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Summary In this study, a lactic acid bacteria strain isolated from oats was used as a producer to obtain exopolysaccharide (EPS) using MRS agar plates. The findings indicated that the bacterial strain identified as Weissella cibaria (accession number: PP515290) was shown to have good probiotic activity, adapt better to alkaline than acidic conditions, and efficiently produce EPS at concentrations of up to 7.93 g L−1. SEM images showed a smooth surface and compact particle whereas FTIR confirmed the basic functional groups of polysaccharides in the sample. Meanwhile, XRD analysis demonstrated a non‐crystalline amorphous structure and thermal analysis indicated two‐phase thermal decomposition with maximum rate at 210 °C. Subsequently, EPS from W. cibaria was applied as stabiliser in set‐type yogurt (ex situ) in comparison with yogurt using mixed culture with W. cibaria (in situ) and traditional starter culture (control). The results showed that the addition of 2% EPS significantly improved syneresis (14.31%) compared to co‐culture (27.50%) and control (29.80%) as well as increased hardness of yogurt during 2‐week refrigeration. [ABSTRACT FROM AUTHOR]

This study aimed to determine the sequence type (ST) of Bartonella henselae infecting small Indian mongooses from Saint Kitts via multi-locus sequence typing (MLST). This investigation used stored EDTA blood (n = 22) samples from mongooses previously identified as positive for B. henselae. Chocolate agar plates were enriched with Bartonella alpha-Proteobacteria growth medium (BAPGM) to culture and isolate Bartonella from the blood samples. To perform MLST, DNA was extracted and purified from isolates followed by amplification by conventional PCR (300–500 bp) for eight genes (16S rDNA, batR, gltA, groEL, ftsZ, nlpD, ribC, and rpoB). Bartonella henselae STs were deposited in the PubMLST repository. Out of 22 B. henselae-positive blood samples, isolates were obtained from 12 mongooses (54.5%; 12/22). Each mongoose was infected with one ST. The studied mongoose population was infected with sequence types ST2, ST3, ST8, and a novel ST represented by ST38. Bartonella henselae ST2, ST3 and ST8 infecting mongooses are known to circulate in humans and cats, with ST2 and ST8 associated with Cat Scratch Disease (bartonellosis) in humans. The results presented herein denote the circulation of B. henselae STs with zoonotic potential in mongooses with risk of B. henselae transmission to humans. [ABSTRACT FROM AUTHOR]

The use of repellents for nematode control has not been established. Leaf extracts of Myrtus communis, an evergreen shrub with wide distribution in the Mediterranean and some other regions, were tested for repellence of second‐stage juveniles (J2s) of Meloidogyne species. Extracts obtained with several solvents and water repelled M. javanica J2s on agar plates, with the 60% methanol extract showing the highest repellent activity. M. hapla J2s were also repelled by the aqueous and 60% methanol extracts. Adding the 60% methanol extract to fluopyram, a chemical J2 attractant, reduced and abolished, respectively, the attraction of M. javanica and M. hapla J2s to it. When the methanolic extract was added near lettuce seedling roots on an agar plate, the number of M. javanica, M. incognita and M. hapla J2s attracted to the root tips was reduced by 70.0%–98.2%, infection rates decreased by 50.1%–95.8% and root length increased by 61.8%–186.7% compared to control seedlings grown on the same plates. When the methanol extract was mixed into the agar plate, the three Meloidogyne species' attraction to and infection of lettuce seedlings was reduced by up to 75.4% and 100%, respectively, and root length increased up to 3.4‐fold. The acetone extract mixed into the agar reduced the attraction of M. javanica J2s to fluopyram but did not affect their repellence by KNO3. The results suggest that M. communis leaf extracts have repellent and infection‐inhibitory activity against Meloidogyne J2, offering potential control means for Meloidogyne species. [ABSTRACT FROM AUTHOR]

(1) Background: There is an ongoing discussion on the elution efficacy of antibiotic-impregnated cements. Our experiments were intended to clarify if there are differences in the antibiotic elution of HPLC compared with inhibition zone testing using eluates or PMMA discs. (2) Materials and Methods: Two cement brands with different concentrations of the active ingredient were tested in antimicrobial Kirby–Bauer (disc diffusion) assays. Cement platelets were directly applied on the agar plates and their zone of inhibition was measured. In parallel, the platelets were incubated in phosphate buffered saline (PBS) and at distinct points of time transferred into new buffer. At these time points, 50 µL of the bone cement eluates was used for zone of inhibition testing. Standard gentamicin sulfate solutions served as a control in the same test setup. To verify the microbiological investigations, the antibiotic content of the eluates was also measured via high-performance liquid chromatography (HPLC). (3) Results: The experiments with cement eluates showed better differentiable results than the direct application of the cement discs. The results were also comparable to investigations with HPLC and gentamicin sulfate standard solutions. (4) Conclusions: The results of elution rates are influenced by the test system and the period of observation chosen. The microbial test systems reflect the results of HPLC to the same degree and give evidence of the efficacy of the antibiotics. The HPLC tests on eluates were more suitable in representing differences in release characteristics. [ABSTRACT FROM AUTHOR]

Introduction: Aeromonas hydrophila is particularly harmful to freshwater aquaculture, and the search for phage is an effective biological control method, but reports of possible temperate phages and their mutants are rare in this field. In this study, a virulent phage highly homologous to prophage in the genomes of A. hydrophila was collected and preliminary biological characterization was carried out to understand its nature. Materials and methods: Water samples taken from eel ponds in Fujian, China were combined with the strain. Spot test method and double-layer agar plate assay was used for confirmation and purification. Phage virions were observed using transmission electron microscope. A total of 68 strains of Aeromonas spp. were used to determine the host range. MOI groups of 1,000, 100, 10, 1, 0.1, 0.01, 0.001, 0.0001, 0.00001 were prepared to detect the optimal MOI. The conditions of thermal stability assay were set as 30, 40, 50, 60, 70 and 80°C for 1 h, respectively, and conditions of acid and alkali stability assay were set as 2.0, 4.0, 6.0, 8.0, 10.0 and 12.0 of pH. MOI of 0.01 and 0.1, respectively, are set to determine the inhibitory capacity of phage. Results: A novel virulent A. hydrophila phage designated phiA051 has been isolated from aquaculture water. Electron microscopic observation showed that the phage phiA051 was composed of an icosahedral capsid. The phage phiA051 possesses an optimal multiplicity of infection (MOI) of 0.01, and its burst size was 108 PFU/cell. The phage maintained a high viability at temperatures of 30-50°C or pH 6.0-10.0 for 1 h. Phage phiA051 has certain potentials in rapidly inhibiting the spread of pathogen early in the outbreak, and it has a linear dsDNA with GC content of 60.55% and a total length of 32,212 bp, including 46 ORFs. Discussion: The phage phiA051 behaved as a virulent phage. However, the BLASTN result showed that 23 of the top 25 hits were genomes of Aeromonas strains. It was suggested that phiA051 was probably derived from some prophage in the chromosome of Aeromonas. Further investigation of the mechanism how phage phiA051 transforms from a temperate phage to a virulent phage will provide a unique perspective and idea to explore the potential of prophages. [ABSTRACT FROM AUTHOR]

Studies have used anaerobic-digester sludge and/or effluent as inocula for bioelectrochemical systems (BESs), such as microbial fuel cells (MFCs), for power generation, while limited studies have isolated and characterized electrochemically active bacteria (EAB) that inhabit anaerobic digesters. In the present work, single-chamber MFCs were operated using the anaerobic-digester effluent as the sole source of organics and microbes, and attempts were made to isolate EAB from anode biofilms in MFCs by repeated anaerobic cultivations on agar plates. Red colonies were selected from those grown on the agar plates, resulting in the isolation of three phylogenetically diverse strains affiliated with the phyla Bacillota, Campylobacterota and Deferribacterota. All these strains are capable of current generation in pure-culture BESs, while they exhibit different electrochemical properties as assessed by cyclic voltammetry. The analyses of their cell-free extracts show that cytochromes are abundantly present in their cells, suggesting their involvement in current generation. The results suggest that anaerobic digesters harbor diverse EAB, and it would be of interest to examine their ecological niches in anaerobic digestion. [ABSTRACT FROM AUTHOR]

