Bacterial Mixture Research Articles

Reducing Methane, Carbon Dioxide, and Ammonia Emissions from Stored Pig Slurry Using Bacillus-Biological Additives and Aeration

This study delves into the innovative application of a novel bacterial and enzyme mixture alone or combined with aeration in mitigating emissions from pig slurry storage and explores their impacts on the methane (CH4), carbon dioxide (CO2), and ammonia (NH3) emissions from stored pig slurry. A dynamic chamber was used in this research to assess the efficacy of the treatments. Biological additives (HIPO-PURÍN) of specific microbial strains were tested (a mixture ofof Bacillus subtilis, Bacillus megaterium, Bacillus licheniformis, Bacillus amyloliquefacien, and Bacillus thuringiensis) alone and combined with an aeration system (OXI-FUCH). Controlled experiments simulated storage conditions, where emissions of ammonia, methane, and carbon dioxide were measured. By analyzing the results statistically, the treatment with HIPO-PURÍN demonstrated a significant reduction in CH4 emissions by 67% and CO2 emissions by 60% with the use of biological additives, which was increased to 99% and 87%, respectively, when combined with OXI-FUCH aeration, compared to untreated slurry. Ammonia emissions were substantially reduced by 90% with biological additives alone and by 76% when combined with aeration. The study was driven by the need to develop sustainable solutions for livestock waste management, particularly in reducing emissions from pig slurry. It introduces techniques that significantly lower greenhouse gases, aligning with circular economy goals and setting a new standard for sustainable agriculture. Furthermore, there is a need to validate that farmers can independently manage pig slurry using simple and effective treatments techniques with profound environmental benefits, encouraging broader adoption of climate-conscious practices.

Silver nanoparticles biosynthesis using mixture of Lactobacillus sp. and Bacillus sp. growth and their antibacterial activity

The biosynthesis of nanoparticles offers numerous advantages, including ease of production, cost-effectiveness, and environmental friendliness. In our research, we focused on the bioformation of silver nanoparticles (AgNPs) using a combination of Lactobacillus sp. and Bacillus sp. growth. These AgNPs were then evaluated for their biological activities against multidrug-resistant bacteria. Our study involved the isolation of Bacillus sp. from soil samples and Lactobacillus sp. from raw milk in Dhamar Governorate, Yemen. The synthesized AgNPs were characterized using various techniques such as UV–visible spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The antibacterial properties of the AgNPs were assessed using the modified Kirby Bauer disk diffusion method against multidrug-resistant strains of Staphylococcus aureus and Pseudomonas aeruginosa. Our results demonstrated that the use of a bacterial mixture for biosynthesis led to faster and more effective production of AgNPs compared to using a single bacterium. The UV–visible spectra showed characteristic peaks indicative of silver nanoparticles, while XRD analysis confirmed the crystalline nature of the synthesized particles. FTIR results suggested the presence of capping proteins that contribute to the synthesis and stability of AgNPs. Furthermore, TEM images revealed the size and morphology of the AgNPs, which exhibited spherical shapes with sizes ranging from 4.65 to 22.8 nm. Notably, the antibacterial activity of the AgNPs was found to be more pronounced against Staphylococcus aureus than Pseudomonas aeruginosa, indicating the potential of these nanoparticles as effective antimicrobial agents. Overall, our study highlights the promising antibacterial properties of AgNPs synthesized by a mixture of Lactobacillus sp. and Bacillus sp. growth. Further research is warranted to explore the potential of utilizing different bacterial combinations for enhanced nanoparticle synthesis.

