International Journal of Environmental Health Research

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Surfactants of microbial origin as antibiofilm

agents

Katarzyna Paraszkiewicz, Magdalena Moryl, Grażyna Płaza, Diksha Bhagat,

Surekha K. Satpute & Przemysław Bernat

To cite this article: Katarzyna Paraszkiewicz, Magdalena Moryl, Grażyna Płaza, Diksha
Bhagat, Surekha K. Satpute & Przemysław Bernat (2019): Surfactants of microbial origin
as antibiofilm agents, International Journal of Environmental Health Research, DOI:
10.1080/09603123.2019.1664729

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Published online: 11 Sep 2019.

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INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH
https://doi.org/10.1080/09603123.2019.1664729

ARTICLE

Surfactants of microbial origin as antibiofilm agents

Katarzyna Paraszkiewicza, Magdalena Morylb, Grażyna Płazac, Diksha Bhagatd,
Surekha K. Satputed and Przemysław Bernata
a
Department of Industrial Microbiology and Biotechnology, Faculty of Biology and Environmental Protection,
University of Lodz, Łódź, Poland; bDepartment of Biology of Bacteria, Faculty of Biology and Environmental
Protection, University of Lodz, Łódź, Poland; cInstitute of Production Engineering, Faculty of Organization and
Management, Silesian University of Technology, Zabrze, Poland; dDepartment of Microbiology, Savitribai Phule
Pune University, Pune, India

ABSTRACT ARTICLE HISTORY

The microbial world provides new energy sources and many various ‘green’ Received 4 June 2019
chemicals. One type of chemicals produced by microorganisms is the bio- Accepted 3 September 2019
surfactant group. Biosurfactants are universal molecules, exhibiting surface KEYWORDS
properties often accompanied by desired biological activity. Biosurfactants Antibiofilm agents; biofilm;
are considered to be environmentally ‘friendly’ due to their low toxicity and biosurfactants
biodegradable nature. These compounds have unique features and therefore
they can find potential applications in many different industries, ranging from
biotechnology to environmental remediation technologies. Antibacterial and
antifungal activities make them relevant for applications as inhibitory agents
against microbial biofilm. This review covers the current knowledge and the
recent advances in the field of biosurfactants as antibiofilm agents.

Introduction
Pathogenic microbial biofilm, a consortium of microbial cells protected by a self-roduced polymer
matrix is considered a worldwide challenge due to its high resistance to antimicrobial agents and the
host immune system (Ribeiro et al. 2016). Research teams around the world have developed different
anti-biofilm strategies. Among new therapeutic methods applied against biofilm, a lot of attention is

CONTACT Przemysław Bernat przemyslaw.bernat@biol.uni.lodz.pl Department of Industrial Microbiology and

Biotechnology, Faculty of Biology and Environmental Protection, University of Lodz, Banacha Street 12/16, Łódź 90-237, Poland
© 2019 Informa UK Limited, trading as Taylor & Francis Group
2 K. PARASZKIEWICZ ET AL.

now being paid to biological surface-active agents (biosurfactants). Many of them have been reported
to exhibit anti-adhesive, antimicrobial and biofilm disruption properties (Banat et al. 2014).
The present review deals with the basic aspects of biosurfactants in microbial biofilms, summarises
the recent studies focused on biosurfactants as antibiofilm agents and discusses mechanisms involved
in the biosurfactants role against microbial biofilms.

Microbial biofilms
Biofilm importance and characteristics
Sessile lifestyle is a natural and favorable form of microorganisms’ existence in environment and
in the human body. The last 20 to 30 years have been a period of intensive research on the
biofilms formed by microorganisms. The current state of knowledge allows defining biofilm as
a well-organized structure of bacterial cells, which is surrounded by a hydrated, extracellular
polymeric matrix and may (or not) be attached to solid biotic or abiotic surfaces (Burmølle et al.
2014; Flemming et al. 2016; Røder et al. 2016).
Sessile cells differ from their planktonic counterparts. Firstly, they are better adopted to stress
conditions e.g. nutrient deprivation, oxygen limitation, pH changes. Secondly, they express different
virulence factors, and are also more resistant to antimicrobial agents and the host immune system
(Berlanga and Guerrero 2016; Flemming et al. 2016; Petrova and Sauer 2016). In the biofilm, we can
also observe improved degradation of organic compounds (Delcaru et al. 2016). Microorganisms in
the biofilm can also communicate with cells of the same or other species. This process, described as
quorum sensing (QS), coordinates and regulates bacterial behavior in environment, e.g. biofilm
formation, antibiotic production, bioluminescence or virulence factors expression. QS is based on
chemical signal molecules – autoinducers (AI) or pheromones. The role of signal molecules in
Gram-negative bacteria is usually played by homoserine lactones (AHLs), and in Gram-positive
bacteria by oligopeptides (autoinducting peptides AIPs). Interspecies communication is allowed by
autoinducter-2. Signaling molecules are released and accumulated in environment and their con-
centration increases as a function of cell density. When a minimal threshold stimulatory concen-
tration of molecules is achieved, the microorganisms detect it and respond collectively e.g. by
altering gene expression. (Rutherford and Bassler 2012; Majumdar and Mondal 2016). The features
of sessile bacteria described above cause that biofilms are extremely difficult to eradicate from
medical devices or implants used in medical practice.
Bacteria biofilms are the most widely studied from the morphological and molecular perspectives.
Both yeast and filamentous fungi can also form biofilms; however, studies of fungal biofilms are
limited compared to those of yeasts (Fanning and Mitchell 2012; Cavalheiro and Teixeira 2018).
Regarding yeasts, Candida albicans is the most studied model of biofilm formation and shows similar
development process to those of bacterial biofilms. Sardi et al. (2015) characterized the biofilm formed
by Paracoccidioides brasiliensis and found that the formation of biofilm was associated with the gene
expression of adhesins and enzymes like GP43, GAPDH, and aspartyl proteinase. In recent years,
studies on fungal biofilms have increased considerably. For example, in the review of Costa-Orlandi
et al. (2017) the biofilm formation by filamentous fungi and some aspects of methodologies are
described. The models of biofilm development in filamentous fungi – dermatophytes (Trichophytum
rubrum and T. mentarophytes) and C. albicans are compared by the authors. The authors proposed
a model for biofilm formation by filamentous fungi. The six stages in development of fungal biofilm
are propsed: (1) propagule adsorption – contact of spores, hyphal fragments or sporangia to a surface,
(2) active adhesion – adhesins are secreted by spores, (3) microcolony formation – elongation and
hyphal branching, a monolayer is formed with the production of extracellular matrix, (4) second
microcolony formation or initial maturation – hyphae networks form covered by an extracellular
matrix and pores or channels are formed, (5) final maturation- fruiting bodies and others survivor
structures are formed, (6) dispersion or planktonic phase – conidia and/or hyphae fragments are
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 3

released, and a new cycle is beginning. Some filamentous fungi produce small proteins known as
hydrophobins. The proteins are involved in the adhesion of hyphae and can be involved in biofilm
formation.
In the paper of Costa-Orlandi et al. (2017) the polymicrobial structure of biofilms are also
described. Most microorganisms live in complex communities, known as polymicrobial biofilms.
In terms of human health, polymicrobial biofilms are prevalent throughout the human body, both
during healthy and disease conditions. Biofilm in human body can be found on the teeth surface,
or intestine and vagina mucosa, and in the most of the examples polymicrobial biofilms are
developed. A serious danger is posed by pathogenic biofilms in the lungs of people with cystic
fibrosis or structures formed on indwelling medical devices, e.g. dental or joint implants, urinary
catheters, prosthetic heart valves. Chronic biofilm-related infections are difficult to treat and
control, and very often the only solution is implant removal, an intensive antimicrobial therapy
and reoperation. They cause a lot of pain for the patient and generate an extremely high cost of
treatment. It is very important to study and properly understand the mechanisms of biofilms
formation and matriculation processes, which is the only way to develop effective strategies to
combat pathological biofilms.

Biofilm matrix
The extracellular polymeric matrix (ECM) is the largest, strongly hydrated part of the biofilm,
which commonly comprises extracellular polysaccharides, proteins, lipids and extracellular DNA
(eDNA) (Whitchurch et al. 2002). In addition, the matrix contains glycoproteins, glycolipids,
lipoteichoic acids, lipopolysaccharide, biosurfactants, and outer membrane vesicles (Absalon et al.
2012; Czaczyk and Myszka 2007; Flemming et al. 2007).
All matrix components form a protective barrier against the surrounding environment (host
factors, antimicrobials) are essential for the processes of cell adhesion, aggregation and biofilm
maturation. They stabilize the biofilm structure, facilitate horizontal gene transfer and are also
a source of nutrients and water for the cells (Moryl et al. 2015; Flemming et al. 2016).

Polysaccharides
Polysaccharides found in the matrix are mainly neutral sugars or uronic acids, usually forming
long chains and having a molecular weight of 0.5–2 × 106 Da (Starkey et al. 2004). The examples
of polysaccharides commonly identified in biofilms formed in the human body are: alginate,
(produced by Pseudomonas aeruginosa), cellulose (Escherichia coli), or poly-N-acetylglucosamine
PNAG (Staphylococcus aureus).

Proteins
Matrix proteins may be derived from both living and dead cells of the biofilm and they can be
transported through the membrane by outer membrane vesicles (OMV) (Schooling and Beveridge
2006; Toyofuku et al. 2012). Proteins isolated from the matrix are usually macromolecules with
the weight 10–200 kDa, which contain from 40 to 60% hydrophobic aminoacids (Czaczyk and
Myszka 2007). Based on their functions, proteins have been divided into enzymes, lectins and
eDNA binding proteins. Enzymes, especially lyases and polysaccharides, are involved in the
biosynthesis of exopolysaccharides, in the detachment and dispersion of cells from the biofilm
and in the degradation of the substances which may be a source of biogenic elements for the
microorganisms in the biofilm. Lectins are proteins able to recognize and bind to specific
oligosaccharide structures. They are located on fimbriae, the outer membrane or the peptidogly-
can and play a role in cells adhesion and the process of biofilm formation (Fong and Yildiz 2015).
4 K. PARASZKIEWICZ ET AL.

