Food Microbiology 113 (2023) 104268

Contents lists available at ScienceDirect

Food Microbiology
journal homepage: www.elsevier.com/locate/fm

Transfer rates of Salmonella Typhimurium, Listeria monocytogenes, and a

human norovirus surrogate impacted by macronutrient composition of food
inks in 3D food printing systems
Allyson N. Hamilton, Kristen E. Gibson *
Department of Food Science, Center for Food Safety, University of Arkansas System Division of Agriculture, Fayetteville, AR, 72704, USA

A R T I C L E I N F O A B S T R A C T

Keywords: 3D food printers (3DFPs) allow for the customization of the physiochemical properties of foods in new ways.
3D printed food safety Transfer kinetics of foodborne pathogens between surfaces and food inks have not been evaluated in 3DFPs. This
Foodini study aimed to determine if the macromolecular composition of food inks would impact the transfer rate of
Capsule
foodborne pathogens from the stainless steel food ink capsule to the 3D printed food. Salmonella Typhimurium,
Sugar
Protein powder
Listeria monocytogenes, and a human norovirus surrogate, Tulane virus (TuV), were inoculated onto the interior
Butter surface of stainless steel food ink capsules and dried for 30 min. Subsequently, 100 g of one of the following
prepared food inks was extruded: (1) pure butter, (2) a powdered sugar solution, (3) a protein powder solution,
and (4) a 1:1:1 ratio of all three macromolecules. Pathogen enumeration was completed for both the soiled
capsules and the printed food products and resulting transfer rates were estimated using a generalized linear
model with quasibinomial errors. A significant two-way interaction effect was found between microorganism
type and food ink type (P = 0.0002). Tulane virus was typically transferred the most, and no significant dif­
ferences between L. monocytogenes and S. Typhimurium were observed for any food matrix or across food
matrices. Among food matrices, the complex mixture transferred fewer microorganisms in all instances, while
butter, protein, and sugar were all statistically indistinguishable. This research seeks to further develop the field
of 3DFP safety and to advance the understanding of the role of macromolecular composition in pathogen transfer
kinetics, which have not previously been explored in pure matrices.

1. Introduction
3D food printing is a new technology that will allow consumers and
manufacturers to uniquely customize the texture, color, shape, and other
physiochemical characteristics of food (Sun et al., 2015). Food materials
that have been 3D printed include cheese (Le Tohic et al., 2018; Ross
et al., 2021), cookies (Yang et al., 2018; Vukušić Pavičić et al., 2021),
fish (Wang et al., 2018), chocolate (Lanaro et al., 2017; Mantihal et al.,
2019), cell cultures of meat (Handral et al., 2020; Ramachandraiah,
2021), chicken nuggets (Wilson et al., 2020), pork (Dick et al., 2020)
and edible insects (Severini et al., 2018) among many others. The 3D
food printer (3DFP) used in the present study, the Foodini (Natural
Machines, Spain) as well as a list of other 3DFPs available on the market
has been published by Baiano (2022). The global 3D food printing
market is anticipated to reach USD$525.6 million by 2023 (Lee, 2021).
Industries that may benefit from the adoption of 3DFP include those that
cater to patients with eating and swallowing disorders, such as hospitals
(Hurst, 2016) and long-term care facilities (Dick et al., 2020). In addi­
tion, 3DFP could aid industries with limited storage space or unreliable
access to food, such as space travel (Terfansky and Thangavelu, 2013),
cruise ships, or geographically isolated locations. Finally, artisanal in­
dustries such as pasta (Scott, 2021) and gourmet restaurants may benefit
from the versatility and consumer curiosity surrounding 3D printed food
products (Mantihal et al., 2020).
New food-related technologies need to be thoroughly characterized
for potential microbial risks to protect consumers from foodborne illness
prior to widespread adoption. 3D food printing may pose increased
foodborne illness risk due to elevated handling demands and niches that
could become harborage points in the food ink capsules, as previously
described by the authors (Hamilton and Gibson, 2022). For example, to
3D print any food, all traditional handling steps are required along with
loading the food ink capsule and post-printing handling. In addition to

* Corresponding author. Department of Food Science, University of Arkansas, 1371 West Altheimer Dr., Fayetteville, AR, 72704, USA.
E-mail address: keg005@uark.edu (K.E. Gibson).

https://doi.org/10.1016/j.fm.2023.104268
Received 2 December 2022; Received in revised form 8 February 2023; Accepted 17 March 2023
Available online 20 March 2023
0740-0020/© 2023 Published by Elsevier Ltd.
A.N. Hamilton and K.E. Gibson
cross-contamination risks between the capsule and food ink, potential
microbial harborage points of concern include the two rubber O-rings
and stainless steel threads used to connect the capsule tip to the capsule.
The primary pathogens of concern in 3D printed foods also represent
major causes of foodborne illness in the U.S. With an estimated 9.4
million illnesses and over 1300 deaths attributed to major foodborne
pathogens in the U.S. per year, human norovirus (HuNoV) is responsible
for 58% (5.4 million) of illnesses and 149 deaths; nontyphoidal Salmo­
nella spp. Are linked to 1.03 million illnesses and 378 deaths; and Listeria
monocytogenes is associated with nearly 1600 illnesses and 255 deaths
(Scallan et al., 2011). The combined economic burden of these three
foodborne pathogens is approximately $9.351 B out of a total of
$51.048 B for all foodborne pathogens (Scharff, 2012).
