975 D3000 new imprint Word template


 

Vol 13, No 1 (2025) 
ISSN 2167-8677 (online) 

DOI 10.5195/d3000.2025.975  

 

	

	

http://dentistry3000.pitt.edu	

Impact of Fixed Orthodontic Appliances on Staphylococcus aureus and 
Candida albicans 
 
Maryam Aljbouri, Nebras Al-Dabbagh, Thaer Jaber Al-Khafaji 
 
College of Den*stry, University of Babylon, Babylon, Iraq 

Abstract 

Objec<ve: This study invesEgated the prevalence of salivary S. aureus and C. albicans in pa-
Eents with fixed orthodonEc appliances and assessed their resistance profiles to selected 
anEbacterial and anEfungal agents over three-Eme intervals. Material and Methods: A pro-
specEve cohort of 40 paEents (20 males, 20 females) undergoing orthodonEc treatment was 
followed across three Eme points: baseline (T0), two weeks post-applicaEon (T1), and four 
weeks post-applicaEon (T2). UnsEmulated saliva samples were collected and cultured for mi-
crobial idenEficaEon. Organisms were confirmed via Vitek biochemical tesEng. AnEmicrobial 
suscepEbility was determined using the disc diffusion method against six anEbacterial and 
six anEfungal agents. Data were analyzed using two way-ANOVA and chi-square tests. Re-
sults: Among the 80 total samples analyzed, 73.75% were posiEve for S. aureus and/or C. 
albicans. S. aureus was most prevalent, with Clindamycin and Vancomycin showing the 
strongest anEbacterial acEvity. Resistance to Amoxicillin, Ampicillin, and Erythromycin was 
notably high. For C. albicans, Fluconazole and Amphotericin B demonstrated the highest ef-
ficacy, while Clotrimazole and NystaEn showed poor inhibiEon. Resistance pa^erns might 
suggest a biofilm-associated microbial adaptaEon and reduced suscepEbility over Eme. Con-
clusion: Fixed orthodonEc appliances significantly influence oral microbial ecology by 

facilitaEng colonizaEon of resistant 
strains of S. aureus and C. albicans. The 
findings highlight the importance of rou-
Ene microbial surveillance and personal-
ized anEmicrobial strategies in orthodon-
Ec care to miEgate infecEon risks and 
manage emerging resistance.  

Open Access 
Cita%on: Aljbouri M, et al. (2025) Impact of Fixed 
Ortohon%c Appliances on Staphylococcus aureus and 
Candida albicans. Den%stry 3000. 1:a001 
doi:10.5195/d3000.2025.975 
Received: June 30, 2025  
Accepted: July 7, 2025 
Published: August 21, 2025 
Copyright: ©2025 Aljbouri M, et al. This is an open ac-
cess ar%cle licensed under a Crea%ve Commons ATribu-
%on Work 4.0 United States License. 
Email: mariamaljbori1999@gmail.com

	

Introduc)on 
Orthodontic	appliances	significantly	modify	
the	 physiological	 environment	 of	 the	 oral	
cavity	 by	 hindering	 natural	 cleaning	
mechanisms,	 leading	 to	 increased	 plaque	
accumulation	 and	 impaired	 oral	 hygiene.	
These	 devices	 create	 retention	 sites	 that	
facilitate	 biofilm	 formation	 composed	 of	
diverse	 bacteria	 and	 fungi,	 which	 colonize	
various	 oral	 surfaces.	 Consequently,	
orthodontic	biofilms	contribute	to	microbial	
overgrowth	and	increase	the	risk	of	infection	
if	hygiene	is	not	properly	maintained	[1].	
Fixed	 orthodontic	 components	 such	 as	
bands,	 wires,	 and	 brackets	 complicate	 oral	
hygiene	 practices.	 Their	 complex	 structure	

and	 the	 extended	 surface	 area	 they	 cover	
hinder	 effective	 cleaning,	 encouraging	
plaque	 accumulation.	 Inadequate	 oral	
hygiene	 in	orthodontic	patients	 can	 lead	 to	
gingival	 inflammation	 and	 progressive	
periodontal	disease.	Therefore,	maintaining	
high	 standards	 of	 cleanliness	 during	
orthodontic	 treatment	 is	 crucial	 to	
preventing	 long-term	 oral	 health	
complications	[2].	
The	 oral	 cavity	 hosts	 the	 second	 most	
diverse	microbial	community	 in	 the	human	
body,	 following	 the	 gut.	 It	 comprises	
numerous	 bacteria,	 fungi,	 and	 viruses	 that	
coexist	 in	 a	 balanced	 microbiome	 under	
normal	 conditions.	 This	 ecosystem	 plays	 a	

