.

The exploration of microbial genetic resources for the production of fFen-flavor Baijiu has not only enriched the microbial library for baijiu production but has also laid the foundation for process improvement and strain optimization in baijiu brewing. In this study, a total of 177 fungal isolates were screened, including Saccharomyces cerevisiae, non-Saccharomyces cerevisiae, molds, and some pathogenic bacteria. Among them, Saccharomyces cerevisiae was the most abundant with 119 isolates, playing a major role in the fermentation of baijiu production. A total of 148 bacterial isolates were obtained from the fermentation mash samples, showing greater diversity compared to fungi. Bacillus species were the most abundant, with 94 isolates. Bacillus licheniformis, in particular, can produce a rich enzymatic system and flavor precursors, making it an important contributor to the sensory quality of baijiu. Lactic acid bacteria were the second most abundant, with 16 isolates. Additionally, five pathogenic fungal species were identified, including Candida pelliculosa, Candida lusitaniae, Fusarium oxysporum, Fusarium solani and Talaromyces marneffei. Six pathogenic bacterial species were also isolated, namely Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus xylosus, Moraxella osloensis, Actinomyces meyeri and Stenotrophomonas maltophilia. Finally, two strains of high acetate ethyl ester-producing yeast and lactate-degrading bacteria with good tolerance to temperature, pH, and ethanol concentration were identified as Saccharomyces cerevisiae and Bacillus licheniformis, respectively.

Five mycobacterial isolates from sewage were classified as members of the genus Mycobacterium but presented inconclusive species assignments. Thus, the isolates (MYC017, MYC098, MYC101, MYC123 and MYC340) were analyzed by phenotypical, biochemical, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and genomic features to clarify their taxonomic position. Phenotypic analysis and biochemical tests did not distinguish these isolates from other non-pigmented mycobacteria. In contrast, MALDI-TOF MS analysis showed that isolates were not related to any previously described Mycobacterium species. Comparative genomic analysis showed values of ANI and dDDH between 81.59-85.56% and 24.4-28.8%, respectively, when compared to the genomes of species of this genus. In addition, two (MYC101 and MYC123) presented indistinguishable protein spectra from each other and values of ANI = 98.57% and dDDH = 97.3%, therefore being considered as belonging to the same species. Phylogenetic analysis grouped the five isolates within the Mycobacterium terrae complex (MTC) but in a specific subclade and separated from the species already described and supported by 100% bootstrap value, confirming that they are part of this complex but different from earlier described species. According to these data, we propose the description of four new species belonging to the Mycobacterium genus: (i) Mycobacterium defluvii sp. nov. strain MYC017T (= ATCC TSD-296T = JCM 35364T), (ii) Mycobacterium crassicus sp. nov. strain MYC098T (= ATCC TSD-297T = JCM 35365T), (iii) Mycobacterium zoologicum sp. nov. strain MYC101T (= ATCC TSD-298T = JCM 35366T) and MYC123 (= ATCC BAA-3216 = JCM 35367); and (iv) Mycobacterium nativiensis sp. nov. strain MYC340T (= ATCC TSD-299T = JCM 35368T). [ABSTRACT FROM AUTHOR]

Background: Meningitis can be attributed to bacterial, fungal, or viral agents. In this study, we demonstrate the common bacterial agents causing meningitis along with their antibiotics susceptibility pattern in patients of all age groups. Material and Methods: This retrospective, observational study was carried out in the Department of Microbiology with cerebrospinal fluid (CSF) samples collected from November 2019 to May 2022. We collected 1986 nonrepeat CSF samples from clinically suspected patients of bacterial meningitis, and clinical information about the patients was extracted from the hospital information system. Results: Out of the 1986 CSF samples included in our study, 150 (7.55%) were found to be positive on bacterial culture. Most of our patients were in the age group of 0–20 years. Common clinical manifestations observed in our patients were: high-grade fever, 87 patients (58%); severe headache, 126 patients (84%); neck rigidity, 47 patients (31.3%); altered mental status, 76 patients (50.7%) and photophobia, 83 patients (55.3%). The most commonly isolated bacteria was Acinetobacter species (30%). The mean length of hospitalization (37.76 ± 25.30), the mean total cell count, high levels of protein (mg/dl) and low levels of glucose (mg/dl) of CSF were statistically significant in meningitis caused by multidrug-resistant bacteria. Conclusion: We recognized the spectrum of pathogens causing meningitis at our center along with the antibiotic resistance pattern to guide and facilitate early treatment by primary health care professionals and family medicine practitioners.

