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In this study, microbial siderophore-mediated Li bioleaching from cathode active material (CAM) of spent LIBs was attempted. Pseudomonas aeruginosa was used to produce siderophores. For maximum siderophore production, favourable conditions like initial pH of Fe-free medium (5–9) and glucose (0.5–5 g/L) were assessed. Under optimal siderophore production conditions, Li bioleaching from CAM powder was studied at varying pulp densities (PD: 1–5 g/L), using bacterial cultures cultivated for 48 h and 120 h. Maximum siderophore production of ∼52 psu occurred at pH 7 and 1 g/L glucose after 48 h. Qualitative and quantitative analyses confirmed the production of mixed-type siderophores (catecholate and hydroxamate), which was further corroborated by FTIR analysis. Produced siderophores using 120 h old bacterial culture showed maximum Li extraction of >99 % at 1 g/L PD in 144 h. Artificial neural network model accurately predicted the response (R2=0.9997; RMSE=0.4955). Following the shrinking core model, kinetics analysis revealed the microbial siderophore-mediated Li bioleaching mechanism followed the diffusion of lixiviant into CAM particles and dissolution of product layer away from the particle surface, which further substantiated by XRD and FESEM analyses. This study offers an alternative bioleaching approach for sustainable Li extraction from spent LIBs employing microbial siderophores. [Display omitted] • Siderophore-mediated lithium (Li) extraction from spent Li-ion batteries was studied. • Pseudomonas aeruginosa produced mixed-type siderophores: catecholate and hydroxamate. • Maximum Li extraction of more than 99 % achieved in 144 h at 1 g/L of pulp density. • Diffusion is the rate-controlling step of Li extraction from cathode active material. • Artificial neural network model reliably predicted siderophore-mediated Li extraction. [ABSTRACT FROM AUTHOR]

This study aimed to develop a nano-biosystem by covalently binding recombinant laccase from Bacillus licheniformis (rLAC BL) to PEGylated silica-coated magnetic nanoparticles (EMNPs-PEG) and evaluate its cytotoxicity and anti-inflammatory properties on a pro-monocytic cell line. The active rLAC BL was purified from the bacterial culture with a yield of approximately 14 mg/L and purity of about 91 %. The physicochemical characteristics of EMNP-PEG-rLAC BL were confirmed using DLS, UV-Vis spectrum, FTIR, EDS, and SEM. The immobilized rLAC BL exhibited improved kinetic parameters compared to the bare enzyme. The IC 50 value for EMNP-PEG-rLAC BL (58.7 μg/mL) was lower than that of the bare enzyme after 48 hours of treatment on U937 cells. Both EMNP-PEG-rLAC BL and rLAC BL significantly decreased the expression levels of key cytokines (TNF-α, IL-6, IL-17, and IL-23) in LPS-stimulated cells, indicating promising anti-inflammatory properties of rLAC BL. Given the association between chronic inflammation and cancer, this newly developed enzyme-based nanobiomedicine holds potential as a therapeutic option for inflammatory conditions and malignant diseases. However, further research is needed to validate these findings and fully assess the therapeutic potential of LACs before clinical translation can be considered. [Display omitted] • Recombinant laccase (rLAC BL) was produced from Bacillus licheniformis. • The rLAC B was immobilized on PEGylated silica-coated magnetic nanoparticles. • Immobilization of rLAC BL notably enhanced enzyme affinity for its substrate. • Immobilized rLAC BL exhibited anti-inflammatory effects on the U937 cell line. [ABSTRACT FROM AUTHOR]

Basal stem rot disease caused by Ganoderma boninense is a major problem for oil palm cultivation. The research was conducted to obtain biocontrol agents from rhizosphere of oil palm to control the disease as part of sustainable pest management in oil palm plantation. Rhizosphere bacteria were isolated from rhizospheres of healthy oil palm trees. Isolation of bacteria was done using serial dilution method. The isolated bacteria were then tested for its antifungal activity against G. boninense in vitro using dual culture assay. The ability of the bacteria to produce antifungal compound was also determined by culturing the bacteria on ISP2 liquid media. Once the bacterial cells were removed, the crude metabolites were then tested against G. boninense using agar well diffusion and toothpick colonization. The result showed that several isolates demonstrated strong antifungal activity against G. boninense. Some isolates were also able to degrade chitin and to solubilize phosphate. Furthermore, the crude metabolites produced by the rhizosphere bacteria demonstrated the ability to inhibit the growth of G. boninense in the agar diffusion method. Colonization of the G. boninense on toothpick following soaking in the crude metabolites was also inhibited. The isolated rhizosphere bacteria (BARK7 and BARK15 in which identified as Burkholderia sp.) showed promising ability to be developed as biocontrol agent for basal stem rot disease of oil palm. [ABSTRACT FROM AUTHOR]

Aims: To assess agreement of bacterial culture results from samples taken from nasal discharge, the nasal cavity and nasal biopsy from dogs and cats with nasal disease. Methods: Nineteen dogs and 21 cats with different nasal diseases (chronic rhinitis, n = 30; neoplasia, n = 7; sinonasal aspergillosis, n = 3) were prospectively enrolled in the study. Nasal swabs were taken bilaterally from nasal discharge at the nares, the nasal cavity, and one nasal mucosal biopsy per side. All samples were subjected to aerobic bacterial culture. Kappa statistics were used to evaluate agreement for the most prevalent bacterial species between sampling sites. Results: A positive culture result for at least one bacterial species was detected in 80% of samples from nasal discharge/nares, 92% of nasal cavity samples, and 75% of biopsy samples. The mean agreement between the three sampling sites for positive vs. negative culture results was never greater than moderate and the precision of the estimates of agreement varied widely. The most frequently isolated bacterial species in dogs were Staphylococcus pseudintermedius, Staphylococcus spp. and Streptococcus spp. In cats, Pasteurella spp. and Staphylococcus felis were the bacterial species cultured most frequently. For the most prevalent cultured species, Staphylococcus spp., mean agreement between sites was never greater than fair and the precision again varied widely. Conclusion: This study indicates that bacterial culture results in feline and canine nasal disease are site-specific and there was no evidence from this study for consistency between sites within a patient for many bacterial species. Consequently, if bacterial culture results from nasal swabs are used to guide therapeutic antimicrobial choice, different treatments may be selected depending on the site of culture. As a consequence, there is no evidence from this study that nasal bacterial cultures should be recommended as a routine diagnostic measure. [ABSTRACT FROM AUTHOR]

Introduction: Veterinary clinical microbiology laboratories play a key role in antimicrobial stewardship, surveillance of antimicrobial resistance and prevention of healthcare associated-infections. However, there is a shortage of international harmonized guidelines covering all steps of veterinary bacterial culture from sample receipt to reporting. Methods: In order to gain insights, the European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT) designed an online survey focused on the practices and interpretive criteria used for bacterial culture and identification (C&ID), and antimicrobial susceptibility testing (AST) of animal bacterial pathogens. Results: A total of 241 microbiology laboratories in 34 European countries completed the survey, representing a mixture of academic (37.6%), governmental (27.4%), and private (26.5%) laboratories. The C&ID turnaround varied from 1 to 2 days (77.8%) to 3-5 days (20%), and 6-8 days (1.6%), with similar timeframes for AST. Individual biochemical tests and analytical profile index (API) biochemical test kits or similar were the most frequent tools used for bacterial identification (77% and 56.2%, respectively), followed by PCR (46.6%) and MALDI-TOF MS (43.3%). For AST, Kirby-Bauer disk diffusion (DD) and minimum inhibitory concentration (MIC) determination were conducted by 43.8% and 32.6% of laboratories, respectively, with a combination of EUCAST and CLSI clinical breakpoints (CBPs) preferred for interpretation of the DD (41.2%) and MIC (47.6%) results. In the absence of specific CBPs, laboratories used human CBPs (53.3%) or veterinary CBPs representing another body site, organism or animal species (51.5%). Importantly, most laboratories (47.9%) only report the qualitative interpretation of the result (S, R, and I). As regards testing for AMR mechanisms, 48.5% and 46.7% of laboratories routinely screened isolates for methicillin resistance and ESBL production, respectively. Notably, selective reporting of AST results (i.e. excluding highest priority critically important antimicrobials from AST reports) was adopted by 39.5% of laboratories despite a similar proportion not taking any approach (37.6%) to guide clinicians towards narrower-spectrum or first-line antibiotics. Discussion: In conclusion, we identified a broad variety of methodologies and interpretative criteria used for C&ID and AST in European veterinary microbiological diagnostic laboratories. The observed gaps in veterinary microbiology practices emphasize a need to improve and harmonize professional training, innovation, bacterial culture methods and interpretation, AMR surveillance and reporting strategies. [ABSTRACT FROM AUTHOR]