vital	role	 in	oral	health	but	 is	vulnerable	to	
disruption	through	external	influences	such	
as	orthodontic	treatment	[3].	
Orthodontic	therapy	has	been	shown	to	alter	
the	oral	microbiota	composition,	 increasing	
the	 prevalence	 of	 streptococci,	
staphylococci,	 lactobacilli,	 and	 yeasts.	 The	
physical	presence	of	orthodontic	appliances	
fosters	 bacterial	 plaque	 accumulation,	
exacerbates	 gingival	 inflammation,	 and	
deepens	 probing	 depth.	 These	 conditions	
create	an	ideal	niche	for	pathogenic	bacteria,	
potentially	escalating	to	periodontitis	 if	 left	
unchecked	[4,5].	
Staphylococcus	aureus	is	commonly	found	on	
the	skin	and	mucosal	surfaces,	including	the	



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
2	

oropharynx.	 Though	 typically	 harmless	 in	
carriers,	it	can	become	pathogenic	and	cause	
a	wide	range	of	 infections,	 from	minor	skin	
conditions	 to	 severe	 systemic	 diseases	 like	
osteomyelitis	and	endocarditis.	Around	30%	
of	individuals	are	asymptomatic	carriers,	but	
the	 spread	 of	 antibiotic-resistant	 strains	
such	 as	 methicillin-resistant	 S.	 aureus	
(MRSA)	poses	a	significant	clinical	challenge	
[6-8].	
Antibiotic	 resistance	 in	 S.	 aureus,	
particularly	 MRSA,	 has	 made	 treatment	
increasingly	 difficult.	 These	 strains	 have	
developed	 resistance	 to	 several	 frontline	
antibiotics,	 including	 penicillin,	 methicillin,	
and	 in	 some	 cases,	 daptomycin	 and	
vancomycin.	 This	 resistance	 complicates	
therapeutic	 options	 and	 demands	 the	
development	 of	 novel	 approaches	 to	
bacterial	infection	control	[9,10].	
As	 bacterial	 resistance	 accelerates	 faster	
than	 antibiotic	 development,	 the	 potential	
for	 a	 post-antibiotic	 era	 looms.	
Understanding	 bacterial	 resistance	
mechanisms	 such	 as	 changes	 in	 cell	 wall	
composition	 and	 virulence	 is	 vital	 for	
designing	 new	 treatments.	 The	
peptidoglycan-rich	cell	wall	of	S.	aureus	is	a	
critical	 target	due	to	 its	role	 in	maintaining	
bacterial	integrity	and	pathogenicity	[11].	
Candida	 species,	 particularly	 C.	 albicans,	
naturally	 colonize	 the	 skin,	 mucosa,	 and	
gastrointestinal	tract	without	causing	harm.	
However,	 under	 certain	 conditions	 like	
immunosuppression	 or	 poor	 oral	 hygiene,	
they	 can	 overgrow	and	 lead	 to	 candidiasis.	
Symptoms	vary	depending	on	 the	 infection	
site	[12-14].	Candida	albicans	predominates	
in	the	oral	cavity,	with	colonization	reported	
in	up	to	60%	of	young	adults.	However,	non-
albicans	 species	 such	 as	 C.	 glabrata	 and	 C.	
krusei	 are	 becoming	 more	 common.	 These	
species	 often	 display	 resistance	 to	
commonly	 used	 antifungals	 like	 azoles	 and	
polyenes,	 emphasizing	 the	 importance	 of	
targeted	antifungal	therapy	[15-17].	
This	 study	aimed	 to	evaluate	 the	 impact	of	
Xixed	orthodontic	appliances	on	the	salivary	
prevalence	 of	 Staphylococcus	 aureus	 and	
Candida	albicans,	as	well	as	their	resistance	
to	 various	 antimicrobial	 agents.	 The	 objec-
tives	included	isolating	and	identifying	both	
microorganisms	using	culture	media,	quanti-
fying	their	presence	at	three	different	treat-
ment	 intervals,	 and	 assessing	 their	 re-
sistance	 patterns	 to	 antibacterial	 and	 anti-
fungal	drugs	over	time.	

Subjects and Methods 
A	prospective	cohort	study	was	followed	in	
which	 40	 individuals	 (20:	 females,	 20:	
males)	 have	 undergone	 orthodontic	 treat-
ment	in	orthodontic	department,	Nasser	Al-
mousawi	specialized	center,	Najaf,	Iraq.	

