Monitoring the presence of pathogenic Bacillus spores is important for industrial applications, as well as necessary for ensuring human health. Bacillus thuringiensis is used as a biopesticide ...against several insect pests. Bacillus cereus spores are a significant cause of food poisoning, and Bacillus anthracis is a recognized biosecurity threat. Laboratory-based methods, such as polymerase chain reaction, enzyme-linked immunosorbent assay, or matrix-assisted laser desorption ionization spectroscopy provide sensitive detection of bacteria and spores, but the application of those methods for quasi-continuous environmental monitoring presents a significant challenge requiring frequent human intervention. To address this challenge, we developed a workstation for quasi-autonomous monitoring of water reservoirs for the presence of bacteria and spores, and designed and validated the functionality of a microprocessor-controlled module capable of repetitive collection and pre-concentration of spores in liquid samples tested with fiberglass (FG), polyether sulfone and polyvinylidene fluoride filters. The best results were obtained with FG filters delivering a 20× concentration of B. thuringiensis and B. cereus spores from saline suspensions. The successful 20× pre-concentration of Bacillus spores demonstrated with FG filters could be repeated up to 3 times when bleach decontamination is applied between filtrations. Taken together, our results demonstrate an attractive instrument suitable for semi-automated, quasi-continuous sampling and pre-processing of water samples for biosensing of bacterial spores originating from a complex environment.
Abstract The mechanisms of toxicity related to human hepatocellular carcinoma HepG2 cell exposures to cadmium telluride quantum dots (CdTe-QDs) were investigated. CdTe-QDs caused cytotoxicity in ...HepG2 cells in a dose- and time-dependent manner. Treated cells showed an increase in reactive oxygen species (ROS). Altered antioxidant levels were demonstrated by depletion of reduced glutathione (GSH), a decreased ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) and an increased NF-E2-related Factor 2 (Nrf2) activation. Enzyme assays showed that superoxide dismutase (SOD) activity was elevated whereas catalase (CAT) and glutathione-S-transferase (GST) activities were depressed. Further analyses revealed that CdTe-QD exposure resulted in apoptosis, indicated by changes in levels of caspase-3 activity, poly ADP-ribose polymerase (PARP) cleavage and phosphatidylserine externalization. Extrinsic apoptotic pathway markers such as Fas levels and caspase-8 activity increased as a result of CdTe-QD exposure. Involvement of the intrinsic/mitochondrial apoptotic pathway was indicated by decreased levels of B-cell lymphoma 2 (Bcl2) protein and mitochondrial cytochrome c, and by increased levels of mitochondrial Bcl-2-associated X protein (Bax) and cytosolic cytochrome c. Further, mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinases (Erk1/2), and p38 were all activated. Our findings reveal that CdTe-QDs cause oxidative stress, interfere with antioxidant defenses and activate protein kinases, leading to apoptosis via both extrinsic and intrinsic pathways. Since the effects of CdTe-QDs on selected biomarkers were similar or greater compared to those of CdCl2 at equivalent concentrations of cadmium, the study suggests that the toxicity of CdTe-QDs arises from a combination of the effects of cadmium and ROS generated from the NPs.
•Evidence for genetic variants associated with CIPN is currently inconclusive.•Several SNPs are of interest and warrant further study.•Clear and consistent definition of phenotype is crucial in ...further research.•Research consortia have aided phenotype standardisation in similar fields of study.•Creation of consortia in cancer drug toxicity may be beneficial to future research.
Chemotherapy-induced peripheral neuropathy (CIPN) can adversely affect completion of systemic anti-cancer treatment and cause long-term morbidity. Increasingly pharmacogenetic studies have been performed to explore susceptibility to this important adverse effect.
A systematic review was conducted to identify pharmacogenetic studies, assess their quality and findings and undertake meta-analysis where possible.
93 studies were included. Notable methodological issues included lack of standardisation and detail in phenotype definition and acknowledgement of potential confounding factors. Insufficient data was presented in many studies meaning only a minority could be included in meta-analysis showing mainly non-significant effects. Nonetheless, SNPs in CYP2C8, CYP3A4, ARHGEF10, EPHA and TUBB2A genes (taxanes), FARS2, ACYP2 and TAC1 (oxaliplatin), and CEP75 and CYP3A5 (vincristine) are of potential interest. These require exploration in large cohort studies with robust methodology and well-defined phenotypes.
Seeking standardisation of phenotype, collaboration and subsequently, individual-patient-data meta-analysis may facilitate identifying contributory SNPs which could be combined in a polygenic risk score to predict those most at risk of CIPN.
