Journal of King Saud University - Science 36 (2024) 103118

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Full Length Article

Exploring the potential: Inhibiting quorum sensing through marine red

seaweed extracts – A study on Amphiroa fragilissima
Prakash Piruthiviraj a, *, B.R. Maha Swetha a, Chitra Balasubramanian a,
Rajapandiyan Krishnamoorthy b, Mansour K. Gatasheh c, Anis Ahmad d,
Rengasamy Parthasarathi e, *, Poonguzhali Pandurangan f, V.K. Bhuvaneshwari g,
Natesan Vijayakumar g
a
Department of Biotechnology, Srimad Andavan Arts and Science College (Autonomous), affiliated to Bharathidasan University, Tiruchirappalli 620005, Tamil Nadu,
India
b
Department of Food Science and Nutrition, College of Food and Agriculture Sciences, King Saud University, Riyadh 11451, Saudi Arabia
c
Department of Biochemistry, College of Science, King Saud University, P.O.Box 2455, Riyadh, 11451, Saudi Arabia
d
Department of Radiation Oncology, Miller School of Medicine/Sylvester Cancer Center, University of Miami, Miami, FL, USA
e
Department of Microbiology, Faculty of Agriculture, Annamalai University, Annamalainagar 608002, Tamil Nadu, India; Department of Soil Science and Agricultural
Chemistry, Anbil Dharmalingam Agricultural College and Research Institute, TNAU, Tiruchirappalli 620027, Tamil Nadu, India
f
Department of Microbiology, M.R. Government Arts College, Mannargudi 614001, Tamil Nadu, India
g
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar 608002, Tamil Nadu, India

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

Keywords: Objectives: A great setback in concern with public health is the development of emerging bacterial resistance
Quorum sensing towards conventional antibiotics making it a huge issue in the medical sector. To combat bacterial resistance,
Anti-QS innovative strategies have been developed. One of the significant strategies is to target the bacterial pathoge­
Antibiofilm activity
nicity through inhibition of quorum sensing (QS) mechanism among bacteria. QS is cell density dependent
Amphiroa fragilissima
Staphylococcus aureus
molecular communication prevailing in bacteria trends to express several genes conferring for its virulence,
Klebsiella pneumoniae biofilm formation, antibiotic resistance, motility and plant-bacterial interactions etc. Methods: The current study
Acinetobacter sp. investigates the anti-QS activity of red seaweed, against the targeted test pathogens Staphylococcus aureus,
Klebsiella pneumoniae, Acinetobacter sp. and E. coli. Results and conclusion: At the concentration of 100 mg/mL of
red seaweed extract, the biofilm was effectively reduced in Acinetobacter sp. and E. coli with 36 % and 40 %
respectively. The exo polymeric substance (EPS) quantification from test pathogens in the presence of algal
extract has declined to 40.8 % in Acinetobacter sp. and 14.8 % in E. coli reduction compared to S. aureus and
K. pneumonia. Furthermore, the QS dependent motility activities are also reduced with the effect of algal extract.
The efflux pump expression, being the noteworthy factor for antibiotic resistance has been inhibited by algal
extract. The bioactive compounds from the algal extract A. fragilissima such as Melamine, Silicic acid, diethyl bis
(trimethylsilyl) ester, Trimethyl [4-(1-methyl-1-methoxyethyl) phenoxy] silane, Benzo[h]quinoline, 2,4-
dimethyl, Pyrazol-3(2H)-one, 4-nitro and 1,2-Bis(trimethylsilyl)benzene etc. Among them, Melamine showed
high peak area and may responsible for the anti-QS activity and helps to overcome antibiotic resistance of the
opportunistic pathogens. The QS dependent phenotypic expression among the test pathogens has been inter­
rupted by the bioactive components of A. fragilissima.