Eligible	study	participants:	 included	 indi-
viduals	aged	18	to	35	years	with	Class	I	mal-
occlusion	 and	 mild	 to	 moderate	 dental	
crowding,	characterized	by	an	ANB	angle	be-
tween	 0º	 and	 4º.	 Subjects	 also	 exhibited	 a	
normal	vertical	relationship	(SN-MP	angle	of	
32º	±	2),	a	straight	facial	proXile,	and	a	full	set	
of	teeth,	with	or	without	third	molars.		
Excluding	 criteria:	 Participants	 were	
excluded	if	they	had	a	history	of	orthodontic	
treatment,	 orthognathic	 surgery,	 systemic	
diseases,	 craniofacial	 anomalies,	 or	
traumatic	 injuries.	 Prior	 to	 treatment,	 all	
necessary	patient	information	was	recorded.	
Saliva	samples	were	collected	at	three	time	
points:	 before	 the	 application	 of	 fixed	
orthodontic	 appliances	 (T0),	 two	 weeks	
after	 (T1),	 and	 four	 weeks	 after	 treatment	
began	 (T2).	 Each	 participant	 underwent	
comprehensive	 intraoral	 and	 extraoral	
examinations,	 including	 panoramic	 and	
lateral	 cephalometric	 radiographs,	 to	
establish	 a	 diagnosis	 and	 determine	 the	
appropriate	treatment	plan.	
Sample	 Collection:	 Unstimulated	 saliva	
samples	were	collected	from	participants	be-
tween	9	and	11	a.m.	using	the	passive	drool-
ing	 method.[18]	 Subjects	 abstained	 from	
food,	drink,	and	oral	hygiene	for	at	least	one	
hour	 prior.	 After	 rinsing	 with	 150	 ml	 dis-
tilled	 water	 and	 resting,	 they	 allowed	 ap-
proximately	 3	 ml	 of	 saliva	 to	 collect	 in	 la-
beled	containers.	Samples	were	immediately	
stored	on	ice	and	transported	to	the	microbi-
ology	laboratory	in	Najaf	Governorate	[19].		
Ethical	approval:	This	study	was	conducted	
in	accordance	with	 the	ethical	principles	of	
the	Declaration	of	Helsinki	and	the	scientiXic	
committee	 in	 College	 of	 Dentistry/	 Univer-
sity	of	Babylon	was	approved	under	Protocol	
Number	123,	dated	03/10/2024.	
Isolation	 and	 Identi<ication	of	 S.	 aureus:	
The	specimens	were	transferred	directly	to	
the	 laboratory.	At	37°C	 for	24	hours,	blood	
agar	media	was	used	then	Mannitol	salt	agar	
(MSA)	 for	 the	 cultivation	 of	 S.	 aureus.	 The	
grown	bacterial	 colonies	were	 subjected	 to	
various	biochemical	tests	by	API	system	(API	
20	-Vitek	apparatus)	to	ensure	their	identity	
[20].	
Antibiotic	 Sensitivity	Test:	The	 antibiotic	
sensitivity	test	was	done	following	the	disk	
diffusion	method.[21]	Using	a	sterile	loop,	4-
5	colonies	were	suspended	in	2	mL	of	sterile	
normal	 saline.	 The	 bacterial	 suspension's	
turbidity	was	adjusted	to	a	McFarland	stand-
ard	of	0.5.	The	bacteria	were	then	inoculated	
into	the	mueller-hinton	agar	plate	by	using	a	
sterile	 swab.	 The	 plates	 were	 left	 at	 room	
temperature	 for	 about	 5	 minutes.	 The	 fol-
lowing	antibiotics	were	used:	 	 Clindamycin	
(2μg),	 Erythromycin	 (15µg),	 Azithromycin	
(15µg),	 Ampicillin	 (10µg),	 Amoxicillin	
(10µg)	and	Vancomycin	(30μg).	Using	sterile	