Polymyxin antibiotics are the last resort for treating patients in intensive care units infected with multiple-resistant Gram-negative bacteria. Due to their polycationic structure, their mode of action is based on an ionic interaction with the negatively charged lipid A portion of the lipopolysaccharide (LPS). The most prevalent polymyxin resistance mechanisms involve covalent modifications of lipid A: addition of the cationic sugar 4-amino-L-arabinose (L-Ara4N) and/or phosphoethanolamine (pEtN). The modified structure of lipid A has a lower net negative charge, leading to the repulsion of polymyxins and bacterial resistance to membrane disruption. Genes encoding the enzymatic systems involved in these modifications can be transferred either through chromosomes or mobile genetic elements. Therefore, new approaches to resistance diagnostics have been developed. On another note, interfering with these enzymatic systems might offer new therapeutic targets for drug discovery. This literature review focuses on diagnostic approaches based on structural changes in lipid A and on the therapeutic potential of molecules interfering with these changes.

Background Isepamicin is a 1-N-S-a-hydroxy-b-aminopropionyl derivative of gentamicin B and the spectrum of pathogenic microorganisms covered by it and its effectiveness is similar to that of amikacin except the action of aminoglycoside inhibitor enzymes is ineffectual on it.

Quinoa is an Andean crop that stands out as a high-quality protein-rich and gluten-free food. However, its increasing popularity exposes quinoa products to the potential risk of adulteration with cheaper cereals. Consequently, there is a need for novel methodologies to accurately characterize the composition of quinoa, which is influenced not only by the variety type but also by the farming and processing conditions. In this study, we present a rapid and straightforward method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to generate global fingerprints of quinoa proteins from white quinoa varieties, which were cultivated under conventional and organic farming and processed through boiling and extrusion. The mass spectra of the different protein extracts were processed using the MALDIquant software (version 1.19.3), detecting 49 proteins (with 31 tentatively identified). Intensity values from these proteins were then considered protein fingerprints for multivariate data analysis. Our results revealed reliable partial least squares-discriminant analysis (PLS-DA) classification models for distinguishing between farming and processing conditions, and the detected proteins that were critical for differentiation. They confirm the effectiveness of tracing the agricultural origins and technological treatments of quinoa grains through protein fingerprinting by MALDI-TOF-MS and chemometrics. This untargeted approach offers promising applications in food control and the food-processing industry.

Five mycobacterial isolates from sewage were classified as members of the genus Mycobacterium but presented inconclusive species assignments. Thus, the isolates (MYC017, MYC098, MYC101, MYC123 and MYC340) were analyzed by phenotypical, biochemical, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and genomic features to clarify their taxonomic position. Phenotypic analysis and biochemical tests did not distinguish these isolates from other non-pigmented mycobacteria. In contrast, MALDI-TOF MS analysis showed that isolates were not related to any previously described Mycobacterium species. Comparative genomic analysis showed values of ANI and dDDH between 81.59–85.56% and 24.4–28.8%, respectively, when compared to the genomes of species of this genus. In addition, two (MYC101 and MYC123) presented indistinguishable protein spectra from each other and values of ANI = 98.57% and dDDH = 97.3%, therefore being considered as belonging to the same species. Phylogenetic analysis grouped the five isolates within the Mycobacterium terrae complex (MTC) but in a specific subclade and separated from the species already described and supported by 100% bootstrap value, confirming that they are part of this complex but different from earlier described species. According to these data, we propose the description of four new species belonging to the Mycobacterium genus: (i) Mycobacterium defluvii sp. nov. strain MYC017T (= ATCC TSD-296T = JCM 35364T), (ii) Mycobacterium crassicus sp. nov. strain MYC098T (= ATCC TSD-297T = JCM 35365T), (iii) Mycobacterium zoologicum sp. nov. strain MYC101T (= ATCC TSD-298T = JCM 35366T) and MYC123 (= ATCC BAA-3216 = JCM 35367); and (iv) Mycobacterium nativiensis sp. nov. strain MYC340T (= ATCC TSD-299T = JCM 35368T).