1. Introduction number of issues, especially emergence of antibiotic resistant bacterial

strains. The whole medical sector has been intense on resolving the great
The discovery of antibiotics at the turn of 20th century freed all set back owing to the emerging antibiotic resistant strains. Further,
people from a variety of life-threatening illnesses. However, after the antibiotic resistance has observed to be higher in bacterial biofilm than
mid – twentieth century, overuse of antibiotics have elevated a wide in their planktonic counterparts. Therefore, it is important to investigate

* Corresponding authors.
E-mail addresses: prakash@andavancollege.ac.in (P. Piruthiviraj), parthasangi@yahoo.com (R. Parthasarathi).

https://doi.org/10.1016/j.jksus.2024.103118
Received 30 August 2023; Received in revised form 25 October 2023; Accepted 30 January 2024
Available online 3 February 2024
1018-3647/© 2024 The Authors. Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

the biofilm formation bacterial populations that are associated to at least 2022; Borges and Simões, 2019).
65 % of all major infections. Quorum sensing (QS) is the communica­ In the present research study, the marine red algae, Amphiroa fragi­
tion/ signalling system prevailing among the bacterial population which lissima has employed to reveal the anti-QS activity among the test
pathogenecity. The expression of signalling molecules varies with gram pathogens. In addition, the expression of efflux pumps in bacteria has
positive and gram negative bacteria to extinguish their QS system that also been investigated as it has been a significant factor to confer anti­
regulates bacterial behaviour in biofilms along with other virulence biotic resistance and bacterial biofilm formation. Therefore, the current
production in a cell-dependent manner (Sarkar and Das, 2019). The QS study has undertaken in the aim to explore the anti-QS molecules in
signal concentration rises along with the population, binds to the it’s marine red algae, Amphiroa fragiliss inhibiting the virulence and QS
appropriate receptor and ensure the QS response that takes its upper dependent biofilm formation.
hand upon motility, biofilm formation, EPS (exopolysaccharide) pro­
duction (physical barrier of bio film), plant-bacterial interaction and 2. Material and methods
sporulation etc., (Tran and Hadinoto, 2021). Acyl homoserine lactone
(AHL) is one among the QS signal molecule that has exhibited by the 2.1. Collection and authentication of marine red seaweed
majority of Gram-negative bacteria. In response to population density,
bacteria can express particular genes by using these signalling mole­ The marine red seaweed, Amphiroa fragilissima (100 g) obtained from
cules. The investigation of Kumar et al. (2016) has revealed 2-hydroxy- R.K. Algae project centre, Mandapam, Ramanathapuram, Tamil Nadu,
4-((methylamino)(phenyl)methyl) cyclopentanone as the QS signal India (shown in Fig. 1) and authenticated in Botanical Survey of India,
molecules in Ralstonia solanacearum enhancing the biofilm formation Coimbatore, India.
that has been confirmed by the Confocal laser scanning microscopic
analysis. 2.2. Extraction of red seaweed
The test microorganism employed in the present study include
Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter sp. and The obtained marine seaweed was shade dried and powdered using a
Escherichia coli are antibiotic resistant species and are involved in sterile mechanical blender aseptically. To 100 mL of sterile distilled
nosocomial infections. Indeed, microbes causing nosocomial infections water, 20 g of algal powder was added and kept at room temperature
with antibiotic resistant characteristic makes the treatment extremely with 140 rpm shaking for two days. The mixture was then freeze-dried in
challenging. In fact, the QS mechanism facilitates various gene expres­ a Lyophilizer after being filtered via Whatman no.1 filter paper (Kul­
sions among the population in a niche resulting in the antibiotic resis­ shreshtha et al., 2016).
tance, biofilm formation, sporulation etc. The QS machinery in S. aureus
has been denoted by the accessory gene regulator (agr) system consti­ 2.3. Bacterial strains and culture conditions
tuting of agrACDB cassette (Kannapan et al. 2023). The agr system in