forceps,	the	antibiotic	discs	were	placed	and	
Xixed	on	the	surface	of	the	cultivated	agar.	Fi-
nally,	the	plates	were	inverted	and	incubated	
at	37°C	for	18	h.	
Preparation	of	Specialized	Culture	Media	
for	Candida	albicans	
Sabouraud's	 Dextrose	 Agar	 Medium	
(SDA)		
This	medium	(65	g)	was	dissolved	in	(1000	
mL)	distilled	water	and	heated	to	dissolve	in	
a	glass	beaker,	after	which	the	medium	was	
sterilized	in	an	autoclave,	distributed	to	Petri	
dishes,	 and	 	 (0.05	 g)	 anti-chloramphenicol	
was	 added	 after	 cooling	 the	 medium	 to	
(45°C)	to	prevent	bacterial	growth	[22].		
Chromo	Agar	Candida	Medium	
The	medium	was	prepared	according	to	the	
manufacturer's	 instructions	 by	 suspending	
42.72	g	of	powder	in	1000	mL	of	distilled	wa-
ter,	 and	 the	 solution	was	 heated	 to	 boiling	
point	to	completely	dissolve	the	medium.		
Isolation	and	Identi<ication	of	Candida	Di-
agnosis	using	biochemical	test	(Vitek	ap-
paratus)		
It	is	a	selective	and	differential	chromogenic	
medium	 intended	 for	 the	 identiXication	 of	
Candida	species	and	to	determine	color	and	
colony	 morphology	 well	 when	 chromium	
was	used	for	Candida	isolate	directly	and	Xil-
amentous	fungi	and	of	clinical	specimens.	C.	
albicans	grow	as	green	colonies	after	incuba-
tion	for	48	hours	at	37°C.	This	medium	does	
not	require	sterilization	in	an	autoclave.[23]	
Diagnosed	by	 genus	 and	 species	 using	 bio-
chemical	 tests,	 the	Vitek	System	consists	of	
cassette	and	reagent	Cards	that	contained	64	
pits	 (every	 pit	 represents	 the	 substrate	 to	
conduct	the	test),	and	plastic	pipes,	as	well	as	
the	Densi	Chek	device	and	the	unity	of	the	in-
put	and	output	of	information.[24]	Diagnosis	
of	Candida	species	 isolates	using	biochemi-
cal	 tests	 Diagnosis	 using	 the	 Vitek®	 appa-
ratus	 (VITEK®	 2	 DensiChek;	 bioMérieux,	
France).	Yeast	isolates	were	diagnosed	as	ge-
nus	and	species	using	biochemical	tests.		
Study	the	resistance	of	oral	C.	albicans	to	
the	antifungal	drugs:	A	clinical	 isolates	of	
Candida	 species	 were	 tested	 for	 antifungal	
susceptibility	 against	 six	 antifungal	 drugs	
[Clotrimazole	 (10µg),	 Fluconazole	 (10µg),	
Nystatin	(50µg),	5-Flucytosine	(1µg),	Caspo-
fungin	(5µg),	and	Amphotericin	B	(100µg)];	
the	 antifungal	 susceptibility	 of	 oral	 C.	 albi-
cans	was	tested	by	disc	diffusion.	
Statistical	Analysis:	Data	were	analyzed	us-
ing	SPSS	(version	26,	SPSS	Inc.	Chicago,	Illi-
nois,	 USA).	 Descriptive	 statistics	 (mean,	
standard	 Error),	 as	 the	 results	 was	 ex-
pressed	as	%.	Statistical	analysis	was	carried	
out	using	chi-square.	Differences	w	ere	com-
pared	 by	 two-way	 ANOVA	 at	 p≤0.05	 using	
Duncan’s	Multiple	Range	test	[25].		



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
3	

Results 
Out	 of	 the	 total	 40	 participants	 enrolled	 in	
this	study,	the	culture	results	showed	a	sig-
niXicant	 presence	 of	 Staphylococcus	 aureus	
(80%)	 and	 Candida	 albicans	 (68%)	 among	
individuals	 undergoing	 Xixed	 orthodontic	
treatment.	Among	male	participants	(n	=	20)	
in	three	period	times	(T0,	T1	and	T2),	Staph-
ylococcus	aureus	was	detected	in	16	cases	in	
each	 period	 time	 (40%	 vs	 total	 No.)	 while	
Candida	 albicans	 was	 found	 in	 13	 cases	 in	
each	period	time	(32.5%	vs	total	No.).	In	the	
female	group	(n	=	20)	in	three	period	times	
(T0,	T1	and	T2),	16	participants	(40%	vs	to-
tal	No.)	tested	positive	for	Staphylococcus	au-
reus	in	each	period	time,	while	only	14	cases	
(35%	vs	total	No.)	were	positive	for	Candida	
albicans	in	each	period	time.	Overall,	59	out	
of	80	 samples	 (73.75%)	were	 culture	posi-
tive	 for	 either	 S.	 aureus	 or	 C.	 albicans,	
whereas	 21	 samples	 (26.25%)	 were	 nega-
tive	for	both.	The	prevalence	appeared	con-
sistent	across	genders	(Table	1).	
Antibiotics	Susceptibility	test	for	Staphy-
lococcus	aureus	Isolates	
Vancomycin	 and	 Clindamycin	 showed	 the	
largest	 inhibition	 zones	 (14–15	 mm),	 sug-
gesting	 strong	 antibacterial	 activity	 against	
S.	 aureus.	 The	difference	 is	 statistically	 sig-
niXicant	 (*p	<	0.001)	 compared	 to	 all	 other	
antibiotics	 while	 Azithromycin	 showed	 in-
termediate	inhibition	(10–11	mm),	which	is	
statistically	 less	 than	 Clindamycin	 but	 still	
more	effective	than	Ampicillin	and	Erythro-
mycin.	The	weakest	Activity	were	Amoxicil-
lin,	 Ampicillin,	 and	 Erythromycin	 that	
demonstrated	smaller	inhibition	zones	(5–8	
mm),	indicating	poor	antimicrobial	effective-
ness.	 These	 results	 may	 reXlect	 the	 emer-
gence	of	 resistance	 to	β-lactam	and	macro-
lide	antibiotics	among	oral	S.	aureus	isolates	
(Figure	1).	
Antifungals	 Susceptibility	 test	 for	 Can-
dida	albicans	Isolates	
Fluconazole	 and	 Amphotericin	 B	 produced	
the	largest	inhibition	zones	(13–14	mm),	in-
dicating	 the	 strongest	 antifungal	 activity	
against	C.	albicans.	Their	effect	is	statistically	
signiXicant	(p	<	0.001)	compared	to	all	other	
agents	while	 5-Flucytosine	 showed	moder-
ate	inhibition	(11	mm)	and	was	signiXicantly	
better	 than	 Nystatin	 and	 Clotrimazole	 but	
slightly	 less	 effective	 than	 Fluconazole	 and	
Amphotericin.	 Clotrimazole	 and	 Nystatin	
had	the	smallest	inhibition	zones	(5–6	mm),	
suggesting	 limited	 antifungal	 effectiveness	
against	oral	C.	albicans	isolates.	The	statisti-
cal	differences	between	Clotrimazole,	Nysta-
tin,	 and	 Caspofungin	 were	 not	 signiXicant	
(Figure	2).	

