Microplastics in beluga whales (Delphinapterus leucas) from the Eastern Beaufort Sea.

Microplastics (MPs, particles <5 mm) represent an emerging global environmental concern, having been detected in multiple aquatic species. However, very little is known about the presence of MPs in higher trophic level species, including cetaceans. We worked with community based monitors and Inuvialuit hunters from Tuktoyaktuk (Northwest Territories, Canada) to sample seven beluga whales (Delphinapterus leucas) in 2017 and 2018. Microplastics were detected in the gastrointestinal tracts in every whale. We estimate that each whale contained 18 to 147 MPs in their GI tract (average of 97 ±â€¯42 per individual). FTIR-spectroscopy revealed over eight plastic polymer types, with nearly half being polyester. Fibres made up 49% of MPs. The diversity of MP shapes and polymeric identities in beluga points to a complex source scenario, and ultimately raises questions regarding the significance and long-term exposure of this pollutant in this ecologically and culturally valuable species.

Filtration capture immunoassay for bacteria: optimization and potential for urinalysis.

A novel assay utilizing immuno-labeling, filtration, and electrochemistry for the rapid detection of bacteria has been optimized for the detection of Escherichia coli O157:H7. Bacteria were specifically labeled with alkaline phosphatase conjugated polyclonal antibodies and captured on a polycarbonate track-etched membrane filter (0.2 microm pore size). The filter was then placed directly against a glassy carbon electrode, incubated with enzyme substrate, and the product detected by square wave voltammetry. The high speed and capture efficiency of membrane filtration and inherent sensitivity of electrochemical detection produced a 25-min assay with a detection limit of 5 x 10(3) E. coli O157:H7 per ml using a filtration volume of 100 microl (i.e. 500 cells filtered). The labeling, filtration, and electrochemical steps were optimized, and the assay performance using electrochemical and colorimetric detection methods was compared. The assay was used to detect E. coli O157:H7 that was spiked into filter-sterilized urine at clinically relevant concentrations.

Filtration capture and immunoelectrochemical detection for rapid assay of Escherichia coli O157:H7.

A new approach for rapid assay of bacteria in liquid samples is described. Cells were labeled by incubation with an enzyme-antibody conjugate and captured by filtration of the sample/conjugate mixture through a 0.2 microm filter. The enzyme-labeled cells were detected by placing the filter on the surface of an electrode, incubating with enzyme substrate, and measuring the current produced by oxidation of the electroactive enzyme product. Assay time was 25 min and a detection limit of approximately 5000 cells/ml was obtained for E. coli O157:H7. Background current due to non-specific binding of conjugate to the filter was the primary factor controlling the detection limit, and fewer than 50 cells could be detected when very small sample volumes (10 microl) were used to minimize background current.

Immunoelectrochemical assays for bacteria: use of epifluorescence microscopy and rapid-scan electrochemical techniques in development of an assay for Salmonella.

Immunoelectrochemical sensors in which the sensor surface functions as both analyte capture phase and electrochemical detector have recently been developed for bacteria analysis. The speed and sensitivity of these devices make them very attractive for applications such as the detection of pathogenic microorganisms in food and water. However, the development and optimization of assays utilizing these sensors can be complicated by undesired interactions between the capture and detection functions. Modification of the sensor to achieve improvements in one function can have deleterious effects on the other function, and such effects can be difficult to diagnose and correct. In the course of investigations on immunoelectrochemical detection of Salmonella, we developed a rapid, nondestructive epifluorescence microscopy method to determine bacteria capture efficiency. This method enabled us to study capture and detection functions independently and efficiently identify performance-limiting factors. Rapid-scan electrochemical methods were used to optimize detection sensitivity and to provide diagnostic information on detection performance.

Enzyme-linked immunomagnetic electrochemical detection of Salmonella typhimurium.

