Bacterial Numbers Research Articles

Laser NIR Irradiation Enhances Antimicrobial Photodynamic Inactivation of Biofilms of Staphylococcus aureus.

Photodynamic inactivation (PDI) is a powerful technique for eradicating microorganisms, and our group previously demonstrated its effectiveness against planktonic cultures of Staphylococcus aureus bacteria using 5,10,15,20-tetrakis[4-(3-N,N-dimethylaminopropoxy)phenyl]porphyrin (TAPP) and visible light irradiation. However, biofilms exhibit a lower sensitivity to PDI, mainly due to limited penetration of the photosensitizer (PS). In the context of emerging antibacterial strategies, near-infrared treatments (NIRTs) have shown promise, especially for combating resistant strains. NIRT can act either through photon absorption by water, causing a thermal effect on bacteria, or by specific chromophores without a significant temperature increase. Our objective was to enhance biofilm sensitivity to TAPP-PDI by pretreatment with NIRT. This combined approach aims to disrupt biofilms and increase the efficacy of TAPP-PDI against bacterial biofilms. In vitro biofilm models of S. aureus RN6390 were utilized. NIRTs involved a 980 nm laser (continuous mode, 7.5 W/cm2, 30 s, totaling 225 J/cm2) post-TAPP exposure to enhance photosensitizer accumulation. Subsequent visible light irradiation at 180 J/cm2 was employed to perform PDI. Colony-forming unit counts evaluated the synergistic effect on bacterial viability. Scanning electron microscopy visualized the architectural changes in the biofilm structure. TAPP was extracted from bacteria to estimate the impact of NIRT on biofilm penetration. Using in vitro biofilm models, NIRT application following biofilm exposure to TAPP increased PS accumulation per bacteria. Under these conditions, NIRT induced a transient increase in the temperature of PBS to 46.0 ± 2.6°C (ΔT = 21.5°C). Following exposure to visible light, a synergistic effect emerged, yielding a substantial 4.4 ± 0.1-log CFU reduction. In contrast, the PDI and NIRT treatments individually caused a decrease in viability of 0.9 ± 0.1 and 0.8 ± 0.2-log respectively. Interestingly, preheating TAPP-PBS to 46°C had no significant impact on TAPP-PDI efficacy, suggesting the involvement of thermal and nonthermal effects of NIR action. In addition to the enhanced TAPP penetration, NIRT dispersed the biofilms and induced clefts in the biofilm matrix. Our findings suggest that NIR irradiation serves as a complementary treatment to PDI. This combined strategy reduces bacterial numbers at lower PS concentrations than standalone PDI treatment, highlighting its potential as an effective and resource-efficient antibacterial approach.

Cover crop monocultures and mixtures enhance bacterial abundance and functionality in the maize root zone

Abstract Cover cropping is an effective method to protect agricultural soils from erosion, promote nutrient and moisture retention, encourage beneficial microbial activity, and maintain soil structure. Re-utilization of winter cover crop root channels by maize roots during summer allows the cash crop to extract resources from distal regions in the soil horizon. In this study, we investigated how cover cropping during winter followed by maize (Zea mays L.) during summer affects the spatiotemporal composition and function of the bacterial communities in the maize rhizosphere and surrounding soil samples using quantitative PCR, 16S rRNA gene amplicon sequencing, and metaproteomics. We found that the bacterial community differed significantly among cover crop species, soil depths, and maize growth stages. Bacterial abundance increased in reused root channels, and it continued to increase as cover crop diversity changed from monocultures to mixtures. Mixing Fabaceae with Brassicaceae or Poaceae enhanced the overall contributions of several steps of the bacterial carbon (C) and nitrogen (N) cycles, especially glycolysis and the pentose phosphate pathway. The deeper root channels of Fabaceae and Brassicaceae as compared to Poaceae corresponded to higher bacterial 16S rRNA gene copy numbers and improved community presence in the subsoil regimes, likely due to the increased availability of root exudates secreted by maize roots. In conclusion, root channel reuse improved the expression of metabolic pathways of the C and N cycles and the bacterial communities, which is beneficial to the soil and to the growing crops.

