Spodoptera frugiperda pheromone lures to avoid nontarget captures of Leucania phragmatidicola.

We confirmed that commercial three- or four-component Spodoptera frugiperda (J.E. Smith) pheromone lures had a high nontarget capture rate for Leucania phragmatidicola Guenée, which compromised monitoring efforts in the northeastern United States. We compiled taxonomic features to distinguish L. phragmatidicola from S. frugiperda, and we compared five new lures. S. frugiperda catch specificity was improved by removing (Z)-11-hexadecen-1-ol acetate (Z11-16:Ac), which attracted L. phragmatidicola. Four lures tracked late-season S. frugiperda immigration, but two of these lures also tracked a bivoltine L. phragmatidicola flight with a second generation coincident with S. frugiperda immigration, and one lure attracted the first, but not the second, generation of L. phragmatidicola. In both low- and high-moth flight conditions, two-component lures had low L. phragmatidicola captures (0.5-1.4%), and although lures with more pheromonal components captured more S. frugiperda, they also had a high percentage of capture of L. phragmatidicola (38-48%). We conclude that although two-component lures captured fewer S. frugiperda, their similar temporal pattern, along with the lower level of L. phragmatidicola, makes them useful for development for monitoring programs in the northeastern United States.

Using the Microcyte flow cytometer to monitor cell number, viability, and apoptosis in mammalian cell culture.

The Microcyte is a novel, portable flow cytometer based on diode laser technology whose use has been established for yeast and bacterial analysis. We present data that demonstrate its suitability for routine mammalian cell counting and viability determination. To extend its range of applications in the field of animal cell culture biotechnology, a test to determine the number of apoptotic cells present has been developed for use with the instrument. Apoptosis was induced in hybridoma cell cultures by treatment with camptothecin. Apoptotic cells were labeled with biotinylated Annexin V and then visualized using a streptavidin-allophycocyanin conjugate. Their numbers were counted, and the cell size of the apoptotic cell population was determined using the Microcyte.

Growth and enumeration of the meat spoilage bacterium Brochothrix thermosphacta.

Brochothrix thermosphacta is a common meat spoilage bacterium. The morphology of this bacterium changes from coccobacilli and short rods to chains during growth, which may give a false estimation in numbers using some enumeration techniques. Methods for the quantification of this bacterium have been compared. Turbidimetric readings showed good agreement with cell dry weight indicating that the former provides a good measure of the change in cell mass during growth. The turbidimetric method also correlated well with bacterial numbers determined by plate counts, flow cytometry and manual counts (by microscope) over a limited range of 10(7)-10(9) cells/ml. Flow cytometry and manual counts gave a linear relationship over a wider range of 10(5)-10(9) cells/ml. The sensitivity of analysis, growth rates and lag time attained using these methods were also compared. As a consequence of changes in bacterial cell size during growth, turbidimetry over-estimated the growth rate. The plate count method proved unable to detect the difference between bacteria existing as chains or single cells. The sensitivity of analysis and the calculated growth related parameters were similar for flow cytometry and manual counts. This suggests that flow cytometry is capable of counting individual cells in a chain. Further investigation showed that passage of B. thermosphacta cells through the flow cytometer resulted in the breakage of chains into single cells. The reliability, low error and rapidity of this technique make it attractive for bacterial enumeration, something which has been demonstrated using B. thermosphacta, a bacterium which exhibits complex morphologies.

Tc(VII) reduction and accumulation by immobilized cells of Escherichia coli.

Resting cells of Escherichia coli, immobilized in a flow-through bioreactor, coupled the oxidation of formate or hydrogen to Tc(VII) reduction and removal from solution. Cells, pregrown anaerobically in a hollow-fiber membrane bioreactor, were challenged with 50 microM Tc(VII) in a carrier solution of phosphate-buffered saline. The radionuclide accumulated within the membrane component of the reactor, corresponding to the localization of the cells. Negligible Tc removal was noted in a reactor containing a mutant deficient in active Tc(VII) reductase, when supplied with formate as an electron donor. Formate or hydrogen was supplied as the electron donor for Tc(VII) reduction to cells immobilized in reactors operated in transverse (crossflow) and direct (dead-end filtration) modes, respectively. Flow-rate activity relationships were used to compare the performance of the reactors. A flow rate of 2.4 mL h(-1) supported the removal of 50% of the Tc from solution in a reactor operated in transverse mode with formate as an electron donor. In contrast, a flow rate of 0.7 mL h(-1), supported comparable Tc removal when hydrogen was introduced to a reactor operated in direct mode. The reduced reactor efficiency, when hydrogen was used as an electron donor, could be attributed, in part, to poor delivery of the gas to the cells. The biocatalyst was highly stable in the reactor; no loss in activity was noted over 200 h of continuous use.