Streptococcus mutans is a major pathogenic habitant of oral caries. Owing to its physiological and biochemical features, it prevails in the form of plaque biofilm together with another important mutans streptococci species, Streptococcus sobrinus. Both species are considered as initiators of cavity lesions, and biofilm is essential to the dental caries process. Compared with the planktonic populations, the biofilm form has higher resistance to environmental conditions and antibiotics. Dental plaques also secure the long-term survival of microorganisms and protection from any stress conditions. To address the need for new antibiofilm agents, we have focused on L-carnitine-fumarate, a fumarate-conjugated quaternary ammonium compound. Using the macro-broth susceptibility testing method, we established its MIC value as 6.0 mg/mL. The MBC value, determined from the broth dilution minimum inhibitory concentration test by sub-culturing it to BHI agar plates, was established as 7.0 mg/mL. Antibiofilm efficacy was tested in 96-well plates coated with saliva using BHI broth supplemented with 1% sucrose as a standard approach. The obtained results allowed us to assess the MIBC as 7.5 mg/mL and the MBBC value as 10.0 mg/mL. The latter concentration also caused approximately 20% eradication of pre-existing biofilm. EPS-rich matrix, forming the core of the biofilm and enabling a confined acidic microenvironment, was also examined and confirmed the effectiveness of 10.0 mg/mL L-carnitine-fumarate concentration in inhibiting EPS formation. Furthermore, the anti-adherent and anti-aciduric impacts of L-carnitine-fumarate were investigated and revealed significant inhibitory effects at sub-MIC concentrations. The influence of L-carnitine-fumarate on the phosphotransferase system was investigated as well. Our results provide a new insight into the antibacterial potential of L-carnitine-fumarate as a valuable compound to be considered for alternative or adjunct anti-caries and antibiofilm preventive approaches. [ABSTRACT FROM AUTHOR]

Introduction: Candida auris is a recently discovered yeast with a multi-drug resistant profile associated with high mortality rates. The rapid identification of Candida auris in hospital settings is crucial to allow appropriate therapeutic and rapid implementation of infection management measures. The aim of this study was to develop a lateral flow immunoassay (LFIA) for the rapid identification of Candida auris. Methods: Highly specific monoclonal antibodies were obtained by immunizing mice with membrane proteins from Candida auris which were then used to develop a LFIA whose performance was assessed by testing 12 strains of Candida auris and 37 strains of other Candida species. Isolates were grown on either Sabouraud dextrose, CHROMagarTM Candida Plus or HardyCHROM™ Candida + auris agar plates. The strains were also cultured on salt sabouraud-dextrose with chloramphenicol or a commercially available Salt-Sabouraud Dulcitol Broth with chloramphenicol and gentamicin, and processed using a simple centrifugation protocol to recover a pellet. Finally, the colonies or yeast extract were transferred to the LFIA to determine the specificity and sensitivity of the assay. Results: The LFIA reached 100% specificity and sensitivity from solid agar plates. For both enrichment broths, some Candida non-auris species were able to grow, but the LFIA remained 100% specific. The use of a dextrose-based sabouraud broth resulted in earlier identification with the LFIA, with most of the Candida auris strains detected at 24 h. Conclusion: The developed LFIA prototype represents a powerful tool to fight the emerging threat of Candida auris. Clinical validation represents the next step. [ABSTRACT FROM AUTHOR]

Mold infestations in buildings pose significant challenges to human health, affecting both private residences and hospitals. While molds commonly trigger asthma and allergies in the immunocompetent, they can cause life-threatening diseases in the immunocompromised. Currently, there is an unmet need for new strategies to reduce or prevent mold infestations. Far-UVC technology can inactivate microorganisms while remaining safe for humans. This study investigates the inhibitory efficacy of far-UVC light at 222 nm on the growth of common mold-producing fungi, specifically Penicillium candidum, when delivered in low-dose on-off duty cycles, a configuration consistent with its use in real-world settings. The inhibitory effect of the low-dose duty cycles was assessed on growth induced by i) an adjacent spore-producing P. candidum donor and ii) P. candidum spores seeded directly onto agar plates. In both setups, the far-UVC light significantly inhibited both vertical and horizontal growth of P. candidum, even when the UV doses were below the Threshold Value Limit of 23 mJ/cm2. These results suggest that far-UVC light holds the potential to improve indoor air quality by reducing or preventing mold growth, also when people are present. [ABSTRACT FROM AUTHOR]

The proliferation and dissemination of antimicrobial-resistant bacteria is an increasingly global challenge and is attributed mainly to the excessive or improper use of antibiotics. Currently, the gold-standard phenotypic methodology for detecting resistant strains is agar plating, which is a time-consuming process that involves multiple subculturing steps. Genotypic analysis techniques are fast, but they require pure starting samples and cannot differentiate between viable and non-viable organisms. Thus, there is a need to develop a better method to identify and prevent the spread of antimicrobial resistance. This work presents a novel method for detecting and identifying antibiotic-resistant strains by combining a cell sorter for bacterial detection and an elastic-light-scattering method for bacterial classification. The cell sorter was equipped with safety mechanisms for handling pathogenic organisms and enabled precise placement of individual bacteria onto an agar plate. The patterning was performed on an antibiotic-gradient plate, where the growth of colonies in sections with high antibiotic concentrations confirmed the presence of a resistant strain. The antibiotic-gradient plate was also tested with an elastic-light-scattering device where each colony's unique colony scatter pattern was recorded and classified using machine learning for rapid identification of bacteria. Sorting and patterning bacteria on an antibiotic-gradient plate using a cell sorter reduced the number of subculturing steps and allowed direct qualitative binary detection of resistant strains. Elastic-light-scattering technology is a rapid, label-free, and non-destructive method that permits instantaneous classification of pathogenic strains based on the unique bacterial colony scatter pattern. Key points: • Individual bacteria cells are placed on gradient agar plates by a cell sorter • Laser-light scatter patterns are used to recognize antibiotic-resistant organisms • Scatter patterns formed by colonies correspond to AMR-associated phenotypes [ABSTRACT FROM AUTHOR]

Ciprofloxacin (CPX), is a broad-spectrum fluoroquinolone antibiotic widely used to treat bacterial infections in humans and animals. In recent years, ecological problems caused by the disposal of CPX-waste product have attracted increasing attention. This study aimed to evaluate the ability of different bacteria to remove CPX from water. The ability of five bacterial strains namely; Acenitobacter sp., Acenitobacter lwofii, Bacillus. pumilus, Burkholderia phytofirmans, and Mesorihizobium sp. were examined in this study. Tests were performed on Luria Bertani (LB) as well as on Mueller-Hinton (MH) agar at different CPX concentrations (20-100 mg L-1). All five tested bacterial strains showed growth on LB as well as on MH agar plates at all concentrations between 20 to 100 mg L-1 CPX. Three bacterial species: Acenitobacter lwofii, Bacillus pumilus, and Mesorihizobium sp. were reported more resistant to CPX of the five strains tested. In a shake flask study, the strain B. pumilus initially showed higher degradation rate (75.53%) followed by Acenitobacter lwofii (72.3%), Mesorihizobium sp. (70.32), Burkholderia phytofirmans (67.10%), and Acenitobacter sp. (63.2%) at low CPX concentration (5 mg L-1) in minimal saline media. However, Mesorihizobium sp. showed higher degradation efficiency (76.32%) at a higher CPX concentration (10 mg L-1). A group of three bacterial strains, A. lowfi, B. pumilus, and Mesorihizobium sp. showed good CPX degradation rate (95.4%), and achieved higher degradation efficiency than the single strains at 20 mg L-1 CPX concentration. Overall, the results suggest that using a combination of bacteria may be a useful tool for the remediation of antibiotic pollution. In addition, this study reveals the ability of previously isolated bacteria to biodegrade CPX as a sole source of carbon, providing new insights into the biodegradation of CPX. [ABSTRACT FROM AUTHOR]

Due to the overuse of antibiotics, the number of multidrug-resistant pathogen bacteria is rising in recent years posing a serious threat to human health. One promising alternative for treatment is the application of phage therapy using highly selective bacteriophages. Because of their selectivity, individual screens called phagograms for each patient are required to select phages from a phage library. Phagograms are mostly performed via bacterial cultivation on double layer agar plates and phage addition causing bacterial lysis. However, these assays are work-intensive and have a low ability for parallelization and automation. Hence, highly parallelizable and automatable microbioreactors in the lowest microliter scale could offer an economic solution increasing the throughput of phagograms. This paper demonstrates the applicability of a novel capillary-wave microbioreactor (cwMBR) to perform phagograms. Due to its small volume of only 7 µL and the open-droplet design, it can be easily automated and parallelized in future. Furthermore, the ability of online biomass measurement makes the cwMBR a perfect phagogram platform in the future. Herein, phagograms with E. coli and different concentrations of the phages MM02 and EASG3 were performed as proof of concept for phagograms in the cwMBR. Thereby, the cwMBR was able to measure differences in lysis kinetics of different phages. Furthermore, the phagograms were compared to those in conventional microtiter plate readers revealing the cwMBR as ideal alternative for phagograms as it combines favorable mixing conditions and a phage repellent hydrophilic glass surface with online biomass measurement in an open-droplet design for future parallelization and automation. [Display omitted] • Phage sensitivity tests called phagograms are mandatory for phage therapy. • State of the art phagograms are not highly parallelizable and automatable. • Novel capillary-wave microbioreactor (cwMBR) can perform parallelized phagograms. • cwMBR combines parallelizability and automatability with online biomass measurement. • cwMBR is able to measure lysis kinetics of phages. [ABSTRACT FROM AUTHOR]

