Viability of Chlamydia trachomatis in different anatomical sites - a systematic review & meta-analysis.

BACKGROUND: Modern assays for the detection of Chlamydia trachomatis (CT) rely on nucleic acid amplification testing (NAAT) of DNA or ribosomal RNA. However, it is also known that both viable ("living") & non-viable ("dead") CT can be detected by NAAT. Multiple laboratory techniques to measure CT viability have emerged. METHODS: We searched PubMed, EMBASE, Scopus and Dimensions as well as conference abstracts for entries between January 2000 to May 2023. We included any studies that measured CT viability among NAAT-positive samples. Viability assays include enhanced cell culture, direct fluorescent antibody (DFA), messenger RNA (mRNA) detection via digital droplet PCR (ddPCR), viability PCR (V-PCR) & real-time PCR measuring RNA-to-DNA ratio (RDR) (e.g. InSignia®). A meta-analysis was performed on the proportions of non-viable CT by anatomical site. RESULTS: We screened 31,342 records and included 16 studies in the analysis. The pooled proportions of non-viable CT by site were: 33% (95%CI 19-47%) in rectal swabs (eight studies), 17% (95%CI 7-27%) in cervical swabs (six studies), 15% (95%CI 6-25%) in vaginal swabs (six studies) and 11% (95%CI 9-17%) in urine/urethral swabs (two studies). CONCLUSION: All included studies found that a proportion of NAAT-detected CT is non-viable. The findings have far-reaching implications for screening programs and studies evaluating new STI tests and antimicrobial regimens.

Effectiveness of Umonium38 against Burkholderia pseudomallei, Escherichia coli, Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus (MRSA).

AIMS: We investigated the antibacterial efficacy of Umonium38 and Virkon® against Burkholderia pseudomallei, Escherichia coli, Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus (MRSA) up to 14 days following treatment. METHODS AND RESULTS: Umonium38 was diluted to 0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 3%, tested against the bacterial strains at various contact times (15 min to 24 h), and incubated for up to 14 days. A minimum concentration of 0.5% Umonium38 with a contact time of 15 min effectively killed approximately 108 CFU/ml of all four bacterial species. No growth was observed on agar plates from day 0 until day 14 for all six concentrations. The bacteria were also inactivated by a 30-minute treatment time using Virkon® 1% solution. CONCLUSIONS: Umonium38 effectively inactivates B. pseudomallei, E. coli, P. aeruginosa and MRSA at a concentration of ≥ 0.5% with a contact time of at least 15 min. The antimicrobial effect of Umonium38 remained for 14 days.

Rapid detection of viable microbes with 5-cyano-2,3-di-(p-tolyl)tetrazolium chloride and 5(6)-carboxyfluorescein diacetate using a fibre fluorescence spectroscopy system.

AIMS: To assess the efficacy of two commercially available viability dyes, 5-cyano-2,3-di-(p-tolyl)tetrazolium chloride (CTC) and 5(6)-carboxyfluorescein diacetate (CFDA), in reporting on viable cell concentration and species using an all-fibre fluorometer. METHODS AND RESULTS: Four bacterial species (two Gram-positive and two Gram-negative) commonly associated with food poisoning or food spoilage (Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Bacillus cereus) were stained with CTC or CFDA and the fibre fluorometer was used to collect full fluorescence emission spectra. A good correlation between concentration and fluorescence intensity was found for Gram-negative bacteria between 107 and 108 colony-forming units (CFU) ml-1. There was no correlation with concentration for Gram-positive bacteria; however, the information in the CTC and CFDA spectra shows the potential to distinguish Gram-negative cells from Gram-positive cells, although it may simply reflect the overall bacterial metabolic activity under staining conditions from this study. CONCLUSIONS: The limit of detection (LoD) is too high in the dip-probe approach for analysis; however, the development of an approach measuring the fluorescence of single cells may improve this limitation. The development of new bacteria-specific fluorogenic dyes may also address this limitation. The ability to differentiate bacteria using these dyes may add value to measurements made to enumerate bacteria using CTC and CFDA.