Multispecies biofilm life cycle

The biofilm life cycle, presented in Figure 1, is a complicated, multi-staged process which depends on
the environment of biofilm growth and the type and number of microorganisms. It consists of a few
stages: (i) adhesion, (ii) microcolony formation and biofilm matriculation, (iii) biofilm dislodging.

Adhesion
In the traditional model, the first step involves the planktonic cells attachment to the solid
surface (Figure 1). This stage is often preceded by the adsorption of proteins to the solid
surface and conditioning film formation, which facilitates bacterial attachment. Bacteria which
are the first to colonize the surface are called primary colonizers, their presence either allows
other microbial species (secondary colonizers) to adhere or disturbs this process. Mechanisms

Figure 1. Multispecies biofilm life cycle (Davies and Marques 2009; Elias and Banin 2012). (i) Adhesion of single cells or bacterial
aggregates to solid surfaces; (ii) Microcolony formation: microcolonies of single species, layering and mixed species micro-
colony; (iii) Biofilm dispersion and detachment: abrasion is bacterial cells removal from biofilm because of collision with liquid
particle; grazing is a biofilm removal in which eukaryotic organisms are involved; erosion is continuous transfer of small
portions of biofilm as a result of fluid shear; sloughing is a removal of large biofilm pieces, embedded in matrix, due to fluid
frictional forces.
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 5

which enable bacteria to achieve (interact with) the surface include: Brownian motion, polar
interactions, Lifshitz-van der Waals forces, sedimentation, convective mass transport and active
transport (mediated e.g. by bacterial flagella and/or chemotaxis) (Palmer et al. 2007; Araujo
et al. 2016).
The most frequently cited theory states that adhesion is a two-step process (in literature also
a three-step and five-step processes are considered) (Hood and Zottola 1995; Stoodley et al. 2002).
The first step – initial adhesion, in which bacteria can be easily removed from the surface, involves
their transport to the surface. Forces involved in this process are: hydrophobic interactions, van
der Waals and electrostatic forces (Van Loosdrecht et al. 1987; Gilbert et al. 1991). The second
step includes irreversible attachment, during which bacteria bind to the solid layer through
specific ligands (e.g. protein adhesions, fimbriae) and cannot be easily removed from the surface.
In this process the production of extracellular polymers is observed.
Adhesion is an extremely important stage in the biofilm formation. Two types of factors are
involved in that process, the first one includes the bacterial cells charge and hydrophobicity, and
the second one is the surface charge, roughness and microtopography. Surface hydrophobicity is
one of the most important factors for adhesion (Araujo et al. 2016). An extremely important role
is also played by growth medium in which the process takes place. It was proved that factors
present in the environment could interfere in the attachment of bacteria because they affect the
charge of the bacterial cell (by changing pH) and its hydrophobicity and could be used as an
effective method to combat bacterial biofilms (Palmer et al. 2007).
Cells which are in the first contact with the solid surface could detach, when influenced by the
environmental signals such as nitric oxide, proteinase K or cis-2-decenoic acid – a molecule
important in cells signaling (Barraud et al. 2009; Davies and Marques 2009; Nguyen and Burrows
2014). This reversion of the initial adhesion is called desorption.

Microcolony formation and biofilm matriculation

Microcolonies are differentiated in mushroom-like or pillar-like structures, which are separated by
channels. The channels are filled with fluid which removes metabolic products and delivers
nutrients to bacterial cells (Donlan 2001).
In multispecies biofilms the spatial organization of the microcolony structure is very interesting
(Figure 1). Positioning of bacterial cells in a microcolony is related with the functions of the
microorganisms and with interspecies interactions (Momeni et al. 2013; Burmølle et al. 2014; Liu
et al. 2016). Elias and Banin (2012) suggest that a microcolony can be organized in three general
forms. The first model includes separate microcolonies formed next to each other by particular
species of bacteria. This model is characteristic for biofilms where noncomensal, competitive or
antagonistic interactions between co-inhabitant organisms are observed, e.g. in mixed biofilm of
Burkholderia sp. and Pseudomonas sp. (Nielsen et al. 2000; Burmølle et al. 2014). In the second
model bacterial cells of various species are mixed in one structure. Bacteria co-aggregate and form
a common microcolony. The third model includes a layered structure, one species is located in the
upper layers of the microcolony and the other species are situated in the lower layers (Elias and
Banin 2012).
Synergistic interactions, co-metabolism or even syntrophy could be the reason why species
form mixed communities, i.e. bacteria arranged according to their metabolic abilities and ener-
getic properties (Seckbach and Oren 2010; Liu et al. 2016). On the other hand, it has also been
shown that competitive interactions between P. aeruginosa and A. tumefaciens are responsible for
their layering organization in microcolonies (An et al. 2006).
In mature biofilm microbial cells change their gene expression and phenotype, which provides
biofilm-specific benefits, e.g. antibiotic resistance, boosting virulence, degradation of a broad
spectrum of compounds (Burmølle et al. 2014).
6 K. PARASZKIEWICZ ET AL.

Biofilm dislodging
Accumulation of waste products, lack of nutrients and large size of the structure induce the sessile
cells to leave the biofilm and settle down in new environments. Sessile cells use a few ways to get
released from the biofilm e.g. detachment or dispersion (Figure 1). Detachment is a relationship
between forces maintaining the biofilm structure and forces acting upon the biofilm. Four
mechanisms of biofilm detachment have been observed: abrasion, grazing, erosion, sloughing.
When bacterial cells are released from biofilm because of collisions with liquid particles, the
process is called abrasion. If eukaryotic organisms are involved in biofilm removal, the process is
referred to as grazing. Erosion is a continuous transfer of small portions of biofilm as a result of
fluid shear. Sloughing is a removal of large biofilm pieces, embedded in the matrix, due to fluid
frictional forces (Davies 2011). Detachment could be initiated by substances present in growth
medium like enzymes degrading the extracellular matrix (Dispersin B, Proteinase K, DNase) and
biosurfactants (Kaplan et al. 2004; Nguyen and Burrows 2014; Bonnichsen et al. 2015).
Dispersion is hypothesized to be an active form of bacterial escape (Davies 2011). Bacteria
receive signals, which are transmitted through the biofilm and cause physiological changes
facilitating cells release from the biofilm structure. Bacteria in biofilm can produce e.g. matrix-
degrading enzymes, hydrolases, lyases, which affect the biofilm structure and allow for easy
detachment of cells. There are two types of dispersion. Native dispersion occurs in the terminal
stages of biofilm development and environmentally induced dispersion is observed when signals
are derived from environment (Petrova and Sauer 2016). Fatty acid signaling molecules e.g. cis-
2-decenoic acid (cis-DA) are responsible for native dispersion, which has been detected in
P. aeruginosa (Davies and Marques 2009). Environmentally induced dispersion occurs as
a result of starvation, lack of carbon sources, oxidative stress and host factors activity (Petrova
and Sauer 2016). There is a hypothesis that biosurfactants (e.g. rhamnolipids) could be involved in
the maintenance of the biofilm matrix structure. They enhance, probably in combination with
enzymes, the degradation of extracellular polymers and facilitate the dispersal of sessile cells
(Davey et al. 2003; Boles et al. 2005; Davies 2011). Dispersal can occur via the separation of single
cells or sloughing of large pieces of biofilm (Purevdorj-Gage et al. 2005). Both motile cells and
aggregates can initiate a biofilm in new ecological niches (Kragh et al. 2016). Cells which escape
from biofilm are distinct from planktonic and sessile cells. They represent a different gene and
protein expression pattern and pathogenicity (e.g. high virulence against macrophages) and
release enzymes degrading the biofilm matrix.

Biosurfactants
Properties and classification
The term surfactant (SURFaceACTiveAgeNT) describes a wide variety of compounds, both
synthetic and biological, all of which exhibit similar tension active properties (Soberon-Chavez
2011; Santos et al. 2016). Biosurfactants are surface-active biomolecules produced by living cells
mainly by microorganisms (bacteria, yeast and microscopic filamentous fungi). They are bio-
chemical compounds containing both a hydrophobic and a hydrophilic group that allow such
amphiphilic (amphipatic) molecules to exist at the interface between polar and nonpolar media.
They are produced on microbial cell surfaces or are secreted. Their structure contains
a hydrophilic moiety (usually a carbohydrate, amino acid, linear or cyclic peptide, phosphate,
carboxylic acid or an alcohol) and a hydrophobic moiety, which is either a long chain fatty acid,
hydroxyl fatty acid or α-alkyl β-hydroxy fatty acid. Most biosurfactants derived from diverse
microbial sources are either anionic or neutral. Their properties of interest regard also wetting and
penetrating actions, spreading, hydrophilicity and hydrophobicity, emulsification, de-
emulsification, detergency, gelling, foaming, flocculation, low critical micellar concentration
(CMC) values, microbial growth enhancement, metal sequestration, and anti-microbial actions
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 7