This study evaluated the antiseptic effect of 0.25% chlorhexidine nanoemulsion (NM-Cl) on cat skin and compare its effect with that of 2.0% chlorhexidine digluconate (CS-Cl). NM-Cl was synthesized using the spontaneous emulsification method, and physical and chemical properties were analyzed. The antiseptic effects of NM-Cl and CS-Cl were randomly tested on the thoracic limbs of 10 healthy male cats. After a wide trichotomy of the thoracic limbs, NM-Cl was randomly applied to the trichotomy area of the right (n = 5) or left (n = 5) thoracic limbs. Then, a catheter was inserted aseptically in the cephalic vein. Subsequently, the same procedure was performed using CS-Cl on the contralateral limb. Cutaneous microbiota swab samples were obtained before antisepsis (Tpre); immediately after antisepsis (Tpost); and 4, 8, and 24 h after antisepsis. The collected samples were immediately inoculated on blood agar plates and incubated at 35 ºC ± 2 ºC in aerobiosis. Colony-forming units (CFUs) were manually counted after 24 h of inoculation. The Kruskal–Wallis and Mann–Whitney U tests were performed between groups and within the same group at different sample times, respectively. The NM-Cl and CS-Cl groups showed a reduction in CFUs between Tpre and Tpost in all animals (P < 0.001). Both formulations presented an antiseptic effect 24 h of antisepsis (P < 0.05), and no difference was observed between formulations at different times (P < 0.05). With a lower concentration of chlorhexidine than CS-Cl, NM-Cl presents effective antiseptic action and prolonged residual effect in antisepsis for cat venipuncture. [ABSTRACT FROM AUTHOR]

In the last decades, biocatalysis has offered new perspectives for the synthesis of (chiral) amines, which are essential building blocks for pharmaceuticals, fine and bulk chemicals. In this regard, amidases have been employed due to their broad substrate scope and their independence from expensive cofactors. To expand the repertoire of amidases, tools for their rapid identification and characterization are greatly demanded. In this work an ultra-high throughput growth selection assay based on the production of the folate precursor p-aminobenzoic acid (PABA) is introduced to identify amidase activity. PABA-derived amides structurally mimic the broad class of commonly used chromogenic substrates derived from p-nitroaniline. This suggests that the assay should be broadly applicable for the identification of amidases. Unlike conventional growth selection assays that rely on substrates as nitrogen or carbon source, our approach requires PABA in sub-nanomolar concentrations, making it exceptionally sensitive and ideal for engineering campaigns that aim at enhancing amidase activities from minimally active starting points, for example. The presented assay offers flexibility in the adjustment of sensitivity to suit project-specific needs using different expression systems and fine-tuning with the antimetabolite sulfathiazole. Application of this PABA-based assay facilitates the screening of millions of enzyme variants on a single agar plate within two days, without the need for laborious sample preparation or expensive instruments, with transformation efficiency being the only limiting factor. Key points: • Ultra-high throughput assay (tens of millions on one agar plate) for amidase screening • High sensitivity by coupling selection to folate instead of carbon or nitrogen source • Highly adjustable in terms of sensitivity and expression of the engineering target [ABSTRACT FROM AUTHOR]

Background: Early and appropriate antibiotic treatment improves the clinical outcome of patients with sepsis. There is an urgent need for rapid identification (ID) and antimicrobial susceptibility testing (AST) of bacteria that cause bloodstream infection (BSI). Rapid ID and AST can be achieved by short-term incubation on solid medium of positive blood cultures using MALDI-TOF mass spectrometry (MS) and the BD M50 system. The purpose of this study is to evaluate the performance of rapid method compared to traditional method. Methods: A total of 124 mono-microbial samples were collected. Positive blood culture samples were short-term incubated on blood agar plates and chocolate agar plates for 5 ∼ 7 h, and the rapid ID and AST were achieved through Zybio EXS2000 MS and BD M50 System, respectively. Results: Compared with the traditional 24 h culture for ID, this rapid method can shorten the cultivation time to 5 ∼ 7 h. Accurate organism ID was achieved in 90.6% of Gram-positive bacteria (GP), 98.5% of Gram-negative bacteria (GN), and 100% of fungi. The AST resulted in the 98.5% essential agreement (EA) and 97.1% category agreements (CA) in NMIC-413, 99.4% EA and 98.9% CA in PMIC-92, 100% both EA and CA in SMIC-2. Besides, this method can be used for 67.2% (264/393) of culture bottles during routine work. The mean turn-around time (TAT) for obtaining final results by conventional method is approximately 72.6 ± 10.5 h, which is nearly 24 h longer than the rapid method. Conclusions: The newly described method is expected to provide faster and reliable ID and AST results, making it an important tool for rapid management of blood cultures (BCs). In addition, this rapid method can be used to process most positive blood cultures, enabling patients to receive rapid and effective treatment. [ABSTRACT FROM AUTHOR]

Introduction: Propolis is a natural substance highly regarded in the medical field, including dentistry. This study is primarily directed towards the examination of the effects of a 5% propolis mouthwash on biofilm growth, bacterial interactions, as well as the sensitivity of cariogenic bacteria (Streptococcus mutans and Streptococcus sanguinis) within an in vitro dual-species biofilm environment. Materials and methods: These bacteria were cultured and subsequently placed in 96-well plates. The 5% propolis mouthwash was introduced into the well plates and subjected to a 24-hour incubation period. Following this, a DNA extraction procedure was performed, enabling the utilization of PCR techniques to assess the relative proportions and sensitivities of S. mutans and S. sanguinis. Crystal Violet assays were conducted to determine biofilm mass and investigate the interactions between the two bacterial species. Results: The antibacterial properties of propolis are efficacious in inhibiting bacterial growth within the biofilm. Furthermore, the presence of sorbitol in the 5% propolis mouthwash is assumed to inhibit glucose utilization by S. mutans and impede acid production from sucrose by sorbitol-grown cells, thereby capable of inhibiting the growth of S. mutans. Streptococcus mutans was more sensitive to 5% propolis mouthwash than Streptococcus sanguinis. Conclusion: There is an antagonistic interaction between Streptococcus mutans and Streptococcus sanguinis bacteria. 5% propolis mouthwash was able to reduce biofilm mass and biofilm growth rate of dual-species Streptococcus mutans and Streptococcus sanguinis biofilms. However, 5% propolis mouthwash could not stop or fully inhibit the growth of the dual-species biofilm. [ABSTRACT FROM AUTHOR]

Background: Actinobacillus pleuropneumoniae is a serious pathogen in pigs. The abundant application of antibiotics has resulted in the gradual emergence of drugresistant bacteria, which has seriously affected treatment of disease. To aid measures to prevent the emergence and spread of drug-resistant bacteria, herein, the kill rate and mutant selection window (MSW) of danofloxacin (DAN) against A. pleuropneumoniae were evaluated. Methods: For the kill rate study, the minimum inhibitory concentration (MIC) was tested using the micro dilution broth method and time-killing curves of DAN against A. pleuropneumoniae grown in tryptic soy broth (TSB) at a series drug concentrations (from 0 to 64 MIC) were constructed. The relationships between the kill rate and drug concentrations were analyzed using a Sigmoid Emax model during different time periods. For the MSW study, the MIC99 (the lowest concentration that inhibited the growth of the bacteria by ≥ 99%) and mutant prevention concentration (MPC) of DAN against A. pleuropneumoniae were measured using the agar plate method. Then, a peristaltic pump infection model was established to simulate the dynamic changes of DAN concentrations in pig lungs. The changes in number and sensitivity of A. pleuropneumoniae were measured. The relationships between pharmacokinetic/pharmacodynamic parameters and the antibacterial effect were analyzed using the Sigmoid Emax model. Results: In kill rate study, the MIC of DAN against A. pleuropneumoniae was 0.016 µg/mL. According to the kill rate, DAN exhibited concentration-dependent antibacterial activity against A. pleuropneumoniae. A bactericidal effect was observed when the DAN concentration reached 4–8 MIC. The kill rate increased constantly with the increase in DAN concentration, with a maximum value of 3.23 Log10 colony forming units (CFU)/mL/h during the 0–1 h period. When the drug concentration was in the middle part of the MSW, drugresistant bacteria might be induced. Therefore, the dosage should be avoided to produce a mean value of AUC24h/MIC99 (between 31.29 and 62.59 h. The values of AUC24h/MIC99 to achieve bacteriostatic, bactericidal, and eradication effects were 9.46, 25.14, and > 62.59 h, respectively. Conclusion: These kill rate and MSW results will provide valuable guidance for the use of DAN to treat A. pleuropneumoniae infections. [ABSTRACT FROM AUTHOR]