Transcriptome of Musca domestica (Diptera: Muscidae) Larvae Induced by Bacteria

Abstract Transcriptome analysis was used to explore the molecular regulation mechanisms of the responses of house fly, Musca domestica L. (Diptera: Muscidae), larvae to mixed solutions of Escherichia coli, Salmonella sp., Staphylococcus aureus, and Streptococcus sp. Sequencing yielded 50,701 genes that were compared with each database. A total of 34,666 (59.06%) transcripts was successfully annotated. In addition, 3,524 differentially expressed genes were screened from the low concentration dose group and the control group, 5,129 were screened from the high concentration dose group and control group, and 7,462 were screened from the high concentration dose group and low concentration dose group. Of those, 1,252, 2,369, and 3,159 differentially expressed genes were annotated in the Gene Ontology database and 306, 676, and 987 differentially expressed genes were annotated in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, respectively. The enrichment analysis of differential gene KEGG showed that the enrichment of insulin signaling pathway, HIF-1 signaling pathway, and chemokine signaling pathway in larvae induced by the high concentration dose of the bacterial mixture was more apparent than that induced by injection with the low concentration dose. Differential gene expression and function further reveal the metabolic pathway of defense response induced by bacteria, which can help in better understanding the related mechanism.

In Situ Cas12a-Based Allele-Specific PCR for Imaging Single-Nucleotide Variations in Foodborne Pathogenic Bacteria.

In situ profiling of single-nucleotide variations (SNVs) can elucidate drug-resistant genotypes with single-cell resolution. The capacity to directly "see" genetic information is crucial for investigating the relationship between mutated genes and phenotypes. Fluorescence in situ hybridization serves as a canonical tool for genetic imaging; however, it cannot detect subtle sequence alteration including SNVs. Herein, we develop an in situ Cas12a-based amplification refractory mutation system-PCR (ARMS-PCR) method that allows the visualization of SNVs related to quinolone resistance inside cells. The capacity of discriminating SNVs is enhanced by incorporating optimized mismatched bases in the allele-specific primers, thus allowing to specifically amplify quinolone-resistant related genes. After in situ ARMS-PCR, we employed a modified Cas12a/CRISPR RNA to tag the amplicon, thereby enabling specific binding of fluorophore-labeled DNA probes. The method allows to precisely quantify quinolone-resistant Salmonella enterica in the bacterial mixture. Utilizing this method, we investigated the survival competition capacity of quinolone-resistant and quinolone-sensitive bacteria toward antimicrobial peptides and indicated the enrichment of quinolone-resistant bacteria under colistin sulfate stress. The in situ Cas12a-based ARMS-PCR method holds the potential for profiling cellular phenotypes and gene regulation with single-nucleotide resolution at the single-cell level.

Aggregation-induced emission silence-mediated pathogen detection using a rapidly degradable nanographene-embedded polymersome.

Typical pathogen detection processes are time-consuming and require expensive equipment and professional operators, limiting their practical applicability. Developing a rapid and easy-to-read method of accurately sensing pathogenic bacteria is critical for reducing the spread and risk of infection in high-risk areas. Herein, the synthesis of nanographene (nanoG) that exhibits aggregation-induced emission (AIE) is described. The nanoG was embedded into a hydrophobic shell of poly(lactic-co-glycolic acid) (PLGA) polymersome in a double-emulsion process, significantly enhancing the nanoG luminescence under irradiation at 330 nm due to the enrichment of nanoG between the inner and outer PLGA shells. Both Gram-positive and Gram-negative bacteria can rapidly degrade the PLGA vesicular structure, leading to dispersal of the nanoG inside the shell and silencing the AIE effect. A linear relationship between the bacterial concentration and emissivity was established, and the detection limit was identified. Moreover, the polymersome has excellent selectivity for methicillin-resistant Staphylococcus aureus (MRSA) detection after a screening pretreatment of a bacterial mixture with suitable antibiotics. The AIE silencing could be observed with the naked eye in an MRSA-infected wound treated with the polymersome after 1 h of incubation, demonstrating a high potential for clinical rapid screening applications.