Transfer of HuNoV, Salmonella spp., and L. monocytogenes has been
investigated outside of 3DFP and 3D printed food products. For example,
HuNoV can be transferred bidirectionally between gloved and ungloved
human finger pads and stainless steel in significant quantities (Sharps
et al., 2012; Tuladhar et al., 2013). Additionally, HuNoV, Salmonella
spp., and L. monocytogenes have documented transfer rates from stainless
steel surfaces to a variety of food products (Moore et al., 2003; Rodri­
guez and McLandsborough, 2007; Stals et al., 2013), yet the risk of
transfer based on macronutrient composition of foods has not been
analyzed, nor has transfer in 3D food printing systems.
Analysis of risk is essential for regulatory agencies to properly con­
trol the spread of foodborne disease and to increase consumer confi­
dence in the safety of 3D printed foods. Therefore, the present study
focuses on evaluating the transfer rates of a HuNoV surrogate (Tulane
virus), Salmonella enterica subsp. Enterica ser. Typhimurium, and
L. monocytogenes based on the macronutrient composition of the food
products. Specifically, the study aimed to evaluate the transfer rates of
each pathogen (or surrogate) from an inoculated, stainless steel food ink
capsule to one of the following food inks: (1) pure butter, (2) a powdered
sugar solution, (2) a protein powder solution, and (4) a 1:1:1 complex
food ink matrix of all three macronutrients when 3D printed.
2. Materials and methods
2.1. Tulane virus propagation
Tulane virus, provided by Dr. Jason Jiang (Cincinnati Children’s
Hospital Medical Center, Cincinnati, OH) was propagated in LLC-MK2
cells (ATCC CCL-7) (American Type Culture Collection, Manassas, VA)
as previously described (Arthur and Gibson, 2015). Briefly, LLC-MK2
cells were maintained in Medium 199 with Earle’s Balanced Salt Solu­
tion (EBSS) (Hyclone, Logan, UT) supplemented with 1% Pen­
icillin/Streptomycin (Hyclone), 1% Amphotericin B (Corning,
Mediatech Inc., Manassas, VA), and 10% Fetal Bovine Serum (FBS)
(Hyclone). TuV stock (at a multiplicity of infection (MOI) of 0.1 in 6 mL
of Opti-MEM (Gibco Life Technologies, Grand Island, NY)) was inocu­
lated in cell monolayers with 90% LLC-MK2 cell confluency and allowed
to incubate with continuous rocking at 5% CO2 and 37 ◦ C for 1 h.
Subsequently, 20 mL of Opti-MEM supplemented with 2% FBS was
added, and the T150 flask was incubated at 5% CO2 and 37 ◦ C until a
complete cytopathic effect (CPE) was observed (approximately 60 h).
Then, the flask was held at − 80 ◦ C overnight, and TuV was harvested via
three freeze-thaw cycles and centrifugation at 3000×g for 15 min at
4 ◦ C. The supernatant was filtered through a Steriflip® 0.45 μm filter
(Millipore, Burlington, MA) and then aliquoted in 1 mL cryovials before
storing at − 80 ◦ C.
2.2. Quantification of TuV
Quantification of TuV was completed as previously described by
Arthur and Gibson (2015). In short, six-well tissue culture-treated plates
were seeded with 4 × 105 LLC-MK2 cells per well and incubated for 24 h
at 37 ◦ C under 5% CO2. The media was removed and 500 μL of 10-fold
2
Food Microbiology 113 (2023) 104268
serially diluted TuV stock (10− 1 to 10− 6) in Opti-MEM supplemented
with 2% FBS was added to each well before the plates were incubated for
1 h at 37 ◦ C under 5% CO2 with gentle rocking. During incubation, an
overlay mixture was prepared by combining liquefied 3% low-melting
NuSieve™ GTG™ Agarose (Lonza, Walkersville, MD) and with an
equal volume of Opti-MEM supplemented with 2% FBS. After incuba­
tion, the sample was aspirated, and 2 mL of overlay mixture was added
to each well. Following 15 min to allow the overlay to solidify, the
six-well plates were incubated at 37 ◦ C and 5% CO2 until plaques
appeared (approximately 96–120 h). Plaque visualization was achieved
by preparing 3% of neutral red stock (Sigma-Aldrich, St Louis, USA) in
1X PBS, adding 2 mL of the stain to each well, and incubating at 37 ◦ C
under 5% CO2 for 3 h. After incubation, the stain was aspirated, and the
plaque forming units (PFU) were counted. Final TuV stock concentra­
tions were approximately 106 PFU/mL.
