Joseph D. Eifert

Inhibition of pathogens on fresh produce by ultraviolet energy

Ultraviolet energy at a wavelength of 253.7 nm (UVC) was investigated for its bactericidal effects on the surface of Red Delicious apples, leaf lettuce and tomatoes inoculated with cultures of Salmonella spp. or Escherichia coli O157:H7. Inoculated samples were subjected to different doses ranging from 1.5 to 24 mW/cm(2) of UVC and enumerated on tryptic soy agar plus 0.05 g/l nalidixic acid to determine effective log reductions of microbial populations. UVC applied to apples inoculated with E. coli O157:H7 resulted in the highest log reduction of approximately 3.3 logs at 24 mW/cm(2). Lower log reductions were seen on tomatoes inoculated with Salmonella spp. (2.19 logs) and green leaf lettuce inoculated with both Salmonella spp. and E. coli O157:H7 (2.65 and 2.79, respectively). No significant statistical difference (p>0.05) was seen in the ability of UVC to inactivate a higher population of either Salmonella spp. or E. coli O157:H7 on the surface of green leaf lettuce. No significant difference was seen among the use of different doses applied to the surface of fresh produce for reduction of E. coli O157:H7 or Salmonella spp. (p>0.05). The use of UVC may prove to be beneficial in protecting the safety of fruits and vegetables in conjunction with Good Agricultural Practices and Good Manufacturing Practices.

Use of a Modified Gompertz Equation to Predict the Effects of Temperature, pH, and NaCl on the Inactivation of Listeria monocytogenes Scott A Heated in Infant Formula

The heat resistance of Listeria monocytogenes was determined in infant formula for all possible combinations of temperature (50, 55, and 60°C), pH level (5, 6, and 7), and NaCl concentration (0, 2, and 4%). Survival curves were fit using nonlinear regression with a Gompertz equation. The Gompertz equation was flexible enough to fit the three most commonly observed survival curves: linear curves, those with an initial lag region followed by a linear region, and sigmoidal shaped. Parameter estimates obtained by the method of nonlinear least squares were used to describe the effect(s) of different heating treatments on the lag region, death rate, and tailing region of survival curves. These estimates were further used to predict single and interactive effects of temperature, pH, and percentage of NaCl on the log of the surviving fraction (LSF) of bacteria. Interactions among these variables significantly (P ≤ .05) affected the LSF. Generally, increased pH or NaCl concentration lead to an increased LSF, whereas increased time or temperature lead to a decreased LSF. All multiple-factor interactions significantly (P ≤ .05) affected the LSF. The correlation of observed LSF versus predicted LSF (R2 = .92) indicated that the estimated Gompertz equation was in close agreement with the observation. This study demonstrated that the Gompertz equation and nonlinear regression can be used as an effective means to predict survival curve shape and response to heat of L. monocytogenes under many different environmental conditions.

Response of Salmonella and Escherichia coli O157:H7 to UV Energy

To determine the efficacy of a UV light treatment at 253.7 nm (UVC light) on microbial growth, plates containing tryptic soy agar plus 50 ppm of nalidixic acid (TSAN) were inoculated with known concentrations of five-strain cocktails of Salmonella and Escherichia coli O157:H7 and subjected to different UVC treatments. The concentration of the cocktail inoculum was determined with TSAN prior to inoculation. Serial dilutions were carried out, and inoculation levels of 10(0) to 10(8) CFU/ ml were tested for each pathogen. Multiple replications of doses of UV light ranging from 1.5 to 30 mW/cm2 were applied to different cocktail concentrations, and doses of > 8.4 mW/cm2 resulted in a 5-log reduction of Escherichia coli O157:H7, while a 5-log reduction of Salmonella was observed with doses of > 14.5 mW/cm2. Results for both organisms yielded sigmoidal inactivation curves. UVC light is effective in reducing microbial populations of pathogens on agar surfaces.