Assessment of amoxicillin biodegradation using a fluidized bed reactor

In recent decades, emerging pharmaceutical contaminants, such as personal care items, pharmaceuticals, and disinfectants, have increased alarmingly in aquatic ecosystems. The efficacy of wastewater treatment plants in eliminating these contaminants has been demonstrated. This study aims to find an alternative method, such as using the fluidized bed reactor (FBR) with bacteria to remove amoxicillin from wastewater. The active bacteria Acinetobacter baumannii and Klebsiella pneumoniae were isolated from one of the pharmaceutical factories in Baghdad, Iraq, from Al-Jazeera wastewater, which was used in FBR technology to decompose the antibiotic amoxicillin. The study examined several operational parameters, including pH, total organic carbon (TOC), dissolved carbon dioxide (CO2) concentration, airflow rate, amoxicillin concentration, and bacterial number, while maintaining a constant temperature. High-performance liquid chromatography (HPLC) was used to find the reaction route. The maximum amoxicillin removal efficiency was 93% and 91% for Acinetobacter baumannii and Klebsiella pneumoniae, respectively. For both bacterial species, the pH decreased due to the formation of amino acids and CO2. The suitable operation condition was 5 mg/L of amoxicillin concentration, flow rate of 30 m/s, and the number of bacteria 47 x 105 cfm/mL and 40 x 105 cfm/mL for Acinetobacter baumannii and Klebsiella pneumoniae, respectively. After treatment, the number of bacteria increased due to the degradation of amoxicillin. The steady-state time is found at 64 hours. The organic matter reaction pathway ends at the aromatic amino acid lysine, which kills gram-negative bacteria. This method was found to be successful in removing amoxicillin in low concentrations by using Acinetobacter baumannii. In the future, treating amoxicillin in wastewater using FBR technology with gram-positive bacteria is recommended.

Diffusive Phyllosphere Microbiome Potentially Regulates Harm and Defence Interactions Between Stephanitis nashi and Its Crabapple Host.

Pear lace bug (Stephanitis nashi) is a significant herbivorous pest, harbouring a diverse microbiome crucial for crabapple (Malus sp.) host adaptation. However, the mutual influence of S. nashi- and plant-associated microbiomes on plant responses to pest damage remains unclear. This study found that S. nashi damage significantly altered bacterial community structure and reduced bacterial evenness in the crabapple phyllosphere. Notably, bacterial diversity within S. nashi was significantly lower than that in the environment, potentially influenced by insect developmental stage, bacterial diffusion stage and endosymbiont species number and abundance. Extensive bacterial correlation and diffusion effect between S. nashi and adjacent plant environments were observed, evident in a gradual decrease in bacterial diversity and an increase in bacterial acquisition ratio from soil to phyllosphere to S. nashi. Correspondingly, S. nashi significantly impacted the metabolic response of crabapple leaves, altering pathways involved in vitamin, amino acid and lipid metabolism and so forth. Furthermore, association analysis linked these metabolic changes to phyllosphere bacterial alterations, emphasizing the important role of diffusive phyllosphere microbiome in regulating S. nashi-crabapple interactions. This study highlights bacterial diffusion effect between insect and plants and their potential role in regulating insect adaptability and plant defence responses, providing new insights into plant-insect-microbiome interactions.

The Decontamination Effect of an Oscillating Chitosan Brush Compared With an Ultrasonic PEEK-Tip: An InVitro Study Using a Dynamic Biofilm Model.

This study aimed to assess the effect of an oscillating chitosan brush (OCB) compared with an ultrasonic device with PEEK tip (US-PEEK) for mechanical implant surface decontamination using an invitro model combining 3D models and a validated dynamic multispecies biofilm. A multispecies biofilm using six bacterial strains (Streptococcus oralis, Veillonella parvula, Actinomyces naeslundii, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans) was seeded on dental implants with machined and sandblasted, large-grit and acid-etched (SLA) surfaces. These were installed in 3D models depicting peri-implant defect. Mechanical decontamination was performed for 120 s using either an OCB or a US-PEEK. A negative control group received no treatment. Scanning electron microscopy (SEM) was used to evaluate the bacterial composition and quantitative PCR (qPCR) analyzed the number of each bacterial species [colony-forming units per milliliter (CFU/mL)]. Well-structured biofilms with a dense microbial distribution were observed on the negative control implants after 72 h. qPCR following mechanical decontamination showed a scarce bacterial reduction in the OCB group. The US-PEEK group exhibited a significant decrease in bacterial species compared to both OCB and control groups (p < 0.05). A biofilm removal effect was also observed in the OCB group for the machined implant surfaces. In vitro assessment using an anatomical 3D model showed that mechanical decontamination effectively reduced biofilm. The US-PEEK group demonstrated biofilm reduction on the SLA surface, while the OCB group showed a reduction on the machined implant surface. Additionally, the US-PEEK group demonstrated greater efficacy in reducing bacterial numbers.