Discussion 
This	study	highlighted	a	significant	increase	
in	 the	 prevalence	 of	 Staphylococcus	 aureus	

and	Candida	albicans	in	the	saliva	of	patients	
undergoing	 treatment	 with	 fixed	
orthodontic	 appliances.	The	 components	of	
these	 appliances,	 such	 as	 brackets,	 wires,	
bands,	and	 ligatures,	create	multiple	niches	
that	 favour	 the	 accumulation	 of	 dental	
plaque	and	biofilm.	These	structures	hinder	
natural	 self-cleansing	 mechanisms	 and	
complicate	routine	oral	hygiene,	fostering	a	
micro-environment	that	promotes	microbial	
colonization	and	growth.	Consequently,	 the	
oral	microbial	load	and	diversity	are	notably	
altered,	creating	conditions	conducive	to	the	
proliferation	 of	 pathogenic	 organisms,	
especially	 during	 the	 early	 phases	 of	
orthodontic	treatment	[1,2,4,5].		
This	 observation	 is	 in	 line	 with	 existing	
research	 indicating	 that	 fixed	 appliances	
function	as	reservoirs	for	microbial	biofilms.	
These	 devices	 hinder	 effective	 plaque	
removal	 and	 increase	 the	 oral	 burden	 of	
opportunistic	pathogens.	Both	S.	aureus	and	
C.	albicans	are	part	of	the	normal	oral	 flora	
but	 can	 become	 pathogenic	 when	 the	
microbial	 balance	 is	 disrupted	 such	 as	 by	
reduced	 immunity,	 increased	 substrate	
availability,	or	structural	changes	in	the	oral	
environment.	This	microbial	shift	can	lead	to	
localized	 infections,	 including	 oral	
candidiasis	and	gingival	inflammation,	as	the	
pathogens	 exploit	 the	 altered	 oral	 ecology	
[3,6,26-28].		
Antibiotic	and	antifungal	susceptibility	tests	
revealed	 patterns	 of	 resistance	 that	 pose	 a	
clinical	 concern.	 Methicillin-resistant	 S.	
aureus	 (MRSA)	 and	 antifungal-resistant	 C.	
albicans	 are	 increasingly	 identified	 in	
orthodontic	 patients,	 raising	 treatment	
challenges.	 Resistance	mechanisms	 such	 as	
gene	mutations,	 efflux	 pumps,	 and	 biofilm-
related	protection	contribute	to	diminished	
therapeutic	 outcomes.	 This	 highlights	 the	
need	 for	 ongoing	 microbiological	
surveillance	 and	 tailored	 antimicrobial	
strategies	in	orthodontic	practice	to	prevent	
complications	 arising	 from	 resistant	
infections	[8,9,29-32].		
The	 antibiotic	 sensitivity	 analysis	 in	 this	
study	showed	limited	efficacy	of	commonly	
used	agents	such	as	Amoxicillin,	Ampicillin,	
and	Erythromycin,	likely	due	to	β-lactamase	
production	 and	 other	 resistance	
mechanisms.	 In	 contrast,	 Clindamycin	 and	
Vancomycin	demonstrated	strong	inhibitory	
effects,	 reflecting	 their	 distinct	 modes	 of	
action.	 Clindamycin	 by	 targeting	 the	 50S	
ribosomal	 subunit	 and	 Vancomycin	 by	
inhibiting	cell	wall	synthesis	with	binding	to	
the	 terminal	D-alanyl-D-alanine	 residues	of	
peptidogly	can	precursors,	specifically	 lipid	
II	[9,29,33].			
Likewise,	 antifungal	 testing	 demonstrated	
varied	 efficacy.	 Clotrimazole	 and	 Nystatin	
showed	 minimal	 inhibition	 zones,	