An experiment was executed to test the hypothesis that supplementation of dietary threonine (d-Thr), above NRC recommendation to diets containing poorly digestible protein source (PS) may compensate ...its detrimental effects on overall performance of broilers. In total, nine hundred 1-day-old mixed sex broilers (Ross-308) were randomly distributed over 6 (2 × 3) experimental diets comprising 5 replicates of 30 broilers each for 35 d. The experimental diets contain either soybean meal (SBM) or canola meal (CM) with 3 levels (100, 110, and 120% of NRC recommendation) of d-Thr. During the course of the trial (0 to 35 D), interactions (P < 0.05) between PS and d-Thr were observed for feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR), carcass, and gut health parameters. The broilers fed recommended level (100%) of d-Thr had 7 and 5% poorer FCR compared with those fed diets with 110 and 120% d-Thr, respectively. For villus height (VH), an interaction (P = 0.007) was found between PS and d-Thr level. Broilers consuming SBM diets had 22% longer villi, 10% deeper crypts, and 30% greater VH to crypt depth ratio (VCR) compared to those fed CM. The broilers fed 110% d-Thr diets had 9% lower crypt depth (CD) and 15% greater VCR compared with those fed diets containing NRC recommended levels. CM resulted in 9% lower protein digestibility with lower (P < 0.05) of some AA, whereas it was improved by 7% in broilers fed 120% d-Thr supplemented diets. The bursa and spleen weights were positively affected (P < 0.001) by PS. Threonine supplementation (10%) resulted in 25% greater thymus, 18% heavier bursa, and 30% greater infectious bursal disease titer. In conclusion, supplementation of d-Thr, above NRC recommendation, resulted in a better growth performance and carcass traits, improved ileal digestibility of protein and amino acids, better gut health, and immunity in broilers.
Bacillus thuringiensis (Bt) is used as a bioinsecticide since it effectively kills insect larvae. Bt is also genetically similar to Bacillus cereus (Bc), a well recognized foodborne human pathogen; ...they are both members of the Bacillus cereus group (BC group). Although approved Bt bioinsecticide products have been confirmed to be non-pathogenic to humans, close monitoring of Bt during dissemination is important for cost considerations and to limit impact on biodiversity towards nontarget organisms. As such, developing rapid, sensitive, and specific tools for quantitative detection of Bt spores during and following spray operations is highly desirable. The goals of this study were to investigate commercially available detection reagents for sensitivity and selectivity in detecting Bt spores, and then functionalize a surface of (001) GaAs used in photonic biosensing. To achieve these goals, we (1) screened commercial antibodies for their capacity to bind recombinant proteins from Bt spores, (2) screened antibodies and aptamers for their sensitivity and selectivity against Bt spores, and (3) tested the efficiency of selected antibodies and aptamers in capturing Bt spores on the surface of functionalized GaAs biochips. Seven genes encoding Bt spore proteins were cloned and expressed in Escherichia coli. The binding of each purified spore antigen was tested by commercially available polyclonal and monoclonal antibodies claimed to exclusively target spores. Of the seven targets, Bacillus collagen-like protein A, was the most abundant protein on Bt spores and demonstrated the strongest binding affinity to all test antibodies. The commercial antibodies (Abs) were also tested for specificity to BC Group versus non-BC Group spores. Three of six commercial antibodies showed selectivity to Bt spores, with recombinant Abs providing the most robust lower range of detection (102 to 6 × 103 spores/mL). The sensitivity and selectivity of three published DNA aptamer sequences demonstrated a wide range of detection sensitivity for Bt spores. Two of the three test aptamers also showed reasonable selectivity towards Bt spores while the third demonstrated reactivity to non-BC Group B. megaterium and B. subtilis. Of the reagents tested, a thiolated aptamer and llama recombinant Ab showed highest Bt spore capture efficiency as measured by spore coverage of the GaAs surface. These results confirm that the selected aptamer and llama rAb can be considered strong candidates for the development of GaAs-based biosensing devices.
Display omitted
•Bacillus thuringiensis (Bt) is a bio-larvicide used globally against insect pests.•Monitoring its environmental dissemination will reduce impact on biodiversity.•Antibodies and aptamers against Bt were screened for sensitivity and selectivity.•Antibody/aptamer functionalization was optimised for GaAs-based biosensors.•Results support development of a field-ready biosensor for monitoring Bt.