Critical septic conditions caused by infectious pathogens are one of the leading causes of death in children glob- ally. The lack of introduced algorithms for rapid assessment of the condition of these patients, the use of inflammatory markers with low specificity and sensitivity and the use of standard microbiological methods pose a risk of delaying the diagnosis and treatment in children with infectious complica- tions. The aim of this scientific review is to create and test a convenient algorithm for early assessment, diagnosis and treatment of septic conditions in children. It will be assessed the physiological condition of patients using pSOFA, PRISM III and IV scores and the risk of mortality will be calculated also. The routine biochemical parameters of inflammation, procalcitonin will be compared with the new ones. Innovative markers of inflammation will be studied - Presepsin, soluble mannose receptor (sMR) by ELISA method for research purposes. An innovative and rapid method with high sensitivity and specificity (MALDI-TOF-MS) for the species identification of the pathogens will be performed, based on microbial protein analysis, incl. direct identification of the causative agent in the blood sample within 1- 2 hours. As a result, it is expected: to create a comprehensive assessment of critical and septic conditions in children by using innovative scores for the first time in Bulgaria; to find a correlation between the effective- ness of new inflammatory laboratory markers with the old ones and to choose the most appropriate combination of markers for an early detection of inflammation in the course of sepsis development in children; to improve the diagnostic process of microbial identification by using an innovative method for early detection of the infectious pathogen associated with the septic condition. More importantly, it will help the preven- tion, prognosis and will improve the therapeutic approach in children with septic and critical conditions. [ABSTRACT FROM AUTHOR]

Background Isepamicin is a 1-N-S-a-hydroxy-b-aminopropionyl derivative of gentamicin B and the spectrum of pathogenic microorganisms covered by it and its effectiveness is similar to that of amikacin except the action of aminoglycoside inhibitor enzymes is ineffectual on it. Material and Methods We performed a prospective study in the Bacteriology section of the Department of Microbiology at a 1,600-bedded hospital in Northern India from Jan 2022 to March 2022. Isepamicin was tested for susceptibility against gram-negative bacteria, identified by routine biochemicals and matrix-assisted-desorption/ionization –time of flight-mass spectrometry (MALDI-TOF-MS) assay. The antibiotic susceptibility testing for each of the isolates was performed by Kirby Bauer's disc diffusion method, according to the CLSI 2019 guidelines. Results The majority of isolates were obtained from blood samples (50, 39.1%). Among the non-inducible Enterobacteriaceae , Escherichia coli was least susceptible to amikacin (8/27, 29.63%) and most susceptible to isepamicin (18/27, 66.67%). Klebsiella pneumoniae followed the same pattern of susceptibility as E. coli and was least susceptible to Amikacin (20/46, 43.48%) and most susceptible to isepamicin (24/46, 52.17%). Enterobacter cloacae (6/7, 85.71%) was most susceptible to both amikacin and isepamicin, followed by 71.43% (5/7, 71.43%) susceptibility to gentamicin and tobramycin each. Enterobacter aerogenes was equally 53.33% (8/15) susceptible to all antibiotics. Pseudomonas aeruginosa was the most susceptible isolate to all antibiotics (18/21, 85.71%). Conclusion Isepamicin is a potential antimicrobial agent for treating an array of gram-negative bacteria-associated infections and shows better in vitro activity than older aminoglycoside agents. [ABSTRACT FROM AUTHOR]

Background: Meningitis can be attributed to bacterial, fungal, or viral agents. In this study, we demonstrate the common bacterial agents causing meningitis along with their antibiotics susceptibility pattern in patients of all age groups. Material and Methods: This retrospective, observational study was carried out in the Department of Microbiology with cerebrospinal fluid (CSF) samples collected from November 2019 to May 2022. We collected 1986 nonrepeat CSF samples from clinically suspected patients of bacterial meningitis, and clinical information about the patients was extracted from the hospital information system. Results: Out of the 1986 CSF samples included in our study, 150 (7.55%) were found to be positive on bacterial culture. Most of our patients were in the age group of 0-20 years. Common clinical manifestations observed in our patients were: high-grade fever, 87 patients (58%); severe headache, 126 patients (84%); neck rigidity, 47 patients (31.3%); altered mental status, 76 patients (50.7%) and photophobia, 83 patients (55.3%). The most commonly isolated bacteria was Acinetobacter species (30%). The mean length of hospitalization (37.76 ± 25.30), the mean total cell count, high levels of protein (mg/dl) and low levels of glucose (mg/dl) of CSF were statistically significant in meningitis caused by multidrug-resistant bacteria. Conclusion: We recognized the spectrum of pathogens causing meningitis at our center along with the antibiotic resistance pattern to guide and facilitate early treatment by primary health care professionals and family medicine practitioners. [ABSTRACT FROM AUTHOR]