Introduction: Protection of horse welfare during transport is crucial. The aim of this study was to determine the effect of head and neck restraint on behavior and airway bacteria. Methods: In a randomized crossover study, six healthy Thoroughbreds were transported by road for 22 h in an individual bay with tight head restraint (50 cm short-rope) or loose head restraint (95 cm long-rope). Behavioral parameters relating to head position, eating, and stress were monitored during transportation. Tracheal wash samples were obtained 6 days before and immediately after transport for bacterial culture and metagenomic analysis. Results and discussion: Compared to before transport, bacterial load (CFU/mL) after transport was significantly increased in the short-rope group (p = 0.04), whereas no changes were observed in the long-rope group. Transport significantly reduced Simpson index at phylum, class, order, and family levels in both groups (p < 0.001) of tracheal microbiota. In both groups, this reduction was associated with increases in the dominant members of relative abundance at phylum (Firmicutes: +24% in long-rope and +14% in short-rope), class (Bacilli: +20% in long-rope and +22% in short-rope) and family (Streptococcaceae: +22% in long-rope and +23% in short-rope) levels. Licking behavior during transportation with short-rope restraint was more frequent than in horses with long-rope restraint. These results suggest loose head restraint during transportation is likely to ameliorate stress and mitigate the associated increased bacterial load in the lower airways associated with transport. Further, head position during transportation is likely a more important determinant of airway hygiene and distress than duration of travel. [ABSTRACT FROM AUTHOR]

The need to address antimicrobial resistance (AMR) through a One Health (OH) approach is now well recognized. There is, however, limited guidance on how AMR surveillance should be implemented across sectors to generate meaningful AMR and AMU data for decision‐making. Using a sympatric approach to cross‐sector sample collection, Nepal adopted the WHO extended‐spectrum beta‐lactamase (ESBL)–producing Escherichia coli (E. coli) Tricycle Project as a step toward OH surveillance for assessing the prevalence of ESBL‐producing E. coli across human, veterinary, and environment sectors. This involved a three‐stage approach: identification of human hotspots (Stage 1) and sample collection sites for poultry (Stage 2) and wastewater (Stage 3). A total of 53 blood cultures from patients with bloodstream infections (BSIs), 100 stool samples from healthy pregnant women, 220 poultry ceca from slaughterhouses and live markets, and 48 wastewater samples were processed for bacterial culture and analyzed for the presence of ESBL‐producing E. coli. The prevalence of ESBL‐producing E. coli among isolated E. coli was the highest in wastewater samples (91%) followed by human BSIs (49%), poultry (38.6%), and fecal carriage isolates from healthy pregnant females (15%). A statistically significant association was seen in the prevalence of multidrug resistance among ESBL producers (52%) and nonproducers (26%). ESBL‐producing E. coli was detected in all wastewater samples tested except for the upstream river. The findings of the study showed a high prevalence of ESBL‐producing E. coli in samples from all three sectors and provided baseline data based upon which strategies for the safe disposal of communal and hospital waste, integrated AMR surveillance, and control strategies could be planned and implemented. [ABSTRACT FROM AUTHOR]

Background and Objectives Materials and Methods Results Conclusion Honduras became the first lower middle‐income country (LMIC) to adopt amotosalen/UVA pathogen‐reduced (PR) platelet concentrates (PCs) as a national platelet safety measure in 2018. The Honduran Red Cross (HRC) produces ~70% of the national platelet supply using the platelet‐rich plasma (PRP) method. Between 2015 and 2018, PCs were screened with bacterial culture and issued as individual, non‐pooled PRP units with weight‐based dosing and 5‐day shelf‐life. PR PCs were produced in six‐PRP pools with a standardized dose (≥3.0 × 1011), no bacterial screening and 7‐day shelf‐life. Gamma irradiation and leukoreduction were not used.PC production and distribution data were retrospectively analysed in two periods. Period 1 (P1) included 3 years of PRP PCs and a transition year (2015–18). Period 2 (P2) included 5 years of PR PCs (2019–23). PC doses were standardized to an equivalent adult dose for both periods. Descriptive statistics were calculated.HRC produced 10% more PC doses per year on average in P2 compared to P1. Mean annual waste at HRC declined from 23.9% in P1 to 1.1% in P2. Two urban regions consumed 96% of PC doses in P1 and 88.3% in P2. PC distributions increased in 14/18 regions.Standardized dosage, PR and 7‐day shelf‐life increased PC availability, reduced waste, eliminated bacterial screening and avoided additional costs for arboviral testing, leukoreduction and irradiation. Access to PC transfusion remains limited in Honduras; however, the conversion to pooled PR PCs illustrates the potential to sustainably expand PC distribution in an LMIC. [ABSTRACT FROM AUTHOR]

Erwinia amylovora bacteriophages are of interest as fire blight control agents. This paper presents data on the biology and molecular genetic properties of 12 E. amylovora tailed bacteriophages. Genome sequences of seven of them were determined and the phages were identified as the representatives of Caudoviricetes; Vequintavirinae, Ounavirinae and Autographiviridae families. The bacteriophages studied were active against E. amylovora, Pantoea agglomerans and Pantoea ananatis strains. The myovirus Hena1 had the narrowest host range lysing 12% of the bacterial cultures tested, the remaining myoviruses—had broader host ranges (56%) for this collection. The phages showed different reactions to the presence of a chelating agent in the cultivation medium. A significant proportion of phage-resistant E. amylovora cells were found both in infected liquid cultures and in the experiments on the plates with the phage agar (more than 20% in case of exposure to podoviruses), which correlates with the data of limited number of previous investigations of the phenomenon for E. amylovora interactions with phages. High prevalence of resistant cells in the host cultures suggest that they are formed not due to spontaneous mutations but result from another phenotypic or genetic dissociation mechanism(s) that remains to be identified. [ABSTRACT FROM AUTHOR]

Background: Suppurative perichondritis of the auricle is a common disease that can easily cause malformations if it develops into an uncontrolled infection. In nearly half of the cases, otolaryngologists cannot identify the pathogens involved. Case presentation: In the present work, we described two cases of pyogenic perichondritis, with negative on conventional culture. However, using metagenomic next-generation sequencing (mNGS), we detected fungal infections in the patients and after the patients were given anti-fungal treatment, the patients achieved a good prognosis. Conclusions: These cases highlighted the possibility that fungi might be the involved pathogens in patients who have had multiple negative bacterial cultures, and mNGS should be applied in these cases. mNGS could be used as a supplement to traditional culture methods. [ABSTRACT FROM AUTHOR]

This ex vivo study evaluated the disinfecting and cleaning effects of root canal preparation using sodium hypochlorite irrigation with 3 different needle designs. Mesial roots from extracted mandibular molars with Vertucci class II configuration were anatomically matched based on micro-computed tomography (micro-CT) analyses and distributed into 3 groups (n = 18/group). The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation using 2.5% sodium hypochlorite irrigation with open-ended flat needle (3 mm short of the working length - WL), closed-ended side-vented irrigation needle (1 mm short of the WL), or a closed-ended plastic needle with 2 lateral outlets (TruNatomy) (1 mm short of the WL). Bacteriological samples were taken from the canals before (S1) and after preparation (S2). After another micro-CT scan, the roots were sectioned and samples were also taken from the apical canal segment (S3). Bacterial reduction was assessed by quantitative real-time polymerase chain reaction. The amount of accumulated hard tissue debris was evaluated by micro-CT. A substantial reduction in bacterial counts was observed in all 3 groups when comparing S1 with S2 (99.95%, 98.93%, and 98.90% in the open-ended, closed-ended, and TruNatomy needle groups, respectively) (P <.01). No significant differences were observed between groups for bacterial reduction in the full and apical canal (P >.05). There were no intergroup differences in the amount of accumulated hard tissue debris either (P >.05). The open-ended needle group showed significantly more specimens with quantitative real-time polymerase chain reaction negative results for bacteria in S3 than TruNatomy (P <.05). When used up to an appropriate insertion depth, the 3 needle types result in similar antibacterial and cleaning performance, provided variables such as needle size and irrigant type, volume and flow rate are controlled. Exclusive analysis of the apical segment, including the isthmus, revealed that the open-ended needle yielded more cases negative for bacteria. [ABSTRACT FROM AUTHOR]

Novel probiotic yoghurt was produced using the combination of bacterial cultures Lactobacillus plantarum HA119 and Bifidobacterium animalis subsp. lactis B94 and yoghurt bacteria Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Its basic nutritional composition, colour, texture, rheological properties, and sensory profile were compared with yoghurt produced using the same technological process and standard yoghurt cultures (control sample), as well as other commercially available yoghurts with different milk fat contents. Despite the fat content of the yoghurt made with the new probiotic cultures being 1.44%, its apparent viscosity was similar to that of high-fat yoghurt (2.99%). Other results from rheological measurements indicate that the new yoghurt had a stronger protein network, presumably due to the higher number of exopolysaccharides compared to both control and commercial yoghurts. Sensory analysis revealed that there were no statistically significant differences between the novel probiotic yoghurt and high-fat yoghurt as perceived by panellists. In conclusion, this combination of probiotic cultures can be used to produce yoghurt with rheological and sensory properties similar to high-fat yoghurts, without the need for hydrocolloids or changes in the production process. [ABSTRACT FROM AUTHOR]

