Food Protection Trends - September/October 2024 - 363

Antimicrobial treatment preparation
SaniDate 15 (PAA; BioSafe Systems, East Hartford,
CT) at a concentration of 75 ± 5 ppm of PAA, and
Ultra Clorox germicidal bleach (Cl; Clorox Professional
Products, Oakland, CA), at a concentration of 25 ± 2
ppm of free available Cl, were used for the treatments,
with sterile deionized (DI) water used as a control (W).
These concentrations were determined by EPA product
labels and the approved concentration limits for use with
produce (32, 35). Before each replication, titrations were
completed following manufacturer instructions to confirm
stock solution concentrations that achieve the required
75 ± 5 ppm of PAA and 25 ± 3 ppm of free available Cl in
the final 1,000-ml treated water samples (treatment details
follow). Sterile aluminum foil-covered flasks were used
to prepare treatments to ensure the treatments were not
affected by light exposure. A free and total Cl high-range
portable photometer (HI96734; Hanna Instruments,
Woonsocket, RI) was used to validate the free available
Cl concentrations. The PAA treatment concentration
was validated using a peracetic acid test kit (BioSafe
Systems, East Hartford, CT) following the manufacturer's
instructions. The concentrations of both treatments were
confirmed at the beginning and the end of each trial day
to ensure the same treatment concentration was used
throughout the duration of the trial. The PAA and Cl
concentrations were not confirmed in each water sample
throughout the trial.
Antimicrobial application and microbial analysis
For time point 0, the sample was collected immediately
after a 10-ml aliquot of the appropriate treatment solution
(Cl, PAA, and W) was added into each 990-ml sample
bottle and swirled to equally distribute the sanitizer for
10 s (hereafter called t = 0). Water sampling for E. coli
enumeration from each treated bottle was also performed 5,
10, 60, 1,440, and 2,880 min after treatment. Enumeration
using EC Petrifilm was previously described in a similar
study conducted by our research team (22) and was one of
two methods used for enumeration in this study. However,
because the EC Petrifilm is not an FDA-approved testing
method for agricultural water, the IDEXX Colilert QuantiTray/2000
was also used to provide an FDA-approved
method in comparison to the previously published EC
Petrifilm method. For the EC Petrifilm method, 1 ml of
each treated sample was neutralized in 9 ml of Dey-Engley
(DE) neutralizing buffer in tubes (Difco, Sparks, MD) and
5 ml of the treated sample was neutralized in 45 ml of DE
neutralizing buffer in Whirl-Pak bags (Nasco, Madison,
WI). Dilutions were prepared using DE-neutralized tubes
in 9 ml of PBDW and plated for enumeration in duplicate
on EC Petrifilm, and colonies (blue and purplish with gas
bubbles) were counted after Petrifilm incubation at 37°C for
48 ± 4 h. The limit of detection (LOD) for the EC Petrifilm
method was 5 CFU/ml. For Colilert enumeration, at each
sampling point, 100 ml of each treated (Cl and PAA)
sample was neutralized in a transparent, nonfluorescing
glass bottle containing 0.2 ml of 10% sodium thiosulfate
(Sigma Aldrich, St. Louis, MO) and then Colilert reagent
was added and samples were shaken to mix for 30 s. For
the W (control) samples, 10 µl from each treated sample
was diluted in 99.99 ml of sterile DI water containing the
Colilert reagent and 0.2 ml of 10% sodium thiosulfate.
The use of 0.2 ml of 10% sodium thiosulfate was validated
following the neutralizer or control validation procedure
of the FDA/EPA protocol (34). Then, all samples were
individually poured in a Colilert Quanti-Tray/2000, sealed,
and incubated at 35°C for 24 h. The LOD for Colilert is
one organism per 100 ml of water. For enrichment and
recovery of E. coli, 50 ml of 2× BHI broth was added to the
50-ml DE-neutralized sample (5 ml of sample + 45 ml of
DE neutralizing buffer) in Whirl-Pak bags, resulting in a 1×
BHI broth dilution, and then the bags were incubated at
37°C for 24 ± 2 h. After incubation, the enriched bags were
streaked on MacConkey agar (MAC; Thermo Scientific
Remel, Lenexa, KS) to determine the presence or absence
of E. coli, and MAC plates with pink colonies following
incubation at 37°C for 18-24 h were interpreted as positive
for E. coli. The enriched samples were considered to have
an LOD of 1 cell per 5 ml, or 20 cells in a 100-ml water
sample. Sample bottles were returned to their respective
temperatures after the 10-min time point and were taken
out for a short period (~5 min or less) for sample collection
at the subsequent time points.
Statistical analysis
The experimental procedures were replicated three times;
however, two replications of data are represented for the
Colilert method because counts were too numerous for
the W (control) samples for one replication. Statistical
analyses were conducted with Statistical Analysis Software
(SAS version 9.4; SAS Institute, Cary, NC). Data for
each combination of water source (creek and rain barrel)
and enumeration method (Colilert and EC Petrifilm)
were analyzed separately (e.g., creek water enumerated
using Petrifilm was analyzed separately from creek water
enumerated using Colilert). All data were subjected to
linear mixed modeling using the PROC MIXED procedure,
with a significance level of 0.05. Because repeated measures
analysis was used in this study, the best covariance structure
for the model was determined. The least squares means (LS
means) were calculated and the Tukey-Kramer adjustment
for multiple comparisons was used to determine statistical
significance between individual treatments. The main effects
of time, temperature, and treatment, as well as the three-way
interaction (time × temperature × treatment) and all twoway
interactions, were evaluated for statistical significance.
September/October Food Protection Trends 363