Pear ring rot caused by Botryosphaeria dothidea severely affects the quality and storage life of pear fruit. Plant endophytes are potential new sources of biocontrol agents due to their abilities to produce novel bioactive products. In this work, we focused on an endophytic strain B1, which was isolated from roots of Dendrobium huoshanense. Notably, strain B1 exhibited strongly inhibitory effects against the growth of Botryosphaeria dothidea with an inhibition rate of 73.2% after 1 week of co-cultivation. Furthermore, it displayed a broad-spectrum inhibitory activity. Strain B1 also effectively delayed the onset of ring rot caused by B. dothidea in pear fruit and reduced the lesion diameter by 73.4%. Strain B1 was identified as Bacillus velezensis based on core-genome phylogeny. Genome mining with antiSMASH revealed 13 potential gene clusters involved in antimicrobial metabolites. Three main groups of lipopeptides (surfactin, iturin and fengycin) were identified with MALDI-TOF-MS, and the lipopeptides in the inhibition zone were greatly upregulated by B. dothidea; especially, fengycin isoforms were detected with higher abundance. These results proved that lipopeptides produced by strain B1 had significant antagonistic effect on B. dothidea. In conclusion, our results suggest that strain B1 has potential applications in the biocontrol of B. dothidea in postharvest fruit. [ABSTRACT FROM AUTHOR]

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Backfat trait is an important economic trait and highly heritable, but difficult to evaluate. Thus, it is of great significance to explore optimal backfat thickness of pigs by using marker-assisted selection (MAS) to speed up its breeding process and improve economic efficiency. This study aimed to investigate the relationship between genetic variations (e.g., SSRs) and backfat of Qinghai Bamei pigs using MALDI-TOF Mass Spectrometry (MALDI-TOF-MS). Herein, five alternative SSR loci (namely V1, V2, V3, V4 and V5) were selected for subsequent detection. The results suggested that 3 (141-, 143- and 145-), 3 (128-, 130- and 132-), 2 (160- and 162-), 2 (136- and 139-) and 3 (170-, 184- and 192-) alleles of V1, V2, V3, V4 and V5 were found, respectively. Subsequent analysis showed that there was linkage equilibrium among five SSRs and Hap19 (13.1%) (141-/132-/160-/139-/192-) had the highest haplotype frequency. Among these five SSR loci, V1, V2 and V3 loci were significantly associated to the backfat of Qinghai Bamei sows. These findings enriched the study of SSRs in Qinghai Bamei pigs, and (AC)n (Chr15:85485851-85485995), (AC)n (Chr10:52724583–52724713) and (TG)n (Chr4:90732644–90732802) could be utilized as the candidate locus for MAS in pig industry. Five novel SSR loci was identified in pigs through MALDI-TOF MS. V1, V2 and V3 loci was were significantly associated to the backfat of pigs. [ABSTRACT FROM AUTHOR]

Escherichia coli (E. coli) is a ubiquitous microorganism with pathogenic and saprophytic clones. The objective of this study was to evaluate the presence, virulence, antibiotic resistance and biofilm formation of E. coli in three industrial farms in Bulgaria, as well as their adjacent sites related to the utilization of manure (feces, wastewater in a separator, lagoons, means of transport, and soils). The isolation of single bacterial cultures was performed via standard procedures with modifications, and E. coli isolates were identified via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and polymerase chain reaction (PCR). The disk diffusion method was used to assess antimicrobial resistance, and PCR was used to detect genes for antibiotic resistance (GAR) (qnr, aac(3), ampC, blaSHV/blaTEM and erm) and virulence genes (stx, stx2all, LT, STa, F4 and eae). The protocol of Stepanović was utilized to measure the biofilm formation of the isolates. A total of 84 isolates from different samples (n = 53) were identified as E. coli. Almost all demonstrated antimicrobial resistance, and most of them demonstrated resistance to multiple antibiotics from different classes. No virulence genes coding the Shiga toxin or enterotoxins or those associated with enteropathogenicity were detected. No GAR from those tested for quinolones, aminoglycosides and macrolides were found. However, all isolates that were resistant to a penicillin-class antibiotic (56) had β-lactamase-producing plasmid genes. All of them had ampC, and 34 of them had blaTEM. A total of 14 isolates formed strongly adherent biofilms. These results in a country where the use of antibiotics for growth promotion and prophylaxis in farms is highly restricted corroborate that the global implemented policy on antibiotics in human medicine and in animal husbandry needs revision. [ABSTRACT FROM AUTHOR]