Assessment of Faecal Microbiota Transplant Stability in Deep-Freeze Conditions: A 12-Month Ex Vivo Viability Analysis.

BACKGROUND: Faecal microbiota transplantation (FMT) is an established treatment for Clostridioides difficile infection and is under investigation for other conditions. The availability of suitable donors and the logistics of fresh stool preparation present challenges, making frozen, biobanked stools an attractive alternative. AIMS: This study aimed to evaluate the long-term viability of bacterial populations in faecal samples stored at -80°C for up to 12 months, supporting the feasibility of using frozen grafts for FMT. METHODS: Fifteen faecal samples from nine healthy donors were processed, mixed with cryoprotectants and stored at -80°C. Samples were assessed at baseline and after 3, 6 and 12 months using quantitative culturing methods to determine the concentration of live bacteria. RESULTS: Quantitative analysis showed no significant decrease in bacterial viability over the 12-month period for both aerobic and anaerobic cultures (p = 0.09). At all timepoints, the coefficients of variability in colony-forming unit (CFU) counts were greater between samples (102 ± 21% and 100 ± 13% for aerobic and anaerobic cultures, respectively) than the variability between measurements of the same sample (30 ± 22% and 30 ± 19%). CONCLUSIONS: The study confirmed that faecal microbiota can be preserved with high viability in deep-freeze storage for up to a year, making allogenic FMT from biobanked samples a viable and safer option for patients. However, a multidonor approach may be beneficial to mitigate the risk of viability loss in any single donor sample.

Cryopreservation of stool samples altered the microbial viability quantitively and compositionally.

Stool is the most commonly used sample for gut microbiota analysis in humans and animals. Cryopreservation of stool at - 80 °C is a feasible and simple method in clinics and researches, especially in large-scale cohort studies. However, the viability of bacteria in stool after freezing has yet well-demonstrated quantitatively and compositionally. This study determined the viable microbiota of samples under cryopreservation at - 80 °C, relative to fresh samples and that stored at ambient. Stool samples were collected from three healthy adults. Propidium monoazide treatment combined with quantitative PCR and 16S rRNA gene sequencing was performed to target viable microbiota. After freezing, the number of viable bacteria decreased, though inter-individual difference existed. Notably, the alpha diversity of viable microbiota after freezing did not change significantly, while its composition changed. Freezing significantly reduced the viable bacteria in Gram-negative genera of Bacteroidetes and Firmicutes, and proportionally increased Gram-positive bacteria in genera of Actinobacteria and Firmicutes, including Bifidobacterium, Collinsella and Blautia, implying that the cell envelope structure associated with the bacterial sensitivity to freezing. On the contrary, the room temperature storage not only decreased the number of viable bacteria, but also decreased the microbial alpha diversity, and remarkably enriched facultative anaerobes of Escherichia-Shigella, Enterococcus and Lactococcus, some of which are opportunistic pathogens. Our findings suggested that changes in viable microbiota in stool samples caused by cryopreservation should be paid enough attention for downstream utilization.

Antibiotic Resistance Influences Growth Rates and Cross-Tolerance to Lactic Acid in Escherichia coli O157:H7 H1730.

Escherichia coli O157:H7-contaminated beef has been implicated in numerous foodborne outbreaks. Contamination occurs despite the use of antimicrobial interventions such as lactic acid (LA). In addition, resistance to antibiotics such as ampicillin and streptomycin among isolates has been frequently reported. The influence of antibiotic resistance (ABR) on growth rates and cross-tolerance of lettuce isolate E. coli O157:H7 H1730 to LA was evaluated. Antibiotic-resistant strain variants were generated by conferring resistance to either ampicillin (ampC) or streptomycin (strepC) or both ampicillin and streptomycin (ampC strepC) through incremental exposure to the antibiotics. Ampicillin resistance was also conferred by plasmid transformation to generate the ampP and ampP strepC strains. The minimum inhibitory concentration of LA on all the strains evaluated was 0.375% v/v. The lag phase duration of all strains except E. coli O157:H7 ampP strepC increased with increasing concentration of LA. The ampP strepC and ampC strains were most tolerant to 5% LA with declines in the cell population of 2.86 and 2.56 log CFU/mL, respectively (p < 0.05). The ampP strepC strain was the most tolerant when evaluated by the live/dead viability assay. The addition of the efflux pump inhibitor, carbonyl cyanide m-chlorophenylhydrazone, with 2.5% LA resulted in a significant increase in sensitivity in the no resistance (NR) wild-type and ampC strains, resulting in 6.62 and 6.65 log CFU/mL reduction, respectively, while the highly tolerant ampP strepC strain had a 2.90 log CFU/mL decrease. Tolerance to LA was significantly influenced by both the ABR profile of the strain and LA concentration. The results from this study indicate that E. coli O157:H7 strains with certain ABR profiles might be more tolerant to LA.

