[FOOD SAFETY & QUALITY]
by Gülhan Ünlü
Bacterial Biofilms: Formation, Prevention, and Control
I
n a recent review article, Niels
Høiby defined microbial biofilms as
“an aggregate of microbial cells
surrounded by a self-produced poly-
mer matrix” (Høiby 2017). Discussing
their history, he said, “The observa-
tion of biofilms is old,” “since both
Leeuwenhoek and Pasteur have
described the phenomenon.” In
the late 1600s, Anthony van
Leeuwenhoek (1632–1723) wrote to
the Royal Society of London about
his observations of the plaque
between his teeth, which he made
with his primitive microscopes. In
the late 1800s, Louis Pasteur (1844–
1895) observed aggregates of
bacterial cells as the source of wine
becoming vinegar. ZoBell and Allen
(1935) introduced the term biofilm in
environmental microbiology in dis-
cussing the significance of marine
Biofilm formation by bacteria in the lumen of a tube. © Blamb/Shutterstock
bacteria in the fouling of submerged Relevance to the Food Industry
surfaces. In the early 1970s, Høiby In food-related environments,
himself observed aggregates of including the meat, dairy, and fresh
Pseudomonas aeruginosa cells in produce sectors, surfaces and
chronically infected cystic fibrosis equipment are habitually populated
patients, by microscopic examina- by microorganisms forming biofilms
tion of mucus and lung tissue, (Pometto and Demirci 2015). Biofilms
initiating the concept of “biofilm can slough off, move to surrounding
infection.” The term biofilm was environments, elevate intermediary
Biofilms can slough off, move to surrounding
environments, elevate intermediary and final product
microbial loads, and reduce shelf life.
introduced into medical microbiol- and final product microbial loads,
ogy and medicine by Nickel et al. and reduce shelf life. Biofilms can
(1985), who demonstrated increased also serve as a source of cross-
antimicrobial resistance of bacterial contamination, consequently
cells growing in biofilms as com- diminishing effectiveness of food
pared with their planktonic processing technologies, compro-
counterparts. mising microbial food quality and
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Bacterial Biofilms: Formation, Prevention, and Control continued...

food safety, and leading to sizable 2018, Matthews et al. 2017, Pometto by food particles or substrates.
economic losses. Microbial cells in and Demirci 2015). Also formed by Biofilms may be formed by a single
biofilms are quite well protected beneficial microorganisms, biofilms microorganism or a group of micro-
from adverse environmental condi- can be advantageous. They may be organisms, through a complex
tions, including sanitizing agents. used to improve human health via the multistage process. Described by
Winkelströter et al. (2014) and
Biofilms can also serve as a source of cross-contamination, McLandsborough (2015), the stages
of formation are as follows: initial
consequently diminishing effectiveness of food processing transfer/adhesion/attachment of
technologies, compromising microbial food quality and food planktonic microbial cells to the sur-
face, production of extracellular
safety, and leading to sizable economic losses. polymeric substance (EPS, also
known as a glycocalyx), microcolony
Thus, prevention and control of bio- the gut microbiota, increase the formation, and maturation into a
films by the food industry is critical quality and the yield of food fermen- 3D-biofilm. These stages may be
and challenging. tations, produce value-added followed by detachment/dispersion/
Biofilms are formed by spoilage products through fermentation, recolonization of microbial cells
microorganisms (e.g., Pseudomonas develop biotechnological applica- involving new preconditioned sur-
spp.) and pathogens, including tions for improving food safety and faces. The first stage in attachment
Aeromonas hydrophila, Bacillus quality, and manage industrial to surfaces is reversible sorption,
cereus, Campylobacter jejuni, waste. thought to be related to van der
Cronobacter sakazakii, Escherichia Waals forces (or other physicochem-
coli O157:H7, Listeria monocyto- Formation ical factors). The second stage deals
genes, Salmonella spp., Shigella Biofilms can form on any type of sur- with irreversible attachment via EPS
spp., Staphylococcus aureus, Vibrio face (e.g., plastic, metal, glass, production, which anchors the cells
cholera, and Yersinia spp. (Bhunia wood, or food) that is preconditioned and protects them against environ-
mental stresses. Many factors can
A Scanning Electron Microscope image of a natural community of bacteria on a grain of sand collected from
influence microbial attachment,
intertidal sediment on a beach. Image courtesy of the Lewis Lab at Northeastern University. Image created by including characteristics of a sur-
Anthony D’Onofrio, William H. Fowle, Eric J. Stewart, and Kim Lewis. face, phase and temperature of
microbial growth, and microbial
motility. This stage is followed by the
formation of microcolonies with
well-defined boundaries, allowing
fluid channels to run through the
biofilm. Quorum sensing, known as
microbial cell-to-cell communica-
tion, plays a role in the formation of
microcolonies and organization of
fluid channels.