Copyright of China Brewing is the property of China Brewing Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Integration of antimicrobial stewardship intervention (ASI) with rapid organism identification has the potential for early customization of antimicrobial therapy and improved clinical outcomes. We aimed to evaluate the impact of this combined approach on antimicrobial therapy-related outcomes in patients with bloodstream infections (BSIs). Materials and methods: A pre–post quasi-experimental study was conducted to analyze the impact of ASI with organism identification via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) among patients with BSIs. Outcomes were compared to a historic pre-intervention group. The 30-day mortality was the primary endpoint. Secondary outcomes included time to first antibiotic modification, length of hospital stay. Results: A total of 1004 adult patients with BSIs were included in the final analysis, 519 patients classified into the intervention group and 485 patients in the preintervention group. The patients in the intervention group were younger (66 vs. 70 years, P = 0.02). The 30-day crude mortality (14.6% vs. 29.9%, P

To capture the structure of assembled hemagglutinin (HA) nanoparticles at single-particle resolution, HA-specific antigen binding fragments (Fabs) were labeled by fluorescent (FLR) dyes as probes to highlight the HA trimers displayed on the assembled tetravalent HA nanoparticles for a qualitative localization microscopic study. The FLR dyes were conjugated to the Fabs through N-hydroxysuccinimide (NHS) ester mediated amine coupling chemistry. The labeling profile, including labeling ratio, distribution, and site-specific labeling occupancy, can affect the imaging results and introduce inconsistency. To evaluate the labeling profile so as to evaluate the labeling efficiency, a combination of intact mass measurement by MALDI-MS and peptide mapping through LC-MS/MS was implemented. At the intact molecular level, the labeling ratio and distribution were determined. Through peptide mapping, the labeled residues were identified and the corresponding site-specific labeling occupancy was measured. A systematic comparative investigation of four different FLR-labeled 1H01-Fabs (generated from H1 strain HA specific mAb 1H01) allowed accurate profiling of the labeling pattern. The data indicate that the labeling was site-specific and semiquantitative. This warrants the consistency of single-particle fluorescent imaging experiments and allows a further imaging characterization of the single nanoparticles. [ABSTRACT FROM AUTHOR]

Rhodococcus hoagie, previously referred to as R. equi, is a Gram-positive intracellular coccobacillus that belongs to the Nocardiaceae family. This multi-host pathogen causes infections in farm animals, particularly foals, but also in immunosuppressed patients, mainly individuals treated with high doses of corticosteroids, subjected to organ transplant, or infected with human immunodeficiency virus Objectives of the study are to report a bloodstream infection in an immunocompromised patient. Immunocompromised patients with advanced HIV who presented bloodstream infection, residing in an urban setting and having undertaken no trips to the countryside or elsewhere during the COVID-19 pandemic. Blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was done in order to identify the bacteria. The immunocompromised female patient presented bloodstream infection by Rhodococcus hoagie, which was identified using MALDI-TOF-MS. R. hoagie can cause a severe infection with a high mortality rate if prompt treatment with a combination of antibiotics is not established. A high level of suspicion is required to establish the diagnosis, as it may be misdiagnosed as pulmonary tuberculosis. On gram stain, R. hoagie may appear as beaded to solid staining coccobacilli, which can be dismissed as a "diphtheroid" contaminant. The infection was identified using MALDI-TOF-MS. [ABSTRACT FROM AUTHOR]

The acylation of a cyclodextrin‐based [3]rotaxane was implemented with anhydride reagents following a facile procedure. Methacrylic anhydride facilitated the introduction of a vinyl‐group into the rotaxane structure toward achieving a crosslinker. Interestingly, only a few methacrylic units were attached to the cyclodextrin at reaction temperatures of up to 60 °C even with an excess amount of the reagent, probably due to steric hindrance. Therefore, the number of vinyl groups on the [3]rotaxane framework was easily controlled to be approximately two units via a simple random acylation, which worked effectively as a facile synthesis route for the rotaxane crosslinker. The acylation behavior was investigated in detail by NMR and MALDI‐TOF‐MS analyses. Such a proposed approach will help overcome the trade‐off between convenience and utility associated with the mono‐modification of cyclodextrins. [ABSTRACT FROM AUTHOR]

Monoclonal immunoglobin (M-protein) is a serum biomarker for the diagnosis of plasma cell dyscrasias. Despite limitation of analytical sensitivity and resolution, serum protein electrophoresis and immunofixation electrophoresis are still the front-line tests for the detection of M-proteins. Herein, we developed a MALDI-TOF Mass spectrometry-based method for the screening test of M-proteins in human serum. Based on the unique mass signature of different immunoglobin isotypes, M-Proteins could be rapidly identified and typed. The method demonstrated with high analytical performance and throughput, rapid and simple, which could be a new choice for the diagnosis of plasma cell dyscrasias. [ABSTRACT FROM AUTHOR]