In this study, poly(lactic acid), poly(ethylene glycol), and benzalkonium chloride with different concentrations (3, 5, 7, and 9%wt.) (PLA/PEG/BCL) composite electrospun mats were produced. PLA is a non‐toxic polymer with high biocompatibility and biodegradability. However, it may be fragile due to its structure. Therefore, in this study, PEG was used as a plasticizer to improve the structural properties of PLA and it was aimed at providing antibacterial properties by adding BCL salt. Its use as an antibacterial composite nanomaterial effective against Gram‐positive Staphylococcus aureus (S. aureus) and Gram‐negative Escherichia coli (E. coli) bacterial cultures and as a dermal wound dressing material has been examined in two different areas. The addition of BCL salt reduced the bead formation in PLA/PEG nanofibers and increased the homogeneity of fiber dispersion. 9% BCL‐doped composite nanofiber was obtained as the smoothest and most homogeneous surface. This mat was reported to have the highest ductility. The low Tm of pure BCL salt enabled the Tg temperature of PLA/PEG/BCL composite nanofibers to be observed. It was observed that as the BCL salt ratio increased, the T5 and T10 temperatures of the nanofibers decreased and then increased. BCL‐doped mats exhibited liquid absorption behavior in the range of 497%–708%. PLA/PEG/BCL composite nanofibers showed high toxicity to the L929 fibroblast cell line. So, it has been reported that it cannot be used as a dermal wound dressing. PLA/PEG/BCL composite nanomaterials were reported to have 99.99% antibacterial activity against E. coli and S. aureus. It was suggested that it could be used in antibacterial coating applications by taking into account modern nanocoating technology. Highlights: Poly(lactic acid), poly(ethylene glycol), and benzalkonium chloride (PLA/PEG/BCL) composite electrospun mats were produced.The addition of BCL salt reduced the bead formation in PLA/PEG nanofibers and increased the homogeneity of fiber dispersion.9% BCL‐doped composite nanofiber was obtained as the smoothest and most homogeneous surface.PLA/PEG/BCL composite nanofibers showed high toxicity to the L929 fibroblast cell line.PLA/PEG/BCL composite nanomaterials were reported to have 99.99% antibacterial activity against E. coli and S. aureus. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effects of the cell-free supernatant of Lactiplantibacillus plantarum ATCC® 10241TM on the biofilm-forming capacity of Pseudomonas aeruginosa strains isolated from cystic fibrosis (CF) patients. In addition, the study evaluated the in vivo potential of the cell-free supernatant to modulate inflammation and reduce lung damage in mice infected with P. aeruginosa strains or co-challenged with P. aeruginosa and the Streptococcus milleri group (SMG). The results showed that CF-derived P. aeruginosa strains can infect the respiratory tract of adult mice, inducing local inflammation and lung damage. The severity of these infections was exacerbated when P. aeruginosa was co-administered with SMG. Notably, nebulization with the cell-free supernatant of L. plantarumproduced beneficial effects, reducing respiratory infection severity and inflammatory responses induced by P. aeruginosa, both alone or in combination with SMG. Reduced bacterial loads and lung damage were observed in supernatant-treated mice compared to controls. Although further mechanistic studies are necessary, the results show that the cell-free supernatant of L. plantarum ATCC® 10241TM is an interesting adjuvant alternative to treat P. aeruginosa respiratory infections and superinfections in CF patients. [ABSTRACT FROM AUTHOR]

Introduction: Different initial manifestations of peritoneal dialysis-associated peritonitis (PDAP) may depend on the type of pathogenic organism. We investigated the association between the clinical characteristics of PDAP and susceptibility to vancomycin and investigated the possibility of using vancomycin monotherapy alone as an initial treatment regimen for some PDAP patients to avoid unnecessary antibiotic exposure and secondary infection. Methods: Patients with culture-positive PDAP were retrospectively analyzed and divided into two groups: peritonitis with only cloudy effluent (PDAP-cloudy) or with cloudy effluent, abdominal pain and/or fever (PDAP-multi). The bacterial culture of PD effluent and antibiotic sensitivity test results were compared between groups. Logistic regression was used to investigate factors predicting susceptibility to vancomycin. Results: Of 162 episodes of peritonitis which had a positive bacterial culture of PD fluid, 30 peritonitis were in the PDAP-cloudy group, and 132 peritonitis were in the PDAP-multi group. Thirty (100%) peritonitis in the PDAP-cloudy group had gram-positive bacterial infections, which was significantly greater than that in the PDAP-multi group (51.5%) (P < 0.001). Twenty-nine (96.7%) peritonitis in the PDAP-cloudy group were susceptible to vancomycin, compared to 67 (50.8%) in the PDAP-multi group (P < 0.001). The specificity of PDAP-cloudy for vancomycin-sensitive peritonitis was 98.48%. Only one patient (3.3%) in the PDAP-cloudy group experienced vancomycin-resistant peritonitis caused by Enterococcus gallinarum, which could neither be covered by vancomycin nor by the initial antibiotic regimen recommended by the current ISPD guidelines. The presence of only cloudy effluent was an independent predictor of susceptibility to vancomycin according to multivariate analysis (OR = 27.678, 95% CI 3.191-240.103, p = 0.003), in addition to PD effluent WBC counts (OR = 0.988, 95% CI 0.980–0.996, p = 0.004), diabetes mellitus (OR = 3.646, 95% CI 1.580–8.416, p = 0.002), first episode peritonitis (OR = 0.447, 95% CI 0.207–0.962, p = 0.039) and residual renal creatinine clearance (OR = 0.956, 95% CI 0.918–0.995, p = 0.027). Addition of these characteristics increased the AUC to 0.813 (95% CI 0.0.749–0.878, P < 0.001). The specificity of presenting with only cloudy effluent for vancomycin-sensitive peritonitis was 98.48%. Conclusions: Cloudy dialysate, as the only symptom at PDAP onset, was an independent predictor of vancomycin-sensitive PDAP, which is an important new insight that may guide the choice of initial antibiotic treatment. [ABSTRACT FROM AUTHOR]

Background Hypothesis/Objectives Animals Materials and Methods Results Conclusions and Clinical Relevance Bacterial identification and antimicrobial susceptibility testing is an important step in timely therapeutic decisions for canine superficial bacterial folliculitis (SBF), commonly caused by Staphylococcus pseudintermedius. Next‐generation sequencing (NGS) offers the appeal of potentially expedited results with complete detection of bacterial organisms and associated resistance genes compared to culture. Limited studies exist comparing the two methodologies for clinical samples.To compare and contrast genotypic and phenotypic methods for bacterial identification and antimicrobial susceptibility from cases of canine SBF.Twenty‐four client‐owned dogs with lesions consistent with SBF were enrolled.A sterile culturette swab was used to sample dogs with SBF lesions. The swab was rinsed in 0.9 mL of sterile phosphate‐buffered saline and vortexed to create a homogenous solution. Two swabs for NGS laboratories (Labs) and one swab for culture (Culture Lab) were randomly sampled from this solution and submitted for bacterial identification and antimicrobial susceptibility.No statistical difference regarding turnaround time for NGS Labs compared to Culture Lab was found. NGS Lab 1 identified more organisms than NGS Lab 2 and Culture Lab, which were both statistically significant. There was no statistical difference in detection frequency for Staphylococcus spp. among all laboratories. There was poor agreement for the presence of meticillin resistance and most antimicrobials among all laboratories.Utilisation of NGS as a replacement for traditional culture when sampling canine SBF lesions is not supported at this time. [ABSTRACT FROM AUTHOR]

Introduction: Urinary tract infections (UTIs) pose a significant health concern, particularly among pregnant women, for whom accurate diagnosis is essential. However, the use of Urine flow cytometry (UF) for detecting UTIs in this demographic often results in misdiagnosis. The objective of this study was to explore the reasons behind these diagnostic errors and to develop a strategy to minimize the rate of UTI misdiagnosis in pregnant women. Material and methods: The study enrolled 1,200 women aged 18 to 40 years, categorized into pregnant and non-pregnant groups. UTIs were diagnosed using urine bacterial culture, microscopic examination, and UF, followed by statistical analysis to identify any discrepancies in diagnosis between the groups. Following the calibration of UF analyzer's parameters, the most effective CR(WBC)-CW-FSC-P Gain setting for diagnosing UTIs in pregnant women through UF was ascertained by applying the Youden index. Results: The clinical diagnosis rate of UTIs was significantly higher in pregnant women (40.91%) compared to non-pregnant women (20.26%). However, urine microscopy and bacterial culture showed no significant difference in the rates of UTIs between the two groups, suggesting a potential for misdiagnosis. The false-positive rate for WBCs detected by UF was 30.43%, and adjusting the CR(WBC)-CW-FSC-P Gain value of UF reduced the false-positive rate to 9.45%. Conclusion: The incidence of UTIs in pregnant women may be overestimated because of the limitations inherent to UF. Adjusting the parameters of the UF analyzer, particularly the CR(WBC)-CW-FSC-P Gain value, can significantly reduce the rate of UTI misdiagnosis in pregnant women. [ABSTRACT FROM AUTHOR]