Escherichia coli in phosphate-buffered saline was treated with high hydrostatic pressures (HHPs) of 400, 500, and 600 MPa at 25 °C for 10 min. The cell suspension was then plated on nonselective agar and incubated at 20, 25, 30, 35, 37, and 40 °C to study the effect of the incubation temperatures on the detection of HHP-injured populations. The number of detected cells was maximum when incubated at 25 °C for 72 h. Cold storage at 4 °C for 7 d after HHP-treatment improved the detection efficacies of incubations at 30–40 °C, implicating increased tolerance of HHP-injured cells against the mild heat. Combined incubation at 25 °C for more than 6 h with subsequent incubation at 35 °C shortened total incubation period from 72 h to 24 h while achieving the maximized colony number obtained by the incubation at 25 °C for 72 h. [ABSTRACT FROM AUTHOR]

challenge clinicians due to their complex nature and the diverse patient demographics they affect. Primary stability is key, especially in cases involving osteoporotic bones or younger patients. The Ellis plate with a locking screw mechanism offers a promising solution to achieve this stability, potentially preventing secondary displacement regardless of bone quality. Aims and Objectives: This study aims to rigorously assess the effectiveness of the Ellis volar locking compression plate in treating distal end radius fractures. It focuses on evaluating functional outcomes post-treatment, providing insights into the efficacy of this approach. Materials and Methods: This study evaluated the efficacy of the Ellis plate in managing distal radius fractures, under ethical approval number KMC/IEC/Ortho/2021-5. Twenty adult patients with unstable fractures were included based on specific criteria and provided informed consent. Exclusion criteria included unwillingness for surgery, medical unsuitability, osteoporosis, minors, and compound fractures with vascular injury. Surgical outcomes were assessed using the Gartland and Werley scoring system, with follow-up appointments scheduled over 1 year to monitor progress and adherence to physiotherapy plans. Results: The study encompassed 20 patients, with a gender distribution of 12 males and eight females and an average age of 43.3 years. Utilizing the Gartland and Werley demerit grading system, the study reported 45% superb outcomes, another 45% good outcomes, 10% fair outcomes, and no poor outcomes. This distribution highlights the potential of the Ellis plate technique in achieving positive results in fracture treatment. Conclusion: The study concludes that the Ellis plate technique is a viable and effective option for stabilizing distal end radius fractures. However, achieving the best possible outcomes depends on several factors, including strict adherence to AO principles, maintaining high standards of aseptic practice, following appropriate post-operative rehabilitation protocols, and ensuring comprehensive patient education. This multifaceted approach is crucial for the successful treatment of these fractures. [ABSTRACT FROM AUTHOR]

Sub-high temperature Daqu, a traditional solid fermenting agent used in Chinese strong-aroma Baijiu production, is abundant in diverse microorganisms, including bacteria, yeasts, molds, and actinomycetes. Among these, yeasts are pivotal for ethanol production and flavor formation. However, counting yeasts in Daqu is challenging due to interference from molds and bacteria. Antibiotics are employed to inhibit bacterial growth, but there is no practical way to suppress molds without affecting the growth of yeasts. In this study, short-chain carboxylates (C1-C6) were added to the culture medium at various pH conditions to investigate their effects on the growth of molds and yeasts. The results demonstrated distinct inhibitory effects of the short-chain carboxylates, depending on both pH and concentration. Several tested short-chain carboxylates effectively suppressed mold growth on agar plates while leaving yeast growth unaffected. This suggests a simple and feasible method for enhancing the efficiency of yeast isolation and counting in Daqu. Such an approach is valuable for studying yeasts in diverse and complex habitats. [ABSTRACT FROM AUTHOR]

Brevibacillus thermoruber strain Nabari cells grow as widely spreading dendritic colonies on reasoner's 2A‐agar (1.5%) plates at around 55°C but as small motile colonies at 37°C. Motile colonies can be divided into colonies that move in straight or curved lines over long distances (wandering colonies), and colonies that rotate at a fixed location (rotating colonies). The addition of surfactant to the agar medium greatly increased the frequency of wandering colonies and facilitated the study of such colonies. The morphology of the wandering colonies varied: circular at the tip and pointed at the back, lemon‐shaped with pointed ends, crescent‐shaped, bullet‐shaped, fish‐like, and so on. A single colony may split into multiple colonies as it moves, or multiple colonies may merge into a single colony. The most surprising aspect of the movement of wandering colonies was that when a moving colony collides with another colony, it sometimes does not make a U‐turn, but instead retreats straight back, as if bouncing back. The migration mechanisms of wandering colonies are discussed based on optical microscopic observations of swimming patterns of single cells in water and scanning electron microscopy of the arrangement of bacterial cells in wandering colonies. [ABSTRACT FROM AUTHOR]

Background: Eradication of oral Helicobacter pylori (H. pylori) not only reduces the infection rate from the transmission route but also improves the success rate of intragastric eradication. MAXPOWER Biological Bacteriostatic Liquid, developed in our previous work, is a composite biological preparation with strong antibacterial ability and unique antibacterial mechanism. The present study evaluated the efficacy of the MAXPOWER biocontrol solution on H. pylori and its success rate in eradicating oral H. pylori in clinical patients. Methods: Live-dead cell staining and hemolysis test were used to evaluate the cellular safety of MAXPOWER biocontrol solution; plate spreading, live-dead bacterial staining, and scanning electron microscopy methods were used to evaluate its antimicrobial effect against H. pylori. Transcriptomics was used to analyze the changes in H. pylori genes before and after treatment. After seven days of gavage treatment, H&E staining and mice feces were collected for 16SrDNA sequencing to evaluate the animals' safety. Oral H. pylori-positive patients were randomized to be given a placebo and MAXPOWER Bio-Bacteriostatic Liquid gargle for seven days to evaluate the effect on oral H. pylori eradication. Results: In vitro tests demonstrated that this product has excellent biocompatibility and hemocompatibility and can effectively eradicate oral H. pylori. In vivo tests further showed that it has good biosafety and virtually no adverse effect on intestinal microflora. Transcriptomics analysis revealed that it kills H. pylori cells mainly by disrupting their cell membranes and metabolism. Additionally, the results of randomized controlled trials on humans disclosed that the oral H. pylori eradication rates achieved by MAXPOWER Biological Antibacterial Liquid were 71.4% and 78.9% according to the intention-to-treat and the per-protocol analysis, respectively. Conclusion: MAXPOWER Biological Antibacterial Liquid is both safe and efficacious in the eradication of oral H. pylori. Trial registration: This study was retrospectively registered in the ClinicalTrials.gov Trial Registry on 21/09/2023 (NCT06045832). [ABSTRACT FROM AUTHOR]

Background: the aim of the study was to assess microbiological air quality in operating theatres by determining the level of microbiological contamination of the air and critical surfaces using the passive air sampling method and compliance of the operating theatre staff with infection control measures. Materials and methods: The prospective study was conducted in the surgical block of the University Medical Centre Maribor. For two months continuously, ten operating theatres were assessed for microbial contamination of air and surfaces during quiet and active times of the day. A passive air sampling method with Petri dishes on an agar specially adapted for this purpose (plate count agar) was used. In addition, ten surgical procedures were observed to assess staff compliance with recommended practises. Results: Air samples met microbiological standards in all operating theatres. In both sampling sessions of the day (quiet and active periods), microbial contamination of the air was always within the limit of 10 CFU/m3. The average number of bacterial colonies was zero to two during quiet phases and one to four during active phases. Approximately 60% of the isolates from the operating theatres belonged mainly to the genus Staphylococcus: S. epidermidis (36% of the isolates), S. hominis (17.5%) and S. haemolyticus (5.5%). The rest were identified as Streptococcus anginosus (23%) and Bacillus sp. (18%). Pathogenic bacteria and moulds were not present. In regard to staff compliance with good surgical practise, the former varied by behaviour and function, with non-compliance in pre-operative skin preparation and operating theatre congestion being notable. The cleanliness of the environment was satisfactory. Conclusions: Microbiological air control is extremely important for the safety and success of both surgical and postoperative practises. In spite of good results obtained in the study, further improvements in surgical staff compliance with good surgical practise are essential to reduce surgical site infections. [ABSTRACT FROM AUTHOR]

The present study evaluated the performance of the fungus Trichoderma reesei to tolerate and biodegrade the herbicide diuron in its agrochemical presentation in agar plates, liquid culture, and solid-state fermentation. The tolerance of T. reesei to diuron was characterized through a non-competitive inhibition model of the fungal radial growth on the PDA agar plate and growth in liquid culture with glucose and ammonium nitrate, showing a higher tolerance to diuron on the PDA agar plate (inhibition constant 98.63 mg L−1) than in liquid culture (inhibition constant 39.4 mg L−1). Diuron biodegradation by T. reesei was characterized through model inhibition by the substrate on agar plate and liquid culture. In liquid culture, the fungus biotransformed diuron into 3,4-dichloroaniline using the amide group from the diuron structure as a carbon and nitrogen source, yielding 0.154 mg of biomass per mg of diuron. A mixture of barley straw and agrolite was used as the support and substrate for solid-state fermentation. The diuron removal percentage in solid-state fermentation was fitted by non-multiple linear regression to a parabolic surface response model and reached the higher removal (97.26%) with a specific aeration rate of 1.0 vkgm and inoculum of 2.6 × 108 spores g−1. The diuron removal in solid-state fermentation by sorption on barley straw and agrolite was discarded compared to the removal magnitude of the biosorption and biodegradation mechanisms of Trichoderma reesei. The findings in this work about the tolerance and capability of Trichoderma reesei to remove diuron in liquid and solid culture media demonstrate the potential of the fungus to be implemented in bioremediation technologies of herbicide-polluted sites. [ABSTRACT FROM AUTHOR]