One-pot facile synthesis of enzyme-encapsulated Zn/Co-infinite coordination polymer nanospheres as a biocatalytic cascade platform for colorimetric monitoring of bacteria viability.

A rapid method for colorimetric monitoring of bacterial viability is described. The colorimetric method was carried out based on glucose oxidase-encapsulated Zn/Co-infinite coordination polymer (Zn/Co-ICP@GOx), which was prepared in aqueous solution free of toxic organic solvents at room temperature. The Zn/Co-ICP@GOx was confirmed to be a robust sphere structure with an average diameter of 147.53 ± 20.40 nm. It integrated the catalytic activity of natural enzyme (GOx) and mimetic peroxidase (Co (П)) all in one, efficiently acting as a biocatalytic cascade platform for glucose catalytic reaction. Exhibiting good multi-enzyme catalytic activity, stability, and selectivity, Zn/Co-ICP@GOx can be used for colorimetric glucose detection. The linear range was 0.01-1.0 mmol/L, and the limit of detection (LOD) was 0.005 mmol/L. As the glucose metabolism is a common expression of bacteria, the remaining glucose can indirectly represent the bacterial viability. Hence, a Zn/Co-ICP@GOx-based colorimetric method was developed for monitoring of bacterial viability. The color was intuitively observed with the naked eye, and the bacterial viability was accurately quantified by measurement of the absorbance at 510 nm. The method was applied to determination of bacterial viability in water and milk samples with recoveries of 99.0-103% and RSD of 0.43-7.5%. The method was rapid (less than 40 min) and applicable to different bacterial species irrespective of Gram-positive and Gram-negative bacteria, providing a universal and promising strategy for real-time monitoring of bacterial viability.

Experimental confirmation of antimicrobial effects of GdYVO4:Eu3+ nanoparticles.

Nanotechnology can be applied to design antibacterial agents to combat antibiotic resistance. The aim of the present study was to assess the antimicrobial effects and cytotoxicity of GdYVO4:Eu3+ nanoparticles (NPs). Biofilm inhibition activity, antimicrobial activity, bacterial viability inhibition and DNA cleavage activity of GdYVO4:Eu3+ NPs were studied. In addition, the impact of GdYVO4:Eu3+ NPs on the mitochondrial membrane potential (ΔΨM) of host immune cells and, hence, their apoptosis was analyzed by JC-1 staining using flow cytometry. GdYVO4:Eu3+ NPs demonstrated good antimicrobial, cell viability inhibition and DNA cleavage activities. In addition, GdYVO4:Eu3+ NPs showed good biofilm inhibition activity against S. aureus and P. aeruginosa and inhibition percentages were 89.15% and 79.54%, respectively. However, GdYVO4:Eu3+ NPs promoted mitochondrial depolarization and apoptosis of leukocytes at high concentrations. GdYVO4:Eu3+ nanoparticles are promising antibacterial agents. However, more efforts should be exerted to ensure their safety.

Transformative Approach To Investigate the Microphysical Factors Influencing Airborne Transmission of Pathogens.