Antibacterial Irrigation Solution Cocoa Bean Cuticle Extract (Theobroma cacao L) Against Streptococcus viridans

Background: Pulp necrosis is the demise of the pulp due to persistent infection in the pulp area, such as Streptococcus viridans. Irrigation steps during biomechanical preparation and sterilization play a very important role. The irrigation material often used is 2.5% NaOCl because it has a broad antibacterial spectrum but is irritating and has an unpleasant aroma. The cuticle of cocoa beans (Theobroma cacao L.) might be an alternative root canal irrigation material. The Objective: To determine the antibacterial properties of irrigation solution of cocoa bean cuticle extract against Streptococcus viridans. Materials and Methods: This study was laboratory experimental with a post-test-only control group design by calculating the number of bacterial colonies. Consisting of eight research groups, namely cocoa bean cuticle extract with concentrations of 25% (P1), 12.5% (P2), 6.25% (P3), 3.12% (P4), 1.56% (P6), 0.78% (P6), NaOCl 2.5% (K+), and sterile distilled water. 0.5 ml of each group was taken and then mixed with 0.5 suspension of Streptococcus viridans, diluted 10-4. Next, the extract and bacterial suspension mixture 10-4 were incubated for 24 hours, then 0.1 ml was inoculated on BHIA media and incubated. After that, the colony counter manually counted the number of colonies. Results: The result showed a significance of p<0.5, followed by the Mann-Whitney test, which showed significant differences except in group P1 (25%) with K+ (NaOCl 2.5%). Conclusion: Cocoa bean cuticle extract irrigation solutions with concentrations of 25%, 12.5%, 6.25%, 3.12%, 1.56% and 0.78% presented antibacterial power against Streptococcus viridans. Keywords: Streptococcus viridans, Antibacterial, Cocoa bean epidermis extract, Irrigation solution.

Effect of Usnea longissima ethyl acetate extract on acute oxidative and inflammatory lung damage from Staphylococcus aureus infection in rats.

The role of oxidants and proinflammatory cytokines in the pathogenesis of pneumonia caused by Staphylococcus aureus (S. aureus) has been demonstrated. The present study aims to investigate the protective effect of ethyl acetate extract (EtOAc) obtained from Usnea longissima (UL) against acute oxidative and inflammatory lung damage due to S. aureus infection in rats. Albino Wistar-type male rats were divided into three groups: Healthy (HG), S. aureus inoculated (SaG), and S. aureus inoculated + ULEtOAc administered (SUL). SaG (n = 6) and SUL (n = 6) group rats' left nostrils (excluding HG) were inoculated with 0.1 ml bacterial mixture. After 24 hours, ULEtOAc (50 mg/kg) was administered orally to the SUL group, and the same volume of normal saline was administered orally to the HG (n = 6) and SaG groups. This procedure was performed once a day for seven days. Levels of oxidant and antioxidant parameters such as malondialdehyde (MDA) and total glutathione (tGSH), as well as pro-inflammatory cytokine levels such as nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-one beta (IL-1β), were measured in removed lung tissues. Tissues were also examined histopathologically. Biochemical results showed that ULEtOAc significantly suppressed the increase of MDA, NF-κB, TNF-α, and IL-1β levels and the decrease of tGSH caused by S. aureus in lung tissue. S. aureus inoculation caused severe mononuclear cell infiltration in interstitial areas, severe lymphoid hyperplasia in bronchial-associated lymphoid tissue and severe alveolar edema, histopathologically. Treatment with ULEtOAc had an attenuating effect on these histopathological findings. Experimental results from this study suggest that ULEtOAc may be beneficial in treating S. aureus-induced oxidative and inflammatory lung damage.

