scholarly journals Evaluation of bacterial proliferation with a microfluidic-based device: Antibiochip

2019 ◽  
Author(s):  
Valentina Gallo ◽  
Alessia Ruiba ◽  
Massimo Zanin ◽  
Paolo Begnamino ◽  
Sabina Ledda ◽  
...  

AbstractThe measurement of the proliferation (and the relevant inhibition of proliferation) of microbes is used in different settings, from industry to laboratory medicine. Thus, in this study, the capacity of the Antibiochip (ELTEK spa), a microfluidic-based device, to measure the amount of E. coli in certain culture conditions, was evaluated. An Antibiochip is composed of V-shaped microchannels, and the amount of microparticles (such as microbes) is measured by the surface of the pellet after centrifugation. In the present study, different geometries, volumes and times were analyzed. When the best conditions were identified, serial dilutions of microbial cultures were tested to validate the linearity of the results. Then, with the use of wild E. coli strains isolated from medical samples, the relationship between bacterial susceptibility to antibiotics (gentamicin, amikacin and ceftriaxone) measured by standard methods and that measured by the Antibiochip was evaluated. In this report, the good quality performances of the methods, their linearity and the capacity to identify susceptible microbial strains after 60 minutes of incubation are shown. These results represent a novel approach for ultrarapid antibiograms in clinics.

TECHNOLOGY ◽  
2015 ◽  
Vol 03 (02n03) ◽  
pp. 80-83
Author(s):  
Mark Polikovsky ◽  
Eshel Ben-Jacob ◽  
Alin Finkelshtein

Cellulose hydrolysis has many industrial applications such as biofuel production, food, paper and textile manufacture. Here, we present a novel approach to cellulose hydrolysis using a consortium of motile bacteria, Paenibacillus vortex, that can swarm on solid medium carrying a non-motile recombinant E. coli cargo strain expressing the β-glucosidase and cellulase genes that facilitate the hydrolysis of cellulose. These two species cooperate; the relationship is mutually beneficial: the E. coli is dispersed over long distances, while the P. vortex bacteria gain from the supply of cellulose degradation products. This enables the use of such consortia in this area of biotechnology.


2021 ◽  
Author(s):  
Thuan Nguyen ◽  
Vinay Bharadwaj Tatipamula ◽  
Tran Thanh Viet ◽  
Nguyen Quang Duc Tien ◽  
Nguyen Hoang Loc

Abstract Eriodictyol is a flavonoid in the flavanones subclass. It is abundantly present in a wide range of medicinal plants, citrus fruits, and vegetables. In addition, eriodictyol owns numerous importantly medicinal bioactivities such as inhibition of proliferation, metastasis and induction of apoptosis in glioma cells or inhibition of glioblastoma migration, and invasion. This study described the heterologous production of eriodictyol by E. coli based co-culture engineering system from the initial substrate as D-glucose. Notably, the upstream module was composed of genes for synthesis of p-coumaric acid (pCA) from D-glucose. The downstream module consisted of genes for the synthesis of eriodictyol from p-coumaric acid. The maximal result of eryodictyol was achieved 51.5 mg/L using optimal culture conditions, while mono-culture was only achieved 21.3 mg/L. In conclusion, co-culture was the efficiently alternative approach for the synthesis of eriodictyol and other natural products.


1995 ◽  
Vol 31 (5-6) ◽  
pp. 19-26 ◽  
Author(s):  
G. J. Medema ◽  
I. A. van Asperen ◽  
J. M. Klokman-Houweling ◽  
A. Nooitgedagt ◽  
M. J. W. van de Laar ◽  
...  