2.3. Bacteria preparation
Listeria monocytogenes (FSL R9-5506, serotype 4 b environmental
isolate from 2016 packaged salad outbreak; Food Safety Laboratory,
Cornell University, Ithaca, NY) (Burall et al., 2017) and Salmonella
enterica subsp. enterica serovar Typhimurium strain CDC 6516–60
(ATCC 14028; American Type Culture Collection, Manassas, VA) were
cultivated and prepared as described by Gibson et al. (2019). Briefly,
L. monocytogenes and S. Typhimurium were streaked from 50% glycerol
stocks onto brain heart infusion (BHI) agar (Hardy Diagnostics, Santa
Monica, CA) and tryptic soy agar (TSA) (Becton, Dickson, and Company,
Sparks, MD), respectfully. The streak plates were incubated at either
35 ◦ C (for L. monocytogenes) or 37 ◦ C (for S. Typhimurium) overnight. A
single colony of L. monocytogenes or S. Typhimurium was selected and
inoculated into 10 mL of BHI broth (Hardy Diagnostics) or 10 mL of
tryptic soy broth (TSB) (Acumedia, Lansing, MI), respectively. Both
cultures were shaken overnight at 125 rpm and 35 ◦ C. The cultures were
centrifuged at 4000×g for 10 min, the supernatant was decanted, and
the pellets were each resuspended in 2 mL of 1X phosphate-buffered
saline (PBS). This washing process was repeated three times, except
after the final centrifugation when the two organisms were resuspended
together into 2 mL of 1X PBS to create a bacterial cocktail.
For enumeration from the cocktail, both L. monocytogenes and S.
Typhimurium were serially diluted and plated on CHROMagar™ Listeria
chromogenic media (CHROM) (CHROMagar, Paris, France) or Xylose
Lysine Tergitol™4 (XLT4) (XLT4 Agar Base; Hardy Diagnostics) fol­
lowed by incubation at 35 ◦ C for 48 h or 37 ◦ C for 24 h, respectively.
Viable cells were reported as log colony forming units per mL (log CFU/
mL), and the final concentration in the cocktail was approximately 9 log
CFU/mL for each bacterial species.
2.4. Food Ink Preparation
Food ink preparation details are identical to the procedure previ­
ously described by the authors (Hamilton and Gibson, 2022, 2023).
Briefly, unsalted butter (Member’s Mark Unsalted Sweet Cream Butter;
Sam’s West, Inc, Midwest City, OK), protein powder (Sports Research
Collagen Powder Supplement; Sports Research, San Pedro, CA), and
powdered sugar (Domino Confectioners Sugar; Domino Food, Yonkers,
NY) were procured and used to prepare a 1:1:1 mixture (based on
amount of macromolecule per gram of product) of fat, protein, and
carbohydrate (25 g butter, 20 g powdered sugar, 21 g protein powder,
and 50 mL DI water). Individual food components were also tested
separately and used in pure form (butter) or mixed with DI water to form
aqueous preparations (protein powder and powdered sugar) suitable for
printing by the 3DFP.
2.5. Preparation and inoculation of the food ink capsules
The Foodini 3DFP has already been described, at length, by the
A.N. Hamilton and K.E. Gibson
authors (Hamilton and Gibson, 2023). In brief, the Foodini is a
counter-top 3DFP marketed for use both in the home and in high-end
restaurants. The main food contact surface is the stainless steel food
ink capsule that takes on the basic shape of a cylinder (ℓ = 11 cm, d = 4
cm), and the food ink is the food material that is extruded from the
capsule. The user manual for the Foodini indicates that the maximum
volume of food ink that should be added in a single capsule is 100 mL
(https://static.naturalmachines.com/images/Natural-Machines-Foo
dini-Brochure.pdf).
All stainless steel capsules were prepared following the methods
described by Hamilton and Gibson (2022). Briefly, all capsules were
washed with dish soap (Dawn Ultra; Proctor and Gamble, Cincinnati,
OH) before being rinsed in DI water and sprayed with 70% ethanol. After
air drying, the capsules were autoclaved for 15 min at 121 ◦ C in sealed
sterilization pouches.
The inoculation method was repeated from prior publication
(Hamilton and Gibson, 2022, 2023). Specifically, 100 μL of ~109
CFU/mL of the bacterial cocktail (containing both S. Typhimurium and
L. monocytogenes) or 250 μL of ~106 PFU/mL of TuV stock were spot
inoculated and distributed evenly with a sterile inoculation loop along
the interior capsule surface. After being allowed to dry in a biosafety
cabinet for 30 min, ~100 g of food ink material (see Food Ink Prepa­
ration) was added, and the capsule cap was placed. The capsule cap was
pushed to the bottom of the capsule, and the extruded food ink was
caught in a stomacher bag (Filtra-bag® with tear-off top, 7 × 12 in.,
VWR).
2.6. Sampling of the food ink capsules and extruded food ink
Sampling of the food ink capsule is replicated from prior publication
(Hamilton and Gibson, 2022, 2023). Briefly, a polyurethane foam (PUF)
swab (PUR-Blue™ Dry Swab; World Bioproducts, Woodinville, Wash­
ington) was premoistened with 3 mL of 1X PBS and used to swab the
entirety of the capsule interior for 60 s after extrusion had been
completed. The swab was vortexed for 1 min, and the sample was
transferred to a 2 mL microcentrifuge tube and serial diluted in 1X PBS
(for bacteria) or Opti-MEM supplemented with 2% FBS, 1% Pen­
icillin/Streptomycin, and 1% Amphotericin B (for TuV).