Simple Summary: The dissemination of antimicrobial resistance (AMR) genes in economic animals affects food safety in our life cycle. The pig gut microbiome can be a reservoir of antimicrobial-resistant bacteria. As a result of the comparison of antimicrobial susceptibility profiles and the existence of ESBL genes, we found no difference between antibiotic- and nonantibiotic-using farms. We suggest a basal level of ESBL E. coli persistence on pig farms, which may not depend on antibiotic usage. Direct detection of antimicrobial-resistant genes from pig fecal samples reduced cumbersome bacterial culture processes. The enhanced sensitivity of the nPCR technique facilitates the surveillance of AMR genes, leading to effective control. The commensal Escherichia coli in the gut of pigs is a major reservoir of antimicrobial resistance and can result in possible transmission to humans through the food chain. Direct detection of E. coli from fecal samples is challenging and can be used as a bioindicator of antimicrobial resistance. This study aimed to compare the antimicrobial susceptibility profiles in commensal E. coli from antibiotic- and nonantibiotic-using pig farms and developed the direct detection of ESBL genes in pig fecal samples using nested PCR (nPCR) and multiplex PCR (mPCR) techniques. All direct genotypic results were validated with the results of PCR sequencing of isolated E. coli colonies. The ESBL-producing E. coli were found in 98.6% (145 isolates) and 96.6% (144 isolates) of antibiotic-using and nonantibiotic-using farms, respectively, predominantly CTX-M-55. The nPCR decreased the limit of detection (LOD) from sPCR about 100 times, and the lower LODs of 102, 101, and 1 CFU/mL were reached after incubating samples in an enrichment medium for 2, 4, and 8 h, respectively. The mPCR, sPCR, and nPCR techniques showed sensitivities of 30.15%, 69.85%, and 91.91%, respectively, compared to PCR sequencing. The stability and recycling of ESBL genes were independent of antibiotic usage in commensal E. coli originating in pig farms. [ABSTRACT FROM AUTHOR]

Centella asiatica, a traditional herb, is widely recognized for its pharmacologically active components, such as asiaticoside, madecassoside, asiatic acid, and madecassic acid. These components render it a highly sought-after ingredient in various industries, including cosmetics and pharmaceuticals. This study aimed to enhance the production and activity of these pharmacological constituents of C. asiatica using the plant growth-promoting rhizobacterium Priestia megaterium HyangYak-01 during its cultivation. To achieve this goal, the researchers conducted field experiments, which revealed an increase in the production of pharmacologically active compounds in C. asiatica cultivated with a P. megaterium HyangYak-01 culture solution. Additionally, quadrupole time-of-flight mass spectrometry (Q-TOF MS) confirmed that the composition ratios of the C. asiatica extract treated with the P. megaterium HyangYak-01 culture solution differed from those of the untreated control and type strain-treated groups. Skin cell experiments indicated that the C. asiatica extract treated with the P. megaterium HyangYak-01 culture solution exhibited greater skin barrier improvement and less pronounced inflammatory responses than those from plants grown without the bacterial culture solution. This study demonstrates that microbial treatment during plant cultivation can beneficially influence the production of pharmacological constituents, suggesting a valuable approach toward enhancing the therapeutic properties of plants. [ABSTRACT FROM AUTHOR]

Objective Animals Studied Procedures Results Conclusions To establish baseline ophthalmic parameters for an endangered, semi‐wild population of healthy whooping cranes (Grus americana) (WHCR) and Mississippi sandhill cranes (Grus canadensis pulla) (SACR).Eighteen WHCR and 16 SACR.Ophthalmic examination was performed by a single observer, followed by conjunctival swab collection for aerobic bacterial culture and measurement of tear production (phenol red thread test, PRTT) and corneal diameter (CD) as tolerated. Measurement of the axial globe (AG) length, anterior chamber (AC) depth, lens thickness, vitreous chamber (VC) depth, and pecten length was performed via ocular ultrasound (OUS) as tolerated.Eyelid cicatrization (n = 1 WHCR), keratitis (n = 2 WHCR), incipient cataracts (n = 1 WHCR, n = 4 SACR), and uveal cysts (n = 1 SACR) were identified. Twenty‐one bacterial species were cultured from SACR, while 18 bacterial species were cultured from WHCR. SACR under 6 months old had increased PRTT values compared to older SACR (p = .0432). AG length and VC depth of male WHCR were greater than in female WHCR (p = .0045 and p = .0008, respectively). WHCR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .001 and p = .0013, respectively). SACR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .0001 and p < .0001, respectively).WHCR and SACR are amenable to complete ophthalmic examination. Age‐related differences in PRTT in SACR, sexual dimorphism in WHCR, and age‐related differences in AC depth and lens thickness in WHCR and SACR were identified. [ABSTRACT FROM AUTHOR]

Aim Methods Results Conclusion To assess the duration of antimicrobial treatment; hospital length of stay; and invasive bacterial infections rates in hospitalised infants following the adoption of a management guideline.Faculty agreed to a standard of 24 h of antibiotic treatment for well‐appearing febrile infants with proven viral infection and no growth on bacterial cultures. The outcomes were the duration of hospitalisation and antibiotic treatment of febrile infants less than 8 weeks of age who have enterovirus, parechovirus, respiratory viruses detected. We monitored re‐admissions and missed invasive infections.Of the total 1696 infants studied, the median antibiotic treatment duration decreased from 31.5 to 24.8 h in virus‐infected infants ≤21 days of age (p = 0.02) and from 26 to 19.7 h in infants 22–56 days of age (p < 0.001). The decrease was less in infants not infected with a virus. No patient had an invasive infection identified after discharge.The implementation of our care standard resulted in reduction in antibiotic treatment duration without known delayed diagnosis of bacterial infections. Infants without a proven viral aetiology may need further study to inform management decisions. [ABSTRACT FROM AUTHOR]

Necrotizing fasciitis (NF) is a life-threatening soft-tissue infection affecting the deep fascia and subcutaneous tissue. It is characterized by a fulminant course and high mortality rates. NF of the head and neck is very rare, with most cases being odontogenic in origin. The purpose of this study is to comprehensively review the most important features of cervical necrotizing fasciitis (CNF) in adults and add our experience in the management of this entity. The most common isolated organisms are Streptococcus spp. and Staphylococcus spp. If the infection progresses to descending mediastinitis, the prognosis becomes very poor. Since the initial clinical features can be similar to those of a non-necrotizing deep cervical infection, a high degree of suspicion is critical for an early diagnosis. A computed tomography scan is essential for the diagnosis and to define the extent of the infection/rule out descending mediastinitis. Early and aggressive surgical debridement of all compromised tissue and antibiotic therapy and fluid resuscitation are essential and should not wait for bacterial culture results. Despite prompt and adequate treatment, the mortality of CNF can be as high as 35%. [ABSTRACT FROM AUTHOR]

Background: Recurrent bacterial cystitis, often referred to as recurrent urinary tract infection (UTI), can be difficult to manage and alternative treatments are needed. Hypothesis/Objective: Intravesicular administration of asymptomatic bacteriuria (ASB) E. coli 212 will not be inferior to antimicrobial treatment for the management of recurrent UTI in dogs. Animals: Thirty‐four dogs with >1 UTI in the 12 months before presentation. Methods: All dogs were deemed normal otherwise based on absence of abnormalities on physical examination, CBC, serum biochemical panel, and abdominal ultrasonography. Dogs were randomized to 1 of 2 treatment groups: Group 1 antimicrobials for 7 days or group 2 intravesicular administration of ASB E. coli 212. Owners were provided a voiding questionnaire regarding their dogs' clinical signs, which was completed daily for 14 days to assess clinical cure. Dogs were examined on days 7 and 14 to assess clinical cure, and urine specimens were submitted for urinalysis and bacterial culture. Results: Clinical cure rates for ASB E. coli 212–treated dogs were not inferior to 7 days of antimicrobial treatment with a 12% margin of difference to determine non‐inferiority. No significant difference was found between the treatment groups on days 7 and 14 in the proportion of dogs achieving ≥50% or ≥75% reduction in their clinical score compared with baseline. Conclusions and Clinical Importance: These data suggest that intravesicular administration of ASB E. coli 212 is not inferior to antimicrobials for the treatment of recurrent UTI in dogs. This biotherapeutic agent could help alleviate the need for antimicrobials for some dogs with recurrent UTI, improving antimicrobial stewardship. [ABSTRACT FROM AUTHOR]

Septic arthritis of the hip is a serious infection in children. However, blood and joint fluid cultures are often negative, which makes the diagnosis and treatment challenging. We analyzed the clinical features of children with septic arthritis of the hip with negative bacterial cultures and identified the risk factors for poor outcomes. The clinical data of children with septic arthritis of the hip with negative bacterial cultures who were treated at our hospital from January 2010 to December 2020 were retrospectively analyzed. The clinical characteristics and outcomes of the culture-negative cohort were compared with those of children with positive bacterial cultures treated during the same period. Culture-negative patients were divided into a group with good outcomes and a group with poor outcomes. The differences between the two groups were compared. Thirty-nine children with culture-negative septic arthritis of the hip were compared with 37 children with culture-positive sepsis. Compared with the culture-positive group, the culture-negative group had a significantly younger mean age and a significantly lower mean serum C-reactive protein concentration. Logistic regression analysis of culture-negative patients with good versus poor outcomes revealed that the independent risk factors for poor outcomes were an increased serum C-reactive protein concentration and prolonged time from onset to surgery. The cut-off values for predicting a poor outcome in the culture-negative group were a time from onset to surgery of greater than 21 days and a C-reactive protein concentration of greater than 23 mg/L. Culture-negative septic arthritis of the hip has similar clinical features to culture-positive septic arthritis of the hip and can result in sequelae of varying severity. Therefore, active anti-infective and hip drainage therapy should be performed when children present with clinical symptoms, inflammatory marker concentrations, and imaging findings that are clinically diagnostic for septic arthritis of the hip. Level II, retrospective study. [ABSTRACT FROM AUTHOR]