suggesting	 possible	 resistance	 due	 to	
frequent	 topical	 use.	 Conversely,	
Fluconazole	and	Amphotericin	B	were	highly	
effective	against	C.	albicans,	supporting	their	
continued	 use	 in	 oral	 candidiasis.	
Amphotericin	 B’s	mechanism	 of	 binding	 to	
ergosterol	and	disrupting	fungal	membranes	
underpins	 its	 potency,	 especially	 in	 drug-
resistant	 strains	 [16,28,3034,35].	 5-
Flucytosine,	 though	 moderately	 effective,	
remains	 valuable	 in	 combination	 therapies	
due	 to	 its	 rapid	 resistance	 development	
when	used	alone	[36,37].	
Overall,	 these	 findings	 emphasize	 the	
importance	 of	 incorporating	 routine	
susceptibility	 testing	 and	 individualized	
treatment	approaches	into	orthodontic	care.	
As	 fixed	 appliances	 significantly	 alter	 the	
oral	 microenvironment,	 they	 increase	 the	
risk	of	harboring	resistant	microbial	species.	
Regular	microbial	assessments	and	targeted	
antimicrobial	interventions	are	essential	for	
preventing	 and	 managing	 persistent	 or	
recurrent	 oral	 infections	 in	 orthodontic	
patients	[1].	

Acknowledgements 
We	would	like	to	extend	our	heartfelt	thanks	
to	 the	 orthodontic	 department,	 Nasser	 Al-
mousawi	specialized	center,	Najaf,	Iraq,	and	
special	thanks	to	Orthodontist	Dr.	Almustafa	
Alhumadi	 for	 their	 support	 and	 the	 assis-
tance	provided. 

Conflict of Interest Statement 
None to declare. 

References 
1. Lucchese	A,	Bondemark	L.	The	influence	
of	 orthodontic	 treatment	 on	 oral	 microbiology.	
Biological	mechanisms	of	tooth	movement,	2021:	
139–158.		
2. Mazin	H,	Salman	SA,	Salah	R.	The	effect	of	
fixed	 orthodontic	 appliances	 on	 gingival	 health	
diode	 laser	 versus	 scalpel	 gingivectomy	 view	
project	periodontology	view	project.	IOSR	Journal	
of	 dental	 and	 medical	 sciences	 (IOSR-JDMS),	
15(11),	 2016:	 82–88.	 Available	 at:	
https://doi.Org/10.9790/0853-1517078288.		
3. Deo	PN,	Deshmukh	R.	Oral	microbiome:	
unveiling	 the	 fundamentals.	 journal	 of	 oral	 and	
maxillofacial	 pathology.	 JOMFP.	 2019;	 23(1),	 p.	
122.		
4. Kado	I,	Hisatsune	J,	Tsuruda	K,	Tanimoto	
K,	 Sugai	 M.	 The	 impact	 of	 fixed	 orthodontic	
appliances	 on	 oral	 microbiome	 dynamics	 in	
Japanese	patients.	Sci	Rep.	2020;10(1):21989.	doi:	
10.1038/s41598-020-78971-2.	
5. Kim	 SH,	 Choi	 DS,	 Jang	 I,	 Cha	 BK,	 Jost-
Brinkmann	PG,	Song	JS.	Microbiologic	changes	 in	
subgingival	 plaque	 before	 and	 during	 the	 early	
period	 of	 orthodontic	 treatment.	 Angle	 Orthod.	
2012	;82(2):254-60.	doi:	10.2319/030311-156.1.		
6. Rojo	 P	 et	 al.	 (2010).	 Community-
associated	S.	Aureus	infections	in	children.	Expert	
Rev	Anti	Infect	Ther,	8,	541–554.		
7. Ryu,S.;	 Song,	 P.I.;	 Seo,	 C.H.;	 Cheong,	 H.;	
Park,	Y.	Colonization	and	infection	of	the	skin	by	S.	