S. aureus has found to be active in the presence of a certain threshold The test pathogens employed in the present study were Staphylo­
extracellular AIP (autoinducing peptide) concentration. Further, the AIP coccus aureus strain ATCC 1690, Klebsiella pneumoniae strain ATCC
molecules bind with AgrC of AgrC-AgrA complex to phosphorylate 13883, Acinetobacter sp. strain ATCC 49139, and Escherichia coli strain
AgrA, which in turn induces the promoters (P2 and P3) of agrACDB ATCC 15224 respectively. All bacterial culture work was done from
cassette. Therefore, the expression of the structural genes such as AgrB, Bishop Heber College in Trichy, Tamil Nadu, India.
AgrD, AgrC, AgrA have accomplished along with the aid of RNAII and
RNAIII to promote P2 and P3 respectively (Arunachalam et al., 2023; 2.4. Antibacterial activity
Vasquez et al., 2017). LuxR/LuxI homologs that are encoded in several
gram-negative bacteria exhibit to regulate a variety of biological activ­ To evaluate the antibacterial efficacy of the algal extract among the
ities (Srinivasan et al., 2021). But only the LuxR homolog SdiA is test pathogens such as S. aureus, K. pneumoniae, Acinetobacter sp. and
encoded in most of the gram negative bacteria including Klebsiella E. coli, the overnight test cultures were swabbed on Muller–Hinton agar
pneumoniae, Escherichia coli, and Salmonella (Ahmed et al., 2021). (MHA) following Mc.Farland standard. The sterile disc was then placed
However, the earlier study of Sun et al. (2021) has insisted on abaI/abaR over the medium and to which 20 mg of algal extract was added indi­
QS system in Acinetobacter sp. such as A. baumannii and A. nosocomialis vidually on each plate aspectically. The positive control antibiotics
regulating its virulence, biofilm formation, antibiotic resistance, energy ampicillin was loaded separately. The plates were then incubated for 24
metabolism, and lipid metabolism. AbaI is the synthase of AHL signal h at 30 ◦ C (Packiavathy et al., 2012) respectively.
while AbaR encodes for the receptor to bind with the auto-inducers
during the activation of abaI/abaR based QS system (Choe et al., 2.5. Anti-biofilm activity
2022). On the other hand, finding innovative approache to treat in­
fections and targeting virulence through QS mechanism has become The anti-biofilm activity among of algal extracts the test pathogens
crucial to resolving the issues (Seleem et al., 2020). However, the in­ were investigated using the 96 well containing micro-titre plates at
hibition of QS mechanism has accomplished by receptor protein various concentrations (20–100 mg/mL). In the sterile, fresh nutrient
degradation, AIs synthase inhibition and signal degradation (Paluch broth medium containing algal extract, 2 % of test organisms are inoc­
et al., 2020). ulated. The Uninnoculated culture broth without algal extract was kept
QS inhibitors have been categorized under natural and synthetic as the control. After 24 h incubation at 37 ◦ C, the media were taken out
inhibitors. Certain natural inhibitors include the secondary metabolites from the micro-titre plates and rinsed with phosphate-buffered saline
of marine environmental organisms especially microalgae /seaweed, (PBS) to eliminate planktonic cells. The wells were then treated with
plant leaves, bark, fruits, fungal and bacterial enzymes. Furthermore, in 100 μl of 0.1 % crystal violet for 10 min. The wells were then rinsed with
recent years ample of bioactive substances from marine algae has been sterile distilled water to remove the excess stain. In order to solubilize
identified to be a good solution in the hunt for alternative medication to the biofilm, 100 μl of 95 % ethanol were added to each well. The biofilm
manage the pathogens with QS and multidrug resistance. Interestingly, was then quantified using a microplate reader at a wavelength of 570 nm
identification of distinct seaweeds excerting certain biological chemicals (Al-kafaween et al., 2019). The percentage of biofilm inhibition was
with anti-QS property has become the pioneer among the researchers calculated by the formula,
and therefore, bioactive component from macroalgae have been the
Biofilm inhibition (%) = [(Control OD − Test OD)/Control OD ] × 100
current trend by numerous researchers to explore anti-QS molecules
(Boominathan et al., 2022; Muthukrishnan et al., 2023; Rima et al.,