Prevention and Control

The structural and physiological
features of microbial cells within
biofilms, such as reduced diffusion,
anaerobic growth, reduced growth
rate, and the viable but noncultur-
able (VBNC) state, cause them to be
significantly more resistant to
adverse environmental conditions
than their planktonic counterparts.
Use of various sanitizers and bio-
cides in cleaning and sanitation is
not sufficient to remove cells within
biofilms. Proper cleaning is impor-
tant for their removal and eradication

pg 74 10.20 • www.ift.org
before they become recalcitrant in
mature biofilms (Matthews et al.
2017).
There has been a great deal of
interest to develop innovative,
efficient, and inexpensive biofilm
inhibition and removal strategies.
One promising approach uses bio-
nanotechnology to modify surface
materials. These include develop-
ments inspired by naturally occurring
antimicrobial surfaces and sub-
stances (e.g., antimicrobial
peptides), generation of artificial
antimicrobial surfaces with both
traditional and advanced surface
modification techniques (e.g., via use
of silver, gold, or titanium dioxide,
and physical topographical modifica-
tions involving functionalization,
derivatization, and polymerization)
(Birkenhauer and Neethirajan 2015).
Nonthermal atmospheric plasmas
are an innovative control technology
of interest to the food industry due to Development of new environ-
their contact-free, waterless nature mentally friendly (“green”) sanitizers
and their effectiveness against a capable of removing biofilms in food
wide range of microorganisms processing environments is growing
(Niemira et al. 2014, Puligundla and in research interest. Enzymes such
Mok 2017, Ziuzina et al. 2015a, b). as alpha-amylase, beta-glucanase,
Other inactivation technologies being DNase, proteinase K, and lipases are
investigated and applied are photody- considered in formulations of enzy-
namic inactivation using pulsed matic detergents due to their ability
Use of microorganisms, such as bacteriocin-producing
lactic acid bacteria and their metabolites,
for competitive exclusion or inactivation of undesirable
microorganisms is an area of research.
ultraviolet light (Montgomery and to degrade the EPS (Araújo et al.
Banarjee 2015), steam heating (Ban 2017, Brown et al. 2015, Nguyen and
and Kang 2016), light (405 nm) Burrows 2014, Seghal Kiran et al.
(McKenzie et al. 2013), ozone 2014, Zetzmann et al. 2015). Plant-
(Nicholas et al. 2013), ultrasound based compounds, such as essential
(Axelsson et al. 2013), and gaseous oils, are also being considered in
chlorine dioxide (Nam et al. 2014). new “green” sanitizer formulations

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[FOOD SAFETY & QUALITY]

Bacterial Biofilms: Formation, Prevention, and Control continued...

(Ashraf et al. 2014).
Use of microorganisms such as
bacteriocin-producing lactic acid
bacteria and their metabolites for
competitive exclusion or inactivation
of undesirable microorganisms is an
area of research. A variety of bacte-
riocins, including enterocin AS-48
(Caballero Gómez et al. 2013),
enterocin B3A-B3B (Al-Seraih et al.
2017), lichenicidin (Mathur et al.
2018), nisin and nisin derivatives
(Field et al. 2016), sonorensin
(Chopra et al. 2015), and subtilomycin
(Mathur et al. 2018) are effective.
Other bacterial metabolites, includ-
ing the biosurfactant lichenysin
(Coronel-León et al. 2016), endogly-
cosidases such as glycosyl
hydrolase (Yu et al. 2015), and unsat-
urated fatty acids such as
cis-2-decenoic acid (Sepehr et al.
2014) are also useful for biofilm pre-
vention and control.
Lytic bacteriophages have been
used as biocontrol agents against
biofilms of specific microorganisms,
such as C. sakazakii (Enderson et al.
2017), E. coli O157:H7 (Sadekuzzaman
et al. 2017), L. monocytogenes
(Nannapaneni and Soni 2015), P.
aeruginosa (Shafique et al. 2017),
Salmonella spp. (Islam et al. 2019,
Sadekuzzaman et al. 2018), and S.
aureus (González et al. 2017). Moye
et al. (2018) comprehensively
reviewed bacteriophage applica-
tions for food production and
processing. They indicated that in
recent years, several bacterio-
phages against a number of specific
foodborne pathogenic bacteria are
considered “approved” by the U.S.
Food and Drug Administration and
U.S. Department of Agriculture.
Need for Additional Research
Biofilms affect disparate processes
in our lives, from oral hygiene, to
safe water and food, bioremediation
of toxic compounds, and wastewater
treatment. Although biofilms have
received a great deal of attention
over the years, there continues to be
a need for additional research. In the
context of the food system, better
understanding of biofilm structure,
microbial physiology within biofilms,
and microbial interactions (e.g., quo-
rum sensing) would make positive
contributions to effective biofilm
prevention and control. A number of
conventional and modern techniques
have been used to detect and evalu-
ate microorganisms in biofilms in
vitro. Development of innovative
techniques for assessing complex
biofilm communities in food process-
ing environments would allow in situ
characterization of biofilms, and lead
to successful prevention and con-
trol. A plethora of physical, chemical,
and biological approaches capable
of preventing biofilm formation and/
or removing them exists. Neverthe-
less, research into simultaneous use
of two or more of these approaches
in combination would enhance
efficacy. FT
Gülhan Ünlü, PhD, Contributing Editor
• Associate Professor; Dept. of
Animal, Veterinary, and Food
Science, The University of Idaho
• gulhan@uidaho.edu

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