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
4	

Aureus:	immune	system	evasion	and	the	response	
to	cationic	antimicrobial	peptides.	 Int.	 J.	Mol.	Sci.	
2014,	15,	8753–8772.			
8. Kale,	P.,	Dhawan,	B.	(2016).	The	changing	
face	 of	 community-acquired	methicillin-resistant	
staphylococcus	 aureus.	 Indian	 J.	Med.	Microbiol.,	
34,	275–285.		
9. Casanova,	 G.N.;	 Ruiz,	 S.M.;	 Bellido,	M.J.L.	
Mechanisms	 of	 resistance	 to	 daptomycin	 in	
staphylococcus	 aureus.	 Rev.	 Española	 quimioter.	
(2017),	30,	391–396.	
10. Sirichoat,	 A.;	 Lulitanond,	 A.;	 Kanlaya,	 R.;	
Tavichakorntrakool,	 R.;	 Chanawong,	 A.;	
Wongthong,	 S.;	 Thongboonkerd,	 V.	 Phenotypic	
characteristics	 and	 comparative	 proteomics	 of	
staphylococcus	 aureus	 strains	 with	 different	
vancomycin-resistance	 levels.	 Diagn.	 Microbiol.	
Infect.	Dis.	2016,	86,	340–344.	
11. Romaniuk,	 J.A.;	Cegelski,	L.	Bacterial	 cell	
wall	composition	and	the	 influence	of	antibiotics	
by	cell-wall	and	whole-cell	NMR.	Philos.	Trans.	R.	
Soc.	B	biol.	Sci.	2015,	370.	
12. Al-Tekreeti	AR,	Al-Halbosiy	MMF,	Dheeb	
BI,	 Hashim	 AJ,	 Al-Zuhairi	 AFH.	 Molecular	
identification	of	clinical	candida	isolates	by	simple	
and	 randomly	 amplified	 polymorphic	 DNA-PCR.	
Arab	J.	Sci.	Eng.	(2017);	doi	10.1007/s13369-017-
2762-1.		
13. Hussain	 AF,	 Sulaiman	 GM,	 Dheeb	 BI,	
Hashim	 AJ,	 Seddiq	 SH.	 Improving	 conditions	 for	
gliotoxin	 production	 by	 local	 isolates	 of	
aspergillus	 fumigatus.	 Journal	 of	 biotechnology	
research	center.	(2017);	11(2):14-24.			
14. Ayesha	 A.	 Prevalence,	 antifungal	
susceptibility	 and	 biofilm	 characterization	 of	
candida	 species	 isolated	 from	 tertiary	 care	
hospitals,	 2016;	 Quaid-I-Azam	 University,	
Islamabad,	Pakistan	(2018);	1-22.		
15. Khanpayeh	 E,	 Jafari	 AA,	 Tabatabaei	 Z.	
Comparison	of	salivary	Candida	profile	in	patients	
with	fixed	and	removable	orthodontic	appliances	
therapy.	Iran	J	Microbiol.	2014	Aug;6(4):263-8.	
16. Al-Harba,	 Huda	 Reyad;	 Nasir-Alla,	
Nebrass;	 Jabir,	 Azher	 Abdulhafidh.	 The	
Correlation	 between	 Oral	 Candida	 albicans	 and	
Interlukine-23	 in	 Diabetic	 Patients	 in	 Hilla	 City,	
Iraq.	Medical	Journal	of	Babylon	21(4):p	915-920,	
October-December	 2024.	 |	 DOI:	
10.4103/MJBL.MJBL_517_23.	
17. Alhamadi	W,	Al-Saigh	RJ,	Al-Dabagh	NN,	
Al-Humadi	 HW.	 Oral	 Candida	 in	 Patients	 with	
Fixed	 Orthodontic	 Appliance:	 In	 Vitro	
Combination	 Therapy.	 Biomed	 Res	 Int.	 (2017);	
2017:1802875.	doi:	10.1155/2017/1802875.		
18. Navazesh,	 M.,	 Kumar,	 S.K.	 2008.	
Measuring	 salivary	 flow:	 challenges	 and	
opportunities.	The	journal	of	the	american	dental	
association,	139,	35S-40S.	
19. Kaushik,	 R.,	 Yeltiwar,	 R.	 K.,	 Pushpanshu,	
K.	2011.	Salivary	interleukin-1β	levels	in	patients	
with	 chronic	 periodontitis	 before	 and	 after	
periodontal	phase	I	therapy	and	healthy	controls:	
A	 case-control	 study.	 Journal	 of	 periodontology,	
82,	1353-1359		
20. James	 C,	 Natalie	 S.	 Microbiology.	 A	
laboratory	 manual.	 Pearson	 education,	 2014.	
Available	 from:	
https://lib.Hpu.Edu.Vn/handle/123456789/289
98			
21. Hudzicki	 J.	 Kirby-bauer	 disk	 diffusion	
susceptibility	 test	 protocol.	 American	 society	 for	
microbiology.	2009;15:55-63.	

