Multiple Microorganisms Research Articles

Performance of a hybrid capture-based target enrichment next-generation sequencing for the identification of respiratory pathogens and resistance-associated genes in patients with severe pneumonia.

Severe pneumonia remains the leading infectious cause of death worldwide. The time-consuming nature and suboptimal sensitivity of sputum cultures hamper prompt pathogen detection for tailored treatments. Advanced techniques such as polymerase chain reaction (PCR) and next-generation sequencing (NGS) offer rapid genetic pathogen detection and identification of antimicrobial resistance (AMR) genes. However, the performance of hybrid capture-based target enrichment NGS, e.g., Respiratory Pathogen ID/AMR Enrichment Panel (RPIP), for pathogen detection in patients with severe pneumonia remains uncertain. A prospective study involving adults with severe pneumonia was conducted. Respiratory samples from the lower respiratory tract were collected via bronchoalveolar lavage, bronchial washing, or endotracheal tube suction. The performance of RPIP in pathogen and AMR-associated gene detection was compared to that of conventional culture methods and the multiplex PCR-based FilmArray Pneumonia Panel (FilmArray-PN). A total of 83 subjects were enrolled. The most prevalent pathogens detected by RPIP were Rothia mucilaginosa, Stenotrophomonas maltophilia, Pseudomonas aeruginosa; herpes simplex virus-1, cytomegalovirus, and Epstein-Barr virus, and Pneumocystis jirovecii. Overall, the positive and negative agreement rates for bacterial detection were 63.6% and 97.5% between RPIP and culture methods, respectively, and 55.8% and 99.4% between FilmArray-PN and culture methods, respectively. Compared to FilmArray-PN, RPIP exhibited significantly better detection rates for bacteria (P = 0.029), viruses (P < 0.001), and fungi (P < 0.001) and identified additional blaOXA, blaCMY as extended-spectrum β-lactamase genes and blaOXA, blaSHV as carbapenemase genes. In conclusion, RPIP can sensitively profile respiratory pathogens and is a promising tool for detecting multiple microorganisms and AMR-associated genes in patients with severe pneumonia.IMPORTANCESensitive pathogen detection is pivotal for timely treatment by tailoring adequate antimicrobial agents. Unlike conventional phenotypic approach, novel measures using molecular interrogation appear promising. This study aimed to elucidate the efficacy of a hybrid capture-based target enrichment next-generation sequencing technique (Respiratory Pathogen ID/AMR Enrichment Panel, RPIP) as exemplified in a cohort with severe pneumonia. Pathogen landscape in the population was illustrated by these three methodologies. As compared with multiplex polymerase chain reaction-based FilmArray Pneumonia Panel and conventional culture, RPIP demonstrated significantly improved sensitivity in identifying bacteria, viruses, and fungi. The RPIP also exhibited better performance in identifying different pathogens in patients co-infected with multiple microorganisms. Additionally, the genotypes contributing to antimicrobial resistance were determined by RPIP. The study facilitated the implementation of molecular diagnosis by presenting real-world data, whereas future studies are mandated to generalize such an approach toward different clinical settings.

Recurrent association between Trichodesmium colonies and calcifying amoebae

Abstract Colonies of the N2-fixing cyanobacterium Trichodesmium spp. constitute a consortium with multiple microorganisms that collectively exert ecosystem-level influence on marine carbon and nitrogen cycling, shunting newly fixed nitrogen to low nitrogen systems, and exporting both carbon and nitrogen to the deep sea. Here we identify a seasonally recurrent association between puff colonies and amoebae through a two-year survey involving over 10 000 Trichodesmium colonies in the Red Sea. This association was most commonly found in near-shore populations during spring. Microscopic observations revealed consistent amoebae morphology throughout the study, and both morphological characteristics and 18S rRNA gene sequencing suggested that these amoebae are likely to belong to the species Trichosphaerium micrum, an amoeba that forms a CaCO3 shell. Co-cultures of Trichosphaerium micrum and Trichodesmium grown in the laboratory suggest that the amoebae feed on heterotrophic bacteria and not Trichodesmium, which adds a consumer dynamic to the complex microbial interactions within these colonies. Sinking experiments with fresh colonies indicated that the presence of the CaCO3-shelled amoebae decreased colony buoyancy. As such, this novel association may accelerate Trichodesmium sinking rates and facilitate carbon and nitrogen export to the deep ocean. Amoebae have previously been identified in Trichodesmium colonies in the western North Atlantic (Bermuda and Barbados), suggesting that this type of association may be widespread. This association may add a new critical facet to the microbial interactions underpinning carbon and nitrogen fixation and fate in the present and future ocean.