During 2018–2021, eight septic transfusion reactions occurred from transfusion of platelet units contaminated with Acinetobacter spp., Staphylococcus saprophyticus, Leclercia adecarboxylata, or a combination of those environmental organisms. Whether biofilm formation contributed to evasion of bacterial risk mitigations, including bacterial culture, point-of-care testing, or pathogen-reduction technology, is unclear. We designed a 12-well plate-based method to evaluate environmental determinants of single-species and multispecies biofilm formation in platelets. We evaluated bacteria isolated from septic transfusion reactions for biofilm formation by using crystal violet staining and enumeration of adherent bacteria. Most combinations of bacteria had enhanced biofilm production compared with single bacteria. Combinations involving L. adecarboxylata had increased crystal violet biofilm production and adherent bacteria. This study demonstrates that transfusion-relevant bacteria can produce biofilms well together. More work is needed to clarify the effect of biofilms on platelet bacterial risk control strategies, but US Food and Drug Administration– recommended strategies remain acceptable. [ABSTRACT FROM AUTHOR]

The diversity of metabolites produced in fermented pine needles at different stages of fermentation has rarely been investigated. In the present study, untargeted metabolomic analysis using GC-MS and LC-MS was performed to detect metabolites in fermented pine needles at different fermentation stages. A total of 30 samples of pine needles fermented first with yeast (Saccharomyces cerevisiae) and then with a mixed bacterial culture of Lactobacillus fermentum CECT5716 and Bifidobacterium Breve M16V, were used to detect differential metabolites at different stages of fermentation. Pearson's correlation analysis was used to determine correlations between metabolites and key microbial communities. A total of 708 differential metabolites (430 from LC-MS and 278 from GC-MS analysis) were identified. The PCA and OPLS-DA revealed distinct differences between metabolites at different fermentation stages. Key differential metabolites identified through GC-MS analysis included; Phosphoric acid, D-Fructose, 2-O-alpha-mannosyl-D-glycerate, 1,3-dihydroxyacetone dimer, Galactosylglycerol, 2-Isopropylmalic acid, alpha-D-Galactose, Citrate, 4-Hydroxycinnamic acid, and Shikimate. Similarly, key differential metabolites identified through LC-MS included; 2-Phenlyethanol, Dimethlglycine, 2-Hydroxybenzaldehyde, 3-Aminoisobutanoic acid, p-Cresol, Triethylamine, 2-Ketobutyric acid, Cytosine, Benzaldehyde, and Creatinine. Annotation of differential metabolites to KEGG pathway enrichment analysis revealed the association of these metabolites with phenylpropanoid, flavonoid, and secondary metabolite biosynthesis. Furthermore, the results showed that three bacterial (Firmicutes, Actinobacteria, and Lactobacillus) and three fungal genera (Penicillium, Candida, and Basidiomycota) significantly correlated with differential metabolites showing synergistic effects. Our study reveals a comprehensive comparison of metabolites at different fermentation stages and provides practical insights into the mechanism of metabolite enrichment in fermented pine needles. [ABSTRACT FROM AUTHOR]

Despite scarcity of data, in recent years, human parvovirus B19 (PVB19) has been emerging as an important pathogen in acute encephalitis syndrome (AES). But, PVB19 virus is mostly looked for only after the exclusion of other common pathogens implicated in AES. Hence, this study was conducted to correlate clinical, radiological, and sequencing data to establish the crucial role of PVB19 in AES. Cerebrospinal fluid and/or serum samples were collected from AES patients as per WHO criteria and tested by ELISA, real‐time PCR and bacterial culture sensitivity for various pathogens. PVB19 positive samples were subjected to sequencing. PVB19 attributed to 5% of total AES cases in the present study with fatalities in two of eight cases. Two isolates of PVB19 belonged to Genotype 1 A whereas one belonged to Genotype 3B. On multivariate analysis of predictive symptoms of PVB19 AES cases, blurring of vision (odds ratio [OR] 20.67; p = 0.001) was found to be significant independent predictor of PVB19 AES. Six of eight patients (two encephalitis specific and four nonspecific) had abnormal radiological findings. Hence, being an emerging viral pathogen, PVB19 should be included in the diagnostic algorithm of AES for prompt diagnosis and definitive management to prevent undesired neurological sequelae. [ABSTRACT FROM AUTHOR]

BACKGROUND: Aedes aegypti is a widespread mosquito in tropical and subtropical regions that causes significant mortality and morbidity in humans by transmitting diseases, such as dengue fever and Zika virus disease. Synthetic insecticides, such as pyrethroids, have been used to control Ae. aegypti, but these insecticides can also affect nontarget organisms and contaminate soil and water. This study aimed to investigate the mosquitocidal activity of Pseudomonas mosselii isolated from pond sludge against larvae of Ae. aegypti. RESULTS: Based on the initial results, similar time‐course profiles were obtained for the mosquitocidal activity of the bacterial culture and its supernatant, and the pellet resuspended in Luria–Bertani (LB) medium also showed delayed toxicity. These results imply that the toxic component can be released into the medium from live bacteria. Further research indicated that the toxic component appeared in the supernatant approximately 4 h after a 3‐mL stock was cultured in 200 mL of LB medium. The stabilities of the P. mosselii culture and supernatant stored at different temperatures were also evaluated, and the best culture stability was obtained at 28 °C and supernatant stability at 4 °C. The bacterial culture and supernatant were toxic to larvae and pupae of not only susceptible Ae. aegypti but also pyrethroid‐resistant strains. CONCLUSION: This study highlights the value of the mosquitocidal activity of P. mosselii, which has potential as an alternative insecticide to control pyrethroid‐resistant Ae. aegypti in the field. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Two distinct bacterial strains were combined in a mixed culture to investigate the degradation of phenol. The mixed cultures consisting of species like Rhodococcus and Bacillus, were isolated from the waste effluent collected from Dankuni coal complex (DCC) in West Bengal. The experimental phenol biodegradation were performed batch wise at an optimized temperature and pH conditions of 37°C and 7.0 for a wide range of phenol concentrations initially, with saline solutions which is not supplemented with nutrients initially. Experimental results concluded a highest possible specific growth rate at 150 mg/L for phenol biodegradation of 2400 mg/L within a span of 72 h. Several significant physicochemical parameters were investigated in order to determine the ideal conditions for phenol breakdown. It was also observed that a spectroscopic study of the intermediate at 260 nm of the phenol biodegradation commenced through the formation of a cis, cis intermediate, indicating ortho‐cleavage pathway. A simulated substrate utilization and growth profiles was obtained by using substrate inhibition model equation using MATLAB R2017a were used to compare for the validation of the obtained experimental growth and substrate characterization data. [ABSTRACT FROM AUTHOR]

Bacterial infections pose significant global health challenges, often underestimated due to difficulties in accurate diagnosis, especially when culture-based diagnostics fail. This study assesses the effectiveness of 16S-based metagenomic next generation sequencing (NGS) for identifying pathogens in culture-negative clinical samples across various medical settings. Overall, 48% of samples were collected from orthopedics, 15% from neurosurgery, and 12% in cardiac surgery, among others. The detection rate of monomicrobial infections was 68.6%, and 5.7% for polymicrobial infections. In addition, NGS detected bacteria in all samples from the lungs, head and neck, and eye specimens. Cutibacterium acnes (11%, 12/105) was the most frequent microorganism, followed by Staphylococcus epidermidis (10.4%, 11/105), and Staphylococcus aureus (9.5%, 10/105). In conclusion, 16S-targeted metagenomic sequencing enhances pathogen detection capabilities, particularly in instances where traditional cultures fail. By the combination of NGS and bacterial cultures, microbiologists might provide a more accurate diagnosis, guiding more effective treatments and potentially reducing healthcare costs associated with empirical treatments. [ABSTRACT FROM AUTHOR]

The most accurate method for detecting the pathogen of orthopedic implant-associated infections (OIAIs) is sonication fluid (SF). However, the frequency and duration of ultrasound significantly influence the number and activity of microorganisms. Currently, there is no consensus on the selection of these two parameters. Through this study, the choice of these two parameters is clarified. We established five ultrasonic groups (40kHz/10min, 40kHz/5min, 40 kHz/1min, 20kHz/5min, and 10kHz/5min) based on previous literature. OIAIs models were then developed and applied to ultrasound group treatment. Subsequently, we evaluated the efficiency of bacteria removal by conducting SEM and crystal violet staining. The number of live bacteria in the SF was determined using plate colony count and live/dead bacteria staining. The results of crystal violet staining revealed that both the 40kHz/5min group and the 40kHz/10min group exhibited a significantly higher bacterial clearance rate compared to the other groups. However, there was no significant difference between the two groups. Additionally, the results of plate colony count and fluorescence staining of live and dead bacteria indicated that the number of live bacteria in the 40kHz/5min SF group was significantly higher than in the other groups. 40kHz/5min ultrasound is the most beneficial for the detection of pathogenic bacteria on the surface of orthopedic implants. [ABSTRACT FROM AUTHOR]

Abstract Objective The need for innovative techniques to preserve microbiota for extended periods, while maintaining the species composition and quantitative balance of the bacterial community, is becoming increasingly important. To address this need, we propose an efficient approach to cryopreserve human gut microbiota using a two-component cryoprotective composition comprising fetal bovine serum (FBS) and 5% dimethyl sulfoxide (DMSO). Fetal serum is a commonly utilized component in the freezing media for eukaryotic cells, however, its effects on prokaryotic cells have not been extensively researched. Results In our study, we demonstrated the high efficiency of using a two-component cryoprotective medium, FBS + 5% DMSO, for cryopreservation of human gut microbiota using three different methods. According to the obtained results, the intact donor microbiota was preserved at a level of 85 ± 4% of the initial composition based on fluorescent analysis using the LIVE/DEAD test. No differences in survival were observed when comparing with pure DMSO and FBS media. The photometric measurement method for growth of aerobic bacteria (A. johnsoni), facultative anaerobes (E. coli, E. faecalis), microaerophilic (L. plantarum), and obligate anaerobic bacterial cultures (E. barkeri, B. breve) also demonstrated high viability rates of 94–98% in the two-component protective medium, reaching intact control levels. However, for anaerobic microflora representatives, serum proved to be a more suitable cryoprotectant. Also, we demonstrated that using cryoprotective media with 50–75% FBS content is enough to preserve a significant level of bacterial cell viability, from an economic standpoint.