Herein, we isolated the pathogenic strain ZZ051 from hemorrhagic channel catfish (Ictalurus punctatus). Physiological and biochemical identification, 16S rRNA gene sequence analysis, and MALDI-TOF-MS showed that the ZZ051 strain was Aeromonas veronii. After artificial infection, the diseased fish showed symptoms similar to the natural disease, and the characteristics of the bacteria reisolated from the tissues were the same as those of the original infection, indicating that the isolated strain ZZ051 was the pathogen responsible for the channel catfish disease. The ZZ051 isolate was highly sensitive to enrofloxacin but resistant to florfenicol. This study provided a theoretical basis for preventing and controlling hemorrhagic disease in channel catfish. [ABSTRACT FROM AUTHOR]

The occurrence of pathogenic bacteria has emerged as a plausible key component of summer mortalities in mussels. In the current research, four bacterial isolates retrieved from moribund Greenshell࣪ mussels, Perna canaliculus , from a previous summer mortality event, were tentatively identified as Vibrio and Photobacterium species using morpho-biochemical characterization and MALDI-TOF MS and confirmed as V. celticus, P. swingsii, P. rosenbergii , and P. proteolyticum using whole genome sequencing. These isolates were utilized in a laboratory challenge where mussels were injected with cell concentrations ranging from 105 to 109 CFU/mussel. Of the investigated isolates, P. swingsii induced the highest mortality. Additionally, results from quantitative polymerase chain reaction analysis, focusing on known virulence genes were detected in all isolates grown under laboratory conditions. Photobacterium rosenbergii and P. swingsii showed the highest expression levels of these virulence determinants. These results indicate that Photobacterium spp. could be a significant pathogen of P. canaliculus , with possible importance during summer mortality events. By implementing screening methods to detect and monitor Photobacterium concentrations in farmed mussel populations, a better understanding of the host–pathogen relationship can be obtained, aiding the development of a resilient industry in a changing environment. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) are multipotent cells which are popular in human regenerative medicine. These cells can renew themselves and differentiate into several specialized cell types including osteoblasts, adipocytes, and chondrocytes under physiological and experimental conditions. MSCs can secret a lot of components including proteins and metabolites. These components have significant effects on their surrounding cells and also can be used to characterize them. This characterization of multipotent MSCs plays a critical role in their therapeutic potential. The metabolic profile of culture media verified by applying matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF-MS) techniques. Also, the differentiation and development of MSCs have monitored through culture media metabolome or secretome (secreted metabolites). Totally, 24 potential metabolites were identified. Between them 12 metabolites are unique to BM-MSCs and 5 metabolites are unique to AD-MSCs. Trilineage differentiation including chondrocytes, osteoblasts, and adipocytes, as well as metabolites that are being differentiated, have been shown in different weeks. In the present study, the therapeutic effects of MSCs analyzed by decoding the metabolome for MSCs secretome via metabolic profiling using MALDI-TOF-MS techniques. [ABSTRACT FROM AUTHOR]

Pear ring rot caused by Botryosphaeria dothidea severely affects the quality and storage life of pear fruit. Plant endophytes are potential new sources of biocontrol agents due to their abilities to produce novel bioactive products. In this work, we focused on an endophytic strain B1, which was isolated from roots of Dendrobium huoshanense. Notably, strain B1 exhibited strongly inhibitory effects against the growth of Botryosphaeria dothidea with an inhibition rate of 73.2% after 1 week of co-cultivation. Furthermore, it displayed a broad-spectrum inhibitory activity. Strain B1 also effectively delayed the onset of ring rot caused by B. dothidea in pear fruit and reduced the lesion diameter by 73.4%. Strain B1 was identified as Bacillus velezensis based on core-genome phylogeny. Genome mining with antiSMASH revealed 13 potential gene clusters involved in antimicrobial metabolites. Three main groups of lipopeptides (surfactin, iturin and fengycin) were identified with MALDI-TOF-MS, and the lipopeptides in the inhibition zone were greatly upregulated by B. dothidea; especially, fengycin isoforms were detected with higher abundance. These results proved that lipopeptides produced by strain B1 had significant antagonistic effect on B. dothidea. In conclusion, our results suggest that strain B1 has potential applications in the biocontrol of B. dothidea in postharvest fruit.

Mycoplasma species (spp.) are predominantly found in the human oropharynx, and extracavity infections are rare. Conventional culture limitations hinder Mycoplasma spp. recovery, potentially causing overlooked infections. Molecular techniques reveal their roles in various infections. Mycoplasma pneumoniae causes pneumonia, while Mycoplasma salivarium (M. salivarium) in empyema is scarcely reported. We present a case of a 61-year-old man who suffered from tonsillitis, deep neck infection, necrotizing mediastinitis, and bilateral pleural infections. Mixed pathogens, mainly M. salivarium, were implicated.