Emerging outbreaks of airborne pathogenic infections worldwide, such as the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, have raised the need to understand parameters affecting the airborne survival of microbes in order to develop measures for effective infection control. We report a novel experimental strategy, TAMBAS (tandem approach for microphysical and biological assessment of airborne microorganism survival), to explore the synergistic interactions between the physicochemical and biological processes that impact airborne microbe survival in aerosol droplets. This innovative approach provides a unique and detailed understanding of the processes taking place from aerosol droplet generation through to equilibration and viability decay in the local environment, elucidating decay mechanisms not previously described. The impact of evaporation kinetics, solute hygroscopicity and concentration, particle morphology, and equilibrium particle size on airborne survival are reported, using Escherichia coli MRE162 as a benchmark system. For this system, we report that (i) particle crystallization does not directly impact microbe longevity, (ii) bacteria act as crystallization nuclei during droplet drying and equilibration, and (iii) the kinetics of size and compositional change appear to have a larger effect on microbe longevity than the equilibrium solute concentration.IMPORTANCE A transformative approach to identify the physicochemical processes that impact the biological decay rates of bacteria in aerosol droplets is described. It is shown that the evaporation process and changes in the phase and morphology of the aerosol particle during evaporation impact microorganism viability. The equilibrium droplet size was found to affect airborne bacterial viability. Furthermore, the presence of Escherichia coli MRE162 in a droplet does not affect aerosol growth/evaporation but influences the dynamic behavior of the aerosol by processing the culture medium prior to aerosolization, affecting the hygroscopicity of the culture medium; this highlights the importance of the inorganic and organic chemical composition within the aerosolized droplets that impact hygroscopicity. Bacteria also act as crystallization nuclei. The novel approach and data have implications for increased mechanistic understanding of aerosol survival and infectivity in bioaerosol studies spanning the medical, veterinary, farming, and agricultural fields, including the role of microorganisms in atmospheric processing and cloud formation.

Immobilization of Phosphatidylserine by Ethanol and Lysozyme on the Cell Surface for Evaluation of Apoptosis-Like Decay in Activated-Sludge Bacteria.

Accurate determination of microbial viability can be crucial in microbe-dominated biosystems. However, the identification of metabolic decay in bacterial cells can be elaborate and difficult. We sought to identify apoptosis-like bacterial processes by using annexin V-fluorescein isothiocyanate (FITC) (AVF), a probe typically used to stain phosphatidylserine (PS) on exposed cell membranes. The bacterial cell wall provides a barrier that is responsible for low efficiency of direct PS staining of decayed bacterial cells. This can be overcome by pretreatment of the bacteria with 70% ethanol, which fixates the bacteria and preserves the PS status, combined with lysozyme treatment to hydrolyze the cell wall. That treatment improved the efficiency of AVF staining considerably, as shown for pure strains of an Ochrobactrum sp. and a Micrococcus sp. Using this method, decayed bacterial cells (induced by starvation) were more strongly stained, indicating externalization of PS to a greater extent than seen for cells harvested at logarithmic growth. A multispecies microbial sludge was artificially decayed by heat treatment or alternating anoxic-oxic treatment, which also induced increased AVF staining, again presumably via decay-related PS externalization. The method developed proved to be efficient for identification of bacterial decay and has potential for the evaluation of multispecies bacterial samples from sources like soil matrix, bioaerosol, and activated sludge.IMPORTANCE Since the externalization of phosphatidylserine (PS) is considered a crucial characteristic of apoptosis, we sought to identify apoptosis-like decay in bacterial cells by PS staining using AVF. We show that this is possible, provided the bacteria are pretreated with ethanol plus lysozyme to remove a physical staining barrier and preserve the original, decay-related externalization of PS. Our work suggests that PS externalization occurs in starved bacteria and this can be quantified with AVF staining, providing a measure of bacterial decay. Since PS is the common component of the lipid bilayer in bacterial cell membranes, this approach also has potential for evaluation of cell decay of other bacterial species.

Bacterial survival and adhesion for formulating new oral probiotic foods.