Identification and comparison of protein composition of biofilms in response to EGCG from Enterococcus faecalis and Staphylococcus lugdunensis, which showed opposite patterns in biofilm-forming abilities

Bacterial biofilm is resistant to conventional antibiotic treatments, leading to complications associated with many infection-related human diseases. Epigallocatechin Gallate (EGCG), a phenolic catechin enriched in green tea, is recognized for its anti-bacterial and anti-biofilm activities. In this study, we examined the protein components of the biofilms formed in the absence or presence of EGCG using Enterococcus faecalis and Staphylococcus lugdunensis, which had shown opposing patterns in biofilm formation. A clustering heatmap revealed that the two microorganisms expressed the different protein sets in response to EGCG. Proteins that were noticeably upregulated included those associated with stress responsiveness and gluconeogenesis in E. faecalis, and gene modification in S. lugdunensis. Conversely, downregulated proteins were related to tRNA-modifying enzyme activity in E. faecalis, and anabolic metabolism in S. lugdunensis. Among the proteins identified only in EGCG-responsive biofilms, enzymes involved in de novo purine biosynthesis were enriched in E. faecalis, while proteins likely to cause DNA instability and pathogenicity changes were abundantly present in S. lugdunensis. The classification based on gene ontology (GO) terms by microorganism exhibited that metabolic process or catabolic activity was at the top rank in E. faecalis with more than 33 proteins, and in S. lugdunensis, localization or transport was highly ranked with 4 proteins. These results support the hypothesis that EGCG might cause different cellular programs in each microorganism. Finally, comparison of the proteomes between two groups that form biofilms to similar extents discovered that 2 proteins were commonly found in the weak biofilm-forming groups (E. faecalis and EGCG-responding S. lugudunensis), whereas 9 proteins were common among the strong biofilm-forming groups (S. lugdunensis and EGCG-responding E. faecalis). It was suggested that these proteins could serve as potential indicators to detect the presence and predict the extent of biofilm formation by multiple microorganisms. Taken all together, proteomics data and analyses performed in this study provided useful and new information on the proteins embedded in the biofilms formed at the specific conditions, which can aid in diagnosis and the development of tailored treatment strategies.

The Protective Role of Baicalin in the Regulation of NLRP3 Inflammasome in Different Diseases.

The NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) inflammasome consists of pro-caspase-1, NLRP3 and apoptosis-related speckle-like protein (ASC). It can detect multiple microorganisms, endogenous danger signals and environmental stimulus including adenosine triphosphate (ATP), urate, cholesterol crystals, and so on, thereby forming activated NLRP3 inflammasome. During the course of the activation of NLRP3 inflammasome, pro-caspase-1 is transformed into activated caspase-1 that results in the maturation and secretion of interleukin-1beta (IL-1β) and IL-18. The dysfunction of NLRP3 inflammasome participates in multiple diseases such as liver diseases, renal diseases, nervous system diseases and diabetes. Baicalin is the primary bioactive component of Scutellaria baicalensis, which has been used since ancient times. Baicalin has many types of biological functions, such as anti-bacterial, anti-tumor and antioxidant. More and more evidence suggests that baicalin regulation of NLRP3 inflammasome is involved in different diseases. However, the mechanism is still elusive. Here, we reviewed the progress of baicalin regulation of NLRP3 inflammasome in many kinds of diseases to lay a foundation for future researches.