Sampling of the food ink was completed by adding 3 mL of 1X PBS to
the stomacher bag containing the food ink, stomaching for 1 min at 230
rev/min, and serially diluting the resulting mixture. For negative con­
trol, 1X PBS, Opti-MEM, and each food ink were tested alongside a
sterile PUF swab on a sterile capsule. For positive control, a capsule was
inoculated, soiled similar to previous studies (Hamilton and Gibson,
2022, 2023), allowed to dry, and immediately swabbed as described
above. An inoculated capsule without food ink was also tested. The
bacteria samples were enumerated in log CFU/mL by the spread plate
method on selective media (XLT-4 or CHROM).
2.7. Statistical analysis
Two experimental trials with three samples each were performed for
each food ink and microorganism combination (n = 6 per combination),
with bacteria being performed simultaneously. Statistical analysis was
computed to determine if macronutrient composition was a significant
predictor of microorganism transfer from the capsule to the food ink.
The proportion transferred was calculated by dividing the CFU recov­
ered from the printed food by the sum of the CFU remaining on the
capsule and the CFU recovered from the printed food. Data were first
analyzed in R Studio (R Core Team, 2022) using a linear model. How­
ever, residual analysis violated the assumptions of normality and ho­
moscedasticity, and the number of observations was not sufficient to
consider that the sample means were approximately normally distrib­
uted (central limit theorem). Data were subsequently analyzed using a
generalized linear model (GLM) with quasibinomial errors to account
for the observed overdispersion. The treatment means and their
3
Food Microbiology 113 (2023) 104268
associated standard errors were calculated using estimated marginal
means, and statistical differences between treatments were determined
using multiple comparisons and visualized using compact letter display.
The data were analyzed in R Studio (R Core Team, 2022) using the
base, ggplot2 (Wickham, 2016), emmeans (Length et al., 2021), tidyverse
(Wickham et al., 2019), ggpubr (Kassambara, 2020), gdata (Warnes et al.,
2022), rstatix (Kassambara, 2021), lme4 (Bates et al., 2015), lmertest
(Kuznetsova et al., 2017), multcomp (Hothorn et al., 2008), and mult­
compView (Graves et al., 2019) packages.
3. Results
All raw data are plotted in Fig. 1, which shows the log of CFUs or
PFUs recovered based on food matrix type, microorganism, and location
of recovery. A significant two-way interaction effect was found between
microbe and food ink (P = 0.0002), therefore, conclusions about the
main effects cannot be made.
A quasibinomial distribution was used to fit the data and estimate
proportional transfer rates from the inoculated food ink capsule to the
printed food product based on the macromolecular composition of the
food ink. The estimated proportion transferred from the capsule to the
3D printed food with the corresponding 95% confidence intervals and
the statistical groupings from the post-hoc analysis is shown in Fig. 2.
No statistical differences in transfer were found between S. Typhi­
murium and L. monocytogenes for any food matrix or across food
matrices, even though S. Typhimurium had higher transfer rates than
L. monocytogenes in all instances. While TuV was not statistically
different from either bacterium in the pure butter matrix, TuV had a
much higher estimated transfer rate 0.75 (95% CI: 0.62, 0.85) compared
to 0.45 (95% CI: 0.33, 0.60) for L. monocytogenes and 0.52 (95% CI: 0.39,
0.65) for S. Typhimurium, which perhaps represents differences of mi­
crobial significance that could further be elucidated with subsequent
studies. Additionally, the pure protein and sugar matrices transferred
significantly more TuV than S. Typhimurium and L. monocytogenes.
Among food matrices, the complex mixture transferred fewer microor­
ganisms in all instances, while butter, protein, and sugar were all sta­
tistically indistinguishable.
4. Discussion
Transfer of foodborne pathogens in homes and in food service es­
tablishments is hypothesized to be a major contributing factor to spo­
radic and epidemic foodborne illnesses (Chen et al., 2001; Beumer and
Kusumaningrum, 2003; Bloomfield, 2003; Pérez-Rodríguez et al., 2008;
Grove et al., 2015). Consumers have been reported to handle and pre­
pare food using suboptimal hygiene practices (Gillespie, O’brien, &
Goutam, 2001; Redmond and Griffith, 2003); therefore, it is not unex­
pected that surface-to-food transfer is a major pathway by which food­
borne illnesses originating in the home are spread (Beumer and
Kusumaningrum, 2003; Bloomfield, 2003; Cardoso et al., 2021). In their
review, Pérez-Rodríguez (2008) and coauthors call for further investi­
gation of surface-to-food transfer in order to reduce foodborne illness
and aid in the development of effective hygiene practices.
Stals and coauthors (2013) investigated transfer of HuNoV GII.4 (~6
log genome copies) from inoculated and dried stainless steel discs to
lettuce, boiled ham, and a sandwich bun with transfer contact time of 10
s at a pressure of 0.2–0.4 kg/cm2 and 90-degree rotation. While lettuce is
not analogous to any food material used in the present study, boiled ham
and a sandwich bun can loosely be compared to protein and sugar,
respectively, but it is difficult to compare food inks to finished food
products. The transfer from stainless steel to the sandwich bun was
similar to transfer to the sugar matrix in the present study with transfer
rates of ~0.6 and 0.78 (95% CI: 0.65, 0.87), respectively (Stals et al.,
2013). The transfer from stainless steel to the boiled ham was different
from transfer to the protein matrix in the present study with transfer
rates of <0.10 and 0.78 (95% CI: 0.65, 0.87), respectively (Stals et al.,
A.N. Hamilton and K.E. Gibson Food Microbiology 113 (2023) 104268

Fig. 1. Raw data of log colony forming units (CFU) or plaque forming units (PFU) recovered in capsule post-extrusion based on food matrix type, microorganism, and
location of recovery. Each boxplot corresponds to six independent samples.