The Effect of the Interaction Between PGPR Bacteria, Mycorrhizae, and Oyster Mushroom Residues on the Proportions of Active Ingredients in Coriander Plant Oil (Coriandrum sativum L.) using the Technique of GC/MS-QP2010 Plus

A factorial experiment was carried out in a sandy mixture soil using a completely randomized design (CRD) in the field of the Department of Soil Sciences and Water Resources, College of Agriculture, University of Diyala, in the agricultural season 2022-2023 in order to study the effect of adding Azotobacter, Pseudomonas, Mycorrhizae, and oyster mushroom residues in proportions of the active ingredients of the medicinal coriander plant oil. The results showed a difference in the number and proportions of the active compounds according to the different treatments of the experiment. Where the treatments of comparison and fertilizer recommendation achieved a decrease in the number of oil-forming active compounds, all treatments were superior by giving the highest number of oil compounds and their proportions compared to the comparison and fertilizer recommendation. The treatment of the bacterial mixture and oyster mushroom residues with mycorrhizae recorded the highest number of components, reaching 171 compounds, with an increase in the proportions of all compounds. The compound 9-Octadecenoic acid, 1,2,3-propanetriyl ester, recorded the highest percentage, which amounted to 20.71%, followed by the compound (E)-9-Octadecenoic acid ethyl ester with a percentage of 16.69%, compared to the comparison and fertilizer recommendation treatments, which recorded (13.33, 14.33%), respectively, and the percentage of Linalool was 2.30% compared to the comparison and fertilizer recommendation treatments, which gave (2.13, 0.95%), respectively.

104 Development of an Experimental Model to Induce Liver Abscesses in Steers Using an Acidotic Diet Challenge and Oral Bacterial Inoculation

Abstract The objective of this study was to develop a model for liver abscesses (LA) in Holstein steers by using an acidotic diet or acidotic diet plus intraruminal inoculation of Fusobacterium necrophorum, Salmonella enterica serotype Lubbock, and Truperella pyogenes. Holstein steers (n = 40; initial BW = 84.9 + 7.1 kg) were individually housed and randomly assigned to either a negative control diet (CON), an acidotic diet (AD), or an acidotic diet plus intraruminal inoculation with the bacterial mixture [ADB; (1 × 109 colony forming unit (CFU)/mL of each bacteria]. Steers in AD and ADB were cycled on (3 d) and off (2 d) a high-starch diet 20 days before intraruminal inoculation of ADB. Following inoculation, AD and ADB remained on the acidotic diet for the remainder of the study. Ruminal pH boluses were randomly inserted into 19 steers across treatments to monitor ruminal pH throughout the study. Steers were euthanized and necropsied to record gross pathology on the lung, rumen, liver, and colon. Culture techniques were used to determine the prevalence inoculated bacteria within LA. Quantitative polymerase chain reaction methods were used to quantify F. necrophorum in rumen tissue. Data were analyzed as a completely randomized design with animal as the experimental unit, where an α of ≤ 0.05 determined significance. Liver abscess prevalence was 43% in ADB vs. 0% in AD and CON treatments (P < 0.01). Ruminal damage was 51.1% greater in ADB than in AD and 78.6% greater than in CON (P ≤ 0.05). There were no pathological differences in the lungs and colons among treatment groups (P ≥ 0.14). There was a treatment × time interaction for ruminal pH, where pH decreased below 5.6 in AD and ADB calves during each cycle of the acidotic diet (P < 0.01). The ADB contained a greater concentration of F. necrophorum subsp. Necrophorum and subsp. Fundliforme within rumen tissue (P ≤ 0.03). Of the LA cultured, 100% contained F. necrophorum, 50% S. enterica, and 0% contained T. pyogenes. The F. necrophorum subsp. Necrophorum was phylogenetically typed and identical to the strain (FN-8LI) infused in the bacterial inoculation. These data suggest that acidotic diet in combination with intraruminal inoculation of the bacterial mixture is sufficient to induce liver abscesses and provides insight on possible mechanisms and origins of LA. Further studies are needed to validate these results and determine if this model may be used to study novel intervention methods.

Isolation, characterization, and application of bacteriophage cocktails for the biocontrol of Pseudomonas fluorescens group strains in whole and skimmed milk.