&lt;i&gt;In vitro&lt;/i&gt; studies on the development of microbial consortia for the management of major diseases in coconut and citrus

Microbial consortia for disease suppression involve combining multiple beneficial microorganisms to enhance their effectiveness in plant disease management. In present study, development of microbial consortia for the management of major diseases in coconut and citrus was carried out using bacteria Pseudomonas fluorescens, Pseudomonas putida (striata), Bacillus subtilis and fungi – Trichoderma reesei, T. harzianum, T. asperellum against major pathogens viz. Ganoderma lucidum, Thielaviopsis paradoxa, Phytopthora palmivora, Lasiodiplodia theobromae isolated from the coconut rhizosphere, and Fusarium solani isolated from the citrus rhizosphere. The promising fungal and bacterial antagonists were identified and studied for compatibility. Non-volatile compounds of consortia inhibited the test pathogens with an increase in concentration from 10 % to 75% with fungal consortia and bacterial consortia and also with mixed consortia which is composed of bacterial consortia + fungal consortia. Superior growth suppression was recorded with mixed consortia even at 10% concentration (59.44% to 65.83%) against the test pathogens in the ascending order of L. theobromae (59.44%) T. paradoxa (63.89%), G. lucidum (65.83%), P. palmivora (63.61%) and F. solani (62.78%). A similar trend was observed in 75% concentration where inhibition observed in the order of Thielaviopsis paradoxa (90.28%), G. lucidum (89.44%), F. solani (82.50%), L. theobromae (81.94%) and P. palmivora (81.39%). Volatile effect by bacterial consortia recorded the superior inhibition on test pathogens in the order of Ganoderma (85.28%), F. solani (75.28%), T. paradoxa (71.94%), P. palmivora (71.67%) and L. theobrome (67.50%) compared to the individual bioagents. Similarly, the fungal consortia showed the superior inhibitory effect on test pathogens in the order of G. lucidum (83.25%), P. palmivora (82.50%), L. theobromae (83.06%), F. solani (80.56%) and T. paradoxa (73.61%). Since there was no zone of inhibition between the strains, the interactions between Pseudomonas and Bacillus strains of Trichoderma spp. were compatible with one another. Neem cake recorded superior CFU population from 9.43 X 106 CFU at seven days by T. asperellum. Shelf life study on mixed consortia with bacterial + fungal bioagents in talc formulation indicated that all the bacterial and fungal CFU count recorded in 106 dilution for 90 days.

Metagenomics-based insights into the microbial community dynamics and flavor development potentiality of artificial and natural pit mud

Strong-flavor Baijiu (SFB) production has relied on pit mud (PM) as a starter culture. The maturation time of natural PM (NPM) is about 30 years, so artificial PM (APM) with a shorter maturation time has attracted widespread attention. This study reveals the microbial and functional dissimilarities of APM and NPM, and helps to elucidate the different metabolic roles of microbes during substrate degradation and flavor formation. Significant differences in the microbial community were observed between APM and NPM, manifesting as variations in the abundance of core microorganisms. Total of 187 high-quality metagenome-assembled genomes (MAGs) were obtained based on the metagenomic binning technology, mainly including Firmicutes (n = 106), Bacteroidota (n = 15) and Chloroflexota (n = 14). Furthermore, the relative concentration of flavor compounds in 4-year APM was similar to those in 30-year NPM, but different from those in 100-year NPMs. Methanosarcina, Methanobacterium, Methanoculleus, Anaerolineae bacterium and Aminobacterium were the key bacteria responsible for the flavor differences. From a functional perspective, amino acid and carbohydrate metabolism were key functions of PM microbial, and showed differences between APM and NPM. Finally, substrate degradation and flavor generation pathways were found to exist in multiple microorganisms. Combine the relative abundance of microorganisms with the absolute abundance of enzymes, Clostridium, Lactobacillus, Petrimonas, Methanoculleus, Prevotella, Methanobacterium, Methanosarcina, Methanothrix, Proteiniphilum, Bellilinea, Anaerolinea, Anaeromassilibacillus, Syntrophomonas and Brevefilum were identified as the key microorganisms in APM and NPM.