(Rosenberg and Ron 1999; Santos et al. 2016; Fait et al. 2019). Functional properties of biosurfac-
tants are mainly determined by their structure, e.g. location and size of functional groups. There is
a growing interest in the investigations of physicochemical and biological properties of biosur-
factants because of their various applications.
In literature there are found two terms ‘biosurfactants’ and ‘bioemulsifiers’, which are often used
interchangeably. However, Rosenberg and Ron (1999) classified biosurfactants into two groups: (1)
low-molecular-weight surface active agents called biosurfactants, which efficiently lower the surface
and interfacial tension, and (2) high molecular-weight polymers named bioemulsifiers, more effective
as emulsion-stabilizing agents. The first group includes: glycolipids, lipopeptides and phospholipids,
whereas the second one consists of polymeric and particulate biosurfactants such as emulsan, alasan
and dispersan. Another group of bio-based surfactants includes phytogenic surfactants such as
saponins, lecithins and humic acids. Phytogenic surfactants are released from roots, and are found
in large amounts in the rhizosphere. Biosurfactants offer many advantages over their chemical
counterparts, including the following: (1) biodegradability – owing to the low toxicity and specific
chemical structure, these compounds do not persist in the environment and are degraded preventing
problems of accumulation in environments; (2) biocompatibility and digestibility, which allow their
unabated application in cosmetics, pharmaceuticals and as functional food additives; (3) availability of
raw materials – biosurfactants can be produced from relatively cheap raw materials which are available
in large quantities; (4) acceptable production economics – biosurfactants can be produced even from
different industrial wastes and by-products, which remains a promising area for bulk production (e.g.,
use in petroleum-related technologies like microbial enhanced oil recovery-MEOR); (5) use in
environmental biotechnology as agents accelerating biodegradation and detoxification of industrial
effluents, bioremediation of contaminated soils, controlling of oil-spills processes or for stabilization
of industrial emulsions; (6) specificity – due to specific functional groups in biosurfactants molecules,
they are used in detoxification of specific pollutants, de-emulsification of industrial emulsions,
development of specific cosmetic, pharmaceutical and food applications; (7) effectiveness at extreme
temperatures, pH and salinity (Soberon-Chavez 2011; Naughton et al. 2019; Patel et al. 2019).
Considering the advantages, some authors suggest that biosurfactants become increasingly attractive
as multifunctional materials for future utilisation in various environmental and health fields
(Paraszkiewicz 2016; Karlapudi et al. 2018).

Antibiofilm potential of biosurfactants

Biosurfactants have been observed to inhibit biofilm formation. They are capable of modifying the
physico-chemical properties of the surface to reduce adhesion (Janek et al. 2012). Besides, some
biosurfactants down-regulate the expression of bacterial genes involved in biofilm formation.
Salehi et al. (2014) observed that the expression of glucosyltransferases (gtfs) and fructosyltrans-
ferase (ftf) genes, which play a critical role in the initial adhesion of Streptococcus mutans to the
tooth surface, was inhibited in the presence of biosurfactants purified from Lactobacillus reuteri.
On the other hand, rhamnolipids of P. aeruginosa could be involved in the process of
detachment, through which bacteria use active mechanisms to leave biofilms and return to the
planktonic (free-living) state. Biosurfactants affect also biofilm architecture by creating cavities
within the centre of biofilm (Boles et al. 2005). The role of biosurfactant molecules against biofilm
forming pathogens is illustrated in Figure 2. Microbial biofilms can be affected through various
following mechanisms: 1) disruption of the membrane structure caused by membrane degrada-
tion, 2) impediment in the electron transport chain and hence restriction in energy needs, 3)
altered cell permeability causing loss of intracellular content, 4) altered cell adhesion as
a consequence of reduced cell surface hydrophobicity, 5) distorted protein structure and 6)
interference in quorum sensing consequently leading to decreased biofilm formation (Satpute
et al. 2016a).
8 K. PARASZKIEWICZ ET AL.

Figure 2. Role of different biosurfactant molecules on microbial biofilm formation (Satpute et al. 2016a).

Various studies have shown that the prior adsorption of biosurfactants on surfaces (surface
conditioning) reduced the adhesion of microbial cells and was efficient in reducing the surface
colonization (Das et al. 2009; Do Valle Gomes and Nitschke 2012; Araujo et al. 2016).
In general, the process of microbial cells adhesion to foreign surfaces is strongly influenced by three
kinds of factors. The first one includes physical and chemical properties of the surfaces (e.g. roughness,
microarchitecture, porosity, charge and hydrophobicity). The second one involves such environmen-
tal parameters as temperature, flow conditions, pH, presence of nutrients, antibiotics (or other
compounds which might influence bacterial adhesion). The third group of factors is connected with
microorganism’s properties e.g. cell surface hydrophobicity, flagella presence and extracellular poly-
meric substance (especially exopolysaccharides) production (Pavithra and Doble 2008).
The relationship between modifications of surface physicochemical properties caused by biosur-
factants as well as the adhesion and biofilm formation by Listeria monocytogenes and P. fluorescens
strains were evaluated by Araujo et al. (2016). It was established that polystyrene conditioned by
surfactin did not show any significant modification of surface properties as opposed to the stainless
steel surface, which exhibited a strong decrease in surface hydrophobicity. On the other hand, crude
and purified rhamnolipids reduced the surface hydrophobicity of polystyrene.
Medical implants used in oral and orthopedic surgery are fabricated using mainly alloys such as
stainless steel and titanium (Veerachamy et al. 2014). In the study of Ciandrini et al. (2016)
biosurfactants produced by Lactobacillus spp. significantly inhibited the adhesion and biofilm
formation on the titanium surface of S. mutans and S. oralis. Biosurfactants produced by
Lactobacillus fermentum RC-14 reduced the adherence of S. aureus to the surgical implants
(silicone) in rats (Gan et al. 2002). On the other hand, Prabhawathi et al. (2014) demonstrated
that biosurfactants secreted by strainsof B. subtilis and P. aeruginosa improved the attachment of
those bacteria to the medical implant surface (made of polyethylene) followed by the improve-
ment of cell proliferation and biofilm formation.

Biosurfactants as antibiofilm agents: examples

The functionality of various types of biosurfactants as antibiofilm agents is given in detail as
follows. Additionally, Table 1 summarizes the latest results of the research on the biosurfactants
activity against various microbial biofilms.
Table 1. Different types of microbial biosurfactants and their activity against various biofilm forming microorganisms.
Biosurfactant
producing Activity against microbial Biosurfactant
organism Biosurfactant type biofilms concentration Specific remark Reference
Coculture of Glycolipid biosurfactant V. harveyi – Coculture improved production of biosurfactant by Hamza et al.
Staphylococcus (BSSLSZ2) Staphylococcus lentus SZ2 and thus enhanced antibiofilm 2018
lentus SZ2 activity.
and Vibrio
harveyi
Bacillus amyloliquefaciens Lipopeptides (surfactin, iturin Agrobacterium 40 μg mL−1 (strain C58) and 80 μg mL−1 (strain B6) –
A and fengycin) tumefaciens
strains C58 and B6
Abdallah et al.
2018
– Affinity surfactant peptide Escherichia coli O157:H7 125 mg mL−1 1. 75% biofilm inhibition observed using surfactant peptide at Sun et al.
(synthesized in laboratory) 125 mg/L. 2018
2. The affinity surfactant peptide works by adsorbing on the
biofilms.
Bacillus sp. Mixture of surfactin and Streptococcus mutans and 44 µg mL−1 (for less concentration of TP combined with surfactin is needed for Bucci et al.
Terpinen-4-ol (TP) an Candida albicans. C. albicans) antimicrobial activity (synergistic effect) 2018
essential oil from Melaleuca
alternifolia plant.
B. subtilis Biosurfactant TIM96 Trichosporon 78.125 to 312.5 μg mL−1 TIM96 as antifungal agent acts by reducing the total content of Cordeiro
(containing surfactin, sterols at sub-inhibitory concentration. et al. 2018
fengicin and iturin as some
of its components)
S. lentus Glycolipid biosurfactant (BS- V. harveyi, P. aeruginosa 10 to 20 μg mL−1 1. The efficacy of glycolipid biosurfactant (BS-SLSZ2) owing to Hamza et al.
SLSZ2) its antiadhesive properties was tested to protect Artemia 2017
salina against infections. 2. A.salina is a major live feed used
in aquaculture.
1. P. aeruginosa Mixed rhamnolipid JBR425 Streptococcus mutans, . oralis, 1. Anti-adhesion assay: Combination of these biosurfactants and standard Elshikh et al.
2. Starmerella (MR) and lactonic Actinomyces naeslundii, MR (0.8–12.5 mg antimicrobial agents further has low MIC values and thus 2017
bombicola sophorolipids (LSLs) Neisseria mucosa mL−1) and LSLs good anti-biofilm activity.
and Candida and Streptococcus sanguinis (0.025–0.4 mg mL−1)
bastistae 2. Anti-biofilm assay: MR
(0.1–1 mg mL−1) and
LSLs (0.1–1.0 mg mL−1)
Pseudomonas Rhamnolipids (RL) Staphylococcus aureus 0.1% concentration at 25° Rhamnolipids when grown in skim milk rather than in nutrient Silva et al.
INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH

spp. C (in nutrient broth) broth reduces biofilm ability to 86.9% at 25°C compared to 2017
only 35% reduction achieved using nutrient broth.
(Continued)
9
 10
K. PARASZKIEWICZ ET AL.

Table 1. (Continued).
Biosurfactant
producing Activity against microbial Biosurfactant
organism Biosurfactant type biofilms concentration Specific remark Reference
Lactobacillus Glycolipid E. coli, S. aureus, Proteus Antimicrobial, antibiofilm and Satpute
acidophilus vulgaris, antiadhesive potential on catheter and polydimethyl et al.,
NCIM 2903 B. subtilis, P. aeruginosa, siloxane surfaces 2016b;
P. putida Satpute
et al. 2018
Staphylococcus Cell-associated biosurfactants: S. haemolyticusMD29 and 0.012–25.0 mg mL−1 Biosurfactant (BS29) has a stronger and broader spectrum of Rossi et al.
haemolyticus BS29 and BS49 MD49, Enterococcus faecium, (concentrations used) activity. 2016
MD29 and P. aeruginosa, S. aureus and
MD49 S. epidermidis
Starmerella Sophorolipid (SL) C. albicans 1. MIC 80–60 μg mL−1 Sophorolipid (SL) downregulates the expression of hypha Haque et al.
species 2. BIC 80–120 μg mL−1 specific genes HWP1, ALS1, ALS3, ECE1 and SAP4 thus, 2017
3. BEC 80–480 μg mL−1 explaining anti-biofilm activity and inhibition of hyphae.
*abbreviations used: BEC – biofilm effect concentration; BIC – biofilm inhibitory concentration; ED50 – dose (or concentration) causing 50% of maximum biofilm inhibition; IC50 – the half maximal
inhibitory concentration reacted on biofilm; MAD – multiple ascending dose; MFC – minimum effective concentration; MIC -minimum inhibitory concentration.
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 11

Glycolipids
Molecules of glycolipids are built from a carbohydrate moiety linked to a fatty acid chain. That
class of biosurfactants includes among others rhamnolipids, trehalolipids, mannosylerythritol-
lipids (MELs) and cellobiose lipids. Many glycolipids have been shown to be able to form pores
and destabilize biological membranes. Antibacterial, antifungal, antitumor and anti-biofilm activ-
ities of glycolipid biosurfactants have been reviewed by Inès and Dhouha (2015).