Catheter-associated urinary tract infections (CAUTI) are the most common secondary cause of bloodstream infection. CAUTI is particularly prevalent in critical care departments and developing countries, where the duration of catheterization remains the most significant risk factor. This study focused on the characteristics, risk factors, and outcomes of CAUTI patients in a tertiary care hospital setting. It also provides the incidence rate of CAUTI in an ICU setting in Palestine. The study adopted a retrospective observational design at a tertiary care hospital in Palestine. The data were collected from patient records as well as from nursing flow charts. Variables are reported as frequencies, percentages and means + standard deviations. Independent t-tests was used for numerical variables, while Pearson's chi-square or Fisher's exact test were used for categorical variables. Multivariate analysis was performed to adjust for confounders using binary logistic regression. Mortality risk factors were assessed using the proportional Cox regression model. Of the 377 patients included in the study, 33 (9%) developed CAUTI. Among CAUTI patients, 75% had Candida species isolated, with non-albicans Candida predominating (72%) fungal isolates. On the other hand, 25% of the patients had bacterial isolates in their urine, with a predominance of Escherichia coli growing in 36% of bacterial cultures. Multivariate regression analysis revealed that female gender, longer catheterization days, and corticosteroid use were associated with an increased risk of CAUTI. On the other hand, developing CAUTI, having a malignant disease, developing kidney injury, and developing shock were associated with increased mortality. This study highlighted the emerging presence of fungal and resistant bacterial CAUTI. It also emphasized that the risk of CAUTI was associated with a longer duration of urinary catheterization. The findings of this study may help formulate antimicrobial management and stewardship plans as well as emphasize the risk of urinary catheterizations. [ABSTRACT FROM AUTHOR]

Background: The spread of Carbapenemase-producing Organisms (CPO) remains a major threat globally. Within clinical settings, the existing method of determining gene load involves traditional culture to determine bacterial load and polymerase-chain-reaction-based Xpert Carba-R Assay to determine carbapenemase gene type. However, there is a need for a fast and accurate method of quantifying CPO colonisation to study the risk of persistent CPO carriage. Objective: This study evaluated the accuracy of Xpert Carba-R Ct value in estimating carbapenamase producing bacterial loads in stool samples. Methods: Stool samples were obtained from an ongoing study investigating the household transmission of CPO in Singapore. Stool samples lacking carbapenemase producing organisms were spiked with organism carrying a single carbapenemase gene (blaKPC, blaNDM, blaVIM, blaOXA-48(-like) or blaIMP-1) and serially diluted before being subjected to Xpert Carba-R assay and traditional culture. Standard curves with regression lines showing correlation between Ct values and plate counts were generated. The standard curves were validated with stool samples collected from patients. Results: The limit of detection of blaNDM, blaKPC, and blaOXA-48 was approximately 103 cfu/mL, while that of blaIMP-1 and blaVIM was approximately 104 cfu/mL. Validation of the blaNDM and blaOXA-48 curves revealed average delta values of 0.56 log(cfu/mL) (95% CI 0.24–0.88) and 0.80 log(cfu/mL) (95% CI 0.53–1.07), respectively. Conclusions: Our validation data for stool positive for blaNDM and blaOXA-48-type suggests that bacterial loads can be estimated within a reasonable range of error. [ABSTRACT FROM AUTHOR]

Simple Summary: Milk or mammary tissue swab samples were collected from both defective and normal udder halves in three different studies to identify the bacterial species involved. Samples were collected at different physiological time points: pre-mating, throughout lactation, at weaning and post-weaning. Numerous bacterial species were identified, with Staphylococcus species, Mannheimia haemolytica and Streptococcus being the most frequently isolated. Bacteria were isolated from at least one third (udder lumps) or more than half (hard udder) of defective udder halves, whereas no bacteria were isolated from more than two thirds of normal udder halves. Frequently isolated bacterial species tended to persist longer, whereas less frequently isolated bacterial species showed less stability over time. Bacterial species more frequently identified from defective udder halves and which appear more stable over time should be considered as a factor for making culling decisions. The objectives of these studies were to identify associations between udder half defects (hard or lump) and bacteria isolated from milk or mammary tissue swabs, to compare with samples from normal udder halves at different physiological time points and to compare bacterial species isolated via milk and swabs of mammary tissue from within the same udder halves. A total of 1054 samples were aseptically collected from each udder half of 199 non-dairy breed (Romney) ewes from three different studies (Study A, n = 77; Study B, n = 74; and Study C, n = 48). Conventional bacterial culture and MALDI-ToF mass spectrometry were used for bacterial identification. Of the 225 samples from which bacteria were isolated, Mannheimia haemolytica and Streptococcus uberis were the dominantly identified species from defective udder halves, whereas coagulase-negative staphylococcus (CNS) species, mostly Staphylococcus simulans and Staphylococcus chromogenes, were more frequently isolated from normal udder halves. The ongoing presence of bacterial species over time was variable, although less frequently identified species showed less stability over time. A very high agreement (91.5%) of bacterial species identified was observed between the mammary tissue swab and udder half milk samples during post-weaning. In summary, palpable udder half defects were associated with bacterial positivity, and the ongoing presence of the bacteria over time was dependent on the species involved. Hence, culling ewes with palpable udder half defects that had more stable bacterial species could contribute to reducing the recurrence of palpable defects or mastitis. [ABSTRACT FROM AUTHOR]

Exposure to the space microenvironment has been found to disrupt the homeostasis of intestinal epithelial cells and alter the composition of the microbiota. To investigate this in more detail and to examine the impact of ginsenoside Rb1, we utilized a mouse model of hindlimb unloading (HU) for four weeks to simulate the effects of microgravity. Our findings revealed that HU mice had ileum epithelial injury with a decrease in the number of intestinal stem cells (ISCs) and the level of cell proliferation. The niche functions for ISCs were also impaired in HU mice, including a reduction in Paneth cells and Wnt signaling, along with an increase in oxidative stress. The administration of Rb1 during the entire duration of HU alleviated the observed intestinal defects, suggesting its beneficial influence on epithelial cell homeostasis. Hindlimb unloading also resulted in gut dysbiosis. The supplementation of Rb1 in the HU mice or the addition of Rb1 derivative compound K in bacterial culture in vitro promoted the growth of beneficial probiotic species such as Akkermansia. The co-housing experiment further showed that Rb1 treatment in ground control mice alone could alleviate the defects in HU mice that were co-housed with Rb1-treated ground mice. Together, these results underscore a close relationship between dysbiosis and impaired ISC functions in the HU mouse model. It also highlights the beneficial effects of Rb1 in mitigating HU-induced epithelial injury by promoting the expansion of intestinal probiotics. These animal-based insights provide valuable knowledge for the development of improved approaches to maintaining ISC homeostasis in astronauts. [ABSTRACT FROM AUTHOR]

Aim Methodology Results Conclusions To evaluate the 6‐year outcome of root canal treatment irrigated with 0.5% or 3% sodium hypochlorite (NaOCl).The baseline trial was designed as a quasi‐randomized clinical trial. Patients referred for root canal treatment to an endodontic specialist clinic were recruited to the study (n = 298). The concentration of NaOCl was allocated quasi‐randomized to 271 subjects (0.5% [n = 139], 3% [n = 132]). Bacterial sampling was performed immediately before root canal filling. Samples were cultured and evaluated as growth or no growth. Patients were invited to a clinical and radiological follow‐up >5 years postoperatively. The clinical outcome measurements were tooth survival, cumulative incidence of endodontic retreatments, patients' assessment of pain, clinical findings and radiological signs of apical periodontitis (AP).Tooth survival was 85.6% in the 0.5% NaOCl group and 81.1% in the 3% NaOCl group (p = .45). There was no record of retreatment in 94.4% in the 0.5% NaOCl group and in 92.2% in the 3% NaOCl group (p = .76). The percentage of asymptomatic cases were 87.8% in the 0.5% group and 85.3% in the 3% NaOCl group (p = .81). Absence of clinical signs of AP was seen in 86.6% in the 0.5% NaOCl group and in 83.6% in the 3% NaOCl group (p = .80). Absence of radiological signs of AP was seen in 74.0% in the 0.5% NaOCl group and 64.1% in the 3% NaOCl group (p = .20). Subjects with positive culture before root filling reported subjective pain with a significantly higher frequency as compared to negative‐culture subjects (p = .014).The use of 0.5% or 3% NaOCl for irrigation during root canal treatment resulted in similar clinical outcomes 5–7 years postoperatively. Persisting bacteria immediately before root filling may predict future episodes of subjective pain. [ABSTRACT FROM AUTHOR]