Microbiota profile in organs of the horseflies (Diptera: Tabanidae) in Northeastern China.

Tabanids, commonly known as horseflies and belonging to the family Tabanidae, are blood-feeding arthropods (BFA) found worldwide. They are known for their ability to mechanically and biologically transmit various animal pathogens. Tabanids are potential vectors for diseases such as Francisella tularensis, Anaplasma marginale, Theileria spp., and contributors to lumpy skin diseases. Despite their involvement in common BFA studies, tabanids have not been extensively explored in microbiome research. In this study, the microbiota structure and composition in various organs of four distinct genera of tabanids: Atylotus, Haematopota, Tabanus, and Hybomitra were examined. High-throughput sequencing of the bacterial 16S rRNA gene was performed to gain insights into the microbial communities associated with the different tabanid species. Result display that microbiota composition and diversity, including Firmicutes, Proteobacteria, and Bacteroidetes, varied significantly among the different organs, with the ovaries exhibiting significantly higher diversity. Apart from the Haematopota genus, Tenericutes were enriched in the midgut of other tabanid species, whereas the Malpighian tubules exhibited a higher abundance of Bacteroides. Notably, the ovarian microbiota structure was conserved among the four tabanid species, indicating its potential association with reproductive development. Evaluation of the potential pathogen risk revealed putative pathogens in over 100 genera associated with these tabanid commensal organisms. Twenty genera were annotated as zoonotic agents with a high abundance of Citrobacter and Brucella, highlighting the presence of this important group of zoonotic pathogens. Functional predictions of vector-microbiota interactions indicate that microbiota significantly affects vector biological traits and can influence pathogen transmission via direct interactions or by regulating host immunity and nutrition. For the first time, the distribution characteristics and functions of four genera of horsefly microbiota were analyzed, revealing the presence of multiple potential pathogenic microorganisms. These findings provide valuable insights for future research and the development of symbiotic-based strategies to control insect-borne diseases among tabanids.

Intestinal microbiota and high-risk antibiotic resistance genes in wild birds with varied ecological traits: Insights from opportunistic direct sampling in Tianjin, China

Within One Health framework, the dissemination of antibiotic resistance genes (ARGs) and pathogenic bacteria by wild birds has attracted increasing attention. In this study, gut samples of wild birds opportunistically collected in Tianjin, China, situated along the East Asian-Australasian Flyway, were used to ascertain the realistic distribution of bacteria and ARGs in their intestinal tracts. These birds have different dietary habits (herbivore, carnivore, and omnivore) and residency statuses (resident and migratory birds). Using 16S rRNA gene sequencing and qPCR, we analyzed microbial communities and the abundance of high-risk ARGs and mobile genetic elements (MGEs). Birds with distinct ecological traits exhibited significant variations in gut bacterial composition, yet similar microbial diversity. Shigella sp. emerged as the core intestinal pathogen, with a mean relative abundance 2.57 to 1466 times higher than that of other pathogenic bacteria, and its concentration correlated with the host's trophic level as indicated by the δ15N values. The distribution of ARGs and MGEs also varied with bird ecological traits. All 10 targeted high-risk ARGs were detected in carnivores or passage migrants, while migratory birds carried significantly greater abundance of intI1 than residents (p < 0.05). The potential of migratory birds to harbor and disseminate pathogenic bacteria and ARGs cannot be ignored. Network analysis revealed blaTEM-1 presence in multiple core microorganisms, positively associated with Clostridioides difficile, emphasizing its risk potential. Positive dfrA12-intI1 correlation across trophic levels suggests potential for intI1-mediated transmission. Our study underscores the high potential risk posed by wild birds in carrying ARGs and pathogenic microorganisms, emphasizing the importance of further research and surveillance in this field.

Breast Milk-Derived Antibodies Impair Vaccine Immunity in Suckling Mice.