Rhamnolipids
Rhamnolipids are extracellular secondary metabolites secreted by various Pseudomonas strains
(predominantly by the opportunistic pathogen P. aeruginosa) and involved in different stages of
biofilm formation by these bacteria. Rhamnolipids are successfully exploited in environmental
technologies especially in water and soil remediation processes due to their ability to accelerate the
removal of various organic and inorganic pollutants (Pacwa-Płociniczak et al. 2011; Bustamante
et al. 2012; Lawniczak et al. 2013). On the other hand, rhamnolipids exhibit anti-adhesive and
disrupting activity against biofilms formed by several pathogenic microorganisms. The involve-
ment of rhamnolipids in microbial cell adhesion and biofilm development has been exhaustively
reviewed by Nickzad and Déziel (2014).
Recently, De Rienzo and Martin (2016) used mono-rhamnolipids (Rha-C10-C10) produced by
P. aeruginosa ATCC 9027 and di-rhamnolipids (Rha-Rha-C14-C14) synthesized by non-pathogenic
Burkholderia thailandensis E264 to disrupt biofilm of B. subtilis BBK006. Biosurfactants were found to
differ in their antibiofilm activity and the biofilm produced by the examined Bacillus strain was more
sensitive to di-rhamnolipids (0.4 g l−1) synthetized by B. thailandensis.
Long chain rhamnolipids from B. thailandensis E264 were also successfully used against some
oral pathogens (Streptococcus oralis, Actinomyces naeslundii, Neisseria mucosa and Streptococcus
sanguinis). Suggested mechanisms of rhamnolipid antimicrobial activity included an increase in
cell membrane permeability and the production of reactive oxygen species (ROS) in treated cells
leading to oxidative stress. Moreover, studied rhamnolipids displayed a good potency against oral-
bacteria biofilms in combination with lauryl sodium sulphate. Therefore, Elshikh et al. (2017)
suggested a potential utility of studied rhamnolipids as ingredients in different oral-related
applications including cosmetic products and medical devices.

Sophorolipids
Sophorolipids are secreted by non-pathogenic yeast (e.g. Candida bombicola and C. apicola)
and molecules of these biosurfactants contain sugar sophorose linked to the fatty acid.
Antimicrobial properties and biofilm disruption activity of sophorolipid biosurfactants were
investigated by De Rienzo et al. (2015) against both Gram-negative and Gram-positive strains.
The obtained data revealed that sophorolipids at a concentration of 5% v/v inhibited bacterial
growth of Cupriavidus necator ATCC 17699 and B. subtilis BBK006 and also disrupted biofilms
of single as well as of mixed B. subtilis BBK006 and S. aureus ATCC 9144 cultures. Haque
et al. (2017) showed that sophorolipids inhibited C. albicans biofilm formation and reduced the
viability of preformed biofilms. Moreover, the mixture of sophorolipids with amphotericin B or
fluconazole acted synergistically against Candida biofilms. Observations performed by scanning
electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) revealed the
absence of hyphae and alterations in the morphology of biofilm cells. It was also found that
studied biosurfactants down regulated the expression of hypha specific genes including HWP1,
ALS1, ALS3, ECE1 and SAP4.

Other glycolipids
Pradhan et al. (2014) used a novel glycolipid biosurfactant produced by Lysinibacillus fusiformis
strain S9 (an isolate from a river bank) to inhibit biofilms formed by pathogenic strains of E. coli
12 K. PARASZKIEWICZ ET AL.

and S. mutans (responsible among others for urinary tract infections and dental caries, respec-
tively). The biosurfactant at a concentration of 40 μg ml−1 did not show any bactericidal activity
but in the case of both pathogenic strains, it completely restricted the biofilm formation on
hydrophilic as well as hydrophobic surfaces. In conclusion these properties were suggested to be
potentially important in biomedical applications, where the antibiofilm agents are expected not to
have a toxic influence on the biotic system.

Lipopeptides
Lipopeptides constitute a structurally diverse class of bacterial and fungal extracellular metabolites.
Compounds from the surfactin, iturin, and fengycin families produced by various Bacillus strains
and Pseudomonas lipopeptides (classified into four major groups: viscosin, amphisin, tolaasin and
syringomycin) are among the most popular lipopeptide biosurfactants (Raaijmakers et al. 2010).
In the most recent study, pontifactin, a novel biosurfactant produced by the marine bacterial
strain Pontibacter korlensis SBK47, was structurally characterized as a lipopeptide containing
palmitic acid and a short heptapeptide with a Ser-Asp-Val-Ser-Ser sequence. The biosurfactant
was shown to exhibit promising antimicrobial and antiadhesive activities against different patho-
gens. Data obtained from the pre-adhesion inhibition assay (with the crystal violet staining)
revealed high anti-biofilm activity of pontifactin used at the concentration of 2 mg ml−1 against
strains belonging to B. subtilis, S. aureus, Salmonella typhi and Vibrio cholera (Balan et al. 2016).
Song et al. (2016) reported that lipopeptide biosurfactants produced by Bacillus amyloliquefaciens
anti-CA, disrupted cells of P. aeruginosa PAO1 and Bacillus cereus, inhibited the biofilm forma-
tion and dispersed pre-formed biofilms of tested bacteria. The obtained data also revealed the
inhibition of the PslC gene expression (related to exopolysaccharide production in P. aeruginosa
PAO1). Dalili et al. (2015) described another novel biosurfactant (called coryxin) secreted by
Corynebacterium xerosis strain NS5. Combined data from IR spectrometric, NMR as well as LC–
MS analysis allowed identifying coryxin as a lipopeptide consisting of the heptapeptide (Asn–Arg–
Asn–Gln–Pro–Asn–Ser) linked to the β-hydroxy fatty acid chain (with the length of 11 carbon
atoms). Besides its excellent surface and emulsifying properties, coryxin was found to have anti-
bacterial, anti-adhesive and anti-biofilm activities against strains of S. aureus, S. mutans, E. coli
and P. aeruginosa. The highest anti-biofilm and anti-adhesive activity (with the inhibition
percentage of up to 82.5 and 77.4%, respectively) was observed for coryxin at the concentration
of 100 mg ml−1 against the test strain of S. aureus.
Nalini et al. (2016) described the structure and anti-biofilm activities of another novel biosur-
factant produced by B. cereus strain SNAU01 (an isolate from hydrocarbons contaminated soil). The
biosurfactant structure was studied by FTIR, TLC and GC–MS techniques and the obtained data
indicated the presence of a lipopeptide built from octadecanoic acid methyl ester and a short peptide
with a predicted sequence of Pro-Leu-Gly-Gly. The obtained data revealed that the studied
lipopeptide at the concentration of 250 mg ml−1 disrupted the formation of biofilms of pathogenic
bacteria (P. aeruginosa MTCC 2453 and E. coli MTCC 2939) by up to 56 and 62%, respectively.
Lichenysin, a lipopeptide from the surfactin family and obtained from a Bacillus licheniformis
culture was demonstrated to effectively prevent biofilm formation by methicillin-resistant
S. aureus (MRSA) (68.7%) and C. albicans (74.4%), with ED50 values of 8.3 and 17.2 μg ml−1,
respectively. It was also efficient in a surface post-treatment at removing biofilms of MRSA
(55.7%) and Yersinia enterocolitica (51.5%) with an ED50 of 2.8 and 4.1 μg ml−1, respectively
(Coronel-León et al. 2016).
Ceresa et al. (2016) used a lipopeptide mixture (containing compounds from the surfactin and
fengycin families) produced by B. subtilis AC7 against adhesion and biofilm formation of
C. albicans. Co-incubation with 2 mg ml−1 of AC7 biosurfactants significantly reduced adhesion
and biofilm formation of three C. albicans strains on medical-grade silicone elastomeric disks
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 13

(SEDs). The obtained results suggested a potential role of the used lipopeptide mixture as a coating
agent for preventing fungal infection associated with silicone medical devices.
Moryl et al. (2015) investigated bacterial planktonic growth, processes of biofilm formation and
dislodging in the presence of a biosurfactant mixture obtained from the B. subtilis strain I’1a. That
strain synthetized compounds from the surfactin, iturin A and fengycin families (Bernat et al. 2016;
Paraszkiewicz et al. 2018). Lipopeptides antimicrobial activity was tested against 32 uropathogenic
strains belonging to 12 different species of Gram-negative and Gram-positive bacteria. It was
established that none of the tested strains was sensitive to pure surfactin at the concentrations
ranging from 0.1 to 0.4 mg ml−1, but the used lipopeptide mixture inhibited the metabolic activity of
planktonic cells by 88.1% in the case of 21 of the studied uropathogens. Therefore, it was suggested
that synergistic application of surfactins, iturins and fengycin could be efficient in the eradication of
biofilms formed by various bacterial uropathogens.
The iturin and fengycin mixture (extracted from the culture of the B. amyloliquefaciens
strain AR2), when used at the sub-minimum growth inhibition concentration, decreased
C. albicans cell surface hydrophobicity, hindered germ tube formation and reduced the
mRNA expression of hyphae-specific gene HWP1 and ALS3. Depending upon the concentra-
tion and kind of the tested Candida strain, the studied lipopeptide mixture inhibited biofilm
formation in the range of 46-100% and dislodged 25-100% of preformed biofilm from poly-
styrene plates (Rautela et al. 2014).
Kuiper et al. (2004) described two novel lipopeptides biosurfactants produced by
Pseudomonas putida PCL1445, referred to as putisolvin I and putisolvin II with 12 amino
acids and a hexanoic acid lipid. Isolated lipopeptides inhibited the formation of Pseudomonas
biofilms on polyvinyl chloride (PVC). Moreover, they broke down biofilms of various
Pseudomonas spp. which had been formed on PVC. The authors suggest that the possible
mode of action of putisolvin I and II during the inhibition of biofilm formation could be
binding to the cell surface or to components of the cell surface thereby influencing the outer
membrane hydrophobicity. In addition, the change in cell surface hydrophobicity might also
influence the interaction between the individual cells in a biofilm, making the formation of
a dense and stable biofilm more difficult.