A double ttrA and pduA knock-out mutant of Salmonella Typhimurium is not attenuated for mice (Mus musculus).

Despite numerous studies on Salmonella enterica subsp. enterica serovar Typhimurium, the underlying mechanisms of several aspects of its virulence are still under investigation, including the role of the pdu and ttrA genes, associated with the metabolism of 1,2-propanediol using tetrathionate as an electron acceptor respectively. Our objective was to contribute to an understanding of the role of these genes inbacterial virulence for mice (Mus musculus) using an S. Typhumirum ΔttrApduA mutant. The experiment was conducted with a group infected by the S. Typhimurium mutant and a control group infected with a wild-type strain. The mutant was not attenuated compared with the parent strain. There were no differences in the bacterial numbers recovered from the mesenteric lymph nodes and Peyer's patches but at 8-day after oral infection higher numbers were recovered from the spleen, liver, and cecum. Unlike the single pduA and ttrA mutants, the double ΔttrApduA mutation did not affect invasion and survival in mice, which highlights the need for further studies to clarify the role of these important metabolism genes under reduced redox conditions linked to Salmonella virulence.

Mathematical modeling suggests heterogeneous replication of Mycobacterium tuberculosis in rabbits.

How quickly Mycobacterium tuberculosis ( Mtb ) replicates and dies in lungs of infected individuals is likely to determine the outcome of the infection - either control/clearance of the bacteria by the host immune response or progression to active disease, tuberculosis ( TB ). And yet, only a few studies, primarily in mice, rigorously estimated the rates of Mtb replication and death during an in vivo infection. We infected rabbits with a novel clinical isolate of Mtb carrying an unstable, "replication clock" plasmid and followed the dynamics of bacteria over time. Interestingly, previous methods developed to estimate Mtb replication and death rates using similar data for Mtb infection of mice failed to describe our novel data on Mtb dynamics in rabbits; we showed that heterogeneous dynamics of Mtb in semi-independent subpopulations in lungs of Mtb-infected rabbits may be one explanation of this failure of the method. Our results highlight potential differences in Mtb dynamics in different mammalian hosts and suggest ways to evaluate heterogeneity of Mtb replication and death rates in vivo.

The efficacy of antimicrobial solutions against multispecies bacterial biofilm with or without negative pressure wound therapy in an in vitro wound model.

Biofilm is the major challenge in chronic wound management. Instilling a wound cleansing solution aids in wound bed cleaning and infectious pathogen elimination. Negative pressure wound therapy (NPWT) improves the wound-healing process. This study investigated the efficacy of two antimicrobials (Vashe Wound Cleanser and Prontosan Wound Irrigation Solution) against a multispecies bacterial biofilm with or without NPWT in an in vitro wound model. A mixed multispecies biofilm containing Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, and Acinetobacter baumannii was developed and verified by scanning electron microscopy and fluorescent in situ hybridization. The efficacy of Vashe and Prontosan against multispecies biofilm with or without NPWT was evaluated by colony-forming unit (cfu) of each species and total bacterial number, and visually confirmed by live/dead stain and confocal microscopy. Prontosan reduced biofilm cell numbers significantly: 6 instils over 24 h resulting in 3.86 ± 0.14 cfu log10 reduction without NPWT and 4.75 ± 0.13 cfu log10 reduction combined with NPWT (P < 0.01) and 12 instils over 48 h resulting in 5.24 ± 0.11 cfu log10 reduction without NPWT and biofilm eradication with NPWT (P < 0.001). NPWT alone or combined with Vashe failed to reduce multispecies biofilm numbers significantly over 24 or 48 h. Prontosan significantly reduced biofilm cell numbers, with better efficacy over 48 than 24 h, emphasizing the necessity for persistent and robust treatment. NPWT enhanced the effectiveness of Prontosan instillation. However, NPWT alone or combined with Vashe showed limited efficacy and difficulty when combating the multispecies biofilm in vitro.