Breast milk confers multiple benefits to the neonate, including passive immunity against multiple microorganisms via Abs. However, it remains unclear whether breast milk-derived Abs affect vaccine-induced immunity in the neonate. We evaluated in C57BL/6 and BALB/c mice whether breastfeeding from an mRNA-SARS-CoV-2-vaccinated dam affects vaccine-induced immunity in neonate mice. Using an experimental model that allows the distinction of maternal Abs and neonate Abs based on their allotype, we show that breastfeeding from an immune dam is associated with reduced vaccine immunity in the neonate. Importantly, mice that breastfed from an immune dam showed reduced numbers of plasma cells after vaccination, relative to mice that breastfed from a naive dam. Our subsequent studies using an mRNA-luciferase reporter system show that passive transfer of Abs through breastfeeding accelerates the clearance of vaccine Ag in suckling mice, resulting in reduced Ag availability. Altogether, maternal Abs transferred through breast milk can protect against infectious microorganisms, but they may also interfere with the neonate's response to vaccination by accelerating the clearance of vaccine Ag. These findings are important for understanding the effects of maternal Abs on the neonate's response to vaccines and may provide insights for improving neonatal vaccines.

Clinical Metagenomic Next-Generation Sequencing for Diagnosis of Central Nervous System Infections: Advances and Challenges.

Central nervous system (CNS) infections carry a substantial burden of morbidity and mortality worldwide, and accurate and timely diagnosis is required to optimize management. Metagenomic next-generation sequencing (mNGS) has proven to be a valuable tool in detecting pathogens in patients with suspected CNS infection. By sequencing microbial nucleic acids present in a patient's cerebrospinal fluid, brain tissue, or samples collected outside of the CNS, such as plasma, mNGS can detect a wide range of pathogens, including rare, unexpected, and/or fastidious organisms. Furthermore, its target-agnostic approach allows for the identification of both known and novel pathogens. This is particularly useful in cases where conventional diagnostic methods fail to provide an answer. In addition, mNGS can detect multiple microorganisms simultaneously, which is crucial in cases of mixed infections without a clear predominant pathogen. Overall, clinical mNGS testing can help expedite the diagnostic process for CNS infections, guide appropriate management decisions, and ultimately improve clinical outcomes. However, there are key challenges surrounding its use that need to be considered to fully leverage its clinical impact. For example, only a few specialized laboratories offer clinical mNGS due to the complexity of both the laboratory methods and analysis pipelines. Clinicians interpreting mNGS results must be aware of both false negatives-as mNGS is a direct detection modality and requires a sufficient amount of microbial nucleic acid to be present in the sample tested-and false positives-as mNGS detects environmental microbes and their nucleic acids, despite best practices to minimize contamination. Additionally, current costs and turnaround times limit broader implementation of clinical mNGS. Finally, there is uncertainty regarding the best practices for clinical utilization of mNGS, and further work is needed to define the optimal patient population(s), syndrome(s), and time of testing to implement clinical mNGS.

Screening of reference genes for gene expression study in different tissues from the transcriptome data of the vector leafhopper Psammotettix striatus

Selecting appropriate reference genes is crucial for ensuring the accuracy and reliability of gene expression study using reverse transcription-quantitative PCR (RT-qPCR). To screen the optimal reference genes for analyzing gene expression in different tissues of the vector leafhopper Psammotettix striatus which causes extensive damage to a wide range of crops by vectoring multiple plant pathogenic microorganisms, the transcriptome data from Malpighian tubules (MTs) of P. striatus were mined. Twenty alternative candidate reference genes were initially selected for screening, among which seven genes with diverse Gene Ontology (GO) annotations were choosed as candidate reference genes, i.e., ribosomal protein L7A (RPL7A), ribosomal protein S28 (RPS28), ribosomal protein L22 (RPL22), ribosomal protein LP2 (RPLP2), H3 histone family 3A (H3F3A), elongation factor 1γ (EF-1γ), and elongation factor 1α (EF-1α). Gene expression levels in different tissues of P. striatus adults were examined using RT-qPCR, and their expression stability was analyzed using multiple reference gene screening software. This study revealed EF-1α as the most abundantly expressed gene, while RPL22 exhibited the lowest expression levels. EF-1α showed the most stable expression, whereas RPS28 showed the least stability. Various software tools confirmed EF-1α as the most stable single reference gene, and EF-1α and RPLP2 an optimal combination. This study provides a foundation for future investigation of the transmission of pathogenic microorganisms mediated by the vector leafhoppers, the function of the MTs, the biosynthesis of brochosomes, the coevolutionary processes and nutritional interactions of symbionts and host insects, and the gene expression study of other sap-sucking insects.