Fig. 2. Generalized linear model with quasibinomial errors for the transfer of L. monocytogenes, S. Typhimurium, and Tulane virus from the inoculated capsule to the
printed food based on food matrix composition. Compact letter format is used to designate statistical differences between treatments at P = 0.05. Error bars represent
95% confidence intervals.

2013). To the authors’ knowledge, HuNoV transfer has not been inves­
tigated in high-fat food materials, as HuNoV is often studied in foods
such as fresh and frozen produce (Sharps et al., 2012; Verhaelen et al.,
2013; Grove et al., 2015).
Dawson and coauthors (2006) investigated transfer of S. Typhimu­
rium from finished ceramic tile to bologna (21% fat, 11% protein, 7%
carbohydrate and 60% water) and found a transfer rate of 0.69 with 5
min of drying compared to 0.37 (95% CI: 0.26, 0.50) for the complex
mixture in the present study (Dawson et al., 2006). Again, it is difficult
to directly compare food inks to prepared food products because there
are inherent differences in consistency and composition; however, after
2 h of drying, Dawson and coauthors (2006) reported a transfer rate of
0.4, which aligns more closely with the value corresponding to the
present study’s 30 min of drying time. Notably, Moore and coauthors

4
A.N. Hamilton and K.E. Gibson
(2003) studied the effect of drying time (tested at 0, 30, 60, 90, and 120
min) on the transfer of S. Typhimurium from stainless steel to dry lettuce
and found fairly steep drop offs in recovery from the lettuce in the in­
terval between 60 (5.18 log CFU) and 90 min (2.51 log CFU).
Currently, the literature surrounding transfer of L. monocytogenes
centers on its transfer between deli slicers and deli meat. Unfortunately,
these studies typically report CFU/slice transfer instead of transfer rates,
making comparison to the present study difficult (Vorst et al., 2006;
Keskinen et al., 2008; Sheen, 2008). Rodriguez and McLandsborough
(2007) measured efficiencies of transfer of L. monocytogenes in biofilms
on stainless steel to either bologna or American cheese and found that
transfer to bologna was more efficient. Additional studies on the transfer
of L. monocytogenes between surfaces and food have focused on
biofilm-related events (Midelet and Carpentier, 2002; Midelet et al.,
2006). For instance, Midelet and coauthors (2006) investigated 12
consecutive transfer events to a solid model food (solidified tryptone
soya agar) from a two-species biofilm on stainless steel containing
L. monocytogenes and one additional nonpathogenic microorganism (e.
g., Kocuria varians, Pseudomonas fluorescens, and Staphylococcus sciuri,
and an unidentified gram-negative bacterium isolated from a food pro­
cessing environment). The study authors observed both biphasic and
monophasic transfer kinetics (Midelet et al., 2006). The numerous
transfer events are somewhat similar to the extrusion process used in
3DFPs, and a similar model could be developed based on extrusion
volume as a future research direction.
Based on the present study, the authors recommend that 3D food
printing be further investigated to protect the safety and wholesomeness
of the global food supply. Foods that contain a complex mixture of
macromolecules may be less susceptible to viral contamination from
transfer than those that contain pure macronutrients as indicated by
data collected in this study. Moreover, food matrix composition doesn’t
seem to play a role in bacterial transfer. In the future, the impact of
processing conditions on pathogen transfer still remains to be examined.
Additionally, the food ink could be contaminated with pathogens, and
transfer to the capsule could be measured. In this instance, the transfer
rate will likely be correlated to the amount of residual food on the inner
surface of the capsule as opposed to the macromolecular composition of
the inks, and this should be considered a variable during the investi­
gation. Additionally, biofilms could be allowed to form on the interior
capsule surface, and transfer could be further studied under those con­
ditions. Overall, this research seeks to advance the field of 3DFP safety
as it relates to foodborne illness risk assessment and the understanding
of the role of macromolecular composition in bacterial and viral transfer
kinetics which has not previously been explored in pure matrices.
Funding
This work is supported in part by the National Institute of Food and
Agriculture (NIFA), U.S. Department of Agriculturee (USDA) Hatch Act
funding and the University of Arkansas Distinguished Doctoral Fellow­
ship (DDF). K.E. Gibson also received a Faculty Equipment and Tech­
nology grant from the University of Arkansas Honors College used to
support the purchase of the 3D food printer used in the present study.
Author contributions
Conceptualization, A.N.H. and K.E.G.; Methodology, A.N.H. and K.E.
G.; Formal Analysis, A.N⋅H.; Investigation, A.N⋅H.; Resources, K.E.G.;
Writing – Original Draft, A.N⋅H.; Writing – Reviewing and Editing, A.N.
H. and K.E.G.; Visualization, A.N⋅H.; Supervision, K.E.G.; Funding
Acquisition, K.E.G.