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P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

Fig. 1. (a) Percentage of Biofilm inhibition (b) Percentage of EPS inhibition by A. Fragilissima.

2.6. Cover glass based microscopic visualization of biofilm extract (100 mg/mL). It was incubated at 37 ◦ C for 18 h. Following in­
cubation, the cover glass was treated with 0.9 % NaCl (0.5 mL), 5 %
In the 3 mL of fresh nutrient broth medium in test tubes nutrient phenol (0.5 mL) along with 5 volumes of concentrated H2SO4 and left in
broth medium with cover glass (12X24 mm), algal extract (50 mg/mL) the dark for an hour. Then the absorbance was recorded using UV–VIS
was added. To which, 1 % of overnight cultures of the test pathogens spectrophotometer at 490 nm (Favre-Bonté et al., 2003). The percentage
were inoculated individually. The mixture was then incubated for 24 h. of EPS inhibition was calculated using the formula,
After incubation, planktonic cells were removed from the cover glasses
EPS inhibition (%) = [(Control OD − Test OD)/Control OD ] × 100
by washing it with sterile distilled water and finally treated the cover
glasses with 0.2 % CV. The stained cover glasses were then air dried and
visualized the biofilms under light microscope (Packiavathy et al., 2.9. Swimming assay
2012).
Solid agar medium containing 25 mg of concentrated algal extracts
2.7. Biofilm visualization using tube method along with 1 % tryptone and 0.5 % NaCl was prepared. The plate devoid
of algal extract was kept as the control. The test pathogens were then
The test pathogens were inoculated into the test tube containing 25 point-inoculated at the centre of medium. The plates were incubated for
mg of algal extract and 5 mL of nutrient broth. The tubes were then 18 h at 37 ◦ C in an upright position and after incubation the result was
incubated after the addition of 1 % glucose. The control tubes were recorded (Musthafa et al., 2012).
maintained by inoculating overnight cultures without extract. The tubes
were stained by adding 0.1 % crystal violet after being incubated at 2.10. Ethidium bromide–agar cartwheel assay
37 ◦ C for 24 h. To get rid of excess stains the tubes were rinsed with
sterile distilled water, air dried and then manually visualized for bio­ The effect of algal extract upon the efflux pump of the test pathogens
films (Freeman et al., 1989). were assessed by employing Ethidium bromide (EtBr) agar cartwheel
assay. The presence of efflux pump was detected through the accumu­
2.8. EPS quantification lation of EtBr inside the cells resulting in the elevation of fluorescence.
For which, 2 mg/mL of EtBr was added to MH agar plates along with 25
The test pathogens were allowed to develop biofilms on a cover glass mg of algal extract. The test was performed both in the presence and
(size?) in a 6-well microtiter plate in the presence and absence of algal absence of algal extract. Further, the test bacterial cultures were

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P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

swabbed onto agar plates like a cartwheel. The plates were evaluated the control.
with a UV transilluminator after 16 h of incubation at 37 ◦ C to detect
EtBr concentration within the test pathogens as an indication of efflux 3.7. Light microscopic analysis of biofilm
pump (Eleftheriadou et al., 2021).
Though the biofilm formation is reduced in all test pathogens by the
2.11. Identification of anti-QS compounds by GC − MS and FTIR effect of algal extract, Acinetobacter sp. biofilm formation has been
analysis greatly declined (depicted in Fig. 2) comparatively and it has been
visualized with the aid of light microscope.
GC–MS (gas chromatography-mass spectrometry) analysis for
aqueous extract of A. fragilissima was performed at TUV SUD South Asia 3.8. Swimming assay
Pvt. Ltd, Tirupur, Tamil Nadu to identify the bio- active compounds
present in the algal extracts. Along with GC–MS, Fourier transform In the presence of algal extract at a concentration of 25 mg/mL, the
infrared spectroscopy (FTIR) was also used to identify the structure and swimming capacity of the test pathogens has been reduced promisingly
functional groups of those compounds at St. Joseph College, Trichy. except K. pneumonia. As because, the control plate has exhibited the non-
motile nature of the strain K. pneumonia (Fig. 3.a and b.). Other strains
3. Results such as Acinetobacter sp., S. aureus, and E. coli treated with the algal
extracts has exhibited the greatest suppression in QS dependent swim­
3.1. Authentication of marine red seaweed ming migration (Fig. 3.c and d.).