A Highly Sensitive Dual-Drive Microfluidic Device for Multiplexed Detection of Respiratory Virus Antigens.

Conventional microfluidic systems that rely on capillary force have a fixed structure and limited sensitivity, which cannot meet the demands of clinical applications. Herein, we propose a dual-drive microfluidic device for sensitive and flexible detection of multiple pathogenic microorganisms antigens/antibodies. The device comprises a portable microfluidic analyzer and a dual-drive microfluidic chip. Along with capillary force, a second active driving force is provided by a removable self-driving valve in the waste chamber. The interval between these two driving forces can be adjusted to control the reaction time in the microchannel, optimizing the formation of antigen-antibody complexes and enhancing sensitivity. Moreover, the material used in the self-driving valve can be changed to adjust the active force strength needed for different tests. The device offers quantitative analysis for respiratory syncytial virus antigen and SARS-CoV-2 antigen using a 35 μL sample, delivering results within 5 min. The detection limits of the system were 1.121 ng/mL and 0.447 ng/mL for respiratory syncytial virus recombinant fusion protein and SARS-CoV-2 recombinant nucleoprotein, respectively. Although the dual-drive microfluidic device has been used for immunoassay for respiratory syncytial virus and SARS-CoV-2 in this study, it can be easily adapted to other immunoassay applications by changing the critical reagents.

Antibacterial, Phytochemical and Toxicological Activities of Garcinia kola Extracts against Multidrug Resistant Clinical Bacteria

Objective: The rise in multiple antibiotic resistant microorganisms has led to a decline in the efficacy of many antibiotics, prompting the investigation of alternative solutions such as medicinal plants. To tackle this concerning issue, this study was undertaken to evaluate the phytochemical composition, antibacterial activity, and toxicological characteristics of Garcinia kola extracts against multidrug-resistant bacteria commonly found in clinical settings. Material-Method: The cold maceration technique was employed to extract the root and leaf of Garcinia kola using water and methanol. The extracts were then subjected to phytochemical screening. To evaluate the extracts' potential to inhibit bacterial growth, five multidrug-resistant isolates were utilized in this study. Agar well assay was used to determine the zones of inhibition. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined using the broth macro dilution technique. Additionally, Brine shrimp lethality assay was conducted to determine the lethal concentration (LC50) of the extracts. The extracts were also combined for synergism. Results: The investigation found that the root extract in methanol had the highest yield at 25.58%. All four extracts contained ten out of the fourteen tested phytochemicals. The methanol leaf and root extracts exhibited the highest and lowest inhibition zones of 20mm and 9mm against Staphylococcus aureus, respectively. The MIC values ranged from 250 to 600 mg/ml, indicating broad-spectrum antibacterial activity. The cytotoxicity test showed a range of 33.03 to 126.3µg/mL for the aqueous and methanol extracts. Conclusion: Although Garcinia kola shows potential as a source of antibacterial compounds, caution should be exercised due to its toxic effects.

Cooperative Fermentation Using Multiple Microorganisms and Enzymes Potentially Enhances the Nutritional Value of Spent Mushroom Substrate

Cooperative Fermentation Using Multiple Microorganisms and Enzymes Potentially Enhances the Nutritional Value of Spent Mushroom Substrate

Tumour organoids and assembloids: Patient-derived cancer avatars for immunotherapy.