Here, we demonstrate the beneficial effect of surfactant-producing pseudomonads on Pantoea eucalypti 299R. We conducted a series of experiments in environments of increasing complexity. P. eucalypti 299R (Pe299R), and Pseudomonas sp. FF1 (Pff1) or Pe299R and surfactant-production deficient Pseudomonas sp. FF1::ΔviscB (Pff1ΔviscB) were co-inoculated in broth, on swarming agar plates, and on plants. In broth, there were no differences in the growth dynamics of Pe299R when growing in the presence of Pff1 or Pff1ΔviscB. By contrast, on swarming agar plates, Pe299R was able to co-swarm with Pff1 which led to a significant increase in Pe299R biomass compared to Pe299R growing with Pff1ΔviscB or in monoculture. Finally in planta, and using the single-cell bioreporter for reproductive success (CUSPER), we found a temporally distinct beneficial effect of Pff1 on co-inoculated Pe299R subpopulations that did not occur in the presence of Pff1ΔviscB. We tested three additional surfactant-producing pseudomonads and their respective surfactant knockout mutants on PE299R on swarming agar showing similar results. This led us to propose a model for the positive effect of surfactant production during leaf colonization. Our results indicate that co-motility might be common during leaf colonization and adds yet another facet to the already manyfold roles of surfactants. [ABSTRACT FROM AUTHOR]

Background: The precise influence of plate position on clinical outcomes in the context of volar fixed-angle plating for distal radius fractures is not fully understood. This article aims to investigate the influence of plate position on clinical results, and functional outcomes in patients treated with volar fixed plating for distal radius fractures. Methods: A total of 58 patients with 64 distal radius fractures were included in the study. Patient demographics, fracture characteristics, surgical details, and radiographic data were collected. Post-operative AP and Lat views of all patients taken on the first day after surgery were evaluated. Volar Tilt, Radial Inclination and Radial Height measurements were used as reduction criteria. In the follow-up, the patients were called for their last control, flexion and extension angles of the wrist and Mayo Wrist Scores, the distance of the plate to the joint line and the angle between the plate and the radial shaft were measured and recorded. Results: A total of 64 distal radius fractures, with a mean age of 46.9 years, and the mean follow-up period 24.9 months were included in this study. There was a significant relationship between the Radial Inclination and Plate-Shaft Angle variables and the Mayo Wrist Score at a 99% confidence interval. Additionally, a relationship was observed between the Radial Height variable and the Mayo Score at a 90% confidence interval. A significant positive association was observed between radial inclination and achieving a Good-to-Excellent Mayo score (OR = 1.28, 95% CI [1.08–1.51], p = 0.004). Plate distance to joint line demonstrated a marginally significant positive association with a Good-to-Excellent Mayo score (OR = 1.31, 95% CI [0.97–1.77], p = 0.077). Univariate analysis revealed a significant negative association between plate-shaft angle and achieving a Good-to-Excellent Mayo score (OR = 0.71, 95% CI [0.52–0.99], p = 0.045). This negative association remained statistically significant in the multivariate analysis (p = 0.016). Conclusion: Radial inclination, plate distance to joint line, and angle between plate and radius shaft were identified as significant factors associated with improved Mayo Wrist Scores. [ABSTRACT FROM AUTHOR]

Background: There is an urge for traditional herbal remedies as an alternative to modern medicine in treating several diseases. A significant number of modern pharmaceutical drugs are based on or derived from medicinal plants or their extracts. These drugs derived from the plant origin have various antimicrobial, antioxidant, anticancer, anti-inflammatory activities. Alangium salviifolium belongs to Cornaceae family and is well known for its medicinal properties. The present study was carried out to evaluate the antibacterial, antioxidant effect and possible bioactive components present in the chloroform, acetone, ethanol, methanol and aqueous extract of Alangium salviifolium leaves. Methodology: Dried leaves of Alangium salviifolium were subjected to serial solvent extraction using increasing polarity of solvents, i.e., chloroform, acetone, methanol, ethanol, and distilled water. Crude extracts were further tested for qualitative analysis of phytochemicals using standard procedure, while GCMS analysis was performed to identify the probable phytocompounds. Antibacterial activity was performed against bacterial pathogens using agar well method, whereas antioxidant activity was performed using in vitro PM, DPPH and FRAP assays. Results: Phytochemical analysis of the extracts revealed the presence of key phytochemical classes. Using gas chromatography-mass spectrometry, several high and low molecular weight chemical compound kinds were discovered. These chemical substances are regarded as having significant biological and pharmacological effects. All crude extracts had considerable and comparable in vitro antioxidant and antibacterial properties. Conclusions: According to the findings of this study, Alangium salviifolium leaves are a rich source of phytoconstituents that are crucial in stopping the advancement of numerous disorders. [ABSTRACT FROM AUTHOR]

Background: Marginal failure at the resin dentin interface promotes biofilm formation, which further leads to secondary caries and hypersensitivity. This likelihood also increases if residual bacteria are present following cavity preparation. In order to achieve a proper biological seal without jeopardizing bonding, efforts were made to functionalize the adhesive system with antibacterial activity. Aim and objectives: To appraise the antibacterial activity of a total-etch adhesive system against S. mutans with and without incorporation of antibiotics Vancomycin and Cefotaxime. Materials and Method: A commercially available 5th-generation total-etch bonding agent (Te-Econorm) was used. S. mutans broth had been standardized and streaked over Muller-Hinton agar culture medium and round wells about 6 mm in diameter were made in the centre of the agar plates. Each experimental group comprised 10 samples, which include: Group 1 - 30µg Cefotaxime, Group 2-30µg Cefotaxime + Bonding agent, Group 3-30µg Vancomycin, Group 4-30µg Vancomycin + Bonding agent, Group 5- Bonding agent, and Group 6- No material. Inoculated culture plates were examined for the zone of inhibition after incubation at 37° C for 24 hours. Results: There was a significant difference in the mean diameter of zone of inhibition (p=0.000), with the maximum exhibited by Group 4, followed by Group 3 and Group 2. The least zone of inhibition was exhibited by Groups 1 and 5. The negative control showed no zone of inhibition. Conclusion: The combination of Vancomycin and bonding agent had superior antibacterial activity against S. mutans in comparison to cefotaxime and bonding agent. [ABSTRACT FROM AUTHOR]

The flagellar components of Vibrio spp., PomA and PomB, form a complex that transduces sodium ion and contributes to rotate flagella. The transmembrane protein PomB is attached to the basal body T‐ring by its periplasmic region and has a plug segment following the transmembrane helix to prevent ion flux. Previously we showed that PomB deleted from E41 to R120 (Δ41–120) was functionally comparable to the full‐length PomB. In this study, three deletions after the plug region, PomB (Δ61–120), PomB (Δ61–140), and PomB (Δ71–150), were generated. PomB (Δ61–120) conferred motility, whereas the other two mutants showed almost no motility in soft agar plate; however, we observed some swimming cells with speed comparable for the wild‐type cells. When the two PomB mutants were introduced into a wild‐type strain, the swimming ability was not affected by the mutant PomBs. Then, we purified the mutant PomAB complexes to confirm the stator formation. When plug mutations were introduced into the PomB mutants, the reduced motility by the deletion was rescued, suggesting that the stator was activated. Our results indicate that the deletions prevent the stator activation and the linker and plug regions, from E41 to S150, are not essential for the motor function of PomB but are important for its regulation. [ABSTRACT FROM AUTHOR]

In the present study, we searched Halobacteriovorax strain preying upon Salmonella from the seawater of the Adriatic Sea. The Halobacteriovorax strain, named M7, was identified using 16S rRNA analysis. The M7 strain predation efficiency was tested against different Salmonella and non- Salmonella strains, all isolated from food matrices obtained from the Adriatic Sea. Finally, the M7 strain was exposed to Salmonella enterica subsp. enterica serovar Napoli in challenge tests to evaluate the killing of this specific prey over time. Double -layer agar plating technique was used to enumerate Halobacteriovorax and to evaluate its host specificity and predation efficiency. In the 103 predator/103 prey challenge test, M7 caused a decrease of Salmonella by about 2 log at 24 h compared to the control. In the 107 predator/104 prey challenge test, M7 caused a decrease of Salmonella by about 5 log at 24 h compared to the control, and good levels of decrease were obtained even at shorter times. Halobacteriovorax strains active against Salmonella are rarely present in the Adriatic Sea, Italy seawater. However, the isolate M7 showed high predatory efficiency towards a wide range of Salmonella strains. The presence of Salmonella in bivalves affects food safety since current decontamination processes are not always effective. M7 may represent a potential candidate for reducing and controlling Salmonella contamination in bivalves from harvesting to trade. [ABSTRACT FROM AUTHOR]