Pseudomonas fluorescens group strains can lead to spoilage of milk as well as loss of quality in dairy products through their heat-resistant enzymes. Phages are important alternatives for combating spoilage bacteria in food industry and used successfully in many applications. The aim of this study was the isolation and characterization of phages and to assess the efficiency of a phage cocktail in whole and skimmed milk. For this purpose, phages effective against Pseudomonas fluorescens (L23.2), Pseudomonas tolaasii (P22.1), and Pseudomonas rhodesiae (A11.1) were isolated. Their host range was found to be highly specific, and the transmission electron micrographs indicates that they belonged to Tectiviridae family. Their genome sizes were found to be vary between 38.3 and 53.5 kb. The latent periods and burst sizes were determined as 15, 10, 15 min and 91, 20, 80 PFU/infected cell for L23.2, P22.1, and A11.1, respectively. All three phages were found to be sensitive to low pH and high temperature. The effect of the phage cocktail was monitored in milk with different fat contents during storage at 4 °C for 5 days. As a result, bacterial reductions up to 4.09 and 5.29 log-units were observed for the whole and skimmed milk, respectively. Thus, the efficacy of a phage cocktail against a bacterial mixture of different P. fluorescens strains was tested in milk samples with different fat contents in accordance with real-life scenarios for the first time.

Electrospinning Microencapsulation of Lactobacillus fermentum K73 Using Gelatin as the Main Component of a Food-Grade Matrix.

This work aimed to establish the conditions that improve the viability of Lactobacillus fermentum K73 during and after the electrospinning process. A mixture of experimental designs were performed to select the formulation (gelatin and bacterial culture) that improves the probiotic viability after blending and under simulated gastrointestinal conditions. A Box-Behnken design was performed to improve the encapsulation yield and survival during the electrospinning process. For the Box-Behnken design, the factors were soy lecithin and bacteria culture concentration at the blend and collector distance for electrospinning. It was hypothesized that soy lecithin improved the electrospinnability, acting as a surfactant in the mixture and allowing lower voltage to be used during the process. The selected volume ratio of the gelatin (25%)/bacterial culture mixture was 0.66/0.34. The physicochemical parameters of the selected blend were in the recommended range for electrospinning. The conditions that improved the encapsulation yield and survival during electrospinning were 200 g/L of bacterial culture, 2.5% (w/v) soy lecithin, and 7 cm collector distance. The experimental encapsulation yield and survival was 80.7%, with an experimental error of 7.2%. SEM micrographs showed the formation of fibers with gelatin/bacterial culture beads. Encapsulation improved the viability of the probiotic under simulated gastrointestinal conditions compared to free cells.

Plant growth-promoting bacterial mixture enhanced growth of barley under salt stress

Plant growth-promoting bacterial mixture enhanced growth of barley under salt stress

Single-Cell Genotyping of Single-Nucleotide Mutations Using In Situ Allele-Specific Loop-Mediated Isothermal Amplification.

Single-nucleotide mutations (SNMs) in the bacterial genome may cause antibiotic resistance. The visualization of SNMs can indicate antibiotic resistance phenotypes at the single-cell level but remains challenging. Herein, we proposed an in situ allele-specific isothermal amplification proceeded inside cells, allowing us to image bacterial genes with single-nucleotide resolution. The primer for loop-mediated isothermal amplification (LAMP) was designed with artificial mismatch bases to serve as an allele-specific probe, endowing LAMP to specifically amplify genes with SNMs. Due to the high amplification efficiency of LAMP, the method termed AlleLAMP can generate high gain for imaging SNMs and precisely quantify mutated quinolone-resistant Salmonella in bacterial mixture. We utilized AlleLAMP to survey the selection of antibiotic resistance under the preservative stress and found that the mutant quinolone-resistant strain owned a survival advantage over the wild-type quinolone-sensitive strain under the stress of preservatives. AlleLAMP can serve as a single-cell tool for analyzing the relationship between bacterial genotype and phenotype.