Lag Plot: A Novel Method to Determine Viable Bacterial Cell Number via a Single OD Measurement.

Bactericidal activity is a valuable parameter which is considered and measured for antimicrobial compounds. The available standard protocol to evaluate bactericidal activity is based on the direct colony count. Colony counting requires serial dilution, plating, overnight incubation, and direct counting, which is time-and labor-intensive. In regard to eliminate direct plate count, novel techniques were developed based on the real-time growth monitoring which can come with some limitations and drawbacks. These drawbacks encourage us to develop a novel technique with simple procedure to determine viable bacterial cell count. In this procedure, real-time growth monitoring is not required. In fact, after an incubation time, the number of viable bacteria can be determined with a single OD measurement and through an equation. In this regard, four standard bacterial strains, including Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), and Bacillus subtilis (ATCC 23857), were cultured with descending inoculum densities (1.5 × 107 to 15CFU/mL) and growth curves were drawn. As expected, growth in the samples with lower inoculum densities recorded with longer lag phase. Also, a direct relation was observed between the recorded turbidities and initial cell counts. A logarithmic curve (lag plot) was obtained by plotting the OD, after 12-18h incubation, against initial cell counts. In all examined strains, R2 was calculated in the range of 0.96-0.99 which is acceptable value for coefficient of determination. Equation corresponding to the lag plot was obtained and called lag equation. This equation is applicable to calculate the number of viable cells in unknown samples simply by inoculation, incubation, and single OD measurement. Developed technique can be introduced as a simple substitution for labor- and time-intensive direct colony counting.

The infectious capacity of Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus saprophyticus in a porcine model of urinary tract infection.

The purpose of this study was to establish a porcine model of urinary tract infection (UTI) with gram-positive uropathogens. Ten female domestic pigs were experimentally inoculated with human UTI isolates of Enterococcus faecalis (n = 3), Staphylococcus saprophyticus (n = 3), or Staphylococcus aureus (n = 4) and followed with regular urine samples. Bladders and kidneys were aseptically removed at termination (5-7 days post infection) and assessed by gross pathology and bacterial enumeration. Enterococcus faecalis (n = 3 of 3) and S. aureus (n = 2 of 4) successfully colonized the pig bladders. Inoculation with S. saprophyticus never resulted in detectable bacteriuria. All infected pigs had cleared the infection spontaneously before termination. Surprisingly, three (of four) pigs inoculated with S. aureus led to spontaneous infection with opportunistic pathogens. Also, one pig colonized with E. faecalis resulted in spontaneous infection with E. coli. In conlusion, the pig supports experimental UTI with E. faecalis for up to 24 h but not prolonged infection. S. aureus and S. saprophyticus fails to cause UTI in pigs and other animals should be considered for studying these pathogens.

PSV-12 Evaluating the effect of lyoprotectants in preserving community structure and bacterial viability in the lyophilized fecal community of pigs

Abstract Fecal microbiota transplantation (FMT) stands as a potential method for enhancing pig health. FMT has progressed from the basic delivery of fresh stool, to lyophilized preparations which is beneficial for sample storage, but can damage bacterial cells. This has necessitated the need for lyoprotectants to preserve bacterial viability. However, the ideal lyoprotectant for retaining community structure and viability remains unclear. Furthermore, DNA-based microbial profiling methods such as 16S rRNA and metagenomics sequencing do not distinguish live and dead bacteria, prompting the need for other methods of distinguishing live and dead members of the bacterial community after lyophilization. PMA-seq (propidium monoazide treatment followed by 16S rRNA gene amplicon sequencing) has been shown to differentiate between live and dead bacteria in microbial communities. To refine FMT in pigs, we sought to ascertain the impact of four lyoprotectants on bacterial viability and fecal microbiome structure. Fecal samples from six pigs were treated with mannitol, maltodextrin, trehalose, maltodextrin-trehalose mix (2.5% each) as lyoprotectants or left untreated. Lyophilized fecal samples underwent PMA treatment prior to 16S rRNA sequencing to capture the active community, and total and viable bacterial cells number were quantified. One-way ANOVA was used to analyze total and viable bacterial cells, and richness, while PERMANOVA was used to compare beta diversity. Results revealed no initial differences in total, viable bacteria, or community structure (P &amp;gt; 0.05). However, untreated, and mannitol-treated groups showed significantly reduced total and viable bacterial cells post-lyophilization (P &amp;lt; 0.01), unlike the other lyoprotectants (P &amp;gt; 0.05). Mannitol and the untreated group also displayed decreased richness and beta diversity shifts (P &amp;lt; 0.05). Overall, lyophilization with mannitol or no lyoprotectant notably impacted pig fecal microbiota community structure, while maltodextrin, trehalose, and maltodextrin-trehalose effectively preserved bacterial viability and structure.

