Grady W. Chism

Pressure Death and Tailing Behavior of Listeria monocytogenes Strains Having Different Barotolerances

The objectives of this study were to investigate the variability among Listeria monocytogenes strains in response to high-pressure processing, identify the most resistant strain as a potential target of pressure processing, and compare the inactivation kinetics of pressure-resistant and pressure-sensitive strains under a wide range (350 to 800 MPa) of pressure treatments. The pressure resistance of Listeria innocua and nine strains of L. monocytogenes was compared at 400 or 500 MPa and 30 degrees C. Significant variability among strains was observed. The decrease in log CFU/ml during the pressure treatment was from 1.4 to 4.3 at 400 MPa and from 3.9 to >8 at 500 MPa. L. monocytogenes OSY-8578 exhibited the greatest pressure resistance, Scott A showed the greatest pressure sensitivity, and L. innocua had intermediate resistance. On the basis of these findings, L. monocytogenes OSY-8578 is a potential target strain for high-pressure processing efficacy studies. The death kinetics of L. monocytogenes Scott A and OSY-8578 were investigated at 350 and 800 MPa. Survivors at 350 MPa were enumerated by direct plating, and survivors at 800 MPa were enumerated by the most-probable-number technique. Both pressure-resistant and pressure-sensitive strains exhibited non-first-order death behavior, and excessive pressure treatment did not eliminate the tailing phenomenon.

Inactivation of Barotolerant Listeria monocytogenes in Sausage by Combination of High-Pressure Processing and Food-Grade Additives

Food-grade additives were used to enhance the efficacy of high-pressure processing (HPP) against barotolerant Listeria monocytogenes. Three strains of L. monocytogenes (Scott A, OSY-8578, and OSY-328) were compared for their sensitivity to HPP, nisin, tert-butylhydroquinone (TBHQ), and their combination. Inactivation of these strains was evaluated in 0.2 M sodium phosphate buffer (pH 7.0) and commercially sterile sausage. A cell suspension of L. monocytogenes in buffer (10(9) CFU/ml) was treated with TBHQ at 100 ppm, nisin at 100 IU/ml, HPP at 400 MPa for 5 min, and combinations of these treatments. Populations of strains Scott A, OSY-8578, and OSY-328 decreased 3.9, 2.7, and 1.3 log with HPP alone and 6.4, 5.2, and 1.9 log with the HPP-TBHQ combination, respectively. Commercially sterile sausage was inoculated with the three L. monocytogenes strains (10(6) to 10(7) CFU/g) and treated with selected combinations of TBHQ (100 to 300 ppm), nisin (100 and 200 ppm), and HPP (600 MPa, 28 degrees C, 5 min). Samples were enriched to detect the viability of the pathogen after the treatments. Most of the samples treated with nisin, TBHQ, or their combination were positive for L. monocytogenes. HPP alone resulted in a modest decrease in the number of positive samples. L. monocytogenes was not detected in any of the inoculated commercial sausage samples after treatment with HPP-TBHQ or HPP-TBHQ-nisin combinations. These results suggest that addition of TBHQ or TBHQ plus nisin to sausage followed by in-package pressurization is a promising method for producing Listeria-free ready-to-eat products.

Assay of enzymes involved in cytokinin metabolism by means of reversed-phase high-performance liquid chromatography

Abstract Assay of enzymes involved in cytokinin metabolism can be rapidly and conveniently carried out by using reversed-phase high-performance liquid chromatography (HPLC) to quantitate either the formation of product or the loss of substrate. The initial rates of loss of added 6-(3-methylbut-2-enylamino)purine or 6-benzylaminopurine can be determined in a variety of plant tissues. Asay of nucleosidase activity with 6-(3-methylbut-2-enylamino)-9-β- d -ribofuranosylpurine, 6-(4-hydroxy-3-methylbut-2-enylamino)-9-β- d -ribofuranosylpurine or adenosine as substrates can be accomplished by quantitating the loss of riboside or the formation of 8-OH cytokinins can be readily accomplished and the subsequent formation of 2,8-diOH cytokinin can be measured simultaneously if dual-wavelength monitoring is used. HPLC with a variety of solvent systems facilitates the assay of very small quantities of enzymes and the detection of multiple products.