Beneficial effects of indigenous Bacillus spp. on growth, antioxidants, immunity and disease resistance of Rhynchocypris lagowskii

This study aimed to investigate the effect of Bacillus aryabhattai (LSG3-7) and Bacillus mojavensis (LSG3-8) on growth performance, antioxidant capacity, and immune response in Rhynchocypris lagowskii (Dybowski, 1869), at the trial and challenge periods. A 630 healthy fish (10.76 ± 0.05) were randomly divided into six groups: control group (D1) was fed the basal diet, D2 and D3 were supplemented with LSG 3–7 and LSG3-8 (1 × 108 CFU/g) for both of them, whereas D4 was supplemented with a mixture of both bacteria (0.5 × 108 CFU/g each), and D5 was supplemented with LSG3-7 0.75 × 108 CFU/g + LSG3-8 0.25 × 108 CFU/g, and D6 supplemented with LSG3-7 0.25 × 108 CFU/g + LSG3-8 0.75 × 108 CFU/g. After the trial, Aeromonas hydrophila was used in a challenge test for 14 days. Treatments showed significant differences (p < 0.05) in growth performance and antioxidant capacity (CAT, CuZn-SOD, GPX) in the liver and intestine compared to the control. The antioxidant-related genes CAT, CuZn-SOD, GPX, and Nrf2 in the liver and intestine showed upregulation compared with the control group. Serum IgM, LZM, C3, C4, and AKP showed a favorable superiority (p < 0.05) in treatments (D2 – D6) at the trial and challenge test compared to controls. In parallel, immune-related genes (IgM, NF-κB, TLR-1, TLR-2, and MyD88) showed an up-regulated level (p < 0.05) in treatments (D2 – D6) compared to the control. In addition, pro-inflammatory cytokines (IL-1, TNF-α) showed a downregulated level in treatments (D2 – D6). After the challenge test, the immune-related genes in the liver and muscle showed an up-regulated level in treatments compared to the controls. The survival rate showed a significant increase (p < 0.05) in the treatment groups (D2 – D6) compared to the control. Overall, individuals and the bacterial mixture of B. aryabhattai and B. mojavensis could improve the growth performance, antioxidant capacity, immune capacity, and survival rate of R. lagowskii and prevent side effects of A. hydrophila. However, B. mojavensis showed a slight improvement compared to B. aryabhattai without a significant difference between them.

Moving towards in pouch diagnostics for ostomy patients: exploiting the versatility of laser induced graphene sensors

The development of a 3D printed sensor for direct incorporation within stoma pouches is described. Laser induced graphene scribed on either side of polyimide film served as the basis of a 2 electrode configuration that could be integrated within a disposable pouch sensor for the periodic monitoring of ileostomy fluid pH. The graphene sensors were characterised using electron microscopy, Raman spectroscopy, DekTak profilometry with the electrochemical properties investigated using both cyclic and square wave voltammetry. Adsorbed riboflavin was employed as a biocompatible redox probe for the voltammetric measurement of pH. The variation in peak position with pH was found to be linear over pH 3–8 with a sub Nernstian response (43 mV/pH). The adsorbed probe was found to be reversible and exhibited minimal leaching through repeated scanning. The performance of the system was assessed in a heterogeneous bacterial fermentation mixture simulating ileostomy fluid with the pH recorded before and after 96 h incubation. The peak profile in the bacterial medium provided an unambiguous signal free from interference with the calculated pH before and after incubation (pH 5.3 to 3.66) in good agreement with that obtained with commercial pH probes.Graphical abstract

A magnetic field augmented ultra-thin layer chromatography coupled surface enhanced Raman spectroscopy separation of hemozoin from bacterial mixture