Data availability
Data will be made available on request.
5
Food Microbiology 113 (2023) 104268
Acknowledgements
The authors would like to thank Anna Rechtin for her assistance in
bacterial media preparation and general assistance throughout the
study.
References
Arthur, S.E., Gibson, K.E., 2015. Physicochemical stability profile of Tulane Virus: a
human norovirus surrogate. J. Appl. Microbiol. 119, 868–875. https://doi.org/
10.1111/jam.12878.
Baiano, A., 2022. 3D printed foods: A comprehensive review on technologies, nutritional
value, safety, consumer attitude, regulatory framework, and economic and
sustainability issues. Food Rev. Int. 38, 986–1016. https://doi.org/10.1080/
87559129.2020.1762091.
Bates, D., Mächler, M., Bolker, B., Walker, S., 2015. Fitting linear mixed-effects models
using lme4. J. Stat. Software 67, 1–48. https://doi.org/10.18637/jss.v067.i01.
Beumer, R.R., Kusumaningrum, H., 2003. Kitchen hygiene in daily life. Int. Biodeterior.
Biodegrad. 51, 299–302. https://doi.org/10.1016/s0964-8305(03)00041-6.
Bloomfield, S.F., 2003. Home hygiene: a risk approach. Int. J. Hyg Environ. Health 206,
1–8. https://doi.org/10.1078/1438-4639-00193.
Burall, L.S., Grim, C.J., Datta, A.R., 2017. A clade of Listeria monocytogenes serotype 4b
variant strains linked to recent listeriosis outbreaks associated with produce from a
defined geographic region in the US. PLoS One 12 (5), e0176912. https://doi.org/
10.1371/journal.pone.0176912.
Cardoso, M.J., Ferreira, V., Truninger, M., Maia, R., Teixeira, P., 2021. Cross-
contamination events of Campylobacter spp. in domestic kitchens associated with
consumer handling practices of Raw Poultry. Int. J. Food Microbiol. 338, 108984
https://doi.org/10.1016/j.ijfoodmicro.2020.108984.
Chen, Y., Jackson, K.M., Chea, F.P., Schaffner, D.W., 2001. Quantification and variability
analysis of bacterial cross-contamination rates in common food service tasks. J. Food
Protect. 64, 72–80. https://doi.org/10.4315/0362-028x-64.1.72.
Dawson, P., Han, I., Cox, M., Black, C., Simmons, L., 2006. Residence time and food
contact time effects on transfer of Salmonella Typhimurium from tile, wood and
carpet: testing the five-second rule. J. Appl. Microbiol. 102, 945–953. https://doi.
org/10.1111/j.1365-2672.2006.03171.x.
Dick, A., Bhandari, B., Dong, X., Prakash, S., 2020. Feasibility Study of hydrocolloid
incorporated 3D printed pork as Dysphagia Food. Food Hydrocolloids 107, 105940.
https://doi.org/10.1016/j.foodhyd.2020.105940.
Gibson, K.E., Almeida, G., Jones, S.L., Wright, K., Lee, J.A., 2019. Inactivation of bacteria
on fresh produce by batch wash ozone sanitation. Food Control 106, 106747.
https://doi.org/10.1016/j.foodcont.2019.106747.
Gillespie, I.A., O’Brien, S.J., Adak, G.K., 2001. General outbreaks of infectious intestinal
diseases linked with private residences in England and Wales, 1992-9: questionnaire
study. BMJ 323, 1097–1098. https://doi.org/10.1136/bmj.323.7321.1097.
Graves, S., Piepho, H.P., Selzer, L., 2019. multcompView: Visualizations of Paired
Comparisons. R package version 0.1-8. https://CRAN.R-project.org/package=mul
tcompView.
Grove, S.F., Suriyanarayanan, A., Puli, B., Zhao, H., Li, M., Li, D., Schaffner, D.W.,
Lee, A., 2015. Norovirus cross-contamination during preparation of fresh produce.
Int. J. Food Microbiol. 198, 43–49. https://doi.org/10.1016/j.
ijfoodmicro.2014.12.023.
Hamilton, A.N., Gibson, K.E., 2022. Performance of manufacturer cleaning
recommendations applied to 3D food ink capsules for the control of a human
norovirus surrogate. Food Environ. Virol. https://doi.org/10.1007/s12560-022-
09539-8.
Hamilton, A.N., Gibson, K.E., 2023. Efficacy of manufacturer recommendations for the
control of Salmonella Typhimurium and Listeria monocytogenes in food ink capsules
utilized in 3D food printing systems. J. Food Protect. 86, 100030 https://doi.org/
10.1016/j.jfp.2022.100030 (pending publication).
Handral, H.K., Tay, S.H., Chan, W.W., Choudhury, D., 2020. 3D printing of cultured meat
products. Crit. Rev. Food Sci. Nutr. 62, 272–281. https://doi.org/10.1080/
10408398.2020.1815172.
Hothorn, T., Bretz, F., Westfall, P., 2008. Simultaneous inference in general parametric
models. Biom. J. 50, 346–363. https://doi.org/10.1002/bimj.200810425.
Hurst, E.J., 2016. 3D printing in healthcare: emerging applications. J. Hosp. Librarian.