This pilot study was carried out to determine the relationship between microbiological water quality parameters and the occurrence of health complaints among triathletes. Data were collected at an Olympic distance triathlon (n=314) and a run-bike-run (n=81; controls for exposure to fresh water). At the time of the triathlon, the concentrations of Escherichia coli , thermotolerant coliforms, faecal streptococci, entero- and reoviruses, F-specific RNA phages, Salmonella, Campylobacter, Aeromonas, Plesiomonas shigelloides, Pseudomonas aeruginosa and Staphylococcus aureus were examined over the swimming course. Information on the occurrence of health complaints during the competition and in the week thereafter was collected through a written questionnaire. The results show that triathletes and run-bike-runners are comparable with respect to factors other than water exposure (age, sex, training history, physical stress, lower intestinal health complaints during the competition) that may influence the occurrence of health complaints in the week after the competition. Triathletes and run-bike-runners reported gastro-intestinal (7.7% vs 2.5%), respiratory (5.5% vs 3.7%), skin/mucosal (2.6% vs 1.2%), general (3.5% vs 1.2%) and total symptoms (14.8% vs 7.4%) in the week after the event. The health risks for triathletes for all symptom groups are not significantly higher than for run-bike-runners. The geometric mean concentration of faecal indicator bacteria is relatively low: E. coli 170/100 ml; faecal streptococci 13/100 ml, enteroviruses were present at concentrations of 0.1/l. The group of triathletes was homogeneusly and relatively intensely exposed to water; they all swam in the same body of water at the same time and 75% reported to have swallowed freshwater. It was concluded that this study design is suitable to study the relationship between health complaints and microbiological water quality. In the summers of 1993 and 1994, a study will be carried out concerning several run-bike-runs and triathlons in freshwaters of different quality.


1997 ◽  
Vol 35 (11-12) ◽  
pp. 35-40 ◽  
Author(s):  
B. Genthe ◽  
N. Strauss ◽  
J. Seager ◽  
C. Vundule ◽  
F. Maforah ◽  
...  

Efforts to provide water to developing communities in South Africa have resulted in various types of water supplies being used. This study examined the relationship between the type of water supply and the quality of water used. Source (communal taps, private outdoor and indoor taps) and point-of-use water samples were examined for heterotrophic plate counts (HPC), total and faecal coliforms, E. coli, and coliphages. Ten percent of samples were also analysed for enteric viruses, Giardia and Cryptosporidium. Approximately 320 households were included in a case-control study. In addition, a cross-sectional study was conducted. Both studies examined the relationship between different types of water facilities and diarrhoea among pre-school children. The source water was of good microbial quality, but water quality was found to have deteriorated significantly after handling and storage in both case and control households, exceeding drinking water quality guideline values by 1-6 orders of magnitude. Coliphage counts were low for all water samples tested. Enteric viruses and Cryptosporidium oocysts were not detected. Giardia cysts were detected on one occasion in case and control in-house samples. Comparisons of whether in-house water, after handling and storage, complied with water quality guideline values demonstrated households using communal taps to have significantly poorer quality than households using private outdoor or indoor taps for HPC and E. coli (χ2 = 14.9, P = 0.001; χ2 = 6.6, P = 0.04 respectively). A similar trend (although not statistically significant) was observed for the other microbial indicators. The cross-sectional study demonstrated an apparent decrease in health risk associated with private outdoor taps in comparison to communal taps. This study suggests that a private outdoor tap is the minimum level of water supply in order to ensure the supply of safe water to developing communities.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
María-Eugenia DeCastro ◽  
Michael P. Doane ◽  
Elizabeth Ann Dinsdale ◽  
Esther Rodríguez-Belmonte ◽  
María-Isabel González-Siso

AbstractIn the present study we investigate the microbial community inhabiting As Burgas geothermal spring, located in Ourense (Galicia, Spain). The approximately 23 Gbp of Illumina sequences generated for each replicate revealed a complex microbial community dominated by Bacteria in which Proteobacteria and Aquificae were the two prevalent phyla. An association between the two most prevalent genera, Thermus and Hydrogenobacter, was suggested by the relationship of their metabolism. The high relative abundance of sequences involved in the Calvin–Benson cycle and the reductive TCA cycle unveils the dominance of an autotrophic population. Important pathways from the nitrogen and sulfur cycle are potentially taking place in As Burgas hot spring. In the assembled reads, two complete ORFs matching GH2 beta-galactosidases were found. To assess their functional characterization, the two ORFs were cloned and overexpressed in E. coli. The pTsbg enzyme had activity towards o-Nitrophenyl-β-d-galactopyranoside (ONPG) and p-Nitrophenyl-β-d-fucopyranoside, with high thermal stability and showing maximal activity at 85 °C and pH 6, nevertheless the enzyme failed to hydrolyze lactose. The other enzyme, Tsbg, was unable to hydrolyze even ONPG or lactose. This finding highlights the challenge of finding novel active enzymes based only on their sequence.