Malaria is considered as one the most widespread disease with highest possibility of co-infection at all levels of the disease prognosis. Rapid detection and discrimination of malaria from other co-infections remains a challenge. Hemozoin is a metabolic biproduct of malaraia possessing paramagnetic property due to presence of iron at its centre. Here, we report a label free, rapid and highly sensitive magnetic field based ultra-thin layer chromatography (UTLC) coupled with surface enhanced Raman spectroscopy (SERS) technique for detection and separation of hemozoin from a bacterial mixture. Highly optimized silver nanorods chip fabricated using glancing angle deposition (GLAD) is explored for the UTLC-SERS separation. These chips possessing channel like characteristic and high surface to the volume ratio serve as excellent UTLC plates. The magnetic nature of hemozoin has been exploited for its separation from the mixture of P. aeruginosa (Gram-negative) and S. aureus (Gram-positive) by allocating a 0.6 T magnet over the UTLC flow setup. The solvent front migrated approximately to a distance of 13 mm from the sample point due to the magnetic environment. Spatially resolved SERS data was collected along the mobile phase and separation of mixture was confirmed. Further, staining of hemozoin, P. aeruginosa and S. aureus was done using methylene blue, acridine orange and rhodamine 6 G respectively. The separation was confirmed for the stained analytes. The present developed method provides plate height as low as 18 µm and hemozoin detection limit as <10 parasites/mL. Therefore, we establish a highly specific and sensitive technique capable of separating small amounts of bioanalytes, aiding in the removal of co-infections from the disease at a very early stage of infection.

Bioelectrochemical Purification of Biomass Polymer Derived Furfural Wastewater and Its Electric Energy Recovery.

With the increasing environmental pollution caused by waste polymers, the conversion of polymer components in biomass into valuable products is of great significance for waste management and resource recovery. A two-stage microbial fuel cell (MFC) was used to treat furfural wastewater in this study. The maximum output voltage was 240-250 mV and the power generation time in an operation cycle was 286 h. The degradation efficiency of furfural reached 99-100% (furfural concentration at 300-3000 mg/L) and was slightly reduced to 91% at 7000 mg/L. In addition, the BOD/COD ratio of the furfural wastewater increased from 0.31 to 0.48 after MFC processing. The molecular analysis of the anodic bacterial isolates indicated that the phylogenetic bacterial mixture was dominated by five active anaerobic bacteria with a similarity percentage above 99% for each strain: Burkholderia (B. burdella), Clostridium sensu stricto (Cymbidaceae), Klebsiella (Klebsiella), Ethanoligenens (anaerobic genus), and Acidocella (anaerobic genus); the mixture exhibited good properties to carry out bioelectricity generation in the microbial fuel cell. This indicates that the MFC has effectively degraded furfural for pollutant removal and power generation and is a promising clean method to treat furfural pollution in industry wastewater.

User-friendly, signal-enhanced planar spiral coil-based magnetoelastic biosensor combined with humidity-resistant phages for simultaneous detection of Salmonella Typhimurium and Escherichia coli O157:H7 on fresh produce

A user-friendly, signal-enhanced magnetoelastic (ME) biosensor was newly constructed, consisting of three sensors, a movable pencil-type planar spiral coil, and a set of hand-held signal amplifiers. It was combined with humidity-resistant, wild-type phages instead of humidity-susceptible E2 phage to simultaneously detect Salmonella Typhimurium (ST) and Escherichia coli O157:H7 (EC) on the surface of an apple. The set of hand-held signal amplifiers, four times smaller than those in the planar spiral coil-based ME biosensor, was constructed with an optimal DC magnetic field strength of 76 G. These optimized hand-held signal amplifiers enhanced the signal amplitude of the sensor by up to 1.5 times, enabling the distinction of individual peaks from three sensors. Both phage-immobilized sensors detected each target specifically under ambient humidity without the need for a bulky humidifier. The optimized exposure time for phage binding with each target on fresh produce was 16 min, almost twice as fast as the rectangular solenoid coil-based ME biosensor. The signal-enhanced ME biosensor detected its target bacteria from a mixture of ST and EC inoculated on an apple simultaneously with an enhanced detection limit of 1.7 ± 0.4 log CFU/25 mm2 for ST and 1.6 ± 0.3 log CFU/25 mm2 for EC. Notably, the detection limit and sensitivity did not show significant differences between each single analyte and bacterial mixture, indicating no interference with another coexisting pathogens. Our findings highlight the signal-enhanced ME biosensor as a rapid, efficient, specific, and user-friendly method for the simultaneous detection of fresh produce-borne pathogens.

