Food Protection Trends - September/October 2020 - 322
only) were used to collect data at another nine Iowa retail
delicatessen establishments. Similar results to the first pilot
occurred with test kits producing false-positive results.
On the basis of these results, the decision was made to use
traditional sponge stick sampling methodology for the final
sampling assignment.
Final collection
There were 57 small and large retail deli operations in Iowa
that were randomly selected for the final data collection.
The selection was based on a quota sampling technique with
approximately 75% (n = 43) of selected operations being small
and the remaining approximately 25% (n = 14) being large
operations. Sampling at more small operations was intentional,
given the higher frequency of violations in small delis. All data
were collected during routine health inspections.
Microbial sample collection
Three trained individuals visited and collected microbial
samples by using a traditional gloved sponge stick technique.
Inspectors were trained in aseptic sample collection
techniques, which is the standard method for all FDA
consumer safety officers during sampling assignments
(48). Each sampling required the inspector to collect five
to seven samples from clean food contact areas used for
RTE delicatessen meat preparation or storage. Sampling
site examples were as follows: slicer, meat scale, cutting
board, walk-in cooler racks, and deli meat cases. Inspectors
labeled each sample bag with the location, sample number,
and sample lot number. Information from each sample was
captured on a sample collection form with location, sample
zone, sample number, and sample lot code. The operator was
instructed to reclean any sampled equipment. All samples
were shipped the same day in soft-sided coolers to the State
Hygienic Laboratory for analysis with the designated chain
of custody forms.
Microbiological analyses
For the final data collection activity, World Bioproducts
EZ Reach™ sterile polyurethane sponges that contained
10 mL of Dey-Engley neutralizing broth (EZ-10DE-PUR,
World Bioproducts, Bothell, WA) were used for collection
and sent to the State Hygienic Laboratory for Listeria species
testing and culture confirmation. The microbiological
procedures used for the environmental sample processing
were from the FDA Bacteriological Analytical Manual (49)
and Hygiena BAX System Real-Time PCR Assay for Genus
Listeria (D15131113, Hygiena, LLC). Upon receipt in the
laboratory, the sponges were homogenized (vigorously
hand massaged) with prewarmed University of Vermont
media (modified Listeria enrichment broth; Difco, BD,
Sparks, MD) broth and incubated at 30°C for 20 to 26 h.
After primary incubation, 0.1 mL of primary enrichment
was transferred into 10 mL of morpholinepropanesulfonic
322
Food Protection Trends September/October
acid-buffered Listeria enrichment broth (BD Bacto; BD)
with antibiotics (0.5% acriflavine, 0.5% nalidixic acid, and
1% cycloheximide solutions, Sigma-Aldrich Corporation,
St. Louis, MO) and incubated at 35°C for 18 to 24 h.
Real-Time PCR by using Hygiena BAX System Real-Time
PCR Assay for Genus Listeria (D15131113, Hygiena,
LLC) was performed first. If the PCR test was positive,
aliquots from morpholinepropanesulfonic acid-buffered
Listeria enrichment broth were transferred to two agar
media: CHROMagar Listeria (CHROMagar, Paris, France)
and modified Oxford Listeria selective agar (Difco, BD)
containing modified Oxford antimicrobic supplement (Difco,
BD). Suspect Listeria colonies were isolated on sheep blood
agar plates (Difco, BD) and the Listeria genus and species
were confirmed by using matrix-assisted laser desorption-
ionization time of flight (Daltonics Microflex LT, Bruker
Corporation, Boston, MA) mass spectrometry.
All five Listeria monocytogenes isolates recovered for this
project were submitted for whole-genome sequencing at
the State Hygienic Lab (Table 1). Sequencing material and
methods are given in the following.
DNA isolation
DNA isolation was performed on a QIAcube (Qiagen,
Hilden, Germany) instrument by using the QIAamp DNA
Mini Kit (catalog #51306, Qiagen) according to the manufacturer's protocol. Prior to loading bacterial cells on the
instrument, an enzymatic digestion was performed by using
lysozyme (20 mg/mL; catalog #L4919, Millipore Sigma, St.