Background: Candida species, commonly present in the human body, can result in severe infections. Non-albicans Candida (NAC) species, with increasing antifungal resistance, pose a growing concern. Aims and Objectives: This study aims to evaluate candidiasis prevalence and antifungal susceptibility in isolated Candida species among suspected patients at tertiary care hospitals in Butwal, emphasizing early identification and appropriate treatment to avoid unnecessary use of toxic antifungal drugs. Materials and Methods: In a descriptive study on 303 patients with clinical symptoms of Candida infections, specimens underwent direct microscopic examination and culture on Sabouraud dextrose agar. Species identification involved phenotypic methods such as chromogenic character on CHROMagar Candida media, germ tube examination, and microscopic characteristics. Subsequently, isolated species were tested for antifungal susceptibility using the disc diffusion method. Results: Among the 303 samples tested, 80 (26.4%) were positive for candidal infection. NAC species were the most commonly isolated, with Candida albicans and Candida krusei being the most virulent. The isolates exhibited the highest sensitivity to fluconazole (77.5%) followed by itraconazole (75%), whereas amphotericin B showed the lowest effectiveness with 63.75% resistance. Conclusion: The rising prevalence of NAC species, particularly their growing resistance to Amphotericin B, has become a significant concern, as these species are frequently detected in various clinical samples. These findings underscore the importance of diligent monitoring and judicious selection of antifungal agents to ensure effective treatment strategies. [ABSTRACT FROM AUTHOR]

This article discusses a study on the impact of preoperative oral hygiene treatment on bacterial transport and colonization during orthopedic surgery. The study found that patients who received oral hygiene treatment had lower rates of pathogen detection and bacterial colonization in the oropharynx, trachea, and endotracheal tube. However, there was no significant difference in the incidence of pneumonia between the treatment group and the control group. The study suggests that while oral hygiene treatment can reduce bacterial transport and colonization, further research is needed to determine its effectiveness in preventing pneumonia. The authors recommend promoting preoperative oral hygiene as a cost-effective treatment for patients undergoing general anesthesia. [Extracted from the article]

The continuing emergence of new strains of antibiotic-resistant bacteria has renewed interest in phage therapy; however, there has been limited progress in applying phage therapy to multi-drug resistant Mycobacterium tuberculosis (Mtb) infections. In this study, we show that bacteriophage strains D29 and DS6A can efficiently lyse Mtb H37Rv in 7H10 agar plates. However, only phage DS6A efficiently kills H37Rv in liquid culture and in Mtb-infected human primary macrophages. We further show in subsequent experiments that, after the humanized mice were infected with aerosolized H37Rv, then treated with DS6A intravenously, the DS6A treated mice showed increased body weight and improved pulmonary function relative to control mice. Furthermore, DS6A reduces Mtb load in mouse organs with greater efficacy in the spleen. These results demonstrate the feasibility of developing phage therapy as an effective therapeutic against Mtb infection. Bacteriophage strain DS6A efficiently eradicates intracellular Mtb in human macrophages, while humanized mice treated with DS6A show improved pulmonary function and reduced Mtb load in mouse organs with greater efficacy in the spleen. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB) contribute to human health, and LAB functionality has been studied using Caenorhabditis elegans as an alternative host. However, many studies have focused on the efficacy of a single strain of LAB, and few reports have compared various LAB strains. In this study, we examined the effects of 15 strains of LAB isolated from vegetables, meat, and fermented foods on nematode longevity and healthy lifespan. To reduce the frequency of laborious survival observations, we performed a lifespan assay on agar plates containing 2′-deoxy-5-fluorouridine (FUdR), which inhibits egg hatching and prevents generation mixing. Four beneficial strains showed significant lifespan extension and increased spontaneous nematode mobility, regardless of treatment with or without FUdR and the frequency of survival observation. These results suggested increased longevity and an extended healthy lifespan, confirming the reliability of our method. The four strains are expected to show anti-ageing effects besides longevity and have effects on age-related degenerative diseases. Our labor-saving method can be used as an alternative to conventional methods and enable simultaneous screening of multiple strains. Future research could explore factors contributing to lifespan regulation by comparing and verifying differential strain effects on lifespan. [ABSTRACT FROM AUTHOR]

Various traditional methods, including Dhoopana and Havana, have been employed in fumigation procedures to reduce microbial levels in the environment to nonpathogenic thresholds. Although the use of formaldehyde in combination with potassium permanganate is widely acknowledged as the predominant fumigation method, it is also associated with various health risks. In the present study the herbal dhoop stick was manufactured. In the manufacturing process of Dhoop sticks, various ingredients such as neem, tulsi, Kapoor, cow dung, cow urine, cow ghee, and guggul were meticulously combined in different quantities. The evaluation of the anti-microbial properties of the prepared dhoop stick by dhoopan was done on the nutrient agar plates in the kitchen area of hospital for three days. An assessment of number of colonies before and after the administration of dhoopstick revealed reduction in the mean score from 2480.66±72.94 cfu/100cm2 to 1938.66±144.57 cfu/100cm2 which was statistically significant (p-p-0.020447). Additionally, it was found that There was no significant difference found between after scores of both dhoopstick and formaldehyde (p-0.103734). Both the Dhoop Stick and formaldehyde exposures show a consistent reduction in microbial colony counts on agar plates in the hospital kitchen over the three-day period. [ABSTRACT FROM AUTHOR]

China possesses abundant wild germplasm resources and a wide range of cultivated strains of Stropharia rugosoannulata. However, issues such as genetic diversity and unclear genetic relationships have had an impact on the classification and preservation of germplasm resources, the breeding of new varieties, and the promotion of superior strains. There is an urgent need for genetic diversity analysis and assessment of germplasm resources. In this study, we conducted whole-genome resequencing of 50 cultivated and wild strains collected from various regions across the country. After applying a series of filtering parameters, we obtained 888,536 high-quality Single Nucleotide Polymorphism (SNP) markers. Using these SNP markers, we performed principal component analysis, population structure analysis, and phylogenetic clusters analysis on the 50 strains. Most cultivated strains exhibited high genetic similarity, while significant genetic diversity was observed among wild strains. Based on factors such as marker distribution throughout the entire genome and marker quality, we selected 358 core SNP markers to construct SNP fingerprints. Two-dimensional barcodes were generated for each strain to enable specific identification. Additionally, the mycelial growth rate of strains was evaluated on Potato Dextrose Agar (PDA) and substrate culture media. We also assessed their lignin degradation capability using guaiacol agar plates assay. It was observed that the mycelial growth rate on PDA and substrate culture medium exhibited a significant correlation with the diameter of the mycelial colony on guaiacol agar medium. Additionally, the correlation between the mycelial growth rate on the substrate medium and that on the guaiacol agar plate was stronger than that on PDA medium. This study provided molecular-level identification and assessment of germplasm resources. It clarified the genetic relationships among strains and the characteristics of mycelium growth-related agronomic traits of each strain. This research contributed to the enrichment and development of utilizable germplasm resources and breeding materials for S. rugosoannulata, offering a scientific basis for further research. [ABSTRACT FROM AUTHOR]

With rising infection rates in recent years, Vibrio vulnificus poses an increasing threat to public safety in the coastal brackish Baltic Sea. It is therefore important to monitor this organism and assess the V. vulnificus infection risk on a more regular basis. However, as the coastline of the Baltic Sea is 8000 km long and shared by nine nations, a convenient, fast, inexpensive, yet efficient V. vulnificus identification method is essential. We evaluated the effectiveness of a two-step agar-based approach consisting of successive Vibrio isolation and cultivation on thiosulphate-citrate-bile salt sucrose (TCBS) agar and CHROMagar™ Vibrio for V. vulnificus in comparison with V. cholerae, V. parahaemolyticus, and V. alginolyticus. Our study contains isolates from water and sediment across a broad expanse of the Baltic Sea including 13 locations and two different summers, the time of year during which Vibrio infections are usually much more frequent. Confirmation of isolate species identity was carried out using molecular analyses. The two-step agar plating method performed well across different locations and timeframes in correctly identifying V. vulnificus by more than 80%, but the sensitivity in other Vibrio species varied. Thus, our approach yielded promising results as a potential tool for early V. vulnificus detection across a broad timeframe and transect of the Baltic Sea and potentially other brackish environments. [ABSTRACT FROM AUTHOR]

Aims: We investigated the antibacterial efficacy of Umonium38 and Virkon® against Burkholderia pseudomallei, Escherichia coli, Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus (MRSA) up to 14 days following treatment. Methods and results: Umonium38 was diluted to 0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 3%, tested against the bacterial strains at various contact times (15 min to 24 h), and incubated for up to 14 days. A minimum concentration of 0.5% Umonium38 with a contact time of 15 min effectively killed approximately 108 CFU/ml of all four bacterial species. No growth was observed on agar plates from day 0 until day 14 for all six concentrations. The bacteria were also inactivated by a 30-minute treatment time using Virkon® 1% solution. Conclusions: Umonium38 effectively inactivates B. pseudomallei, E. coli, P. aeruginosa and MRSA at a concentration of ≥ 0.5% with a contact time of at least 15 min. The antimicrobial effect of Umonium38 remained for 14 days. Significance: and impact of the study: The study provides evidence supporting the efficacy of Umonium38 as a bactericidal agent against E. coli and P. aeruginosa, specifying the required concentrations and contact time. Our investigation adds to the existing knowledge by demonstrating the bactericidal activity of Umonium38 against B. pseudomallei and MRSA. A limitation of the study is that the results presented have been produced under optimal laboratory conditions, and additional studies are required to determine the effectiveness of Umonium38 and Virkon® under sub-optimal conditions, including high levels of organic matter or variable pH. [ABSTRACT FROM AUTHOR]