Drug Research ◽  
2020 ◽  
Author(s):  
Pinki Yadav ◽  
Kashmiri Lal ◽  
Ashwani Kumar

AbstractThe in vitro antimicrobial properties of some chalcones (1a–1c ) and chalcone tethred 1,4-disubstituted 1,2,3-triazoles (2a–2u) towards different microbial strains viz. Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger and Candida albicans are reported. Compounds 2g and 2u exhibited better potency than the standard Fluconazole with MIC values of 0.0063 µmol/mL and 0.0068 µmol/mL, respectively. Furthermore, molecular docking was performed to investigate the binding modes of two potent compounds 2q and 2g with E. coli topoisomerase II DNA gyrase B and C. albicans lanosterol 14α-demethylase, respectively. Based on these results, a statistically significant quantitative structure activity relationship (QSAR) model was successfully summarized for antibacterial activity against B. subtilis.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Weiwei Wang ◽  
Yan Gao ◽  
Yanting Tang ◽  
Xiaoting Zhou ◽  
Yuezheng Lai ◽  
...  

AbstractCytochromes bd are ubiquitous amongst prokaryotes including many human-pathogenic bacteria. Such complexes are targets for the development of antimicrobial drugs. However, an understanding of the relationship between the structure and functional mechanisms of these oxidases is incomplete. Here, we have determined the 2.8 Å structure of Mycobacterium smegmatis cytochrome bd by single-particle cryo-electron microscopy. This bd oxidase consists of two subunits CydA and CydB, that adopt a pseudo two-fold symmetrical arrangement. The structural topology of its Q-loop domain, whose function is to bind the substrate, quinol, is significantly different compared to the C-terminal region reported for cytochromes bd from Geobacillus thermodenitrificans (G. th) and Escherichia coli (E. coli). In addition, we have identified two potential oxygen access channels in the structure and shown that similar tunnels also exist in G. th and E. coli cytochromes bd. This study provides insights to develop a framework for the rational design of antituberculosis compounds that block the oxygen access channels of this oxidase.


1998 ◽  
Vol 333 (3) ◽  
pp. 565-571 ◽  
Author(s):  
Casey E. BRECHTEL ◽  
Steven C. KING

A previous study [Ferson, Wray and Fisher (1996) Mol. Microbiol. 22, 693–701] has shown that transposon-mediated disruption of a protein 47% identical to the Escherichia coli GABA (4-aminobutyrate) transporter abolishes the ability of nitrogen-limited culture conditions to induce expression of a GABA transport activity in Bacillus subtilis. Here it is demonstrated directly that the B. subtilis GABA permease (gabP) gene can complement the transport defect in the gabP-negative E. colistrain. Unexpectedly, the ligand-recognition profile of the B. subtilis GabP was found to differ substantially from that of the highly homologous E. coli GabP. Unlike the E. coli GabP, the B. subtilis GabP: (i) exhibits approx. equal preference for the 3-carbon (β-alanine, Km = 9.6 µM) and the 4-carbon (GABA, Km = 37 µM) amino acids, and (ii) resists inhibition by bulky, conformationally constrained compounds (e.g. nipecotic acid, guvacine), which are active against GABA transporters from brain. The present study shows additionally that the B. subtilis GabP can translocate several open-chain GABA analogues (3-aminobutyrate, 3-aminopropanoate, cis-4-aminobutenoate) across the membrane via counterflow against [3H]GABA. Thus, consistent with the idea that the ligand-recognition domain of the B. subtilis GabP is less spacious than that of the close homologue from E. coli, the former exhibits more stringent requirements than the latter for substrate recognition and translocation. These distinct functional characteristics of the E. coli and B. subtilis GABA transporters provide a basis by which to identify ligand-recognition domains within the amine-polyamine-choline transporter superfamily.


2021 ◽  
Vol 11 (5) ◽  
pp. 1980
Author(s):  
Kazimierz Józefiak ◽  
Artur Zbiciak ◽  
Karol Brzeziński ◽  
Maciej Maślakowski

The paper presents classical and non-classical rheological schemes used to formulate constitutive models of the one-dimensional consolidation problem. The authors paid special attention to the secondary consolidation effects in organic soils as well as the soil over-consolidation phenomenon. The systems of partial differential equations were formulated for every model and solved numerically to obtain settlement curves. Selected numerical results were compared with standard oedometer laboratory test data carried out by the authors on organic soil samples. Additionally, plasticity phenomenon and non-classical rheological elements were included in order to take into account soil over-consolidation behaviour in the one-dimensional settlement model. A new way of formulating constitutive equations for the soil skeleton and predicting the relationship between the effective stress and strain or void ratio was presented. Rheological structures provide a flexible tool for creating complex constitutive relationships of soil.


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