High-pressure resistance variation of Escherichia coli O157: H7 strains and Salmonella serovars in tryptic soy broth, distilled water, and fruit juice

The effect of high pressure on the log reduction of six strains of Escherichia coli O157:H7 and five serovars of Salmonella enterica was investigated in tryptic soy broth, sterile distilled water, and commercially sterile orange juice (for Salmonella) and apple cider (for E. coli). Samples were subjected to high-pressure processing treatment at 300 and 550 MPa for 2 min at 6 degrees C. Samples were plated onto tryptic soy agar directly after pressurization and after being held for 24 h at 4 degrees C. At 300 MPa, little effect was seen on E. coli O157:H7 strains, while Salmonella serovars varied in resistance, showing reductions between 0.26 and 3.95 log CFU/ml. At 550 MPa, E. coli O157:H7 strains exhibited a range of reductions (0.28 to 4.39 log CFU/ml), while most Salmonella populations decreased beyond the detection limit (> 5-log CFU/ml reduction). The most resistant strains tested were E. coli E009 and Salmonella Agona. Generally, bacterial populations in fruit juices showed larger decreases than did populations in tryptic soy broth and distilled water. E. coli O157:H7 cultures held for 24 h at 4 degrees C after treatment at 550 MPa showed a significant log decrease as compared with cultures directly after treatment (P < or = 0.05), while Salmonella serovars did not show this significant decrease (P > 0.05). All Salmonella serovars tested in orange juice treated at 550 MPa for 2 min at 6 degrees C and held for 24 h showed a > 5-log decrease, while E. coli O157:H7 strains require a higher pressure, higher temperature, longer pressurization, or a chemical additive to achieve a 5-log decrease.

Efficacy of detergents in removing Salmonella and Shigella spp. from the surface of fresh produce.

Fresh produce has been implicated in several foodborne disease outbreaks. Produce surfaces can be primary sites of contamination during production and handling. One approach to reduce contamination is to treat fresh produce with rinsing agents. In this study, different detergent agents were used at 22 and 40 degrees C to determine their efficacy in removing Salmonella and Shigella spp. from the surfaces of strawberries, tomatoes, and green-leaf lettuce. Produce was inoculated at 22 degrees C with a cocktail of nalidixic acid-resistant organisms (6 to 6.5 log CFU/ml). After air drying for 1 h, samples were rinsed with either 0.1% Tween 80, 0.1% sodium lauryl sulfate (SLS), or water (control) at 22 or 40 degrees C. Rinse solutions were spiral plated onto tryptic soy agar supplemented with 50 mg of nalidixic acid per liter. In trials involving strawberries and lettuce, Salmonella and Shigella were removed at levels of 4 and 3 log CFU/ml, respectively, except from Salmonella-inoculated strawberries rinsed with SLS, for which minimal removal rates were 1.5 log CFU/ml at 22 degrees C and < 1 log CFU/ml at 40 degrees C. When whole strawberries were analyzed after rinsing with SLS, few organisms were recovered. This result suggests that SLS may have a lethal or sublethal effect on Salmonella, especially when a 40 degrees C solution is used. Salmonella and Shigella removal rates for tomatoes were 1 and 1.5 log CFU/ml lower, respectively, than those for strawberries or lettuce. Overall, detergents were no more effective in removing organisms from produce than water was. The detergents examined would not constitute effective overall produce rinse treatments.

Pathogenic and Fecal Escherichia coli Strains from Turkeys in a Commercial Operation

SUMMARY. The biochemical phenotypes and antimicrobial susceptibility patterns of 105 clinical Escherichia coli isolates from flocks with colibacillosis in a turkey operation were compared with 1104 fecal E. coli isolates from 20 flocks in that operation. Clinical isolates and 194 fecal isolates with biochemical phenotypes or minimum inhibitory concentrations for gentamicin and sulfamethoxazole similar to clinical isolates were tested for somatic antigens and the potential virulence genes hylE, iss, tsh, and K1. The predominant biochemical phenotype of clinical isolates contained 21 isolates including 14 isolates belonging to serogroup O78 with barely detectable β-d-glucuronidase activity. Thirty-five fecal isolates had biochemical phenotypes matching common phenotypes of clinical isolates. Sixty-six (63%) clinical isolates exhibited intermediate susceptibility or resistance to gentamicin and sulfamethoxazole compared with 265 (24%) fecal isolates (P < 0.001). Seventy-seven clinical isolates reacted with O-antisera, of which 51 (66%) belonged to the following serogroups: O1, O2, O8, O25, O78, O114, and O119. In comparison, 8 of 35 (23%) fecal isolates subtyped on the basis of biochemical phenotype belonged to these serogroups and four of 167 (2%) fecal isolates subtyped on the basis of their antimicrobial resistance patterns belonged to these serogroups. Iss, K1, and tsh genes were detected more often among clinical isolates than these fecal isolates (P < 0.05). In summary, a small subgroup of E. coli strains caused most colibacillosis infections in this operation. These strains existed at low concentration in normal fecal flora of healthy turkeys in intensively raised flocks. The data suggest that colibacillosis in turkey operations may be due to endogenous infections caused by specialized pathogens.