A study on bacterial self-healing concrete encapsulated in lightweight expanded clay aggregates

Concrete is a widely used construction material susceptible to cracking due to internal and external stresses, affecting the long-term strength and durability of the structure. Self-healing concrete appears to be a promising solution to address this issue. For concrete, self-healing is naturally achievable to a certain extent by utilising unhydrated cement particles. However, to heal significant internal and external cracks over a long period, we need to develop new techniques to reach a practical level of self-healing. This study investigates autonomous self-healing concrete using Microbially Induced Calcite Precipitation (MICP). The concrete mix is created by incorporating Lightweight Expanded Clay Aggregate (LECA) encapsulated with Lysinibacillus sphaericus and Bacillus coagulans bacteria. Encapsulation is used to prolong the life of the bacteria and shield them from the highly alkaline environment of the concrete. A bacterial concrete mixture was made by replacing 20% coarse aggregate by volume with the bacteria-encapsulated LECA. The properties of self-healing concrete specimens were compared with conventional concrete specimens of grade M30. After curing, the cracked bacterial specimens underwent a 14-day wet and dry cycle to examine how well the bacteria healed the cracks. Then, the durability property was analysed based on water permeability and chloride penetration tests. The microstructure of concrete was examined using SEM and EDX analysis. Bacillus coagulans and Lysinibacillus sphaericus bacteria specimens showed favourable results compared to conventional concrete for hardened and durability tests performed. The two bacteria-encapsulated concrete also exhibited positive crack healing, and the macrocracks in the range of 0.1 to 0.5 mm were healed. A salt analysis confirmed the presence of Calcium Carbonate (CaCO3) in the healed portion of the concrete. Lysinibacillus sphaericus bacteria showed slightly better results than Bacillus coagulans, and both bacteria can be used for self-healing concrete applications.

Immobilization of bacterial mixture of Klebsiella variicola FH-1 and Arthrobacter sp. NJ-1 enhances the bioremediation of atrazine-polluted soil environments.

In this study, the effects of the immobilized bacterial mixture (IM-FN) of Arthrobacter sp. NJ-1 and Klebsiella variicola strain FH-1 using sodium alginate-CaCl2 on the degradation of atrazine were investigated. The results showed that the optimal ratio of three types of carrier materials (i.e., rice straw powder, rice husk, and wheat bran) was 1:1:1 with the highest adsorption capacity for atrazine (i.e., 3774.47 mg/kg) obtained at 30°C. On day 9, the degradation efficiency of atrazine (50 mg/L) reached 98.23% with cell concentration of 1.6 × 108 cfu/ml at pH 9 and 30°C. The Box-Behnken method was used to further optimize the culture conditions for the degradation of atrazine by the immobilized bacterial mixture. The IM-FN could be reused for 2-3 times with the degradation efficiency of atrazine maintained at 73.0% after being stored for 80 days at 25°C. The population dynamics of IM-FN was explored with the total soil DNA samples specifically analyzed by real-time PCR. In 7 days, the copy numbers of both PydC and estD genes in the IM-FN were significantly higher than those of bacterial suspensions in the soil. Compared with bacterial suspensions, the IM-FN significantly accelerated the degradation of atrazine (20 mg/kg) in soil with the half-life shortened from 19.80 to 7.96 days. The plant heights of two atrazine-sensitive crops (wheat and soybean) were increased by 14.99 and 64.74%, respectively, in the soil restored by immobilized bacterial mixture, indicating that the IM-FN significantly reduced the phytotoxicity of atrazine on the plants. Our study evidently demonstrated that the IM-FN could significantly increase the degradation of atrazine, providing a potentially effective bioremediation technique for the treatment of atrazine-polluted soil environment and providing experimental support for the wide application of immobilized microorganism technology in agriculture.