Probiotics are defined as live microorganisms, which, when administered in adequate amounts, confer health benefits to the host. Traditionally, probiotic food research has heavily focused on the genera Bifidobacteria and Lactobacilli, along with their benefits for gut health. Recently with the identification of new probiotic strains specifically intended for oral health applications, the development of probiotic foods for oral health benefits has garnered interest, with a renewed focus on identifying new food formats for delivering probiotics. The development of novel oral probiotic foods is highly complex, as the composition of a food matrix dictates: (1) bacterial viability during production and shelf life and (2) how bacteria partition with components within a food matrix and subsequently adhere to oral cavity surfaces. At present, virtually no information is available on oral probiotic strains such as Streptococcus salivarius; specifically, how orally-derived strains survive under different food parameters. Furthermore, limited information exists on the partition behavior of probiotics with food components, governed by physico-chemical interactions and adhesion phenomena. This review aspires to examine this framework by providing a foundation with existing literature related to the common probiotic genera, in order to inform and drive future attempts of designing new oral probiotic food formats.

Development and evaluation of a quantitative polymerase chain reaction for aquatic Streptococcus agalactiae based on the groEL gene.

AIMS: The aim of this study was to develop a TaqMan quantitative polymerase chain reaction (qPCR), based on the Streptococcus agalactiae groEL gene, to specifically quantify levels of bacteria within samples derived from aquatic sources, particularly aquaculture. Enumeration of bacteria by qPCR was compared with culture-based methods. METHODS AND RESULTS: The qPCR was sensitive to 33 isolates of S. agalactiae, representing 11 clonal complexes from aquatic, bovine and human hosts. The specificity of the assay was 92·5% at a threshold Cq value of 35. No cross-reaction with Streptococcus iniae was noted and of the 22 comparator species screened to test assay specificity, Streptococcus porcinus had a Cq value of 33·7 S, while Streptococcus gallolyticus subsp. macedonicus and Streptococcus ictaluri had one replicate value above the Cq threshold of 35 (34·5 and 34·4 respectively), while only S. agalactiae were detected with a Cq value of 30. The limit of detection of the assay was 1·7 copies per µl at Cq 35. Discrepancies between molecular and culture-based methods of enumeration were noted. CONCLUSIONS: The qPCR was able to detect a diverse range of S. agalactiae isolates from different clonal complexes (CCs) and could distinguish between S. agalactiae and closely related species, notably S. iniae. The results suggest that a Cq 30 would be a very meaningful cut-off, allowing the detection of infected fish while ruling out all false positives. SIGNIFICANCE AND IMPACT OF THE STUDY: This rapid and sensitive qPCR assay is useful to quantify DNA copy number in the laboratory and could prove useful for detecting low levels of S. agalactiae in aquaculture systems, including Oreochromis niloticus culture.

Interdigitated and Wave-Shaped Electrode-Based Capacitance Sensor for Monitoring Antibiotic Effects.

Label-free and real-time monitoring of the bacterial viability is essential for the accurate and sensitive characterization of the antibiotic effects. In the present study, we investigated the feasibility of the interdigitated and wave-shaped electrode (IWE) for monitoring the effect of tetracycline or kanamycin on Staphylococcus aureus (S. aureus) and methicillin-resistant S.aureus (MRSA). The electrical impedance spectra of the IWE immersed in the culture media for bacterial growth were characterized in a frequency range of 10 Hz to 1 kHz. The capacitance index (CI) (capacitance change relevant with the bacterial viability) was used to monitor the antibiotic effects on the S. aureus and MRSA in comparison to the traditional methods (disk diffusion test and optical density (OD) measurement). The experimental results showed that the percentage of change in CI (PCI) for the antibiotic effect on MRSA was increased by 51.58% and 57.83% in kanamycin and control, respectively. In contrast, the PCI value decreased by 0.25% for tetracycline, decreased by 52.63% and 37.66% in the cases of tetracycline and kanamycin-treated S. aureus, and increased 2.79% in the control, respectively. This study demonstrated the feasibility of the IWE-based capacitance sensor for the label-free and real-time monitoring of the antibiotic effects on S.aureus and MRSA.

Implications of Chemical Reduction Using Hydriodic Acid on the Antimicrobial Properties of Graphene Oxide and Reduced Graphene Oxide Membranes.