Organoid technology is an emerging and rapidly growing field that shows promise in studying organ development and screening therapeutic regimens. Although organoids have been proposed for a decade, concerns exist, including batch-to-batch variations, lack of the native microenvironment and clinical applicability. The concept of organoids has derived patient-derived tumour organoids (PDTOs) for personalized drug screening and new drug discovery, mitigating the risks of medication misuse. The greater the similarity between the PDTOs and the primary tumours, the more influential the model will be. Recently, 'tumour assembloids' inspired by cell-coculture technology have attracted attention to complement the current PDTO technology. High-quality PDTOs must reassemble critical components, including multiple cell types, tumour matrix, paracrine factors, angiogenesis and microorganisms. This review begins with a brief overview of the history of organoids and PDTOs, followed by the current approaches for generating PDTOs and tumour assembloids. Personalized drug screening has been practised; however, it remains unclear whether PDTOs can predict immunotherapies, including immune drugs (e.g. immune checkpoint inhibitors) and immune cells (e.g. tumour-infiltrating lymphocyte, T cell receptor-engineered T cell and chimeric antigen receptor-T cell). PDTOs, as cancer avatars of the patients, can be expanded and stored to form a biobank. Fundamental research and clinical trials are ongoing, and the intention is to use these models to replace animals. Pre-clinical immunotherapy screening using PDTOs will be beneficial to cancer patients. The current PDTO models have not yet constructed key cellular and non-cellular components. PDTOs should be expandable and editable. PDTOs are promising preclinical models for immunotherapy unless mature PDTOs can be established. PDTO biobanks with consensual standards are urgently needed.

The synergistic effect of microorganisms and multiple admixtures on improving the self-healing of cracks in biogenic mortar exposed to different marine environments

Existing research has inadequately addressed the synergistic effects of microorganisms and admixtures in promoting self-healing behavior and mechanisms of cracks in marine environments, including the atmospheric zone, tidal-splash zone, and submerged zone. This study primarily involved the incorporation of multiple admixtures and microorganisms into cement to prepare two types of biogenic mortars: Bacillus pasteurii-based mortar (BPM) and Bacillus subtilis-based mortar (BSM), with ordinary mortar (OM) serving as the control group. The mechanical properties, electrical flux, and pore structure of the mortars were investigated. Additionally, the self-healing behavior and mechanisms of cracks were studied under atmospheric, tidal-splash, and submerged conditions. The results indicate that with increasing curing age, the synergistic effect of admixtures and microorganisms continuously refined the pore structure of the specimens and enhanced their flexural strength, compressive strength, and resistance to chloride ion penetration. The atmospheric environment was found to be unfavorable for the self-healing of cracks in the specimens. However, in the submerged and tidal-splash zones, the combined action of microorganisms and admixtures led to more pronounced self-healing phenomena in the biogenic mortars compared to OM. Particularly, the self-healing effect of BSM cracks was more pronounced in the tidal-splash zone, with a 100% self-healing efficiency observed after 53 days. This was attributed to the ability of microorganisms in the tidal-splash zone to induce calcium carbonate precipitation on the surface of cracks in admixture-modified cementitious materials, thereby reducing the penetration of seawater, CO2, and O2 into the cracks, lowering the carbonation reaction of Ca(OH)2 crystals, and reducing the formation of brucite crystals inside the cracks, consequently enhancing the water permeability resistance of the specimens. These research findings hold significant engineering implications for improving the durability and service life of coastal self-healing concrete structures.

An integrated sample-to-answer graphene microchip for ultrafast detection of multiple microorganisms

The integrated detection of nucleic acids is a molecular biological technology integrating the entire process of nucleic acid detection into a “sample-to-answer” system, which is widely used in microbial identification and disease diagnosis. Although the integrated detection of nucleic acids has the advantages of fully-enclosed system and simple operation, it still faces challenges like dependence on bulky equipment, complex design, and long detection time. To address these challenges, we have proposed and fabricated an integrated graphene microchip for ultra-fast detection of multiple microorganisms. This microchip is composed of two main regions: the electrolysis-extraction region and the isothermal amplification-detection region. In the electrolysis-extraction region, the microchip was equipped with the micron-structured graphene interdigital electrode and a regular battery for power supply, and the nucleic acids of H. pylori were extracted only in 1.0 s by electrolysis. Then in the isothermal amplification-detection region, the nucleic acids of H. pylori were detected in less than 9 min at a concentration of 10 CFU/µL. As a result, the detection time of the whole process were controlled in merely 9 min. In addition, this integrated graphene microchip has three parallel channels for simultaneous detection of oral microorganisms, e.g., C. albicans, S. aureus and H. pylori. These characteristics make our integrated graphene microchip promising to contribute to the development in integrated technology of nucleic acid detection and be widely used at various medical structures or on-site environments such as home testing.