Aim Materials and Methods Results Conclusions This meta‐analysis aimed to compare the antibacterial efficacy of chitosan/chitosan nanoparticles (Ch/Ch‐NPs) versus sodium hypochlorite/chlorhexidine (NaOCl/CHX).A search was performed in four electronic databases until December 08, 2023. Studies with missing, unclear, and insufficient data sets were excluded. The included studies were assessed by two independent reviewers using the Joanna Briggs Institute Critical Appraisal Checklist for Quasi‐Experimental Studies. The meta‐analysis of standardized mean difference was performed using a random effects model. Additionally, funnel plots as well as Egger's regression intercept test were used to evaluate potential publication bias.A total of 426 samples were used in nine included studies. There was no difference in antibacterial efficacy between Ch/Ch‐NPs‐NaOCl (SMD: 0.005; 95% CI: −0.844–0.854; p = 0.990). However, the antibacterial efficacy of NaOCl was statistically more effective than Ch/Ch‐NPs (SMD: 0.807; 95% CI: 0.015–1.599; p = 0.046) using the bacterial culture method, and Ch/Ch‐NPs was statistically higher than NaOCl (SMD: −1.827; 95% CI: −2.720, −0.934; p < 0.000) using confocal laser scanning microscopy.Ch/Ch‐NPs may be an alternative to NaOCl against Enterococcus faecalis. The methods used in the in vitro studies evaluating the antibacterial efficacy of irrigation solutions against E. faecalis may have had an impact on the results. [ABSTRACT FROM AUTHOR]

Background: Cupriavidus gilardii is an aerobic, gram-negative, motile, glucose-nonfermenting bacillus, first described in 1999. Typically, it exhibits low pathogenicity in humans, causing opportunistic infections primarily in individuals with compromised immune systems. This bacterium has been also found in various environmental sources such as plants and contaminated soils. Notably, there have been no documented cases of C. gilardii infections in animals. Case presentation: This case report outlines a bovine neonatal diarrhea outbreak that occurred in Northern Greece, during which C. gilardii was isolated. Faecal samples from 5-day-old calves were collected and transported to the laboratory for further examination. Bacterial culture and next generation sequencing techniques were employed to confirm the presence of this bacterium in the samples. Following the isolation and identification of C. gilardii from the samples, an autogenous vaccine was produced and administered to the cows within the farm. Subsequent to vaccination, a progressive reduction in calf diarrhea and deaths was observed, leading to their eventual complete resolution. To the best of our knowledge, this represents the first documentation of C. gilardii isolation from cases of bovine neonatal diarrhea. Conclusion: This case report presents the first isolation case of C. gilardii from animal samples and more specifically from calf faecal samples. It represents an important observation, providing evidence that this opportunistic human pathogen could contribute to clinical symptoms in animals. [ABSTRACT FROM AUTHOR]

Introduction: This study assessed the risk of first treatment for bovine respiratory disease (BRD) given detection of nasopharyngeal bacteria (Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni) and corresponding likelihood of antimicrobial susceptibility (C/S) at two time points during the early feeding period. Relationships between C/S results and later treatment for BRD were evaluated at both the calf-level and pen-level. The association between calf-level and pen-level C/S findings during the early feeding period and subsequent C/S results at BRD treatment were also reported. Methods: Auction-sourced, recently-weaned beef calves (n = 1,599 steers) were placed in adjacent feedlot pens (8 × 100 calves) in two subsequent years. Deep nasopharyngeal (DNP) swabs were collected from all calves at time of arrival processing (1DOF) and before metaphylaxis administration with either tulathromycin or oxytetracycline, 12 days later (13DOF), and at the time of first treatment for BRD. All samples were tested for C/S. Results: Several pen-level and individual calf-level C/S measures of interest were associated with future treatment for BRD and C/S at the time of treatment. The median DOF for first BRD treatment was 24 days following tulathromycin metaphylaxis and 11 days following oxytetracycline. Overall, sampling at 13DOF resulted in the best fit for more models of subsequent treatment for BRD and C/S results at BRD treatment than for sampling at arrival. In individual calves, recovery of M. haemolytica, P. multocida, or H. somni at 13DOF was associated with subsequent treatment for BRD within 45DOF. Pen-level prevalence of Pasteurellacea bacteria with tetracycline or macrolide resistance at arrival and 13DOF were associated with detection of bacteria with antimicrobial resistance (AMR) at BRD treatment, as were individual calf results at 13DOF. Discussion: These findings suggest that the bacteria and AMR outcomes recovered from cattle near two weeks on feed can inform the prediction of future BRD risk and concurrent antimicrobial susceptibility results at time of first BRD treatment. Notably, the associations between pen-level C/S results from previous testing and corresponding findings in calves with BRD from the same pen suggested potential testing strategies to inform antimicrobial use protocols for feedlot cattle. [ABSTRACT FROM AUTHOR]

Higher cost of production and functioning efficiency are the two most important aspects of cellulolytic enzymes, as they directly affect the bioconversion of cellulosic waste into fermentable sugars. Thus, the current study aims to investigate the production of cellulolytic enzymes from low-cost waste biomass for use in the sustainable hydrolysis of biomass to yield fermentable sugars. Since coconut waste is a major contributor to biodegradable waste, two different parts of coconut, including the hard shell of coconut (CS) and the fibers of the shell (FS), have been used for enzyme production as feedstock. Further, enzyme production has been carried out on co-fermentative and acid-pretreated coconut substrates (CS and FS) at different quantities using mixed bacterial cultures. Following this, a crude enzyme with a maximum 20 U/gds FPA, 62 U/gds BGL, and 167 U/gds EG has been produced by mixed Bacillus cultures under co-fermentation of 0.5 % acid-pretreated CS and FS substrates at a quantity of 5:5 (g:g) at 35 °C and pH 5.0. The crude enzyme showed its thermal stability for 16 h at 50 °C and released ~34 g/L of sugars from the hydrolysis of untreated FS biomass in 24 h. This work may have significant applications in the area of biomass-based biorefining as well as waste management. [ABSTRACT FROM AUTHOR]

Background • Nosocomial infections pose a significant global health challenge. Effective nursing management plays a pivotal role in hospital administration, with the quality of nursing closely linked to nosocomial infection rates. Objective • This study explores the impact of implementing high-quality nursing practices on nosocomial infection control. Design • A randomized controlled experiment was conducted. Setting • The study was conducted at the Fourth Affiliated Hospital of Nanjing Medical University. Participants • Between December 2021 and December 2022, 120 hospitalized patients were selected and randomly assigned to the control and research groups, each comprising 60 patients. Interventions • The control group received routine nursing, while the research group received high-quality nursing. Primary Outcome Measures • (1) Infection incidence rate, (2) psychological state of patients, (3) bacterial culture qualified rate, (4) health knowledge awareness, (5) nursing quality, (6) quality of life, and (7) patient satisfaction. Results • The research group exhibited a significantly lower infection incidence rate compared to the control group (P < .05). No significant differences were observed between the groups before nursing (P > .05). Post-nursing, SAS, and SDS scores decreased in both groups, with a more pronounced reduction in the research group (P < .05). The research group demonstrated an increased qualified rate of bacterial culture in various environments compared to the control group (P < .05). After nursing, both groups showed higher scores in disease knowledge, nosocomial infection knowledge, treatment and rehabilitation knowledge, and self-care management knowledge, with the research group scoring higher than the control group (P < .05). Quality of life scores, nursing quality scores, and nursing satisfaction were all higher in the research group compared to the control group (P < .05). Conclusions • High-quality nursing practices emerge as the preferred choice in nosocomial infection control and prevention. It prioritizes patients’ wishes and needs, offers comprehensive services, and respects patient autonomy; these measures contribute to a substantial reduction in nosocomial infections. [ABSTRACT FROM AUTHOR]

A lot of methodological studies have been carried out on sulfate-reducing bacteria (SRB) at home and abroad. However, these methods often have varying application conditions and limitations, leading to imprecise measurement results. This paper reviews the progress in the identification, quantification and application of sulfate-reducing bacteria, and analyse their application in various situations. According to the theoretical basis of the methodology, we classify the theory of SRB concentration detection, which is mainly based on the principles of four categories, namely, bacterial culture, genetic genes, specific bio-enzymes, and bio-sensors, and analyse their respective strengths and weaknesses, and analyse the problems faced in their application. On this basis, the research direction of sulfate-reducing bacteria content determination and corrosion activity monitoring technology is proposed in response to the real needs of sulfate-reducing bacteria assessment and management in oil and gas fields. [ABSTRACT FROM AUTHOR]