Food Protection Trends - September/October 2024

Table of Contents for the Digital Edition of Food Protection Trends - September/October 2024

Food Safety Needs Assessment for North American Pecan Shellers
Identifying Training Needs in Washington through Insights from Produce Safety Alliance Grower Training
Hazard versus Risk in Perceptions of Food Safety: The Case of Titanium Dioxide
Peroxyacetic Acid and Chlorine Reduce Escherichia coli in Agricultural Surface Water for Potential Produce Postharvest Uses
Beyond the Bio Jeffrey LeJeune
PDG Highlight Food Packaging
General Interest Paper Promoting Food Safety in the Informal Markets of Low- and Middle-Income Countries: The Need for a Rethink
Industry Products
Coming Events
Food Protection Trends - September/October 2024 - Cover1
Food Protection Trends - September/October 2024 - Cover2
Food Protection Trends - September/October 2024 - 329
Food Protection Trends - September/October 2024 - 330
Food Protection Trends - September/October 2024 - 331
Food Protection Trends - September/October 2024 - 332
Food Protection Trends - September/October 2024 - 333
Food Protection Trends - September/October 2024 - 334
Food Protection Trends - September/October 2024 - 335
Food Protection Trends - September/October 2024 - Food Safety Needs Assessment for North American Pecan Shellers
Food Protection Trends - September/October 2024 - 337
Food Protection Trends - September/October 2024 - 338
Food Protection Trends - September/October 2024 - 339
Food Protection Trends - September/October 2024 - 340
Food Protection Trends - September/October 2024 - 341
Food Protection Trends - September/October 2024 - 342
Food Protection Trends - September/October 2024 - 343
Food Protection Trends - September/October 2024 - Identifying Training Needs in Washington through Insights from Produce Safety Alliance Grower Training
Food Protection Trends - September/October 2024 - 345
Food Protection Trends - September/October 2024 - 346
Food Protection Trends - September/October 2024 - 347
Food Protection Trends - September/October 2024 - 348
Food Protection Trends - September/October 2024 - 349
Food Protection Trends - September/October 2024 - 350
Food Protection Trends - September/October 2024 - Hazard versus Risk in Perceptions of Food Safety: The Case of Titanium Dioxide
Food Protection Trends - September/October 2024 - 352
Food Protection Trends - September/October 2024 - 353
Food Protection Trends - September/October 2024 - 354
Food Protection Trends - September/October 2024 - 355
Food Protection Trends - September/October 2024 - 356
Food Protection Trends - September/October 2024 - 357
Food Protection Trends - September/October 2024 - 358
Food Protection Trends - September/October 2024 - Peroxyacetic Acid and Chlorine Reduce Escherichia coli in Agricultural Surface Water for Potential Produce Postharvest Uses
Food Protection Trends - September/October 2024 - 360
Food Protection Trends - September/October 2024 - 361
Food Protection Trends - September/October 2024 - 362
Food Protection Trends - September/October 2024 - 363
Food Protection Trends - September/October 2024 - 364
Food Protection Trends - September/October 2024 - 365
Food Protection Trends - September/October 2024 - 366
Food Protection Trends - September/October 2024 - 367
Food Protection Trends - September/October 2024 - 368
Food Protection Trends - September/October 2024 - 369
Food Protection Trends - September/October 2024 - Beyond the Bio Jeffrey LeJeune
Food Protection Trends - September/October 2024 - 371
Food Protection Trends - September/October 2024 - 372
Food Protection Trends - September/October 2024 - PDG Highlight Food Packaging
Food Protection Trends - September/October 2024 - 374
Food Protection Trends - September/October 2024 - 375
Food Protection Trends - September/October 2024 - General Interest Paper Promoting Food Safety in the Informal Markets of Low- and Middle-Income Countries: The Need for a Rethink
Food Protection Trends - September/October 2024 - 377
Food Protection Trends - September/October 2024 - 378
Food Protection Trends - September/October 2024 - 379
Food Protection Trends - September/October 2024 - 380
Food Protection Trends - September/October 2024 - 381
Food Protection Trends - September/October 2024 - 382
Food Protection Trends - September/October 2024 - 383
Food Protection Trends - September/October 2024 - Industry Products
Food Protection Trends - September/October 2024 - 385
Food Protection Trends - September/October 2024 - 386
Food Protection Trends - September/October 2024 - 387
Food Protection Trends - September/October 2024 - 388
Food Protection Trends - September/October 2024 - 389
Food Protection Trends - September/October 2024 - 390
Food Protection Trends - September/October 2024 - 391
Food Protection Trends - September/October 2024 - Coming Events
Food Protection Trends - September/October 2024 - Cover3
Food Protection Trends - September/October 2024 - Cover4
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