Louis, MO) and RNase A (11 ng/μL final concentration;
catalog #19101, Qiagen). DNA was eluted in 100 L of 10
mM Tris HCl, pH 8. DNA concentrations were measured
on a Qubit 3.0 Fluorometer (Thermo Fisher Scientific Inc.,
Waltham, MA) by using the Qubit dsDNA BR Assay Kit
(catalog #Q32853, Thermo Fisher Scientific Inc.).
Library preparation and sequencing
Libraries were prepared by using the Illumina Nextera
XT DNA Library Prep Kit (catalog #FC-131-1096) and
Nextera XT Index Kit v2 (catalog #FC-131-2003, Illumina
Inc., San Diego, CA) following the FDA Center for Food
Safety and Applied Nutrition protocol "Nextera XT Library
Preparation for Sequencing on a MiSeq" (29). Libraries
quality were inspected by using the Qubit dsDNA HS
Assay Kit (catalog #Q32854, Thermo Fisher Scientific Inc.,
Waltham, MA) and the Agilent Bioanalyzer High Sensitivity DNA kit (catalog #5067-4626, Agilent Technologies Inc.,
Waldbronn, Germany). Libraries for Listeria were pooled
together with other bacterial libraries to reach a total of 160
Mb per run and sequenced on an Illumina MiSeq instrument (Illumina Inc.) for 2 × 250 cycles by using a MiSeq
Reagent Kit v3 (catalog #MS-102-3003, Illumina Inc.).
Food Protection Trends - September/October 2020
Table of Contents for the Digital Edition of Food Protection Trends - September/October 2020
Self-Reported Food Safety Behaviors in Independent Chinese and Mexican Restaurants in Kansas
Microbiological Quality of Bottled Water Obtained from Mexican Small Water Purification Plants: A Pilot Study, Carried Out in Morelia (Central Mexico)
Listeria monocytogenes Occurrence and Adherence to Recommendations: Small and Large Retail Delicatessens in Iowa
Utilization of Quantitative and Qualitative Methods to Investigate the Impacts of a Pilot Media Campaign Targeting Safe Cooking Techniques and Proper Thermometer Use
Defining the Flow and Food Safety Behaviors of Actors in the Cambodian Vegetable Value Chain
Beyond the Bio Yaohua "Betty" Feng
PDG Highlight Meat and Poultry Safety and Quality PDG
General Interest Paper Identifying Vulnerable Populations at Risk of Foodborne Infection: People with Diabetes Mellitus
Industry Products
Coming Events
Food Protection Trends - September/October 2020 - Cover1
Food Protection Trends - September/October 2020 - Cover2
Food Protection Trends - September/October 2020 - 289
Food Protection Trends - September/October 2020 - 290
Food Protection Trends - September/October 2020 - 291
Food Protection Trends - September/October 2020 - 292
Food Protection Trends - September/October 2020 - 293
Food Protection Trends - September/October 2020 - 294
Food Protection Trends - September/October 2020 - 295
Food Protection Trends - September/October 2020 - Self-Reported Food Safety Behaviors in Independent Chinese and Mexican Restaurants in Kansas
Food Protection Trends - September/October 2020 - 297
Food Protection Trends - September/October 2020 - 298
Food Protection Trends - September/October 2020 - 299
Food Protection Trends - September/October 2020 - 300
Food Protection Trends - September/October 2020 - 301
Food Protection Trends - September/October 2020 - 302
Food Protection Trends - September/October 2020 - 303
Food Protection Trends - September/October 2020 - 304
Food Protection Trends - September/October 2020 - 305
Food Protection Trends - September/October 2020 - 306
Food Protection Trends - September/October 2020 - 307
Food Protection Trends - September/October 2020 - 308
Food Protection Trends - September/October 2020 - 309
Food Protection Trends - September/October 2020 - 310
Food Protection Trends - September/October 2020 - 311
Food Protection Trends - September/October 2020 - 312
Food Protection Trends - September/October 2020 - 313
Food Protection Trends - September/October 2020 - Microbiological Quality of Bottled Water Obtained from Mexican Small Water Purification Plants: A Pilot Study, Carried Out in Morelia (Central Mexico)
Food Protection Trends - September/October 2020 - 315
Food Protection Trends - September/October 2020 - 316
Food Protection Trends - September/October 2020 - 317
Food Protection Trends - September/October 2020 - 318
Food Protection Trends - September/October 2020 - 319