The antimicrobial properties of graphene-based membranes such as single-layer graphene oxide (GO) and modified graphene oxide (rGO) on top of cellulose ester membrane are reported in this study. rGO membranes are made from GO by hydriodic acid (HI) vapor treatment. The antibacterial properties are tested after 3 h contact time with selected model bacteria. Complete bacterial cell inactivation is found only after contact with rGO membranes, while no significant bacterial inactivation is found for the control i) GO membrane, ii) the mixed cellulose ester support, and the iii) rGO membrane after additional washing that removes the remaining HI. This indicates that the antimicrobial effect is neither caused by the graphene nor the membrane support. The antimicrobial effect is found to be conclusively linked to the HI eliminating microbial growth, at concentrations from 0.005%. These findings emphasize the importance of caution in the reporting of antimicrobial properties of graphene-based surfaces.

High-resolution novel method for tracking bacteria in a multi-species biofilm.

The aim of this study is to establish a novel high resolution tracking ability of a specific bacterium in multispecies biofilm. A periodontal multispecies biofilm was constructed with Streptococcus sanguis, Actinomyces naeslundii, Porphyromonas gingivalis and Fusobacterium nucleatum. A single species was stained with fluorescein isothiocyanate (FITC). The mature biofilm was stained for viability (propidium iodide) and analysis was performed with flow cytometry. The sensitivity of the assay was compared with colony forming units (CFU) counts. A single cell suspension of P. gingivalis was grown in broth and biofilm to identify the location of these events on side scatter and forward scatter. The sensitivity of the assay was comparable to that of the CFU counts. The assay allows quantification of the ratio of a single bacterium within the biofilm, and its viable proportion. The described method is reproducible and of high resolution, and allows the examination of microbes' composition and viability within a biofilm structure.

Rapid and cost-effective evaluation of bacterial viability using fluorescence spectroscopy.

A rapid and easy method that takes advantage of an inexpensive and portable fibre-based spectroscopic system (optrode) to determine the ratio of live to dead bacteria is proposed. Mixtures of live and dead Escherichia coli with proportions of live:dead cells varying from 0 to 100% were stained using SYTO 9 and propidium iodide (PI) and measured using the optrode. We demonstrated several approaches to obtaining the proportions of live:dead E. coli in a mixture of both live and dead, from analyses of the fluorescence spectra collected by the optrode. To find a suitable technique for predicting the percentage of live bacteria in a sample, four analysis methods were assessed and compared: SYTO 9:PI fluorescence intensity ratio, an adjusted fluorescence intensity ratio, single-spectrum support vector regression (SVR) and multi-spectra SVR. Of the four analysis methods, multi-spectra SVR obtained the most reliable results and was able to predict the percentage of live bacteria in 108 bacteria/mL samples between c. 7 and 100% live, and in 107 bacteria/mL samples between c. 7 and 73% live. By demonstrating the use of multi-spectra SVR and the optrode to monitor E. coli viability, we raise points of consideration for spectroscopic analysis of SYTO 9 and PI and aim to lay the foundation for future work that uses similar methods for different bacterial species.

Impacts of indoor surface finishes on bacterial viability.

Microbes in indoor environments are constantly being exposed to antimicrobial surface finishes. Many are rendered non-viable after spending extended periods of time under low-moisture, low-nutrient surface conditions, regardless of whether those surfaces have been amended with antimicrobial chemicals. However, some microorganisms remain viable even after prolonged exposure to these hostile conditions. Work with specific model pathogens makes it difficult to draw general conclusions about how chemical and physical properties of surfaces affect microbes. Here, we explore the survival of a synthetic community of non-model microorganisms isolated from built environments following exposure to three chemically and physically distinct surface finishes. Our findings demonstrated the differences in bacterial survival associated with three chemically and physically distinct materials. Alkaline clay surfaces select for an alkaliphilic bacterium, Kocuria rosea, whereas acidic mold-resistant paint favors Bacillus timonensis, a Gram-negative spore-forming bacterium that also survives on antimicrobial surfaces after 24 hours of exposure. Additionally, antibiotic-resistant Pantoea allii did not exhibit prolonged retention on antimicrobial surfaces. Our controlled microcosm experiment integrates measurement of indoor chemistry and microbiology to elucidate the complex biochemical interactions that influence the indoor microbiome.