The red seaweed has been identified as Amphiroa fragilissima (L.) J.V. 3.9. Ethidium bromide-agar cartwheel assay
Lamour-LITHOPHYLLACEAE (BSI/SRC/5/23/2022/Tech/468) by Dr.
S.S. Hameed, The Scientist- E & Head of Office, Botanical Survey of The efflux pump inhibitory activity of seaweed extracts (25 mg/mL)
India, TNAU Campus, Coimbatore- 641003. against the test pathogens has been identified from the cart wheel test in
MHA agar plates containing 2 mg/mL of EtBr. Interestingly, the
3.2. Extraction yield A. fragilissima extract has disrupted the active efflux pumps in E. coli, K.
pneumoniae, and S. aureus.
The Amphiroa fragilissima has yielded of 8 % of aqueous extract
which has been quantified in terms of percentage by using the following 3.10. Identification of bioactive compounds
formula adapting Bhuyar et al. (2020).
The aqueous extract of A. fragilissima has been investigated
Dry weight of crude extract
× 100 employing FTIR (Fig. 4) and GC–MS analysis (Fig. 5). The FTIR has
Intial weight of the sample shown the presence of amides, alkanes, phosphines, alkyls, carboxylic
compounds, ether, alcohols, aromatic compounds and alkyl halides
3.3. Anti-bacterial activity functional groups. The presence of functional groups is of great impor­
tance in the prepared compounds. One such functional group is the
While examining the algal extract for its antibacterial activity against carboxyl group, which is indicated by a peak at 3778 and 3424 cm-1.
test pathogens, none of the four test strains of S. aureus, K. pneumoniae, FTIR analysis confirmed the presence of amine and amide as functional
Acinetobacter sp., and E. coli showed any activity. Since then, microbes group the peak at 2918 and 2854 cm− 1. The peak at 1610, 1549, and
including S. aureus and K. pneumoniae have become resistant to the 1425 cm− 1 indicated the presence of amino acid, nitro compound, and
antibiotics like ampicillin. At a concentration of 20 mg/mL of algal aromatic functional group, respectively. The peak at 1385, 1319, and
extract, the zone of inhibition for ampicillin and algal extract has shown 1222 cm− 1 confirms the presence of alkane, aldehyde, fluoride, nitro,
as 10 mm in S. aureus, 16 mm in Acinetobacter sp., and 14 mm in E. coli and alkyl halide group, respectively. The remaining peak at 1093, 861,
respectively. and 601 cm− 1 indicated the presence of alcohol, aliphatic amines, aro­
matic compound, primary or secondary amines, and halogen group,
3.4. Anti-biofilm activity respectively. The GC–MS interpretation was tabulated in Table 1.

The marine seaweed A. fragilissima has exhibited a well profound 4. Discussion

anti-biofilm activity at distinct concentration ranging from 20 to 100
mg/mL. The biofilm formation among the test pathogens have been The QSI potential of the marine red seaweed, Amphiroa fragilissima
much reduced when treated with A. fragilissima extract. The percentage has publicized in the present investigation for the first time against the
of biofilm inhibition by the algal extract has been visualized in Fig. 1.a. QS dependent phenotypic expressions in the test pathogens such as
E. coli, Acinetobacter sp., K. pneumoniae, and S. aureus. Though initially,
3.5. Quantification of EPS the seaweed extract has employed for detecting its antibacterial activity,
based on the result with no appreciable effect over the growth of the test
The EPS production of the test pathogens have declined to an pathogens, the alternative strategy has been pursued in the present
appreciable level owing to the effect of aqueous extract of A. fragilissima study. In fact, even before checking out the tactics of combined action of
comparing with the control. The algal extract at 100 mg/mL has reduced algal extract with the antibiotic ampicillin, precise action of ampicillin
the EPS synthesis upto 40.8 %, 14.8 %, 15 %, and 13 % in Acinetobacter towards the individual test pathogens has been evaluated. Though the
sp., E. coli, K. pneumoniae, and S. aureus, respectively (Fig. 1.b). test pathogens such as K. pneumoniae, and S. aureus has shown resistance
towards ampicillin, while combining with algal extract with the anti­
3.6. Biofilm visualization using tube method biotic ampicillin has interestingly exhibited a tremendous activity of
anti-biofilm and anti-QS effect. It may be due to the reason that certain
Biofilm formation has been visualized from the naked CV stained bioactive compounds or phytochemicals from various sources exerts its
thick film lining appearance at the test tube wall. The algal extract has anti-QS activity against the microbial population in a niche by affecting
remarkable effect to express the anti-biofilm activity by inhibit the its cellular communication system (QS system) rather directly killing the
biofilm formation among all the test pathogens while comparing with pathogens. This view has been in acceptance with the earlier reports of

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Fig. 2. Biofilm visualization under light microscope – (a, c, e, g) control of Acinetobacter sp., E. coli, K. pneumoniae and S. aureus respectively; (b, d, f, h) treated with
A. fragilissima extract.