Impact of different illegal Artisanal Small-Scale Mining techniques on soil properties in a major mining landscape in Ghana

The widespread prevalence of illegal Artisanal Small-Scale Mining has raised significant concerns about its environmental and health impacts. This study investigates the effects of two mining methods (alluvial and chamfi) on soil physical, chemical, and microbial properties four years after mining in three municipalities of the Western Region of Ghana. The study identified four species of Aspergillus fungi (niger, flavus, ochraceous and fumigatus) and Trichoderma species in both mined and control soils. The quantity of A. fumigatus was significantly higher in control plots compared to mined plots. In alluvial sites, bacterial numbers were significantly higher in control plots than in mined plots, while in chamfi sites, bacterial numbers did not differ significantly between mined and control plots. Soil texture in both mined sites was predominantly sandy loam and loamy sand, with a generally high silt to clay ratio. Mined plots at both sites were significant and more acidic (p < 0.05) than the control plots. Additionally, cation exchange capacity (CEC), electrical conductivity (EC), organic carbon (OC), total nitrogen (TN), phosphorus (P), potassium (K+), calcium (Ca2+), and magnesium (Mg2+) levels were significantly higher in control plots compared to mined plots. The control plots also exhibited higher base saturation and lower exchangeable acidity than mined plots. Bacteria count at Chamfi sites was generally higher at mined areas than the control sites, with the reverse being the case for Alluvial sites. Soil chemical parameters were higher at control areas than at mined sites, irrespective of the mining approach used. However, there was no clear pattern of differences in soil chemical properties at Chamfi and Alluvial mined sites. This study provides insights and additional perspectives for improving decision-making and guiding the potential restoration of these areas.

Role of the JAK2/STAT3 pathway on infection of Francisella novicida.

Francisella tularensis is a causative agent of the zoonotic disease tularemia, and is highly pathogenic to humans. The pathogenicity of this bacterium is largely attributed to intracellular growth in host cells. Although several bacterial factors important for the intracellular growth have been elucidated, including the type VI secretion system, the host factors involved in the intracellular growth of F. tularensis are largely unknown. To identify the host factors important for F. tularensis infection, 368 compounds were screened for the negative regulation of F. tularensis subsp. novicida (F. novicida) infection. Consequently, 56 inhibitors were isolated that decreased F. novicida infection. Among those inhibitors, we focused on cucurbitacin I, an inhibitor of the JAK2/ STAT3 pathway. Cucurbitacin I and another JAK2/STAT3 inhibitor, Stattic, decreased the intracellular bacterial number of F. novicida. However, these inhibitors failed to affect the cell attachment or the intrasaccular proliferation of F. novicida. In addition, treatment with these inhibitors destabilized actin filaments. These results suggest that the JAK2/STAT3 pathway plays an important role in internalization of F. novicida into host cells through mechanisms involving actin dynamics, such as phagocytosis.

Aerosolic Application of Phages Against S. infantis on Plates and Chicken Skin.

Phages are known as a promising method to combat antimicrobial resistance (AMR) in the human and veterinary sector. Use of phage aerosols enormously increases the application field, although the impact on the infectivity of phages during nebulization needs to be evaluated. In this study S. infantis was treated on plates and chicken skin with nebulized phage particles of the Myoviridae type, identified by transmission electron microscopy, using a commercial nebulizer primarily used for H2O2 disinfection. The reduction of bacterial number by aerosol applied phage particles was evaluated. It could clearly be shown that the phage particles were able to infect Salmonella after being nebulized using ultrasound technology. Further studies on other types of phages as well as other conditions must be performed to standardize the aerosolic application of phages.