Potential acute effects of suspended aluminum nitride (AlN) nanoparticles on soluble microbial products (SMP) of activated sludge.

The study aims to identify the potential acute effects of suspended aluminum nitride (AlN) nanoparticles (NPs) on soluble microbial products (SMP) of activated sludge. Cultured activated sludge loaded with 1, 10, 50, 100, 150 and 200mg/L of AlN NPs were carried out in this study. As results showed, AlN NPs had a highly inverse proportionality to bacterial dehydrogenase and OUR, indicating its direct toxicity to the activated sludge viability. The toxicity of AlN NPs was mainly due to the nano-scale of AlN NPs. In SMP, AlN NPs led to the decrease of polysaccharide and humic compounds, but had slight effects on protein. The decrease of tryptophan-like substances in SMP indicated the inhibition of AlN NPs on the bacterial metabolism. Additionally, AlN NPs reduced obviously the molecular weight of SMP, which might be due to the nano-scale of AlN.

Efficacy of the novel oxazolidinone compound FYL-67 for preventing biofilm formation by Staphylococcus aureus.

OBJECTIVES: Infections of hospitalized patients caused by biofilms formed by Staphylococcus aureus represent a major problem. Using in vitro and in vivo biofilm models, we evaluated the efficacy of the novel oxazolidinone FYL-67, by using linezolid (the only clinically approved oxazolidinone antibiotic) as a control, for inhibiting S. aureus biofilm formation. METHODS: Antibiofilm activity was determined using strains of methicillin-susceptible S. aureus and methicillin-resistant S. aureus. We studied the mechanism(s) and pharmacodynamics of antibiofilm activity as follows: (i) effects of pre- and post-exposure to FYL-67 or linezolid on biofilm formation; (ii) the effect of FYL-67 on biofilm structure; (iii) the role of FYL-67 in biofilm composition; (iv) effects on cell morphology; and (v) efficacy of FYL-67 and linezolid using an in vivo murine model of catheter infection. RESULTS: FYL-67 effectively inhibited biofilm formation using in vitro and in vivo assays. CONCLUSIONS: Our data suggest that oxazolidinone compounds, such as FYL-67, may serve as antibiofilm agents.

Bacterial viability differentially influences the immunomodulatory capabilities of potential host-derived probiotics in the intestinal epithelial cells of Atlantic cod Gadus morhua.

AIM: This study explored the effect of heat inactivation on the immunomodulatory capabilities of two potential host-derived probiotics (GP21 and GP12) on the intestinal epithelial cells (IEPC) derived from Atlantic cod. METHODS AND RESULTS: The cells were isolated from the four segments of the gut, namely anterior intestine (AI), mid-intestine (MI), posterior intestine (PI) and rectum (RC). The IEPC cultures were exposed to live or heat-inactivated form of GP21 and GP12 for 24 h. The expression profiles of bacterial defence genes and cytokine genes in the probiotics-exposed IEPCs showed differential patterns. Heat inactivation did not drastically affect the immunomodulatory properties of the probiotics, and this was explicitly typified by the stimulated expression of g-type lysozyme, hepcidin, transferrin and metallothionein in both forms of the bacteria. There was no distinct expression pattern of the interleukin genes during bacterial exposure. This was in contrast to the chemokines where the expression of these genes in IEPCs was down-regulated upon exposure to the heat-inactivated probiotics. Although heat inactivation did not drastically affect the immunomodulatory capabilities of the probiotics, the live form elicited higher immune responses in the IEPCs in most cases. CONCLUSION: This study showed that bacterial viability was a contributing influence, but not a major limiting factor on the immune-related functions of the host-derived probiotics in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: GP21 and GP12 are beneficial host-derived bacteria and could be utilized as candidate probiotics in cod aquaculture.