Background: Microbial communities harbor important biotechnological potential in diverse domains, however, the engineering and propagation of such communities still face both knowledge and know-how gaps. More specifically, culturing tools are needed to propagate and shape microbial communities, to obtain desired properties, and to exploit them. Previous work suggested that micro-confinement and segregation of microorganisms using invert (water-in-oil, w/o) emulsion broth can shape communities during propagation, by alleviating biotic interactions and inducing physiological changes in cultured bacteria. The present work aimed at evaluating invert emulsion and simple broth monophasic cultures for the propagation and shaping of bacterial communities derived from raw milk in a serial propagation design. Results: The monophasic setup resulted in stable community structures during serial propagation, whereas the invert emulsion system resulted in only transiently stable structures. In addition, different communities with different taxonomic compositions could be obtained from a single inoculum. Furthermore, the implementation of invert emulsion systems has allowed for the enrichment of less abundant microorganisms and consequently facilitated their isolation on culture agar plates. Conclusions: The monophasic system enables communities to be propagated in a stable manner, whereas the invert emulsion system allowed for the isolation of less abundant microorganisms and the generation of diverse taxonomic compositions from a single inoculum. [ABSTRACT FROM AUTHOR]

Maintaining water quality is a crucial factor for shrimp cultivation. Detecting the vibrio bacteria distribution in the ponds could prove to be a crucial for successful cultivation. Thiosulfate-citrate-bile salts-sucrose (TCBS) agar is a commonly used medium to stimulate the Vibrio colony presence. In this research, water samples from shrimp cultivation were collected, isolated in TCBS agar plates, and incubated for 24 hours, afterwards exported as an image dataset. The YOLOv7 computer vision technique was implemented to detect and count Vibrio colonies with faster and higher precision compared to the tedious manual counting technique done by laboratory researchers. Additionally, this technique is applicable and scalable to large sets of data. From the execution done so far, the proposed system has achieved an average accuracy of 78.99%. [ABSTRACT FROM AUTHOR]

Objective: To study the biomechanical characteristics of each tissue structure when using different 3D printing Cage in osteoporotic patients undergoing interbody fusion. Methods: A finite element model of the lumbar spine was reconstructed and validated with regarding a range of motion and intervertebral disc pressure from previous in vitro studies. Cage and pedicle screws were implanted and part of the lamina, spinous process, and facet joints were removed in the L4/5 segment of the validated mode to simulate interbody fusion. A 280 N follower load and 7.5 N·m moment were applied to different postoperative models and intact osteoporotic model to simulate lumbar motion. The biomechanical characteristics of different models were evaluated by calculating and analyzing the range of motion of the fixed and cephalic adjacent segment, the stress of the screw-rod system, the stress at the interface between cage and L5 endplate, and intervertebral disc pressure of the adjacent segment. Results: After rigid fixation, the range of motion of the fixed segment of model A-C decreased significantly, which was much smaller than that of the osteoporotic model. And with the increase of the axial area of the interbody fusion cages, the fixed segment of model A-C tended to be more stable. The range of motion and intradiscal pressure of the spinal models with different interbody fusion cages were higher than those of the complete osteoporosis model, but there was no significant difference between the postoperative models. On the other hand, the L5 upper endplate stress and screw-rod system stress of model A-C show a decreasing trend in different directions of motion. The stress of the endplate is the highest during flexion, which can reach 40.5 MPa (model A). The difference in endplate stress between models A-C was the largest during lateral bending. The endplate stress of models A and B was 150.5% and 140.9% of that of model C, respectively. The stress of the screw-rod system was the highest during lateral bending (model A, 102.0 MPa), which was 108.4%, 102.4%, 110.4%, 114.2% of model B and 158.5%, 110.1%, 115.8%, 125.4% of model C in flexion, extension, lateral bending, and rotation, respectively. Conclusions: For people with osteoporosis, no matter what type of cage is used, good immediate stability can be achieved after surgery. Larger cage sizes provide better fixation without significantly increasing ROM and IDP in adjacent segments, which may contribute to the development of ASD. In addition, larger cage sizes can disperse endplate stress and reduce stress concentration, which is of positive significance in preventing cage subsidence after operation. The cage and screw rod system establish a stress conduction pathway on the spine, and a larger cage greatly enhances the stress-bearing capacity of the front column, which can better distribute the stress of the posterior spine structure and the stress borne by the posterior screw rod system, reduce the stress concentration phenomenon of the nail rod system, and avoid exceeding the yield strength of the material, resulting in the risk of future instrument failure. [ABSTRACT FROM AUTHOR]

In hospital environments, droplets generated by urination within shared toilets may represent a route of dissemination for bacteria such as vancomycin-resistant Enterococcus faecium (VREfm), which contributes significantly to the burden of hospital-acquired infections. We investigated the potential activity of a foam in preventing the generation of droplets containing Enterococcus spp. during urination. A uniform layer of foam was deposited in the inner walls and at the bottom of an experimental toilet contaminated with suspensions of Enterococcus strains (including a VREfm strain). Human urination was simulated, and colonies of Enterococcus were recovered through a toilet lid where agar plates had been placed. Results showed that the foam was able to suppress production of droplets containing Enterococcus spp. generated by a liquid hitting inner toilet walls. Conversely, Enterococcus colonies were recovered in absence of foam. Moreover, the foam did not show antibacterial activity. We propose a new non-antimicrobial approach aimed at limiting transmission of multidrug-resistant bacteria, particularly in healthcare settings. [ABSTRACT FROM AUTHOR]

Objective(s): We investigated the effect of short-term low-intensity direct current (LIDC) on Staphylococcus aureus. Materials and Methods: The reference strain of S. aureus was used. Experiments were performed in agar culture and on a model of rat's femur osteomyelitis. K-wires were used as electrodes. The exposure to LIDC of 150 µA continued for one minute. In vitro exposure was performed once. In vivo group 1 was a control group. Osteomyelitis was modeled in three groups but only groups 3 and 4 were exposed to LIDC four times: either from day 1 or from day 7 post-surgery. The effect was evaluated on day 21. Microbiological, histological, scanning electron, and light microscopy methods were used for evaluation of the LIDC effect. Results: Bacteria diameter, oblongness, and division increased 15 min after LIDC exposure in the culture around the cathode. After 24 hr, the amount of exomatrix was lower than in the control test, and the cell diameter and roundness increased. Similar changes around the anode were less pronounced. In vivo, biofilm formation on the intramedullary wire cathode was suppressed in group 3. In group 4, detachment and destruction of the biofilm were observed. The formation of S. aureus microcolonies was suppressed, and the adhesion of fibroblasts and immune cells was activated. LIDC did not stop the development of the osteomyelitis process. Conclusion: Short-term exposure to LIDC suppresses S. aureus biofilm formation on the implant cathode surface in the acute and early postoperative period but does not have an impact on the development of osteomyelitis. [ABSTRACT FROM AUTHOR]

Aim: The aim and objective of this in vitro study was to evaluate the antibacterial efficacy of mineral trioxide aggregate, bioactive glass sealer, and epoxy-resin–based sealer. Materials and Methods: In the present study, 22 Mueller Hinton agar (MH agar) plates were employed and equally divided into two groups. Three holes were made by removal of agar at equidistant points and filled with root canal sealers. The strains of the bacteria used in this study were S. aureus (ATCC 6538) and C. albicans (ATCC 10231) and were divided into two groups and root canal sealers were divided into three subgroups: mineral trioxide aggregate (MTA) fillapex Sealer, Nishika Bioactive Glass sealer, and Syntex Epoxy Resin base sealer. For Staphylococcus aureus, peptone water was placed in a 2 mL test tube and bacteria were extracted from blood agar plates using a nichrome wire loop and poured into the peptone water–containing test tube and incubated for 2 hours and for C. albicans, fungi were grown at 37°C for 24 hours in MH Broth and seeded into MH agar to produce turbidity of 0.5 on the McFarland scale, which corresponds to a concentration of 108 CFU/mL. This MH broth was used as a second layer. The seeded agar was then added over the plates immediately after the insertion of sealer cement. After incubation, the diameters of zones of inhibition around the plates were measured. Results: The results of this study showed that the highest inhibition was recorded in Syntex sealer against Staphylococcus aureus followed by MTA fillapex sealer and Nishika sealer, whereas MTA fillapex showed the highest inhibition against C. albicans followed by Syntex sealer and Nishika sealer. Conclusion: Syntex sealer exhibits better antibacterial efficacy against Staphylococcus aureus and MTA fillapex exhibit better antibacterial efficacy against C. albicans. [ABSTRACT FROM AUTHOR]