The objective of our study was to assess the effect of nitrate-reducing bacteria and natural nitrate source on the sensory properties of dried meat biltong. Appearance, colour on the cut, aroma, taste and juiciness, acidity, sweetness, and consistency were evaluated. The members of the panel gave their ratings between 1 and 5 according to the characteristics of the samples, with 1 being the lowest rating and 5 being the best rating. In the sensory evaluation of dried meat samples without oxygen absorber packaging, the group (P2) with bacterial culture, celery powder, and NaCl showed the highest scores for appearance, taste, juiciness, acidity, and consistency. Notably, P3 samples had significantly lower consistency scores compared to P2. Additionally, sample P1, with bacterial culture and NaCl, improved the colour on the cut compared to the negative control (C) with only NaCl. The best aroma and sweetness results were in the P3 group with curing salt, although these differences were not statistically significant (P = 0.05). The sensory evaluation of dried meat samples packaged with an oxygen absorber revealed that the combination of bacterial culture, celery powder, and NaCl (AP2) had the most positive influence on appearance, colour on the cut, and consistency. The group with the absorber and curing salt (AP3) showed improved aroma, taste, and juiciness. Acidity was primarily affected by the addition of bacterial culture and NaCl (AP1), as well as by AP2 and AP3. The most pleasing sweetness was achieved by adding bacterial culture and NaCl to samples with the absorber (AP1). Samples with the best consistency when packaged with the absorber were those with bacterial culture and celery powder (AP2). In summary, among dried meat samples with oxygen absorber, those with added curing salt (AP3) had the best overall sensory evaluation compared to the negative control with NaCl (AC). For samples without an oxygen absorber, the highest overall sensory scores were in the group with bacterial culture and celery powder, surpassing the control with NaCl (C). The use of an oxygen absorber and curing salt proved to be effective in the production of biltong, but replacing the curing salt with NaCl and the addition of celery powder and bacterial culture significantly improved the sensory evaluation compared to the other groups (except AP3). [ABSTRACT FROM AUTHOR]

Objective: To explore the clinical efficacy of platelet-rich technology combined with moist dressings in the treatment of chronic infectious ulcers. Methods: This was a retrospective study. The subjects of the study were 48 patients with chronic infectious ulcers in Sichuan Provincial Orthopedics Hospital from January 2019 to June 2022. Enrolled patients were randomly divided into four groups(n=12), and received different treatment methods respectively. Further analysis and comparison were performed on the changes in wound volume, wound healing status, wound bacterial culture results, and the incidence of adverse reactions among the four groups. Results: Three months after debridement, the wound volume of all four groups of patients was significantly reduced compared with that before debridement, with a statistically significant difference in intra-Group-Comparison(P<0.05). The inter-Group-Comparison revealed a statistically significant difference in wound volume in Group-A, Group-B, and Group-C than that in Group-D(P<0.05). After treatment, the wound healing status of patients in groups A, B, and C was significantly better than that of patients in Group-D, with a statistically significant difference(P<0.05). During treatment, patients in all four groups had decreased count of would bacteria, and showed negative results of wound bacterial culture by the three-month follow-up. No serious adverse reactions were observed in the four groups during treatment, and all improved after management, with no statistically significant difference in the incidence of adverse reactions(P>0.05). Conclusion: Platelet-rich technology combined with moist dressings may effectively promote the repair of chronic infectious ulcer wounds, with good clinical safety. [ABSTRACT FROM AUTHOR]

Introduction: The accuracy of urine culture results can be affected by pre-analytical factors such as transport delays and storage conditions. The objectives of this study were to analyze urine collection practices and assess the impact of introducing boric acid tubes for urine collection on quantitative urinary bacterial cultures of hospitalized patients in medical wards. Methods: A quasi-experimental pre-post study conducted in an acute care facility. In the pre-intervention phase (2020–2021), urine samples were transported without preservatives at room temperature. In 2022 (post-intervention), we transitioned to boric acid transport tubes, evaluating its effect on significant bacterial growth (≥ 105 CFU/ml). Bivariate and multivariate analyses identified predictors of culture positivity. Results: Throughout the duration of the study, a total of 12,660 urine cultures were analyzed. Date and time documentation was complete for 38.3% of specimens. Culture positivity was higher with longer processing times: positivity was 21.3% (220/1034) when specimens were processed within 4 h, 28.4% (955/3364) when processed in 4–24 h, and 32.9% (137/417) when processed after 24 h (p < 0.0001). For 4-24-hour processing, positivity decreased from 30.4% (704/2317) pre-intervention to 24.0% (251/1047) post-intervention (p < 0.001), with no significant changes in < 4 or ≥ 24-hour specimens. Stratified analysis by processing time revealed that the intervention was associated with reduced positivity only in cultures processed within 4–24 h (OR 0.80, 95% CI 0.67–0.94; p = 0.008). Conclusion: The introduction of boric acid transport tubes predominantly influenced cultures transported within a 4–24-hour window. This presents an opportunity to improve urine tract infection diagnostic practices in healthcare settings. [ABSTRACT FROM AUTHOR]

Introduction: Cholesteatoma usually harbors a poly-microbial infection. As the diversity of bacterial pathogens in the Indian COM is unknown, we set out to identify the bacteria associated with cholesteatoma disease in different patients of North India using targeted metagenomic analysis of the 16 S rRNA gene. Methods: We recruited 15 patients of cholesteatomatous chronic otitis media (COM), who underwent surgical disease clearance. We divided these patients into four groups based on the four clinic-radiological stages categorized as per the EAONO/JOS joint consensus statement classification. Representative samples were extracted during the surgery and sent for bacterial culture and sensitivity and 16 S rRNA gene metagenomic analysis. Results: While 12 (80%) of the patients belonged to clinical Stage I/II; one patient had an extracranial complication (stage III) and two patients had an intracranial complication (stage IV). Our detailed bacterial metagenomics analyses showed that while phylum Proteobacteria was most abundant (reads up to ∼ 95%) in specimens from nine patients, phylum Firmicutes was most abundant (up to ∼ 80%) in specimens from four patients. Gamma (γ) Proteobacteria and Epsilon (ε) Proteobacteria were the most abundant class amongst Proteobacteria. Class Tissierellia stood out as the most abundant Firmicutes (40–60%), followed by Clostridia (20%) and Bacilli (10%). There was negligible difference in the bacterial profiles across all four clinical stages. Conclusion: Cholesteatoma is primarily associated with Proteobacteria and Firmicutes phyla, even in complicated disease. Further studies with a larger sample size are required to validate our findings. [ABSTRACT FROM AUTHOR]

Insoluble phosphorous compounds solubilization by soil bacteria is of great relevance since it puts available the phosphorus to be used by plants. The production of organic acids is the main microbiological mechanism by which insoluble inorganic phosphorus compounds are solubilized. In Gram negative bacteria, gluconic acid is synthesized by the activity of the holoenzyme glucose dehydrogenase-pyrroloquinoline quinine named GDH-PQQ. The use of marker genes is a very useful tool to evaluate the persistence of the introduced bacteria and allow to follow-up the effect of biotic and abiotic factors on these beneficial microorganisms in the soil. In previous studies we detected the presence of the pqqE gene in a great percentage of both non-culturable and culturable native soil bacteria. The objective of this study was to analyze the phylogeny of the sequence of pqqE gene and its potential for the study of phosphate solubilizing bacteria from pure and mixed bacterial cultures and rhizospheric soil samples. For this, the presence of the pqqE gene in the genome of phosphate solubilizing bacteria that belong to several bacteria was determined by PCR. Also, this gene was analyzed from mixed bacterial cultures and rhizospheric soil associated to peanut plants inoculated or not with phosphate solubilizing bacteria. For this, degenerate primers designed from several bacterial genera and specific primers for the genus Pseudomonas spp., designed in this study, were used. DNA template used from simple or mixed bacterial cultures and from rhizospheric soil samples was obtained using two different DNA extraction techniques. Results indicated that pqqE gene amplification product was found in the genome of all Gram negative phosphate solubilizing bacteria analyzed. It was possible to detect this gene in the DNA obtained from mixed cultures where these bacteria grew in interaction with other microorganisms and in that obtained from rhizospheric soil samples inoculated or not with these bacteria. The phylogenetic analysis indicated that pqqE gene is a conserved gene within related genera. In conclusion, pqqE gene could be a potential marker for the study of phosphate solubilizing bacterial populations. [ABSTRACT FROM AUTHOR]

Biopolymers are highly desirable alternatives to petrochemical-based plastics owing to their biodegradable nature. The production of bioplastics, such as polyhydroxyalkanoates (PHAs), has been widely reported using various bacterial cultures with substrates ranging from pure to biowaste-derived sugars. However, large-scale production and economic feasibility are major limiting factors. Now, using algal biomass for PHA production offers a potential solution to these challenges with a significant environmental benefit. Algae, with their unique ability to utilize carbon dioxide as a greenhouse gas (GHG) and wastewater as feed for growth, can produce value-added products in the process and, thereby, play a crucial role in promoting environmental sustainability. The sugar recovery efficiency from algal biomass is highly variable depending on pretreatment procedures due to inherent compositional variability among their cell walls. Additionally, the yields, composition, and properties of synthesized PHA vary significantly among various microbial PHA producers from algal-derived sugars. Therefore, the microalgal biomass pretreatments and synthesis of PHA copolymers still require considerable investigation to develop an efficient commercial-scale process. This review provides an overview of the microbial potential for PHA production from algal biomass and discusses strategies to enhance PHA production and its properties, focusing on managing GHGs and promoting a sustainable future. [ABSTRACT FROM AUTHOR]