16, 255–267. https://doi.org/10.1080/15323269.2016.1188042.
Kassambara, A., 2020. Ggpubr: ’ggplot2’ Based Publication Ready Plots. R package
version 0.4.0. https://CRAN.R-project.org/package=ggpubr.
Kassambara, A., 2021. Rstatix: Pipe-Friendly Framework for Basic Statistical Tests. R
package version 0.7.0. https://CRAN.R-project.org/package=rstatix.
Keskinen, L.A., Todd, E.C., Ryser, E.T., 2008. Transfer of surface-dried Listeria
monocytogenes from stainless steel knife blades to roast Turkey breast. J. Food
Protect. 71, 176–181. https://doi.org/10.4315/0362-028x-71.1.176.
Kuznetsova, A., Brockhoff, P.B., Christensen, R.H., 2017. Lmertest package: tests in linear
mixed effects models. J. Stat. Software 82. https://doi.org/10.18637/jss.v082.i13.
Lanaro, M., Forrestal, D.P., Scheurer, S., Slinger, D.J., Liao, S., Powell, S.K., Woodruff, M.
A., 2017. 3D printing complex chocolate objects: platform design, Optimization and
Evaluation. J. Food Eng. 215, 13–22. https://doi.org/10.1016/j.
jfoodeng.2017.06.029.
Le Tohic, C., O’Sullivan, J.J., Drapala, K.P., Chartrin, V., Chan, T., Morrison, A.P.,
Kerry, J.P., Kelly, A.L., 2018. Effect of 3D printing on the structure and textural
properties of processed cheese. J. Food Eng. 220, 56–64. https://doi.org/10.1016/j.
jfoodeng.2017.02.003.
A.N. Hamilton and K.E. Gibson
Lee, J., 2021. A 3D food printing process for the new normal era: a review. Processes 9,
1495. https://doi.org/10.3390/pr9091495.
Length, R.V., Buerkner, P., Herve, M., Love, J., Miguez, F., Riebl, H., Singmann, H., 2021.
Emmeans: Estimated Marginal Means, Aka Least-Squares Means. R package version
1.7.5. https://CRAN.R-project.org/package=emmeans.
Mantihal, S., Kobun, R., Lee, B.-B., 2020. 3D food printing of as the new way of preparing
food: a Review. Int. J. Gastron. Food Sci. 22, 100260 https://doi.org/10.1016/j.
ijgfs.2020.100260.
Mantihal, S., Prakash, S., Bhandari, B., 2019. Textural modification of 3D printed dark
chocolate by varying internal infill structure. Food Res. Int. 121, 648–657. https://
doi.org/10.1016/j.foodres.2018.12.034.
Midelet, G., Carpentier, B., 2002. Transfer of microorganisms, including Listeria
monocytogenes, from various materials to beef. Appl. Environ. Microbiol. 68,
4015–4024. https://doi.org/10.1128/AEM.68.8.4015-4024.2002.
Midelet, G., André, Kobilinsky, Carpentier, B., 2006. Construction and analysis of
fractional multifactorial designs to study attachment strength and transfer of Listeria
monocytogenes from pure or mixed biofilms after contact with a solid model food.
Appl. Environ. Microbiol. 72, 2313–2321. https://doi.org/10.1128/aem.72.4.2313-
2321.2006.
Moore, C.M., Sheldon, B.W., Jaykus, L.-A., 2003. Transfer of Salmonella and
Campylobacter from stainless steel to romaine lettuce. J. Food Protect. 66,
2231–2236. https://doi.org/10.4315/0362-028x-66.12.2231.
Pérez-Rodríguez, F., Valero, A., Carrasco, E., García, R.M., Zurera, G., 2008.
Understanding and modelling bacterial transfer to foods: a Review. Trends Food Sci.
Technol. 19, 131–144. https://doi.org/10.1016/j.tifs.2007.08.003.
R Core Team, 2022. R: A Language and Environment for Statistical Computing. R
Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/.
Ramachandraiah, K., 2021. Potential development of sustainable 3D-printed meat
analogues: a Review. Sustain. Times 13, 938. https://doi.org/10.3390/su13020938.
Redmond, E.C., Griffith, C.J., 2003. Consumer food handling in the home: a review of
food safety studies. J. Food Protect. 66, 130–161. https://doi.org/10.4315/0362-
028x-66.1.130.
Rodriguez, A., McLandsborough, L.A., 2007. Evaluation of the transfer of Listeria
monocytogenes from stainless steel and high-density polyethylene to Bologna and
American cheese. J. Food Protect. 70, 600–606. https://doi.org/10.4315/0362-
028x-70.3.600.
Ross, M.M., Crowley, S.V., Crotty, S., Oliveira, J., Morrison, A.P., Kelly, A.L., 2021.
Parameters affecting the printability of 3D-printed processed cheese. Innovat. Food
Sci. Emerg. Technol. 102730 https://doi.org/10.1016/j.ifset.2021.102730.
Scallan, E., Hoekstra, R.M., Angulo, F.J., Tauxe, R.V., Widdowson, M.-A., Roy, S.L.,
Jones, J.L., Griffin, P.M., 2011. Foodborne illness acquired in the United
States—major pathogens. Emerg. Infect. Dis. 17, 7–15. https://doi.org/10.3201/
eid1701.p11101.