Surface attached communities of microbes grow in a wide variety of environments. Often, the size of these microbial community is constrained by their physical surroundings. However, little is known about how size constraints of a colony impact the outcome of microbial competitions. Here, we use individual-based models to simulate contact killing between two bacterial strains with different killing rates in a wide range of community sizes. We found that community size has a substantial impact on outcomes; in fact, in some competitions the identity of the most fit strain differs in large and small environments. Specifically, when at a numerical disadvantage, the strain with the slow killing rate is more successful in smaller environments than in large environments. The improved performance in small spaces comes from finite size effects; stochastic fluctuations in the initial relative abundance of each strain in small environments lead to dramatically different outcomes. However, when the slow killing strain has a numerical advantage, it performs better in large spaces than in small spaces, where stochastic fluctuations now aid the fast killing strain in small communities. Finally, we experimentally validate these results by confining contact killing strains of Vibrio cholerae in transmission electron microscopy grids. The outcomes of these experiments are consistent with our simulations. When rare, the slow killing strain does better in small environments; when common, the slow killing strain does better in large environments. Together, this work demonstrates that finite size effects can substantially modify antagonistic competitions, suggesting that colony size may, at least in part, subvert the microbial arms race. Author summary: Bacterial colonies are often crowded with many bacteria in direct contact. As a result, the competition for space and resources often turns deadly. Bacteria have evolved many mechanisms with which to kill each other; this bacterial warfare is often studied in large communities on agar plates or in flow cells. However, in nature these colonies are often smaller, due to spatial constraints or shear forces. It is unclear how bacterial warfare proceeds in small systems. We performed individual based model simulations of bacterial warfare comprising two strains, each capable of killing the other on direct contact. We found that the community size played a substantial role in determining the outcome. When at a numerical disadvantage, the slow killing strain survived at much higher rates in small communities. In fact, there were many conditions in which the slow killing strain survives in small spaces but is completely eliminated in large ones. Conversely, when the slow killing strain is more common, it performs better in large spaces. Together, these observations demonstrate that finite size effects aid the strain that is at a disadvantage, and in some conditions, can even flip which strain increases its abundance. Finally, we experimentally tested the results of these simulations. Two mutual killing strains of V. cholerae were grown unconfined on agar plates (i.e., in large spaces), confined within square holes with sides 7.5μm long (i.e., in small spaces), and confined within large square holes with sides 60μ long to control for effects of the grids. In these experiments we found that the slow killing strain survived at significantly higher rates in small confinement, validating simulation results. [ABSTRACT FROM AUTHOR]

Introduction: Neutrophil extracellular traps (NETs) have various beneficial and detrimental effects in the body. It has been reported that some bacteria may evade the immune system when entangled in NETs. Thus, the aim of the current study was to evaluate the effects of a combined DNase and antibiotic therapy in a murine model of abdominal sepsis. Methods: C57BL/6 mice underwent a cecum-ligation-and-puncture procedure. We used wild-type and knockout mice with the same genetic background (PAD4-KO and DNase1-KO). Mice were treated with (I) antibiotics (Metronidazol/Cefuroxime), (II) DNAse1, or (III) with the combination of both; mock-treated mice served as controls. We employed a streak plate procedure and 16s-RNA analysis to evaluate bacterial translocation and quantified NETs formation by ELISA and immune fluorescence. Western blot and proteomics analysis were used to determine inflammation. Results: A total of n=73 mice were used. Mice that were genetically unable to produce extended NETs or were treated with DNases displayed superior survival and bacterial clearance and reduced inflammation. DNase1 treatment significantly improved clearance of Gram-negative bacteria and survival rates. Importantly, the combination of DNase1 and antibiotics reduced tissue damage, neutrophil activation, and NETs formation in the affected intestinal tissue. Conclusion: The combination of antibiotics with DNase1 ameliorates abdominal sepsis. Gram-negative bacteria are cleared betterwhen NETs are cleaved by DNase1. Future studies on antibiotic therapy should be combined with anti-NETs therapies. [ABSTRACT FROM AUTHOR]

Root system architecture (RSA) influences the acquisition of heterogeneously dispersed soil nutrients. Cytokinin and C-TERMINALLY ENCODED PEPTIDE (CEP) hormones affect RSA, in part by controlling the angle of lateral root (LR) growth. Both hormone pathways converge on CEP DOWNSTREAM 1 (CEPD1) and CEPD2 to control primary root growth; however, a role for CEPDs in controlling the growth angle of LRs is unknown. Using phenotyping combined with genetic and grafting approaches, we show that CEP hormone-mediated shallower LR growth requires cytokinin biosynthesis and perception in roots via ARABIDOPSIS HISTIDINE KINASE 2 (AHK2) and AHK3. Consistently, cytokinin biosynthesis and ahk2,3 mutants phenocopied the steeper root phenotype of cep receptor 1 (cepr1) mutants on agar plates, and CEPR1 was required for trans -Zeatin (t Z)-type cytokinin-mediated shallower LR growth. In addition, the cepd1,2 mutant was less sensitive to CEP and t Z, and showed basally steeper LRs on agar plates. Cytokinin and CEP pathway mutants were grown in rhizoboxes to define the role of these pathways in controlling RSA. Only cytokinin receptor mutants and cepd1,2 partially phenocopied the steeper-rooted phenotype of cepr1 mutants. These results show that CEP and cytokinin signaling intersect to promote shallower LR growth, but additional components contribute to the cepr1 phenotype in soil. [ABSTRACT FROM AUTHOR]

Urogenital Mollicutes, that is, Mycoplasma hominis and Ureaplasma spp., can colonize the urogenital tract. While urogenital colonization is frequent, infections are rare but should not be missed. Furthermore, extragenital infections are even rarer. Over the past years, they have been increasingly documented as a cause of hyperammonemia syndrome (HS) and post-surgical infections. We review the literature on studies focused on post-surgical infections and HS involving urogenital Mollicutes after thoracic surgery including lung (LTR) and heart (HTR) transplantation. A systematic review was performed by searching PubMed/Medline case reports, case series, cohort studies, and clinical trials. Cases of infections and HS by urogenital Mollicutes after HTR and LTR transplantations were reported. Overall, urogenital Mollicutes were associated with 15 HS, 31 infections in HTR and LTR, and 18 post-thoracic surgical infections in another context. Post-surgical infections were reported in all contexts. They were mainly due to M hominis , the only species that could cultivate on standard enriched agar forming pinpoint colonies after 3–5 days of incubation. Microbiologists should be prompted to pinpoint colonies even if the examination of Gram-staining is negative. The patients' management required surgical treatment and antimicrobials, almost always tetracyclines and/or fluoroquinolones. Conversely, HS occurred almost exclusively in bilateral LTR and is more likely due to Ureaplasma spp. As Ureaplasma spp. do not cultivate on standard media, the microbiological diagnosis was performed using molecular methods. Infections involving urogenital Mollicute should be considered in LTR with HS. The overall rate of mortality is high and might be due in part to delay in etiologic diagnosis. Post-surgical infections were reported in all contexts. The route of contamination with Mollicutes remains unknown in HTR and non-transplant surgery, but evidence of transmission from donors has been documented for LTR. [ABSTRACT FROM AUTHOR]

We developed a new ligamentoplasty technique using the ulnotriquetral ligament (UTL) via a palmar approach for cases with dorsal instability of the ulna due to a foveal tear of the triangular fibrocartilage complex (TFCC). The study aimed to evaluate the clinical outcomes of this procedure. We retrospectively analyzed 19 cases with foveal tears of TFCC who underwent the ligamentoplasty using UTL. We harvested the UTL from the triquetrum as a pedicle graft, avoiding damage of the connection between the base of the UTL and palmar radioulnar ligament (PRUL) as possible. The graft was flipped proximally and pulled out through the bone tunnel from the ulnar fovea to the ulnar neck, then fixed to the fovea using a tenodesis screw. The function of the TFCC can be reproduced by pulling out the UTL along with the PRUL through the ulnar fovea and re-establishing the PRUL tension. Clinical results were evaluated at least 12 months following surgery. The preoperative and postoperative wrist pain level on a neumerical rating scale (NRS), the radioulnar joint (DRUJ) ballottement test, the range of motion, the grip strength, and the Patient-rated wrist evaluation (PRWE) scores were evaluated. In all cases, the DRUJ ballottement test became negative at the final follow up compared with the normal side on manual examination. The pain levels significantly decreased as reflected by a decrease in NRS from 6.8 to 2.5 at final follow up (p < 0.01). The range of pronation/supination motion was improved from 149° to 157°. The mean PRWE score significantly decreased from 52.1 to 22.8 (p < 0.01). The postoperative pain level on the NRS scale of the three patients poorly improved. One case of the three with the 3 mm positive ulnar variance needed additional ulnar shaft shortening using a plate seven months after the primary procedure. The ligamentoplasty using UTL efficiently restored the ulnar palmar stability in all 19 cases and significantly decreased the wrist pain and the PRWE scores. Clinical, retrospective study. [ABSTRACT FROM AUTHOR]