Curcumin-mediated photodynamic disinfection strategy with specific spectral range for mucoid Pseudomonas Aeruginosa from hospital water

BackgroundHospital water systems represent critical environments for the transmission of pathogens, including multidrug-resistant strains like mucoid Pseudomonas aeruginosa (M-PA). Conventional disinfection methods often struggle to eradicate these pathogens effectively, highlighting the need for innovative approaches. ObjectiveThis study aimed to develop an enhanced photodynamic disinfection strategy targeting M-PA from hospital water systems, using curcumin-mediated photodynamic inactivation (PDI) with specific spectral range. MethodsAn M-PA strain isolated from hospital water was subjected to photodynamic treatment using curcumin as the photosensitizer. The efficacy of different wavelengths of light and varying concentrations of curcumin, with and without Tris-EDTA adjuvants, was evaluated through bacterial enumeration, ROS level measurements, transcriptome analysis, and assessment of virulence factors and biofilm formation. In vivo experiments utilizing a DSS-induced colitis mouse model assessed the protective effects of the photodynamic treatment against M-PA infection. ResultsOur findings demonstrated that the combination of curcumin-mediated PDI with specific spectral range effectively reduced M-PA counts in water, particularly when supplemented with Tris-EDTA. Transcriptome analysis revealed significant downregulation of virulence-related genes under sublethal photodynamic conditions. Furthermore, photodynamic treatment inhibited pyocyanin production and biofilm formation in M-PA, highlighting its potential to disrupt pathogenicity mechanisms. In vivo experiments showed that PDI attenuated M-PA-induced colitis in mice, indicating its protective efficacy. ConclusionThis study presents a promising photodynamic disinfection strategy for combating M-PA from hospital water. By optimizing curcumin-mediated PDI with specific spectral range and adjuvants, our approach demonstrates substantial efficacy in reducing bacterial counts, inhibiting virulence factors, and preventing M-PA-associated colitis.

Antimicrobial Coatings Based on a Photoreactive (Meth)acrylate Syrup and Ferulic Acid-The Effectiveness against Staphylococcus epidermidis.

A novel photopolymerizable (meth)acrylate oligomer syrup modified with ferulic acid (FA) was verified as an antimicrobial coating binder against a biofilm of Staphylococcus epidermidis. A solution-free UV-LED-initiated photopolymerization process of aliphatic (meth)acrylates and styrene was performed to prepare the oligomer syrup. The influence of ferulic acid on the UV crosslinking process of the varnish coatings (kinetic studies using photo-DSC) as well as their chemical structure (FTIR) and mechanical (adhesion, hardness), optical (gloss, DOI parameter), and antibacterial properties against S. epidermidis were investigated. The photo-DSC results revealed that FA has a positive effect on reducing the early occurrence of slowing processes and has a favorable effect on the monomer conversion increment. It turned out, unexpectedly, that the more FA in the coating, the greater its adhesion to a glass substrate and hardness. The coating containing 0.9 wt. part of FA exhibited excellent antimicrobial properties against S. epidermidis, i.e., the bacterial number after 24 h was only 1.98 log CFU/mL. All the coatings showed relatively high hardness, gloss (>80 G.U.), and DOI parameter values (30-50 a.u.).

Long-term measurements reveal a 100-day lag between peaks in phytoplankton chlorophyll and benthic bacterial abundance in the Fram Strait

Abstract Repeated measurements of benthic and pelagic parameters in the rapidly changing Arctic Ocean provide a unique insight into spatial and interannual trends and changes in the ecosystem. Here, we compiled biogenic and biogeochemical measurements collected from sediment cores at the Long-Term Ecological Research Observatory HAUSGARTEN located in the Fram Strait. A total of 21 stations were visited yearly over a period of 18 years (2002–2019). The time series highlighted an increase in bacterial numbers for samples collected 50 days after the peak phytoplankton bloom. Although bacterial abundances were not bathymetric depth-dependent when viewed across all years, we observed a seasonal trend in benthic microbial abundance closely related to the timing of the phytoplankton bloom with a time-lag of 100 days between the surface phytoplankton peak and the peak in bacterial abundance in the sediment. Considering the residence time of phytoplankton in the upper ocean and the water depth, we estimated an average settling velocity for phytodetritus of 30 m.d−1, which is similar to previous observations from Fram Strait. This suggests that settling organic matter promotes vertical microbial connectivity and benthic bacterial abundance in the deep ocean, shaping the microbial biogeography, diversity, and biogeochemical processes.