Biosurfactants synthesised from lactic acid bacteria (LAB)

Lactic acid bacteria (LAB) are a large group of bacteria containing at least 50 species. Moreover,
a number of LAB have been proposed as probiotic strains (Fariq and Saeed 2016). In comparison
to other microbial surfactants like lipopeptides or rhamnolipids, biosurfactants produced by LAB
are still poorly described, due to the lower surface activity, higher CMC values, lower production
levels and frequent association with the LAB cell wall, which makes their isolation and purification
difficult. However, LAB are generally regarded as safe (GRAS) and many of those bacteria are used
in food and pharmaceutical industries (Gudiña et al. 2010). Recently, Satpute et al. (2016b)
discussed the properties and commercial potential of Lactobacilli biosurfactants.
Biosurfactants isolated from Lactobacillus jensenii and Lactobacillus rhamnosus cell surfaces,
when applied at the concentrations between 25 and 100 mg ml−1, showed antibacterial, anti-
adhesive and anti-biofilm activity against clinical Multidrug Resistant (MDR) strains of
Acinetobacter baumannii, E. coli, and S. aureus (MRSA) considered as prominent biofilm formers
on wounds, medical implants and industrial surfaces. Analysis performed by transmission electron
microscopy (TEM) also revealed that studied biosurfactants caused damage to the A. baumannii cell
membrane and S. aureus cell wall. Based on the obtained data, Sambanthamoorthy et al. (2014)
suggested the utility of studied biosurfactants for coating medical surfaces to prevent microbial
colonization and/or to treat hospital-acquired infections.
A cell-bound biosurfactant of Lactobacillus agilis CCUG31450 (reducing the surface tension of
water to 42.5 mN m−1 and exhibiting a high emulsifying activity) was identified by FTIR analysis as
14 K. PARASZKIEWICZ ET AL.

a glycoprotein (Gudiña et al. 2015). The studied biosurfactant exhibited an anti-adhesive activity
against S. aureus, as well as an antimicrobial activity against S. aureus, Streptococcus agalactiae and
P. aeruginosa.
Crude biosurfactants isolated from the cell surfaces of two Lactobacillus casei strains (ATCC 393
and LBl) were reported as potent anti-adhesive and anti-biofilm agents against oral opportunistic
S. aureus strains (Merghni et al. 2017). Moreover, the high efficacy of biofilm eradication accompanied
by proved antioxidant and antiproliferative activities allowed to suggest a potential applicability of the
studied biosurfactants for limiting or preventing oral infections and diseases. Sharma and Saharan
(2016) reported surface and biological properties of biosurfactant produced by Lactobacillus helveticus
MRTL91 – a strain isolated from ethnic fermented food prepared from Yak milk (described as
Chhurpi cheese). The antimicrobial and anti-adhesive activity of the purified biosurfactant was
investigated against a broad range of pathogenic and non-pathogenic strains, including Gram-
positive and Gram-negative bacteria, as well as yeasts. The highest anti-adhesive properties (using co-
incubation of silicon tubes with the biosurfactant) were observed against B. cereus (87%),
S. epidermidis (85%), L. monocytogenes (84.3%), S. aureus (83.1%) and L. innocua (82.1%). On the
other hand, the biosurfactant even at the highest concentration used (25 mg ml−1) was shown to have
weaker (from 69.2 to 87%) anti-adhesive activity against the yeast C. albicans. Ceresa et al. (2015)
reported that the biosurfactant of the Lactobacillus brevis strain CV8LAC did not inhibit C. albicans
growth in both planktonic and sessile form but had an ability to counteract significantly the initial
deposition of C. albicans to silicone surfaces and to effectively slow biofilm growth. Shokouhfard et al.
(2015) described the antiadhesive capability of the biosurfactant produced by the probiotic strain
Lactobacillus acidophilus ATCC 4356, against pathogenic, biofilm-producing strains of Serratia
marcescens (considered to be a nosocomial pathogen responsible for infections in the urinary, vaginal
and gastrointestinal tracts, as well as skin). The FTIR analysis revealed a protein component in the
composition of the derived biosurfactant. When the studied biosurfactant was applied at the con-
centration of 2.5 mg ml−1, it showed anti-adhesive activity against all tested strains of S. marcescens
(including S. marcescens ATCC 13880, S. marcescens ATCC 19180 and six other strains of
S. marcescens isolated from hospitalized patients and exhibiting biofilm formation ability).

Other biosurfactants
Kumar et al. (2014) reported that ochrosin – a novel biosurfactant (characterized as 4-dimethy-
laminobenzaldehyde) from the halophilic bacterial strain Ochrobactrum sp. BS-206 exhibited
multifunctional biological activity including antimicrobial, anti-adhesive, anti-feedant and insec-
ticidal properties. Ochrosin anti-adhesive activity facilitated the disruption of biofilm formation
by different test bacterial and Candida strains. Extracellular polymeric substance (EPS) produced
by the Oceanobacillus iheyensis strain BK6 (a biofilm-forming marine bacterium) was shown to
exhibit biosurfactant properties and an ability to prevent biofilm formation by multidrug-resistant
clinical isolate of S. aureus, with 62.3% biofilm inhibition determined for EPS used at
a concentration of 175 µg ml−1 (Kavita et al. 2014). Another polymeric biosurfactant obtained
from a Trichosporon montevideense CLOA72 culture was shown to be an agent limiting the
adhesion of C. albicans and C. krusei cells. Additionally, the obtained data revealed that the
used biosurfactant changed the cell surface characteristics and modified the chemical composition
(carbohydrate and protein concentrations) of the biofilm matrix and planktonic cell surfaces of
the two studied Candida strains (Ferreira et al. 2016).

Synergistic activity of biosurfactant with nanoparticles against biofilm forming

pathogens
Among various biofilm forming pathogens, P. aeruginosa and S. aureus are a serious menace to the
clinical as well as industrial settings (food, fermentation, breweries). They have an ability to form
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 15

resistant biofilm, leading to a plethora of consequences and ultimately causing an array of health
problems. In this respect, synthesis of rhamnolipid coated silver and iron oxide nanoparticles not only
exhibit anti-bacterial, anti-adhesive, anti-biofilm activity but also show excellent anti-microbial activity
by generating reactive oxygen species (ROS). These results suggest that such complexes could be used to
decrease biofilm formation and consequently the severity of infection (Khalid et al. 2019).
Complexes of rhamnolipid with silver and iron nanoparticles have proved to be effective against
various microorganisms (Salmonella enteritids, S. aureus, Bacillus pumilus, L. monocytogenes and
Yarrowia lipolytica). The size of silver (35 nm) and iron (48 nm) nanoparticles is crucial to exhibit
antibiofilm potential. The antimicrobial activity of these complexes is mainly due to their increased
surface to volume ratio, release of ions and ROS of nanoparticle. Anti-adhesive activity is connected
with an ability to reduce cell surface hydrophobicity. These complexes were found to be effective at
a minimum inhibitory concentration of 1 mg ml−1, being more susceptible to S. aureus than
P. aeruginosa. Moreover, rhamnolipid exhibits antibiofilm activity above 7 mg ml−1. The antiadhe-
sive activity of rhamnolipid functionalized Fe3O4 was found to be effective against both the strains
(Khalid et al. 2019).

Conclusion and future prospects

Many biosurfactants (surface active agents of microbial origin) exhibit valuable antibacterial,
antifungal, antiviral activities as well as anti-adhesive properties and/or an ability to disrupt
microbial biofilms. Therefore, biosurfactants are indicated to be compounds with a great potential
in biomedical and health related areas. Wider applications of biosurfactants as anti-biofilm agents
require overcoming problems with the high production cost and still limited information about:
(1) structures and properties of particular biosurfactants due to the fact that they are usually
synthesized as a mixture of various isoforms and homologues; (2) the mechanisms responsible for
biosurfactant antibiofilm activities and (3) biosurfactants toxicity towards humans. It would be
interesting to continue the study using biosurfactants as coating agents in order to develop novel
biomaterials with an ability to reduce the initial microorganism attachment. More attention
should be paid to exploring the anti-biofilm potential of biosurfactants combined with other
molecules e.g. enzymes, antibiotics, metal nanoparticles, or liposomes. The future study should
alsobe focused to a greater extent on biofilms formed by multidrug resistant microorganisms and
by mixed cultures. It is worth mentioning that the fast progress in biosurfactants commercializa-
tion and extending the knowledge of biosurfactant-producing and biofilm-forming microorgan-
isms has resulted from the advances in ‘omics’ sciences and novel analytical techniques (e.g. mass
spectrometry).

Highlights
(1) Biofilms are extremely difficult to eradicate.
(2) Biosurfactants possess anti-adhesive properties, an ability to disrupt microbial biofilms and/or antibiotic
activity.
(3) Biosurfactants gather at the interface between polar and nonpolar media.
(4) Biosurfactants are indicated to be compounds with a great potential in health care and environment.

