Rapid and sensitive detection of E. coli O157:H7 by lateral flow immunoassay and silver enhancement.

The aim of this study was to develop a sensitive lateral flow immunoassay (LFIA) for the rapid detection of Escherichia coli (E. coli) O157:H7, a pathogen contributor to diseases and fatalities worldwide. Au nanoparticles with high stability, uniform size, and shape were synthesized and coated with heterobifunctional PEG polymer with carboxyl groups, and they were bioconjugated to be used as label in sandwich-LFIA. Then, a silver enhancement strategy was developed as an accessible, rapid, and cost-effective approach for signal amplification to reduce the limit of detection (LOD). The optimal results were achieved when a solution of silver nitrate and hydroquinone/citrate buffer was added to the strips for 4 min. This led to a decrease in the visual LOD from 2 × 106 (CFU mL-1) to 2 × 103 (CFU mL-1), resulting in a threefold improvement in sensitivity compared to the conventional LFIA system. The specificity of the system was evaluated by using non-target bacteria (E. coli BL21 and E. coli T515) and its reliability was determined by testing commercial food samples (milk, tap water, and orange juice), demonstrating its effectiveness for quickly detecting pathogenic bacteria in food products.

Fe3O4@Au Core-Shell Magnetic Nanoparticles for the Rapid Analysis of E. coli O157:H7 in an Electrochemical Immunoassay.

Escherichia coli (E. coli) O157:H7 is a pathogenic bacterium that causes serious toxic effects in the human gastrointestinal tract. In this paper, a method for its effective analytical control in a milk sample was developed. To perform rapid (1 h) and accurate analysis, monodisperse Fe3O4@Au magnetic nanoparticles were synthesized and used in an electrochemical sandwich-type magnetic immunoassay. Screen-printed carbon electrodes (SPCE) were used as transducers, and electrochemical detection was performed by chronoamperometry using a secondary horseradish peroxidase-labeled antibody and 3,3',5,5'-tetramethylbenzidine. This magnetic assay was used to determine the E. coli O157:H7 strain in the linear range from 20 to 2 × 106 CFU/mL, with a limit of detection of 20 CFU/mL. The selectivity of the assay was tested using Listeria monocytogenes p60 protein, and the applicability of the assay was assessed by analyzing a commercial milk sample, demonstrating the usefulness of the synthesized nanoparticles in the developed magnetic immunoassay.

Lipid-Polymer Hybrids Encapsulating Iron-Oxide Nanoparticles as a Label for Lateral Flow Immunoassays.

The feasibility of using Superparamagnetic Iron Oxide Nanoparticles (SPIONs) encapsulated by lipid-polymer nanoparticles as labels in lateral flow immunoassays (LFIA) was studied. First, nanoparticles were synthesized with average diameters between 4 and 7 (nm) through precipitation in W/O microemulsion and further encapsulated using lipid-polymer nanoparticles. Systems formulated were characterized in terms of size and shape by DLS (Nanozetasizer from Malvern) and TEM. After encapsulation, the average size was around (≈20 and 50 nm). These controlled size agglomerates were tested as labels with a model system based on the biotin-neutravidin interaction. For this purpose, the encapsulated nanoparticles were conjugated to neutravidin using the carbodiimide chemistry, and the LFIA was carried out with a biotin test line. The encapsulated SPIONs showed that they could be promising candidates as labels in LFIA test. They would be useful for immunomagnetic separations, that could improve the limits of detection by means of preconcentration.

Recent advancements in the production of rhamnolipid biosurfactants by Pseudomonas aeruginosa.

Rhamnolipid (RL) biosurfactant which is produced by Pseudomonas species is one of the most effective surface-active agents investigated in the literature. Over the years, many efforts have been made and an array of techniques has been developed for the isolation of RL produced strains as well as RL homolog characterization. Reports show that RL productivity by the best-known producer, Pseudomonas aeruginosa, is very diverse, from less than 1 gr/l to more than 200 g L-1. There are some major parameters that can affect RL productivity. These are culture conditions, medium composition, the mode of operation (batch, fed-batch and continuous), bioengineering/gene manipulation and finally extraction methods. The present paper seeks to provide a comprehensive overview on the production of rhamnolipid biosurfactant by different species of Pseudomonas bacteria. In addition, we have extensively reviewed their potential for possible future applications.

Overproduction of rhamnolipid by fed-batch cultivation of Pseudomonas aeruginosa in a lab-scale fermenter under tight DO control.

Rhamnolipids are one of the most well-known classes of biosurfactants having wide applications in various industries due to low toxicity, high biodegradability, and environmentally friendly. Dissolved oxygen (DO) concentration has the crucial effect on rhamnolipids production, particularly through fed-batch cultivation. In this study, the effect of different levels of DO concentrations on rhamnolipid production by Pseudomonas aeruginosa in both batch and fed-batch fermentation was investigated in a lab-scale fermenter under precise DO control. A maximal rhamnolipid production of 22.5 g/l was obtained at a DO concentration of 40% in batch fermentation. In order to achieve the high rhamnolipid production, a fed-batch operation under tight DO control of 40% was conducted. As a result, the overall rhamnolipid production and productivity reached to 240 g/l and 0.9 (g/l h), corresponding to a 10.7 and 4.8-fold improvement compared to the batch experiments. The high level of rhamnolipid production via the fed-batch cultivation can be attributed to both DO concentration and the feeding strategy. This achievement is promising for the production of rhamnolipid in industrial scale.

Overproduction of lipopeptide biosurfactant by Aneurinibacillus thermoaerophilus HAK01 in various fed-batch modes under thermophilic conditions.

An efficient lipopeptide biosurfactant (BS) producer, Aneurinibacillus thermoaerophilus HAK01, was isolated from municipal landfill sites. The strain was able to produce about 4.9 g L-1 lipopeptide at a thermophilic temperature of 45 °C. After optimization of culture component concentrations using the response surface method, the main focus is to find the most appropriate fed-batch strategy to enhance lipopeptide production by the HAK01 strain. For this purpose, four fed-batch strategies including (a) pH-stat mode, (b) constant feeding rate strategy, (c) DO-stat mode, and (d) combined feeding strategy were designed. The production of BS was increased systematically from 4.9 g L-1 in batch mode to 5.9, 7.1, 8.8 and 11.2 g L-1 in each fed-batch mode, respectively. While poor results were obtained in the pH-stat mode, the DO-stat mode showed excellent results in the production of BS. The results of the study confirmed the importance of operational mode, oxygen supply and the kind of feeding strategy in BS production.