Honey is the most famous natural sweet substance produced by honeybees (Apis mellifera). It contains numerous functional compounds, from which health benefits are obtained. In Chile, the production of honey is associated with its unique biodiversity, but it is exported mostly, as bulk honey. This work aimed to characterize the chemical and biological properties of Chilean honey on a large scale. The use of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) technique, combined with multivariate statistical analysis was introduced to study the chemical profiles of polyfloral honey. The use of a complementary mass spectrometry method allowed the identification of 25 different constituents in the studied honey, including hydrocarbons, acids, esters, glycoside isoprenoids, ketones, and a dihydroxyflavanone. The evaluation of biological properties in Chilean honey was measured in a representative number of polyfloral samples. For this purpose, the total phenolics and flavonoid content were measured in the selected samples. The antioxidant activity of the honey was evaluated through the ferric reducing antioxidant power assay (FRAP) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method. On the other hand, the antibacterial activity was assessed by the dilution technique to determine the minimum inhibitory concentration (MIC) of the honey against seven bacterial strains, including Gram-positive and Gram-negative bacteria. The results demonstrated that the antioxidant and antibacterial activities of Chilean honey are variable and they could have similar properties in comparison with other well-recognized bioactive honey.

Glycans are involved in many fundamental cellular processes such as growth, differentiation, and morphogenesis. However, their broad structural diversity makes analysis difficult. Glycomics via mass spectrometry has focused on the composition of glycans, but informatics analysis has not kept pace with the development of instrumentation and measurement techniques. We developed Toolbox Accelerating Glycomics (TAG), in which glycans can be added manually to the glycan list that can be freely designed with labels and sialic acid modifications, and fast processing is possible. In the present work, we improved TAG for large-scale analysis such as cohort analysis of serum samples. The sialic acid linkage-specific alkylamidation (SALSA) method converts differences in linkages such as α2,3- and α2,6-linkages of sialic acids into differences in mass. Glycans modified by SALSA and several structures discovered in recent years were added to the glycan list. A routine to generate calibration curves has been implemented to explore quantitation. These improvements are based on redefinitions of residues and glycans in the TAG List to incorporate information on glycans that could not be attributed because it was not assumed in the previous version of TAG. These functions were verified through analysis of purchased sera and 74 spectra with linearity at the level of R2 > 0.8 with 81 estimated glycan structures obtained including some candidate of rare glycans such as those with the N,N'-diacetyllactosediamine structure, suggesting they can be applied to large-scale analyses. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to analyze the cultivable oral microbiota of patients with obstructive sleep apnea (OSA) and its association with the periodontal condition. Methods: The epidemiology profile of patients and their clinical oral characteristics were determined. The microbiota was collected from saliva, subgingival plaque, and gingival sulcus of 93 patients classified into four groups according to the periodontal and clinical diagnosis: Group 1 (n = 25), healthy patients; Group 2 (n = 17), patients with periodontitis and without OSA; Group 3 (n = 19), patients with OSA and without periodontitis; and Group 4 (n = 32), patients with periodontitis and OSA. Microbiological samples were cultured, classified, characterized macroscopically and microscopically, and identified by MALDI-TOF-MS. The distribution of complexes and categories of microorganisms and correlations were established for inter- and intra-group of patients and statistically evaluated using the Spearman r test (p-value <0.5) and a multidimensional grouping analysis. Result: There was no evidence between the severity of OSA and periodontitis (p = 0.2813). However, there is a relationship between the stage of periodontitis and OSA (p = 0.0157), with stage III periodontitis being the one with the highest presence in patients with severe OSA (prevalence of 75%; p = 0.0157), with more cases in men. The greatest distribution of the complexes and categories was found in oral samples of patients with periodontitis and OSA (Group 4 POSA); even Candida spp. were more prevalent in these patients. Periodontitis and OSA are associated with comorbidities and oral conditions, and the microorganisms of the orange and red complexes participate in this association. The formation of the dysbiotic biofilm was mainly related to the presence of these complexes in association with Candida spp. Conclusion: Periodontopathogenic bacteria of the orange complex, such as Prevotella melaninogenica, and the yeast Candida albicans, altered the cultivable oral microbiota of patients with periodontitis and OSA in terms of diversity, possibly increasing the severity of periodontal disease. The link between yeasts and periodontopathogenic bacteria could help explain why people with severe OSA have such a high risk of stage III periodontitis. Antimicrobial approaches for treating periodontitis in individuals with OSA could be investigated in vitro using polymicrobial biofilms, according to our findings. [ABSTRACT FROM AUTHOR]