Predictive model with improved statistical analysis of interactive factors affecting the growth of Staphylococcus aureus 196E

The growth of pathogenic bacteria in foods is affected by several factors which may interact to enhance or inhibit microbial growth. A model to predict the growth of Staphylococcus aureus 196E in microbiological media was developed using a modified Gompertz function and response-surface methodology. The predictive equation required the estimation of 23 parameters which describe singular and interactive effects of the growth factors studied. S. aureus 196E was inoculated into brain heart infusion broth formulated with either 0.5, 4.5, or 8.5% NaCl, adjusted to pH 5.0, 6.0, or 7.0, and incubated aerobically at 12, 20, or 28°C. Several interactive relationships between time, temperature, pH, and NaCl concentration were significant. The model adequately predicted the growth of S. aureus 196E. Predicted responses to multiple-factor interactions were displayed with three-dimensional and contour plots. A second model developed from a smaller subset of the growth data demonstrated that models could be produced with much less data collection. This methodology can provide important information to food scientists about the growth kinetics of microorganisms and prediction ranges or confidence intervals for growth parameters. Consequently, the effects of food formulations and storage conditions on the growth kinetics of foodborne pathogens or spoilage microorganisms could be predicted.

EFFECT OF FAT CONTENT AND TEMPERATURE ON DIELECTRIC PROPERTIES OF GROUND BEEF

Microwave heating depends on dielectric properties, microwave frequencies, food composition, physical state of water in the food, and product temperature and density. Proper microwave cooking and thawing procedures for ground beef products are needed to ensure food safety. Recent studies showed that the fat content significantly affected the survival of pathogens. The objective of this study was to determine the effect of fat content and temperatures on dielectric properties of ground beef. The dielectric properties were determined using an open-ended, coaxial line with copper conductors, connected to a network analyzer. The relationship among dielectric properties, fat level, and temperature was obtained for 915 MHz and 2450 MHz. Results of the experiment showed that dielectric loss factor and dielectric constant increased with increase in temperature at both frequencies at temperatures below the freezing point. Above the freezing point, dielectric constant decreased with increase in temperature. Dielectric loss factor increased with temperature at 915 MHz. At 2450 MHz, it remained almost constant with varying temperature. Dielectric constant and dielectric loss factor of low-fat ground beef were higher than that of high-fat ground beef. Regression equations were developed for predicting dielectric properties at different temperatures and fat levels.

Biofilms promote survival and virulence of Salmonella enterica sv. Tennessee during prolonged dry storage and after passage through an in vitro digestion system

Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. While the biofilm-forming abilities of Salmonella improve its survival during thermal processing and sanitation it is unclear whether biofilms enhance survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation and storage in dry milk powder on Salmonella survival and gene expression after passage through an in vitro digestion model. Planktonic cells of Salmonella enterica serotype Tennessee were deposited onto membranes while biofilms were formed on glass beads. The cells were subsequently dried at room temperature and stored in dried milk powder (a(w)=0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto Brilliant Green Agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling period both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and Quantitative Real-Time PCR was used to determine relative expression for genes associated with stress response (rpoS, otsB), virulence (hilA, invA, sipC) and a housekeeping gene 16S rRNA. The physiological state and length of storage affected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in vitro digestion system (p<0.05). Larger numbers of S. Tennessee were recovered by plate counts for biofilms compared to planktonic, however, the numbers of Salmonella genomes detected by qPCR were not significantly different suggesting entry of the planktonic cells of S. Tennessee into a viable but non-culturable state. The increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross-protection to low water activity and acid stress. Increased expression of virulence-associated genes was seen in cells exposed to dry storage for short periods, however the largest amount of expression occurred in biofilm cells stored for 30 days at aw 0.3, suggesting increased virulence potential.

Area and volume measurements of objects with irregular shapes using multiple silhouettes

Surface area and volume measurements provide important information for agriculture and food-processing applications. A machine vision system that uses a nondestructive method to measure volume and surface area of objects with irregular shapes is presented in this paper. The system first takes a series of silhouettes of the object from different directions by rotating the object at a fixed angular interval. The boundary points of each image are then extracted to construct a silhouette. A three-dimensional wire-frame model of the object can be reconstructed by integrating silhouettes obtained from different view angles. Surface area and volume can then be measured by means of surface fitting and approximation on the wire-frame model. System calibration and surface approximation were two major challenges for the design of this machine vision system. A unique centerline calibration method is introduced in this paper. Surface approximation and calculation are also discussed. Examples of applications in agriculture and food processing using this vision system for surface area measurement are included, and its accuracy is verified.