Acknowledgments
This paper was prepared in connection with the work conducted under the project no 2013/09/B/NZ9/01759 that
was sponsored by the National Science Center (Poland). Dr. Surekha K. Satpute expresses special thanks to
Department research and development programs (DRDP), Savitribai Phule Pune University, Pune – 411007,
Maharashtra, and Department of Science and Technology- Promotion of University Research and Scientific 618
Excellence (DST-PURSE), Government of India, New Delhi grants for financial support.
16 K. PARASZKIEWICZ ET AL.

Funding
This work was supported by the Narodowe Centrum Nauki [2013/09/B/NZ9/01759].

Conflict of Interest
The authors declare no conflict of interest.

References
Abdallah DB, Tounsi S, Gharsallah H, Hammami A, Frikha-Gargouri O. 2018. Lipopeptides from Bacillus
amyloliquefaciens strain 32a as promising biocontrol compounds against the plant pathogen Agrobacterium
tumefaciens. Environ Sc Poll Res. 25(36):36518–36529. doi:10.1007/s11356-018-3570-1.
Absalon C, Ymele-Leki P, Watnick PI. 2012. The bacterial biofilm matrix as a platform for protein delivery. Mbio. 3
(4):e00127–12. doi:10.1128/mBio.00127-12.
An D, Danhorn T, Fuqua C, Parsek MR. 2006. Quorum sensing and motility mediate interactions between
Pseudomonas aeruginosa and Agrobacterium tumefaciens in biofilm cocultures. Proc Nation Acad Sc USA. 103
(10):3828–3833. doi:10.1073/pnas.0511323103.
Araujo LV, Guimarães CR, Da Silva Marquita RL, Santiago VM, de Souza MP, Nitschke M, Freire DMG. 2016.
Rhamnolipid and surfactin: anti-adhesion/antibiofilm and antimicrobial effects. Food Cont. 63(1):171–178.
doi:10.1016/j.foodcont.2015.11.036.
Balan SS, Kumar C, Ganesh JS. 2016. Pontifactin; a new lipopeptide biosurfactant produced by a marine Pontibacter
korlensis strain SBK-47: purification; characterization and its biological evaluation. Process Biochem. 51
(12):2198–2207. doi:10.1016/j.procbio.2016.09.009.
Banat IM, De Rienzo MA, Quinn GA. 2014. Microbial biofilms: biosurfactants as antibiofilm agents. App Microbiol
Biotechnol. 98(24):9915–9929. doi:10.1007/s00253-014-6169-6.
Barraud N, Storey MV, Moore ZP, Webb JS, Rice SA, Kjelleberg S. 2009. Nitric oxide-mediated dispersal in single-
and multi-species biofilms of clinically and industrially relevant microorganisms. Microbiol Biotechnol. 2
(3):370–378. doi:10.1111/j.1751-7915.2009.00098.x.
Berlanga M, Guerrero R. 2016. Living together in biofilms: the microbial cell factory and its biotechnological
implication. Microbial Cell Fact. 15:165. doi:10.1186/s12934-016-0569-5.
Bernat P, Paraszkiewicz K, Siewiera P, Moryl M, Płaza G, Chojniak J. 2016. Lipid composition in a strain of Bacillus
subtilis, a producer of iturin A lipopeptides that are active against uropathogenic bacteria. World J Microbiol
Biotechnol. 32(10):157. doi:10.1007/s11274-016-2126-0.
Boles BR, Thoendel M, Singh PK. 2005. Rhamnolipids mediate detachment of Pseudomonas aeruginosa from
biofilms. Mol Microbiol. 57(5):1210–1223. doi:10.1111/j.1365-2958.2004.04767.x.
Bonnichsen L, Svenningsen N, Rybtke M, de Bruijn I, Raaijmakers JM, Tolker-Nielsen T, Nybroe O. 2015.
Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms.
Microbiology. 161(12):2289–2297. doi:10.1099/mic.0.000191.
Bucci AR, Marcelino L, Mendes RK, Etchegaray A. 2018. The antimicrobial and antiadhesion activities of micellar
solutions of surfactin, CTAB and CPCl with terpinen-4-ol: applications to control oral pathogens. World
J Microbiol Biotechnol. 34(6):86. doi:10.1007/s11274-018-2472-1.
Burmølle M, Ren D, Bjarnsholt T, Sørensen SJ. 2014. Interactions in multispecies biofilms: do they actually matter?
Trends Microbiol. 22(2):84–91. doi:10.1016/j.tim.2013.12.004.
Bustamante M, Duran N, Diez MC. 2012. Biosurfactants are useful tools for the bioremediation of contaminated
soil: a review. J Soil Sc Plant Nut. 12(4):667–687.
Cavalheiro M, Teixeira MC. 2018. Candida biofilms: threats, challenges, and promising strategies. Front Med. 5:28.
doi:10.3389/fmed.2018.00028.
Ceresa C, Rinaldi M, Chiono V, Carmagnola I, Allegrone G, Fracchia L. 2016. Lipopeptides from Bacillus subtilis
AC7 inhibit adhesion and biofilm formation of Candida albicans on silicone. Antonie Van Leeuwenhoek. 109
(10):1375–1388. doi:10.1007/s10482-016-0736-z.
Ceresa C, Tessarolo F, Caola I, Nollo G, Cavallo M, Rinaldi M, Fracchia L. 2015. Inhibition of Candida albicans
adhesion on medical-grade silicone by a Lactobacillus-derived biosurfactant. J App Microbiol. 118(5):1116–1125.
doi:10.1111/jam.12760.
Ciandrini E, Campana R, Casettari L, Perinelli DR, Fagioli L, Manti A, Palmieri GF, Papa S, Baffone W. 2016.
Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral streptococci
biofilm. App Microbiol Biotechnol. 100(15):6767–6777. doi:10.1007/s00253-016-7531-7.
Cordeiro RA, Weslley Caracas Cedro E, Raquel Colares Andrade A, Serpa R, José de Jesus Evangelista A, Sales de
Oliveira J, Santos Pereira V, Pereira Alencar L, Bruna Leite Mendes P, Cibelle Soares Farias B, et al. 2018.
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 17

Inhibitory effect of a lipopeptide biosurfactant produced by Bacillus subtilis on planktonic and sessile cells of
Trichosporon spp. Biofouling. 34(3):309–319. doi:10.1080/08927014.2018.1437617
Coronel-León J, Marqués AM, Bastida J, Manresa A. 2016. Optimizing the production of the biosurfactant
lichenysin and its application in biofilm control. J App Microbiol. 120(1):99–111. doi:10.1111/jam.12992.
Costa-Orlandi CB, Sardi JCO, Pitangui NS, de Oliveira HC, Scorzoni L, Galeane MC, Medina-Alarcon KP,
Melo WCMA, Marcelino MY, Braz JD, et al. 2017. Fungal Biofilms and polymicrobial diseases. J Fungi
(Basel). 3(2):pii: E22. doi:10.3390/jof3020022.
Czaczyk K, Myszka K. 2007. Biosynthesis of extracellular polymeric substances EPS and its role in microbial biofilm
formation. Pol J Environ Studies. 16(6):799–806.
Dalili D, Amini M, Faramarzi MA, Fazeli MR, Khoshayand MR, Samadi N. 2015. Isolation and structural
characterization of Coryxin; a novel cyclic lipopeptide from Corynebacterium xerosis NS5 having emulsifying
and anti-biofilm activity. Coll Surface B. 135:425–432. doi:10.1016/j.colsurfb.2015.07.005.
Das P, Mukherjee S, Sen R. 2009. Antiadhesive action of a marine microbial surfactant. Coll Surfaces B Biointer. 71
(2):183–186. doi:10.1016/j.colsurfb.2009.02.004.
Davey ME, Caiazza NC, Tootle GAO. 2003. Rhamnolipid surfactant production affects biofilms architecture in
Pseudomonas aeruginosa PA-01. J Bacteriol. 185(3):1027–1036. doi:10.1128/JB.185.3.1027-1036.2003.
Davies DG. 2011. Biofilm dispersion. In: Flemming HC, editor. Biofilm highlights. Heidelberg: Springer-Verlag
Berlin; p. 1–28.
Davies DG, Marques CNH. 2009. A fatty acid messenger is responsible for inducing dispersion in microbial
biofilms. J Bacteriol. 191(5):1393–1403. doi:10.1128/JB.01214-08.
De Rienzo MA, Banat IM, Dolman B, Winterburn J, Martin PJ. 2015. Sophorolipid biosurfactants: possible uses as
antibacterial and antibiofilm agent. New Biotechnol. 32(6):720–726. doi:10.1016/j.nbt.2015.02.009.
De Rienzo MA, Martin PJ. 2016. Effect of mono and di-rhamnolipids on biofilms pre-formed by Bacillus subtilis
BBK006. Curr Microbiol. 73(2):183–189. doi:10.1007/s00284-016-1046-4.
Delcaru C, Alexandru I, Podgoreanu P, Grosu M, Stavropoulos E, Chifiriuc MC, Lazar V. 2016. Microbial biofilms
in urinary tract infections and prostatitis: etiology, pathogenicity, and combating strategies. Pathogens. 5(4):65.
doi:10.3390/pathogens5040065.
Do Valle Gomes ZM, Nitschke M. 2012. Evaluation of rhamnolipid and surfactin to reduce the adhesion and
remove biofilms of individual and mixed cultures of food pathogenic bacteria. Food Con. 25:441–447.
doi:10.1016/j.foodcont.2011.11.025.
Donlan RM. 2001. Biofilm formation: A clinically relevant microbiological process. Clin Infect Dis. 33
(8):1387–1392. doi:10.1086/322972.
Elias S, Banin E. 2012. Multi-species biofilms: living with friendly neighbours. FEMS Microbiol Rev. 36
(5):990–1004. doi:10.1111/j.1574-6976.2012.00325.x.
Elshikh M, Funston S, Chebbi A, Ahmed S, Marchant R, Banat IM. 2017. Rhamnolipids from non-pathogenic
Burkholderia thailandensis E264: physicochemical characterization; antimicrobial and antibiofilm efficacy against
oral hygiene related pathogens. New Biotechnol. 36:26–36. doi:10.1016/j.nbt.2016.12.009.
Fait ME, Bakas L, Garrote GL, Morcelle SR, Mario CN, Saparrat MCN. 2019. Cationic surfactants as antifungal
agents. App Microbiol Biotechnol. 103(1):97–112. doi:10.1007/s00253-018-9467-6.
Fanning S, Mitchell AP. 2012. Fungal biofilms. PLoS Pathog. 8(4):e1002585. doi:10.1371/journal.ppat.1002585.
Fariq A, Saeed A. 2016. Production and biomedical applications of probiotic biosurfactants. Cur Microbiol. 72
(4):489–495. doi:10.1007/s00284-015-0978-4.
Ferreira GF, Dos Santos Pinto BL, Souza EB, Viana JL, Zagmignan A, Dos Santos JR, Santos AR, Tavares PB,
Denadai AM, Monteiro AS. 2016. Biophysical effects of a polymeric biosurfactant in Candida krusei and
Candida albicans cells. Mycopathologia. 181(11–12):799–806. doi:10.1007/s11046-016-0054-z.
Flemming HC, Neu T, Wozniak D. 2007. The EPS matrix: the house of biofilm cells. J Bacteriol. 189
(22):7945–7947. doi:10.1128/JB.00858-07.
Flemming HC, Wingender J, Szewzyk U, Steinberg P, Rice SA, Kjelleberg S. 2016. Biofilms: an emergent form of
bacterial life. Nature Rev Microbiol. 14(9):563–575. doi:10.1038/nrmicro.2016.94.
Fong JNC, Yildiz F. 2015. Biofilm matrix proteins. Microbiol Spectr. 3(2). doi:10.1128/microbiolspec.
Gan BS, Kim J, Reid G, Cadieux P, Howard JC. 2002. Lactobacillus fermentum RC-14 inhibits Staphylococcus aureus
infection of surgical implants in rats. J Inf Dis. 185(9):1369–1372. doi:10.1086/340126.
Gilbert P, Evans DJ, Evans E, Duguid IG, Brown MR. 1991. Surface characteristics and adhesion of E. coli and
Staphylococcus epidermidis. J App Bacteriol. 71(1):72–77. doi:10.1111/j.1365-2672.1991.tb04589.x.
Gudiña EJ, Elisabete C, Fernandes JAT, Lígia RR. 2015. Antimicrobial and anti-adhesive activities of cell-bound
biosurfactant from Lactobacillus agilis CCUG31450. RSC Adv. 110:90960–90968. doi:10.1039/C5RA11659G.
Gudiña EJ, Teixeira JA, Rodrigues LR. 2010. Isolation and functional characterization of a biosurfactant produced
by Lactobacillus paracasei. Coll Surf B. 76(1):298–304. doi:10.1016/j.colsurfb.2009.11.008.
Hamza F, Kumar AR, Zinjarde S. 2018. Coculture induced improved production of biosurfactant by Staphylococcus
lentus SZ2: role in protecting Artemia salina against Vibrio harveyi. Enzyme Microb Technol. 114:33–39.
doi:10.1016/j.enzmictec.2018.03.008.
18 K. PARASZKIEWICZ ET AL.