Truchado et al., (2009); Vattem et al., (2007) who has insisted that those virulence. The antibiofilm action of the marine red algae Amphiroa
bioactive components does not involve in the evolution of resistant fragilissima extract has been well established in the current investigation
strains and has suggested that those bioactive components can express through the reduction of biofilm formation among the test pathogens
its anti-QS effect without even exhibiting its lethal effect upon patho­ E. coli, Acinetobacter sp. Indeed, Sagar et al. (2022) has revealed different
gens respectively. In a similar way, the finding of Packiavathy et al. quorum sensing regulated virulence parameters, such as swarming
(2012) has revealed that Cuminum cyminum did not exhibit anti- motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilms of
bacterial activity but has suppressed QS mechanism. P. aeruginosa, were reduced by Eucalyptus globulus methanol extract.
However, there has been the evidence (Tang et al., 2020) of certain Further, the motility is a significant parameter among the bacterial
components like phlorotannins from the marine algae like Hizikia fusi­ species that produces biofilm (Tang et al., 2020). The motility of E. coli,
forme unveiling its efficacy to inhibit QS system to combat pathogenic Acinetobacter sp. and S. aureus have been much reduced by Amphiroa
bacterial species. In which, the antibacterial action of phlorotannins fragilissima extract. While, in the previous study of Tran and Hadinoto
against specific gram-positive and gram-negative bacteria has been (2021), a plant-derived flavonoid Quercetin has complexed with chito­
demonstrated by insisting that its mode of action would not be specific san nanoparticles and has reduced the swimming activity of
like that of antibiotics. Interestingly, the seaweed compounds utilised in P. aeruginosa. The study also highlights the EPS production in the
this investigation interferes with cell–cell communication and is inef­ presence and absence of red algae Amphiroa fragilissima extract that has
fective against bacterial growth when examined individually. Therefore, effectively reduced the EPS in E. coli, Acinetobacter sp. when comparing
this present study has concentrated more on anti-QS action of the with K. pneumoniae, and S. aureus. In yet another study of Karuppiah
seaweed Amphiroa fragilissima. Moreover, the QS mechanism highly in­ et al. (2021), a similar report has been submitted insisting on the
fluences the biofilm formation, EPS production and efflux pumps in seaweed Musa paradisiaca having a significant effect on the EPS activity
bacteria. On this note, the present study has undertaken afore said QS with the influence of the compound 1,8-cineole. In fact, the biofilm,
related phenotypic expressions, which has also been conferring for its motility and EPS are the pioneer among the virulent factors regulated by

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P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

Fig. 3. Swimming assay (a) control (b) treated with A. fragilissima; Efflux pump inhibition assay (c) control (d) Treated with A. fragilissima.

Fig. 4. FTIR analysis - Aqueous extract of A. fragilissima.

the QS machinery, which may add up to its higher level of pathogenicity ability to reduce QS in the dental biofilm-forming bacteria Alcaligenes
towards the offering bacterial pathogens. Certain study of Abdul Malik faecalis and Pseudomonas gingivalis (Lahiri et al., 2021).
et al. (2020) has insisted on the anti-QS activity by studying the surface The efflux pump has a direct effect on the antimicrobial resistance of
attached bacteria from Mexican red algae, Halymeniafloresii. Further­ pathogens and has been identified to be QS mediated mechanism.
more, in comparison to certain components like quercetin, nimbolide, Therefore, efflux pump suppression lowers the level of antibiotic resis­
nimbin, and azardirachtin, phytocompounds like catechin from Aza­ tance (Rasamiravaka and El Jaziri, 2016). The current study on efflux
dirachta indica has demonstrated the maximum biofilm eradication pump inhibition has employed EtBr as an indicator to fluoresce when
along with the degradation of EPS structural components like carbo­ accumulating EtBr inside the bacterial cell as a result of efflux pump and
hydrates and proteins. Catechin is a significant compound which has the has found that red seaweed extract has efficiently suppressed efflux

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P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

Fig. 5. GC MS analysis of A. Fragilissima.