Food Protection Trends - September/October 2020 - Listeria monocytogenes Occurrence and Adherence to Recommendations: Small and Large Retail Delicatessens in Iowa
Food Protection Trends - September/October 2020 - 321
Food Protection Trends - September/October 2020 - 322
Food Protection Trends - September/October 2020 - 323
Food Protection Trends - September/October 2020 - 324
Food Protection Trends - September/October 2020 - 325
Food Protection Trends - September/October 2020 - 326
Food Protection Trends - September/October 2020 - 327
Food Protection Trends - September/October 2020 - 328
Food Protection Trends - September/October 2020 - 329
Food Protection Trends - September/October 2020 - 330
Food Protection Trends - September/October 2020 - 331
Food Protection Trends - September/October 2020 - Utilization of Quantitative and Qualitative Methods to Investigate the Impacts of a Pilot Media Campaign Targeting Safe Cooking Techniques and Proper Thermometer Use
Food Protection Trends - September/October 2020 - 333
Food Protection Trends - September/October 2020 - 334
Food Protection Trends - September/October 2020 - 335
Food Protection Trends - September/October 2020 - 336
Food Protection Trends - September/October 2020 - 337
Food Protection Trends - September/October 2020 - 338
Food Protection Trends - September/October 2020 - 339
Food Protection Trends - September/October 2020 - 340
Food Protection Trends - September/October 2020 - 341
Food Protection Trends - September/October 2020 - 342
Food Protection Trends - September/October 2020 - 343
Food Protection Trends - September/October 2020 - 344
Food Protection Trends - September/October 2020 - 345
Food Protection Trends - September/October 2020 - 346
Food Protection Trends - September/October 2020 - 347
Food Protection Trends - September/October 2020 - 348
Food Protection Trends - September/October 2020 - Defining the Flow and Food Safety Behaviors of Actors in the Cambodian Vegetable Value Chain
Food Protection Trends - September/October 2020 - 350
Food Protection Trends - September/October 2020 - 351
Food Protection Trends - September/October 2020 - 352
Food Protection Trends - September/October 2020 - 353
Food Protection Trends - September/October 2020 - 354
Food Protection Trends - September/October 2020 - 355
Food Protection Trends - September/October 2020 - 356
Food Protection Trends - September/October 2020 - 357
Food Protection Trends - September/October 2020 - 358
Food Protection Trends - September/October 2020 - 359
Food Protection Trends - September/October 2020 - 360
Food Protection Trends - September/October 2020 - 361
Food Protection Trends - September/October 2020 - 362
Food Protection Trends - September/October 2020 - 363
Food Protection Trends - September/October 2020 - Beyond the Bio Yaohua "Betty" Feng
Food Protection Trends - September/October 2020 - 365
Food Protection Trends - September/October 2020 - 366
Food Protection Trends - September/October 2020 - 367
Food Protection Trends - September/October 2020 - 368
Food Protection Trends - September/October 2020 - 369
Food Protection Trends - September/October 2020 - 370
Food Protection Trends - September/October 2020 - 371
Food Protection Trends - September/October 2020 - 372
Food Protection Trends - September/October 2020 - PDG Highlight Meat and Poultry Safety and Quality PDG
Food Protection Trends - September/October 2020 - General Interest Paper Identifying Vulnerable Populations at Risk of Foodborne Infection: People with Diabetes Mellitus
Food Protection Trends - September/October 2020 - 375
Food Protection Trends - September/October 2020 - 376
Food Protection Trends - September/October 2020 - 377
Food Protection Trends - September/October 2020 - 378
Food Protection Trends - September/October 2020 - 379
Food Protection Trends - September/October 2020 - Industry Products
Food Protection Trends - September/October 2020 - 381
Food Protection Trends - September/October 2020 - 382
Food Protection Trends - September/October 2020 - 383
Food Protection Trends - September/October 2020 - Coming Events
Food Protection Trends - September/October 2020 - Cover3
Food Protection Trends - September/October 2020 - Cover4
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