Hamza F, Satpute S, Banpurkar A, Kumar AR, Zinjarde S. 2017. Biosurfactant from a marine bacterium disrupts
biofilms of pathogenic bacteria in a tropical aquaculture system. FEMS Microbiol Ecol. 93(11):123–138.
doi:10.1093/femsec/fix140.
Haque F, Sajid M, Cameotra SS, Battacharyya MS. 2017. Anti-biofilm activity of a sophorolipid-amphotericin
B niosomal formulation against Candida albicans. Biofouling. 33(9):768–779. doi:10.1080/08927014.2017.1363191.
Hood SK, Zottola EA. 1995. Biofilms in food processing. Food Cont. 6(1):9–18. doi:10.1016/0956-7135(95)91449-U.
Inès M, Dhouha G. 2015. Glycolipid biosurfactants: potential related biomedical and biotechnological applications.
Carbohyd Res. 416(1):59–69. doi:10.1016/j.carres.2015.07.016.
Janek T, Łukaszewicz M, Krasowska A. 2012. Antiadhesive activity of the biosurfactant pseudofactin II secreted by
the Arctic bacterium Pseudomonas fluorescens BD5. BMC Microbiol. 12:24. doi:10.1186/1471-2180-12-24.
Kaplan JB, Ragunath C, Velliyagounder K, Fine DH, Ramasubbu N. 2004. Enzymatic detachment of Staphylococcus
epidermidis biofilms. Antimicrob Agents Chemother. 48(7):2633–2636. doi:10.1128/AAC.48.7.2633-2636.2004.
Karlapudi AP, Venkateswarulu TC, Tammineedi J, Kanumuri L, Ravuru BK, Dirisala VR, Kodali VP. 2018. Role of
biosurfactants in bioremediation of oil-pollution: a review. Petroleum. 4(1):241–249. doi:10.1016/j.petlm.2018.03.007.
Kavita K, Singh VK, Mishra A, Jha B. 2014. Characterisation and anti-biofilm activity of extracellular polymeric
substances from Oceanobacillus iheyensis. Carbohyd Pol. 101:29–35. doi:10.1016/j.carbpol.2013.08.099.
Khalid HF, Tehseen B, Sarwar Y, Hussain SZ, Khan WS, Raza ZA, Bajwa SZ, Kanaras AG, Hussain I, Rehman A.
2019. Biosurfactant coated silver and iron oxide nanoparticles with enhanced anti-biofilm and anti-adhesive
properties. J Hazard Mater. 364:441–448. doi:10.1016/j.jhazmat.2018.10.049.
Kragh KN, Hutchison JB, Melaugh G, Rodesney C, Roberts AE, Irie Y, Jensen PO, Diggle SP, Allen RJ, Gordon V,
et al. 2016. Role of multicellular aggregates in biofilm formation. MBio. 7(2):e00237. doi:10.1128/mBio.00237-16.
Kuiper I, Lagendijk EL, Pickford R, Derrick P, Lamers EM, Thomas-Oates JE, Lugtenberg J, Bloemberg GV. 2004.
Characterization of two Pseudomonas putida lipopeptide biosurfactants, putisolvin I and putisolvin II, which
inhibit biofilm formation and break down existing biofilms. Mol Microbiol. 51(1):97–113. doi:10.1046/j.1365-
2958.2003.03751.x.
Kumar CG, Sujitha P, Mamidyala SK, Usharani P, Das B, Reddy CR. 2014. Ochrosin; a new biosurfactant produced
by halophilic Ochrobactrumsp. strain BS-206 MTCC 5720: purification; characterization and its biological
evaluation. Process Biochem. 49(10):1708–1717. doi:10.1016/j.procbio.2014.07.004.
Lawniczak L, Marecik R, Chrzanowski L. 2013. Contributions of biosurfactants to natural or induced
bioremediation. App Microbiol Biotechnol. 97(6):2327–2339. doi:10.1007/s00253-013-4740-1.
Liu W, Røder HL, Madsen JS, Bjarnsholt T, Sørensen SJ, Burmølle M. 2016. Interspecific bacterial interactions are
reflected in multispecies biofilm spatial organization. Front Microbiol. 7:1366. doi:10.3389/fmicb.2016.01366.
Majumdar S, Mondal S. 2016. Conversation game: talking bacteria. J Cell Commun Sign. 10(4):331–335.
doi:10.1007/s12079-016-0333-y.
Merghni A, Dallel I, Noumi E, Kadmi Y, Hentati H, Tobji S, Ben Amor A, Mastouri M. 2017. Antioxidant and
antiproliferative potential of biosurfactants isolated from Lactobacillus caseiand their anti-biofilm effect in oral
Staphylococcus aureus strains. Microb Pathog. 104:84–89. doi:10.1016/j.micpath.2017.01.017.
Momeni B, Brileya KA, Fields MW, Shou W. 2013. Strong interpopulation cooperation leads to partner intermixing
in microbial communities. Elife 2: e00230.Moryl M. 2015. Extracellular matrix as a microbial virulence factor in
the development of human diseases. Post Hig Med Dos. 69:1485–1498. (in Polish).
Moryl M, Spętana M, Dziubek K, Paraszkiewicz K, Różalska S, Płaza G, Różalski A. 2015. Antimicrobial;
antiadhesive and antibiofilm potential of lipopeptides synthesised by Bacillus subtilis; on uropathogenic
bacteria. Acta Biochem Pol. 62(4):725–732. doi:10.18388/abp.2015_1120.
Nalini S, Parthasarathi R, Prabudoss V. 2016. Production and characterization of lipopeptide from Bacillus cereus
SNAU01 under solid state fermentation and its potential application as anti-biofilm agent. Biocat Agricul
Biotechnol. 5:123–132. doi:10.1016/j.bcab.2016.01.007.
Naughton PJ, Marchant R, Naughton V, Banat IM. 2019. Microbial Biosurfactants: current trends and applications
in agriculture and biomedical industries. J App Microbiol. 127(1):12–28. doi:10.1111/jam.14243.
Nguyen UT, Burrows LL. 2014. DNase I and proteinase K impair Listeria monocytogenes biofilm formation and induce
dispersal of pre-existing biofilms. Int J Food Microbiol. 18(1):26–32. doi:10.1016/j.ijfoodmicro.2014.06.025.
Nickzad A, Déziel E. 2014. The involvement of rhamnolipids in microbial cell adhesion and biofilm development -
an approach for control? Lett App Microbiol. 58(5):447–453. doi:10.1111/lam.12211.
Nielsen AT, Tolker-Nielsen T, Barken KB, Molin S. 2000. Role of commensal relationships on the spatial structure
of a surface-attached microbial consortium. Environ Microbiol. 2(1):59–68.
Pacwa-Płociniczak M, Płaza GA, Piotrowska-Seget Z, Cameotra SS. 2011. Environmental applications of biosur-
factants: recent advances. Intern J Mol Sc. 12(1):633–654. doi:10.3390/ijms12010633.
Palmer J, Flint S, Brooks J. 2007. Bacterial cell attachment, the beginning of a biofilm. J Indust Microbiol
Biotechnol. 34(9):577–588. doi:10.1007/s10295-007-0234-4.
Paraszkiewicz K. 2016. Biosurfactant enhancement factors in microbial degradation processes. In: Długoński J,
editor. From omics to function and application. Norfolk UK: Caister Academic Press; p. 167–182.
 INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH 19