pump in S. aureus, K. pneumoniae, and E. coli. Though the QS machinery for LasR and LuxR can be rendered inactive (Shukla et al., 2020). Indeed,
varies with the regulation of distinct gene expression in gram positive the researchers are concentrating on the QS inhibitors to manage the
and negative bacteria, as a consequence has a major impact in the antimicrobial resistant pathogens. The QS inhibitors extracted from
virulence and pathogenicity. The expression of the QS genes such as lasI, Plumula nelumbini iseffective against P. aeruginosaand exhibits biofilm
lasR, rhlI, rhlR, pqsA and pqsR have decreased and confirmed by qRT- inhibition activity of 44.63 % at 100 mM without impeding bacterial
PCR (Abbas et al., 2020). The organic AHL antagonist from Curcumin growth (Chen et al., 2022). P. aeruginosa infections can be treated with
plant has been demonstrated in an In-silico investigation. Due to specific sitagliptin, a new anti-quorum sensing drug. In the present study,
hydrogen bonding and hydrophobic interactions, the P. aeruginosa genes through the identification FTIR and GCMS analysis, A. fragilissima

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P. Piruthiviraj et al. Journal of King Saud University - Science 36 (2024) 103118

Table 1 Software. B.R. MahaSwetha: Data curation, Formal analysis, Method­

GC MS compounds of A. fragilissima. ology. Chitra Balasubramanian: Visualization. Rajapandiyan Krish­
Peak Retention Peak Peak Compounds namoorthy: Funding acquisition, Writing – review & editing. Mansour
no time area % K.Gatasheh: Funding acquisition. Anis Ahmad: Funding acquisition.
1 4.276 8876 5.47 Benzo[h]quinoline, 2,4-dimethyl Rengasamy Parthasarathi: Investigation, Validation. Poonguzhali
2 4.876 4021 2.48 6,7-Dimethoxy-3-(4- Pandurangan: Investigation, Validation. V.K. Bhuvaneshwari: Re­
trifluoromethoxy-phenylamino)-3H- sources. Natesan Vijayakumar: Writing – review & editing.
isobenzofuran-1
3 5.275 3840 2.36 1,2-Benzenediol, 3,5-bis(1,1-
dimethylethyl)
4 12.230 11,475 7.07 Pentadecanoic acid, 14-methyl-, Declaration of competing interest
methyl ester
5 12.552 66,997 41.26 1,3,5-Triazine-2,4,6-triamine The authors declare that they have no known competing financial
6 16.119 7115 4.38 Pyrazol-3(2H)-one, 4-nitro-
7 16.407 3810 2.35 4-Quinolinecarboxylic acid, 2-chloro
interests or personal relationships that could have appeared to influence
8 17.729 3611 2.22 1,2-Bis(trimethylsilyl)benzene the work reported in this paper.
9 18.974 5192 3.20 Silicic acid, diethyl bis
(trimethylsilyl) ester
10 19.629 6300 3.88 Trimethyl[4-(1-methyl-1-
Acknowledgment
methoxyethyl)phenoxy]silane
11 20.429 3637 2.24 Silane, trimethyl[5-methyl-2-(1- The work was supported by the Researchers Supporting Project
methylethyl)phenoxy]- number (RSP2024R393), King Saud University, Riyadh, Saudi Arabia
12 20.940 5382 3.31 1,2-Bis(trimethylsilyl)benzene
13 21.529 15,730 9.69 Silicic acid, diethyl bis
(trimethylsilyl) ester References
14 21.896 12,014 7.40 Trimethyl[4-(1-methyl-1-
methoxyethyl)phenoxy]silane Abbas, H.A., Shaldam, M.A., Eldamasi, D., 2020. Curtailing quorum sensing in
15 23.662 4387 2.70 Silane, trimethyl[5-methyl-2-(1- Pseudomonas aeruginosa by sitagliptin. Curr. Microbiol. 77 (6), 1051–1060.
methylethyl)phenoxy]- Abdul Malik, S.A., Bazire, A., Gamboa-Muñoz, A., Bedoux, G., Robledo, D., García-
Maldonado, J.Q., Bourgougnon, N., 2020. Screening of surface-associated bacteria
from the Mexican red alga Halymeniafloresii for quorum sensing activity.
Microbiology 89 (6), 778–788.
extract has found to constitute the major bioactive compounds such as
Ahmed, M.Z., Muteeb, G., Khan, S., Alqahtani, A.S., Somvanshi, P., Alqahtani, M.S.,
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