Application of miniaturized most probable number method for bacterial detection in water samples: detection of multi-drug-resistant Ralstonia insidiosa in drinking water.

The detection of bacterial contamination in drinking water is essential for monitoring the spread of foodborne diseases. We developed a simple, portable, and low-cost method of mini most probable number (mini MPN) to semi-enumerate bacterial suspension in water as a drinking water analogue. In this study, there is no significant difference between mini MPN and the standard method, technique plate count (TPC), at 10 and 100 CFU/ml Klebsiella pneumoniae suspension with a P-value of 0.28. For the ease-of-use aspect of this method, we tested several variables to prove it can be mass-applied in society. The usage of a sterile-plastic pipette, sample inoculation conducted in a biosafety cabinet (BSC), the usage of a 3-month storage medium, and incubation temperature conducted at room temperature compared to aseptic standard laboratory technique showed P-value > 0.05. In a trial for this method, we used commercialized drinking water for bacterial enumeration and characterization. We found multi-drug resistant (MDR) Ralstonia insidiosa which was resistant to at least four antimicrobial classes, including aminoglycosides, penicillins, cephalosporin, and carbapenem. Vitek 2 Compact was used for bacterial identification and antimicrobial susceptibility testing. A virulence test in Omphisa fuscidentalis larvae showed R. insidiosa strain D had a low virulence.

Combined treatment with vitamin C, hydrocortisone and thiamine does not attenuate morbidity and mortality of septic sheep

BackgroundSepsis is associated with a highest mortality rate in the ICU. Present study tests the efficacy of combined therapy with vitamin C, hydrocortisone and thiamine (combined therapy) in the ovine model of sepsis induced by Pseudomonas aeruginosa. In this study, sepsis was induced in sheep by instillation of Pseudomonas aeruginosa (1 × 1011 CFU) into the lungs via bronchoscope, under anesthesia. Nine hours after injury, intravenous infusion of vitamin C (0.75 g every 6 h), hydrocortisone (25 mg every 6 h), and thiamine (100 mg every 12 h) or saline was given to the treatment and control groups. Cardiopulmonary variables were recorded.ResultsThe survival rate was 16.7% in control and 33.3% in treatment groups. In the control group, mean arterial pressure dropped from 93.6 ± 8.6 to 75.5 ± 9.7 mmHg by 9 h, which was not affected by the combined therapy. Pulmonary dysfunction was not attenuated by the combined therapy either. The combined therapy had no effect on increased extravascular lung water content and fluid effusion into thoracic cavity. The bacterial number in the bronchoalveolar lavage fluid was significantly increased in the treatment group than the control group. The blood bacterial number remained comparable between groups.ConclusionsCombined vitamin C, hydrocortisone, and thiamine did not attenuate severity of ovine sepsis.

ITO-Coated Vertical Nanowires-Based Biosensors for the Label-Free Detection of Escherichia coli

A major concern in public health is the direct in-situ detection of pathogenic bacteria in food, beverages, and the air [1]. This study utilizes ITO-coated vertical nanowires (ITO-VNWs) as sensing substrates for the detection of Escherichia coli (E. coli). The innovation in this research lies in the direct detection of various concentrations of E. coli using boronic acid (BA)-modified ITO-VNWs connected to a benchtop E4980A precision LCR meter. Impedance spectra were expressed in terms of real and imaginary parts, corresponding to resistance (R) and reactance (X). Serial dilutions of E. coli were prepared in saline for bacterial enumeration and tested using the standard plate count method. Subsequently, 100 μl of each bacterial dilutions (107, 106, 105, 104, 103, 102, and 101 CFU/ml) were applied to BA-modified ITO-VNWs. In summary, BA-modified ITO-VNWs demonstrated accurate and distinct detection of varying concentrations of E. coli. This accomplishment highlights the promising potential of our BA-modified ITO-VNW biosensor for precise bacterial detection across diverse applications. Further exploration and optimization of this platform could lead to advancements in the rapid and specific identification of pathogenic bacteria, making it an invaluable tool in the realm of public health and environmental safety. REFERENCES [1] A. Ahmed, J.V. Rushworth, N.A. Hirst, P.A. Millner, Biosensors for whole-cell bacterial detection, Clin Microbiol Rev 27(3) (2014) 631-46. Figure 1