Paraszkiewicz K, Bernat P, Kuśmierska A, Chojniak J, Płaza G. 2018. Structural identification of lipopeptide

biosurfactants produced by Bacillus subtilis strains grown on the media obtained from renewable natural
resources. J Environ Manag. 209:65–70. doi:10.1016/j.jenvman.2017.12.033.
Patel S, Homaei A, Patil S, Daverey A. 2019. Microbial biosurfactants for oil spill remediation: pitfalls and
potentials. App Microbiol Biotechnol. 103(1):27–37. doi:10.1007/s00253-018-9434-2.
Pavithra D, Doble M. 2008. Biofilm formation, bacterial adhesion and host response on polymeric implants - issues
and prevention. Biomedical Matt. 3:034003. doi:10.1088/1748-6041/3/3/034003.
Petrova OE, Sauer K. 2016. Escaping the biofilm in more than one way: desorption, detachment or dispersion. Cur
Opp Microbiol. 30(1):67–78. doi:10.1016/j.mib.2016.01.004.
Prabhawathi V, Thirunavukarasu K, Doble M. 2014. A study on the long term effect of biofilm produced by
biosurfactant producing microbe on medical implant. Mater Sc Eng C Mater Biol App. 40(1):212–218.
doi:10.1016/j.msec.2014.03.050.
Pradhan AK, Pradhan N, Sukla LB, Panda PK, Mishra BK. 2014. Inhibition of pathogenic bacterial biofilm by
biosurfactant produced by Lysinibacillus fusiformis S9. Bioprocess Biosyst Eng. 37(2):139–149. doi:10.1007/
s00449-013-0976-5.
Purevdorj-Gage B, Costerton WJ, Stoodley P. 2005. Phenotypic differentiation and seeding dispersal in non-mucoid
and mucoid Pseudomonas aeruginosa biofilms. Microbiol. 151(Pt5):1569–1576. doi:10.1099/mic.0.27536-0.
Raaijmakers JM, De Bruijn I, Nybroe O, Ongena M. 2010. Natural functions of lipopeptides from Bacillus and
Pseudomonas: more than surfactants and antibiotics. FEMS Microbiol Rev. 34(6):1037–1062. doi:10.1111/j.1574-
6976.2010.00221.x.
Rautela R, Singh AK, Shukla A, Cameotra SS. 2014. Lipopeptides from Bacillus strain AR2 inhibits biofilm
formation by Candida albicans. Antonie Van Leeuwenhoek. 105(5):809–821. doi:10.1007/s10482-014-0135-2.
Ribeiro SM, Felício MR, Boas EV, Gonçalves S, Costa FF, Samy RP, Santos NC, Franco OL. 2016. New frontiers for
anti-biofilm drug development. Pharmacol Ther. 160:133–144. doi:10.1016/j.pharmthera.2016.02.006.
Røder HL, Sørensen SJ, Burmølle M. 2016. Studying bacterial multispecies biofilms: where to start? Trends
Microbiol. 24(6):503–513. doi:10.1016/j.tim.2016.02.019.
Rosenberg E, Ron EZ. 1999. High- and low-molecular mass microbial surfactants. App Microbiol Biotechnol. 52
(2):154–162. doi:10.1007/s002530051502.
Rossi CC, Santos-Gandelman JF, Barros EM, Alvarez VM, Laport MS, Giambiagi-de Marval M. 2016.
Staphylococcus haemolyticus as a potential producer of biosurfactants with antimicrobial, anti-adhesive and
synergistic properties. Lett App Microbiol. 63(3):215–221. doi:10.1111/lam.12611.
Rutherford ST, Bassler BL. 2012. Bacterial quorum sensing: its role in virulence and possibilities for its control.
Cold Spring Harb Perspect Med. 2(11):pii: a012427. doi:10.1101/cshperspect.a012427.
Salehi R, Savabi O, Kazemi M, Kamali S, Salehi AR, Eslami G, Tahmourespour A. 2014. Effects of Lactobacillus
reuteri-derived biosurfactant on the gene expression profile of essential adhesion genes gtfB, gtfC and ftf of
Streptococcus mutans. Adv Biomed Res. 3:169. doi:10.4103/2277-9175.140622.
Sambanthamoorthy K, Feng X, Patel R, Patel S, Paranavitana C. 2014. Antimicrobial and antibiofilm potential of
biosurfactants isolated from lactobacilli against multi-drug-resistant pathogens. BMC Microbiol. 14:197.
doi:10.1186/1471-2180-14-197.
Santos DKF, Rufino RD, Luna JM, Valdemir A, Santos VA, Sarubbo LA. 2016. Biosurfactants: multifunctional
biomolecules of the 21st Century. Intern J Mol Sc. 17(3):401. doi:10.3390/ijms17030401.
Sardi JC, Pitangui NS, Voltan AR, Braz JD, Machado MP, Fusco Almeida AM, Mendes Giannini MJ. 2015. vitro
Paracoccidioides brasiliensis biofilm and gene expression of adhesins and hydrolytic enzymes. Virulence. 6
(6):642–651. doi:10.1080/21505594.2015.1031437.
Satpute S, Mone N, Das P, Banpurkar A, Banat IM. 2018. Lactobacillus acidophilus derived biosurfactant as a biofilm
inhibitor: a promising investigation using microfluidic approach. Appl Sci. 8:1555. doi:10.3390/app8091555.
Satpute SK, Banpurkar AG, Banat IM, Sangshetti JN, Patil RH, Gade WN. 2016a. Multiple roles of biosurfactants in
biofilms. Curr Pharm Des. 22(11):1429–1448. doi:10.2174/1381612822666160120152704.
Satpute SK, Kulkarni GR, Banpurkar BAG, Banat IM, Mone NS, Patil RH, Cameotra SS. 2016b. Biosurfactant/s
from Lactobacilli species: properties, challenges and potential biomedical applications. J Basic Microbiol. 56
(11):1140–1156. doi:10.1002/jobm.201600143.
Schooling SR, Beveridge TJ. 2006. Membrane vesicles: an overlooked component of the matrices of biofilms.
J Bacteriol. 188(16):5945−5957. doi:10.1128/JB.00698-06.
Seckbach J, Oren A. 2010. Microbial mats: modern and ancient microorganisms in stratified systems. 2nd ed.
Heidelberg: Springer.
Sharma D, Saharan BS. 2016. Functional characterization of biomedical potential of biosurfactant produced by
Lactobacillus helveticus. Biotechnol Rep. 11(1):27–35. doi:10.1016/j.btre.2016.05.001.
Shokouhfard M, Kermanshahi RK, Shahandashti RV, Feizabadi MM, Teimourian S. 2015. The inhibitory effect of
a Lactobacillus acidophilus derived biosurfactant on biofilm producer Serratia marcescens. Iran J Basic Med Sc.
18(10):1001–1007.
20 K. PARASZKIEWICZ ET AL.

Silva SS, Carvalho JWP, Aires CP, Nitschke M. 2017. Disruption of Staphylococcus aureus biofilms using rhamno-
lipid biosurfactants. J Dairy Sc. 100(10):7864–7873. doi:10.3168/jds.2017-13012.
Soberon-Chavez G. 2011. Biosurfactants. From genes to applications. Berlin, Heidelberg: Springer-Verlag.
Song B, Wang Y, Wang G, Liu GL, Li W, Yan F. 2016. The lipopeptide 6-2 produced by Bacillus amyloliquefaciens
anti-CA has potent activity against the biofilm-forming organisms. Marine Poll Bull. 108(1–2):62–69.
doi:10.1016/j.marpolbul.2016.04.062.
Starkey M, Gray K, Chang S, Parsek M. 2004. A sticky business: the extracellular polymeric substance matrix of
bacterial biofilms. In: Ghannoum G, Toole G, editors. Microbial biofilms. Washington DC: ASM Press; p.
174–192.
Stoodley P, Sauer K, Davies DG, Costerton JW. 2002. Biofilms as complex differentiated communities. Ann Rev
Microbiol. 56:87–209. doi:10.1146/annurev.micro.56.012302.160705.
Sun W, Wang Y, Zhang W, Ying H, Wang P. 2018. Novel surfactant peptide for removal of biofilms. Coll Sur B:
Biointer. 172:180–186. doi:10.1016/j.colsurfb.2018.08.029.
Toyofuku M, Roschitzki B, Riedel K, Eberl L. 2012. Identification of proteins associated with the Pseudomonas
aeruginosa biofilm extracellular matrix. J Proteome Res. 11(10):4906–4915. doi:10.1021/pr300395j.
Van Loosdrecht MCM, Lyklema J, Norde W, Schroa G, Zehnder AJB. 1987. Electrophoretic mobility and
hydrophobicity as a measure to predict the initial steps of bacterial adhesion. App Environ Microbiol. 53
(8):1898–1901.
Veerachamy S, Yarlagadda T, Manivasagam G, Yarlagadda PK. 2014. Bacterial adherence and biofilm formation on
medical implants: a review. Proc Inst Mech Eng H. 228(10):1083–1099. doi:10.1177/0954411914556137.
Whitchurch CB, Tolker-Nielsen T, Ragas PC, Mattick JS. 2002. Extracellular DNA required for bacterial biofilm
formation. Science. 295(5559):1487. doi:10.1126/science.295.5559.1487.