Petroleum hydrocarbons are one of the most well-known environmental pollutants not only for plants, and animals but also for humans because of their carcinogenic mutagenic, toxic, and teratogenic effects. Hence, various remediation technologies have been used to overcome these environmental pollutants. Among them, bioremediation is novel promising sustainable approach for treating petroleum and petroleum derivative raw material deleterious components with their effective, low-cost eco-friendly features. In this perspective, the evaluation of soil samples was collected from areas containing petroleum and petroleum derivatives from Kazanlı and Karaduvar refinery regions in Mersin-Turkey. Hence, potential hydrocarbon and petroleum degrading bacteria were identified and characterized by the MALDI-TOF MS method as well as biochemical, physiological, and morphological tests. According to the MALDI-TOF MS analysis; members of Bacillus, Stenotrophomonas, Enterobacter, Pseudomonas, Cupriavidus, Acinetobacter, and Escherichia were successfully identified as hydrocarbon and petroleum degrading bacteria. [ABSTRACT FROM AUTHOR]

Background In 2020, an incident involving spoiled salad dressing from a commercial source occurred. Upon opening the bottle, the contents exploded from gas that seemed to have fermented inside the bottle. For safety concerns, we sought to investigate the bacteria from the salad dressing in order to notify the company that made the product and relevant authorities. Methods Anaerobic and carbon dioxide culture methods were used. To determine species of colonies, MALDI-TOF-MS and 16S rRNA whole sequencing were performed. Results There were no colonies grown in anaerobic condition; however, we obtained three colonies from the carbon dioxide atmosphere. We determined the first colony as Alkalihalobacillus clausii (Bacillus clausii), the second as Bacillus spp. such as B. australimaris, B. safensis or B. safensis subsp. osmophilus and the third as B. paralicheniformis. Phylogenic tree analysis using the16S rRNA sequence revealed these colonies to be in a proximity of known gas-producing species. The NCBI database search revealed that a key gas production pathway gene, pyruvate formate-lyase (pfl), of which the gene product catalyzes pyruvate to formate conversion, exists in B. paralicheniformis. Formate dehydrogenase (FdhH) produces CO2 from formate that the coding gene fdhF positive bacteria can participate in gas production when formate is present in the culture. And we found fdhF from A. clausii, B. australimaris/B. safensis and B. paralicheniformis. Furthermore, under butanediol producing pathway, genes coding two enzymes involved in CO2 production, namely als and ald, existed in B. australimaris/B. safensis and B paralicheniformis, whereas A. clausii possessed als. Conclusion Candidate species A. clausii, B. australimaris/B. safensis and B. paralicheniformis from spoiled salad dressing were thought to produce CO2 gas each from their own enzymes, or in combination, which caused the explosion upon opening. The endospore forming nature of Bacillus should alert us to be cautious when considering food producing process regulations where we need to thoroughly heat any product during manufacture in order to inactivate any bacteria as there is the possibility of this type of dangerous occurrence. [ABSTRACT FROM AUTHOR]

Objective: To understand the virulence genes and molecular epidemiological characteristics of human-infected strains of Streptococcus suis in Rui'an, Zhejiang Province, from 2021 to 2022, and to provide a scientific basis for diagnosis, treatment, prevention, and control., Methods: Three blood-borne strains of Streptococcus suis were analysed by morphological observation, identification, and drug sensitivity tests. We performed polymerase chain reaction (PCR) amplification of their main seven virulence factors and housekeeping genes. This was followed by virulence analysis and multilocus sequence typing. We analysed their relationships with local pathogens from previous years., Results: Three Streptococcus suis strains were isolated from the blood samples of three patients. From these, the virulence genotypes demonstrated that the two strains were orf2+ and ef+/orf2+/sly +, respectively. The Multilocus sequence typing (MLST) typing results demonstrated that the two strains were ST25 and ST7, respectively., Conclusion: The first isolation of ST25 Streptococcus suis in Rui'an was presumed to have a close affinity with the endemic strain in North America. The other strain was an ST7 clone, consistent with the endemic strain in Sichuan, and which may have originated from Sichuan. Virulence genotype analysis demonstrated that different virulence genes of the pathogens resulted in different clinical manifestations., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Qianying Zhu reports financial support was provided by 10.13039/100014587Ruian Municipal Science and Technology Bureau. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)