There is a need for rapid methods to detect pathogenic bacteria in food products as alternatives to the current laborious and time-consuming culture procedures. We report a microbial detection technique that combines the selectivity of antibody-coated superparamagnetic beads with the rapidity and sensitivity of electrochemical detection in a format termed enzyme-linked immunomagnetic electrochemistry. In it, Salmonella typhimurium were sandwiched between antibody-coated magnetic beads and an enzyme-conjugated antibody. With the aid of a magnet, the beads (with or without bound bacteria) were localized onto the surface of disposable graphite ink electrodes in a multi-well plate format. Enzyme substrate was added and conversion of substrate to an electroactive product was measured using electrochemical detection. The electrochemical response was directly proportional to the number of captured bacteria. Using this technique, a minimum detectable level of 8 x 10(3) cells/ml of Salmonella typhimurium in buffer was achieved in ca. 80 min.

Binding geometry, stoichiometry, and thermodynamics of cyclomalto-oligosaccharide (cyclodextrin) inclusion complex formation with chlorogenic acid, the major substrate of apple polyphenol oxidase.

The inclusion complexes of cyclomaltohexaose (alpha-CD), cyclomaltoheptaose (beta-CD), cyclomaltooctaose (gamma-CD), and polymerized beta-CD (beta-CDn) with chlorogenic acid (CA), the major substrate of apple fruit polyphenol oxidase (PPO), were studied with regard to pH, ionic strength, and temperature in model buffer systems and apple juice. The thermodynamics of CD.CA inclusion complex formation, which were studied in solution using UV spectrophotometry, displayed enthalpy-entropy compensation typical of processes driven by solvation phenomena. We also found that the apparent association constants (K) of the CD.CA equilibrium were relatively insensitive to pH for beta-CD, compared to alpha- and gamma-CDs, but were subject to substantial enhancement at low ionic strengths. The beta-CD.CA inclusion complex was also characterized for binding geometry and stoichiometry at 9.4 T and 25 degrees C in 0.05 M Na phosphate buffer by 1H NMR spectroscopy. A 1:1 stoichiometric ratio for the complex was found using the method of continuous variations. 1H Spin-lattice relaxation and chemical-shift data indicate that the phenolic ring of CA docks within the cavity of beta-CD. The Ks for beta-, alpha-, and gamma-CD determined in apple juice, which contains a mixture of PPO substrates, were found to correlate with PPO activity-related data. Apple juice, treated with beta-CDn, did not brown until CA was added back. These latter findings strongly argue that the mechanism for inhibition of juice browning with cyclodextrins was mainly due to the binding of PPO substrates and not some other means such as enzyme inactivation via sequestration of Cu2+ by CDs.

Rapid biorecognition assay for herbicides in biological matrices.

A rapid assay capable of detecting several commonly used herbicides at nanogram per milliliter concentrations in biological fluids is described. The assay is based on inhibition of photosynthetic electron transport in spinach thylakoids by the target compounds with colorimetric detection using a redox dye. Using a microtiter plate format, high throughput assays of water, urine, and homogenized tissue were performed in minutes with minimal sample preparation. Detection limits of 3 ng/mL for atrazine, 3 ng/mL for diuron, and 1 ng/mL for terbutryn were observed.

Evaluation of restricted access media for high-performance liquid chromatographic analysis of sulfonamide antibiotic residues in bovine serum.

Three commercially-available high-performance liquid chromatographic columns packed with restricted access media were evaluated for suitability in multi-residue direct injection analysis at the ng/ml level. The internal surface reversed-phase and shielded hydrophobic phase columns were not sufficiently retentive to separate all analytes from the tail of the matrix peak. Coelution of some of the analytes was also observed with these columns. The semi-permeable surface column was significantly more retentive and selective, providing good separation of analyte and matrix peaks. With this column, an analytical protocol requiring no organic solvents was developed for the assay of six sulfonamides at a detection limit of 25 ng/ml.

Rapid electrodialytic clean-up of biological samples for high-performance liquid chromatography.

A sample clean-up system employing electrodialysis with size-selective and charge-selective membranes is described. When applied to the treatment of 0.5-ml milk samples containing sulfamethazine, the system produced an undiluted, clear solution in 3 min and eliminated the components in untreated milk that caused column fouling and double peaks. In contrast to conventional liquid- and solid-phase extraction procedures, electrodialytic clean-up is readily automated and uses no organic solvents.