Several Acinetobacter strains have utility for biotechnology applications, yet some are opportunistic pathogens. We compared strains of seven Acinetobacter species (baumannii, Ab; calcoaceticus, Ac; ...guillouiae, Ag; haemolyticus, Ah; lwoffii, Al; junii, Aj; and venetianus, Av-RAG-1) for their potential virulence attributes, including proliferation in mammalian cell conditions, haemolytic/cytolytic activity, ability to elicit inflammatory signals, and antibiotic susceptibility. Only Ah grew at 10(2) and 10(4) bacteria/well in mammalian cell culture medium at 37°C. However, co-culture with colonic epithelial cells (HT29) improved growth of all bacterial strains, except Av-RAG-1. Cytotoxicity of Ab and Ah toward HT29 was at least double that of other test bacteria. These effects included bacterial adherence, loss of metabolism, substrate detachment, and cytolysis. Only Ab and Ah exhibited resistance to killing by macrophage-like J774A.1 cells. Haemolytic activity of Ah and Av-RAG-1 was strong, but undetectable for other strains. When killed with an antibiotic, Ab, Ah, Aj and Av-RAG-1 induced 3 to 9-fold elevated HT29 interleukin (IL)-8 levels. However, none of the strains altered levels of J774A.1 pro-inflammatory cytokines (IL-1β, IL-6 and tumor necrosis factor-α). Antibiotic susceptibility profiling showed that Ab, Ag and Aj were viable at low concentrations of some antibiotics. All strains were positive for virulence factor genes ompA and epsA, and negative for mutations in gyrA and parC genes that convey fluoroquinolone resistance. The data demonstrate that Av-RAG-1, Ag and Al lack some potentially harmful characteristics compared to other Acinetobacter strains tested, but the biotechnology candidate Av-RAG-1 should be scrutinized further prior to widespread use.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Pseudomonas species are opportunistically pathogenic to humans, yet closely related species are used in biotechnology applications. In order to screen for the pathogenic potential of strains ...considered for biotechnology applications, several Pseudomonas strains (P.aeruginosa (Pa), P.fluorescens (Pf), P.putida (Pp), P.stutzeri (Ps)) were compared using functional virulence and toxicity assays. Most Pa strains and Ps grew at temperatures between 28°C and 42°C. However, Pf and Pp strains were the most antibiotic resistant, with ciprofloxacin and colistin being the most effective of those tested. No strain was haemolytic on sheep blood agar. Almost all Pa, but not other test strains, produced a pyocyanin-like chromophore, and caused cytotoxicity towards cultured human HT29 cells. Murine endotracheal exposures indicated that the laboratory reference strain, PAO1, was most persistent in the lungs. Only Pa strains induced pro-inflammatory and inflammatory responses, as measured by elevated cytokines and pulmonary Gr-1 -positive cells. Serum amyloid A was elevated at ≥ 48 h post-exposure by only some Pa strains. No relationship was observed between strains and levels of peripheral leukocytes. The species designation or isolation source may not accurately reflect pathogenic potential, since the clinical strain Pa10752 was relatively nonvirulent, but the industrial strain Pa31480 showed comparable virulence to PAO1. Functional assays involving microbial growth, cytotoxicity and murine immunological responses may be most useful for identifying problematic Pseudomonas strains being considered for biotechnology applications.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Coral reefs are the most biodiverse of all marine ecosystems; however, very little is known about prokaryotic diversity in these systems. To address this issue, we sequenced over 1000 bacterial 16S ...rDNAs from 3 massive coral species (Montastraea franksi,Diploria strigosa, andPorites astreoides) in Panama and Bermuda. Analysis of only 14 coral samples yielded 430 distinct bacterial ribotypes. Statistical analyses suggest that additional sequencing would have resulted in a total of 6000 bacterial ribotypes. Half of the sequences shared <93% identity to previously published 16S sequences, and therefore probably represent novel bacterial genera and species; this degree of novelty was substantially higher than that observed for other marine samples. Samples from the Panama corals were more diverse than those from Bermuda, paralleling diversity gradients seen in metazoans. The coral-bacteria associations were non-random. Different coral species had distinct bacterial communities, even when physically adjacent, while bacterial communities from the same coral species separated by time (~1 yr) or space (3000 km) were similar. Analysis of the branching coralPorites furcatashowed that bacterial ribotypes can also be structured spatially within colonies. Therefore, corals and reefs represent landscapes of diverse, ecologically structured prokaryotic communities.
Some recent studies revealed that Himalayan glaciers were shrinking at an accelerated rate since the beginning of the 21st century. However, the climatic causes for this shrinkage remain unclear ...given that surface energy balance studies are almost nonexistent in this region. In this study, a point-scale surface energy balance analysis was performed using in situ meteorological data from the ablation zone of Chhota Shigri Glacier over two separate periods (August 2012 to February 2013 and July to October 2013) in order to understand the response of mass balance to climatic variables. Energy balance numerical modelling provides quantification of the surface energy fluxes and identification of the factors affecting glacier mass balance. The model was validated by comparing the computed and observed ablation and surface temperature data. During the summer-monsoon period, net radiation was the primary component of the surface energy balance accounting for 80 % of the total heat flux followed by turbulent sensible (13%), latent (5%) and conductive (2%) heat fluxes. A striking feature of the energy balance is the positive turbulent latent heat flux, suggesting re-sublimation of moist air at the glacier surface, during the summer-monsoon characterized by relatively high air temperature, high relative humidity and a continual melting surface. The impact of the Indian Summer Monsoon on Chhota Shigri Glacier mass balance has also been assessed. This analysis demonstrates that the intensity of snowfall events during the summer-monsoon plays a key role on surface albedo (melting is reduced in the case of strong snowfalls covering the glacier area), and thus is among the most important drivers controlling the annual mass balance of the glacier. The summer-monsoon air temperature, controlling the precipitation phase (rain versus snow and thus albedo), counts, indirectly, also among the most important drivers.
PDF