Scharff, R.L., 2012. Economic burden from health losses due to foodborne illness in the
United States. J. Food Protect. 75, 123–131. https://doi.org/10.4315/0362-028x.
jfp-11-058.
Scott, C., 2021. Barilla Continues to Develop Pasta 3D Printer, Envisions Gourmet
Customization. The Voice of 3D Printing/Additive Manufacturing. https://3dprint.
com/151348/barilla-pasta-3d-printer/. accessed 10.18.22.
Food Microbiology 113 (2023) 104268
Severini, C., Azzollini, D., Albenzio, M., Derossi, A., 2018. On printability, quality and
nutritional properties of 3D printed cereal based snacks enriched with edible insects.
Food Res. Int. 106, 666–676. https://doi.org/10.1016/j.foodres.2018.01.034.
Sharps, C.P., Kotwal, G., Cannon, J.L., 2012. Human norovirus transfer to stainless steel
and small fruits during handling. J. Food Protect. 75, 1437–1446. https://doi.org/
10.4315/0362-028x.jfp-12-052.
Sheen, S., 2008. Modeling surface transfer of Listeria monocytogenes on salami during
slicing. J. Food Sci. 73 https://doi.org/10.1111/j.1750-3841.2008.00833.x.
Stals, A., Uyttendaele, M., Baert, L., Van Coillie, E., 2013. Norovirus transfer between
foods and food contact materials. J. Food Protect. 76, 1202–1209. https://doi.org/
10.4315/0362-028x.jfp-12-392.
Sun, J., Zhou, W., Huang, D., Fuh, J.Y., Hong, G.S., 2015. An overview of 3D printing
technologies for Food Fabrication. Food Bioprocess Technol. 8, 1605–1615. https://
doi.org/10.1007/s11947-015-1528-6.
Terfansky, M.L., Thangavelu, M., 2013. 3D printing of food for space missions. In: AIAA
SPACE 2013 Conference and Exposition. https://doi.org/10.2514/6.2013-5346.
Tuladhar, E., Hazeleger, W.C., Koopmans, M., Zwietering, M.H., Duizer, E., Beumer, R.R.,
2013. Transfer of noroviruses between fingers and fomites and food products. Int. J.
Food Microbiol. 167, 346–352. https://doi.org/10.1016/j.ijfoodmicro.2013.09.018.
Verhaelen, K., Bouwknegt, M., Carratalà, A., Lodder-Verschoor, F., Diez-Valcarce, M.,
Rodríguez-Lázaro, D., de Roda Husman, A.M., Rutjes, S.A., 2013. Virus transfer
proportions between gloved fingertips, soft berries, and lettuce, and associated
health risks. Int. J. Food Microbiol. 166, 419–425. https://doi.org/10.1016/j.
ijfoodmicro.2013.07.025.
Vorst, K.L., Todd, E.C., Ryser, E.T., 2006. Transfer of Listeria monocytogenes during
mechanical slicing of Turkey breast, Bologna, and salami. J. Food Protect. 69,
619–626. https://doi.org/10.4315/0362-028x-69.3.619.
Vukušić Pavičić, T., Grgić, T., Ivanov, M., Novotni, D., Herceg, Z., 2021. Influence of
flour and fat type on dough rheology and technological characteristics of 3D-printed
cookies. Foods 10, 193. https://doi.org/10.3390/foods10010193.
Wang, L., Zhang, M., Bhandari, B., Yang, C., 2018. Investigation on fish surimi gel as
promising food material for 3D printing. J. Food Eng. 220, 101–108. https://doi.org/
10.1016/j.jfoodeng.2017.02.029.
Warnes, G.R., Bolker, B., Gorjanc, G., Grothendieck, G., Korosec, A., Lumley, T.,
MacQueen, D., Magnusson, A., Rogers, J., 2022. Gdata: Various R Programming
Tools for Data Manipulation. R package version 2.18.0.1. https://CRAN.R-project.
org/package=gdata.
Wickham, H., 2016. ggplot2: Elegant Graphics for Data Analysis. Springer International
Publishing, Cham, Switzerland.
Wickham, H., Averick, M., Bryan, J., Chang, W., McGowan, L., François, R.,
Grolemund, G., Hayes, A., Henry, L., Hester, J., Kuhn, M., Pedersen, T., Miller, E.,
Bache, S., Müller, K., Ooms, J., Robinson, D., Seidel, D., Spinu, V., Takahashi, K.,
Vaughan, D., Wilke, C., Woo, K., Yutani, H., 2019. Welcome to the tidyverse. J. Open
Source Softw. 4, 1686. https://doi.org/10.21105/joss.01686.
Wilson, A., Anukiruthika, T., Moses, J.A., Anandharamakrishnan, C., 2020. Customized
shapes for chicken meat–based products: feasibility Study on 3D-printed nuggets.
Food Bioprocess Technol. 13, 1968–1983. https://doi.org/10.1007/s11947-020-
02537-3.
Yang, F., Zhang, M., Fang, Z., Liu, Y., 2018. Impact of processing parameters and post-
treatment on the shape accuracy of 3D-printed baking dough. Int. J. Food Sci.
Technol. 54, 68–74. https://doi.org/10.1111/ijfs.13904.