MULTIPLE DRUG RESISTANT PATHOGENS IN ACTION AFTER COVID PANDEMIC

The COVID-19 pandemic impacted the unique means for prompting the scientific community to consider developing healthcare infrastructures and logistics that will deal with any future pandemics, may be in a manner comparable to dealing with biological war or an outside attack due to Manmade weapons. Biomedical experts have refocused their efforts on creating innovative vaccines and medicines to halt the deadly spread of these infections using new biological approaches. Only a small number of innovative nations have a healthy security policy on national level in the event of a biological attack that has been bio-weaponized by a human. After the Covid pandemic, It is assumed that the whole world may have a fight against Multiple resistant bacterial pathogens or microorganisms connected to water and food. MDR bacteria (like MRSA and VRSA) in addition to newly mutated microorganism may also infect human beings, birds, and animals.

Evaluation of ready-to-use freezer stocks of a synthetic microbial community for maize root colonization.

Synthetic communities (SynComs) are an invaluable tool to characterize and model plant-microbe interactions. Multimember SynComs approximate intricate real-world interactions between plants and their microbiome, but the complexity and time required for their construction increase enormously for each additional member added to the SynCom. Therefore, researchers who study a diversity of microbiomes using SynComs are looking for ways to simplify the use of SynComs. In this manuscript, we evaluate the feasibility of creating ready-to-use freezer stocks of a well-studied seven-member SynCom for maize roots. The frozen ready-to-use SynCom stocks work according to the principle of "just add buffer and apply to sterilized seeds or seedlings" and thus can save time applied in multiple days of laborious growing and combining of multiple microorganisms. We show that ready-to-use SynCom stocks provide comparable results to those of freshly constructed SynComs and thus allow for significant time savings when working with SynComs.

Viral and bacterial microorganisms in Vietnamese children with severe and non-severe pneumonia

To investigate potential respiratory pathogens in children with community-acquired pneumonia (CAP) and risk factors for severe disease. This prospective study was conducted among 467 children at the Thai Binh Paediatric Hospital, Vietnam between 1 July 2020 and 30 June 2021. Clinical data and laboratory results were collected. Twenty-four respiratory microorganisms were tested from nasopharyngeal swabs using real-time PCR. Logistical regression was used to estimate a factor’s adjusted odd ratios of the severity of disease. Mean age of patients = 15.4 ± 13.3 months, 63.0% were male. Over 97% of patients had a positive PCR result. 87% of patients were positive for multiple (up to eight) microorganisms. Rhinovirus (46%), respiratory syncytial virus (RSV) (24%), enterovirus (17%), and parainfluenza viruses-3 (13%) were the most frequent viruses. H. influenzae (61%), S. pneumoniae (45%) and M. catarrhalis (30%) were the most common bacteria. 128 (27%) cases were classified as severe pneumonia. Presence of smokers at home (aOR 2.11, 95% CI 1.27–3.52, P value = 0.004), CRP level ≥ 50 mg/dL (aOR 6.11, 95% CI 3.86–9.68, P value < 0.0001), RSV (aOR 1.78, 95% CI 1.07–2.96, P value = 0.03) and H. influenzae (aOR 1.66, 95% CI 1.03–2.67, P value = 0.04) PCR detection associated with a higher risk of severe pneumonia; ,. Causative agents of pneumonia in children are complex. Children positive with RSV and H. influenzae need to be closely monitored to prevent severe pneumonia.