22. Scognamiglio	 T,	 Zinchuk	 R,	 Gumpeni	 P,	
Larone	DH.	Comparison	of	inhibitory	mold	agar	to	
Sabouraud	dextrose	agar	as	a	primary	medium	for	
isolation	 of	 fungi.	 J	 Clin	 Microbiol.	 2010	
May;48(5):1924-5.	doi:	10.1128/JCM.01814-09.		
23. Nadeem,	 S.G.,	 Hakim,	 S.T.,	 Kazmi,	 S.U.	
(2010).	 Use	 of	 chromagar	 candida	 for	 the	
presumptive	 identification	 of	 candida	 species	
directly	 from	 clinical	 specimens	 in	 resource-
limited	settings,	Libyan	Journal	of	Medicine,	5(1).		
24. Abdulbaqi	 NJ,	 Dheeb	 BI,	 Irshad	 R.	
Expression	of	 biotransformation	and	antioxidant	
genes	in	the	liver	of	albino	mice	after	exposure	to	
aflatoxin	 B1	 and	 an	 antioxidant	 sourced	 from	
turmeric	 (curcuma	 longa).	 Jordan	 Journal	 of	
Biological	Sciences.	(2018);	11(2)	89	–	93.	
25. Cohen,	 J.	 (2013).	Statistical	 power	
analysis	for	the	behavioral	sciences.	Routledge.	
26. Wade,	 W.G.	 The	 oral	 microbiome	 in	
health	 and	 disease,	 Pharmacological	 Research,	
(2013);	69(1),	pp.	137–143.	
27. Salman,	S.A.,	Salah,	R.	(2016).	The	effect	of	
fixed	 orthodontic	 appliances	 on	 gingival	 health	
diode	 laser	 versus	 scalpel	 gingivectomy	 view	
project	periodontology	view	project,	IOSR	Journal	
of	 Dental	 and	 Medical	 Sciences	 (IOSR-JDMS),	
15(11),	 pp.	 82–88.	 Available	 at:	
https://doi.Org/10.9790/0853-	1517078288.		
28. Moyes	 DL,	 Wilson	 D,	 Richardson	 JP,	
Mogavero	S,	Tang	SX,	Wernecke	J,	Höfs	S,	Gratacap	
RL,	Robbins	J,	Runglall	M,	Murciano	C,	Blagojevic	
M,	Thavaraj	S,	Förster	TM,	Hebecker	B,	Kasper	L,	
Vizcay	G,	Iancu	SI,	Kichik	N,	Häder	A,	Kurzai	O,	Luo	
T,	 Krüger	 T,	 Kniemeyer	 O,	 Cota	 E,	 Bader	 O,	
Wheeler	 RT,	 Gutsmann	 T,	 Hube	 B,	 Naglik	 JR.	
Candidalys	in	is	a	fungal	peptide	toxin	critical	for	
mucosal	 infection.	 Nature.	 2016	 Apr	
7;532(7597):64-8.	doi:	10.1038/nature17625.	
29. Ali	T,	Basit	A,	Karim	AM,	Lee	J-H,	Jeon	J-H,	
Lee	 SH.	 Mutation-based	 antibiotic	 resistance	
mechanism	 in	 methicillin-resistant	
staphylococcus	 aureus	 clinical	 isolates.	
Pharmaceuticals.	
Https://doi.Org/10.3390/ph14050420	 18.	
(2021);14(5):420.		
30. Nobile	 CJ,	 jJhnson	 AD.	 (2015).	 Candida	
albicans	 biofilms	 and	 human	 disease.	 Annu	 rev	
microbiol,	69:71–92.		
31. Parastan	 R,	 Kargar	 M,	 Solhjoo	 K,	
Kafilzadeh	 F.	 A	 synergistic	 association	 between	
adhesion-related	genes	and	multidrug	 resistance	
patterns	 of	 staphylococcus	 aureus	 isolates	 from	
different	patients	and	healthy	individuals.	Journal	
of	 global	 antimicrobial	 resistance.	 2020;22:379-
85.	Https://doi.Org/10.1016/j.Jgar.2020.02.025			
32. Abbasi	 Montazeri,	 E.,	 Khosravi,	 A.D.,	
Khazaei,	 S.	 et	 al.	 Prevalence	 of	 methicillin	
resistance	 and	 superantigenic	 toxins	 in	
Staphylococcus	 aureus	 strains	 isolated	 from	
patients	 with	 cancer.	 BMC	 Microbiol	 21,	 262	
(2021).	 https://doi.org/10.1186/s12866-021-
02319-7	
33. Gardete	 S,	 Tomasz	 A.	 Mechanisms	 of	
vancomycin	resistance	in	Staphylococcus	aureus.	J	
Clin	 Invest.	 2014	 Jul;124(7):2836-40.	 doi:	
10.1172/JCI68834.		
34. Achkar	JM,	Fries	BC.	Candida	infections	of	
the	 genitourinary	 tract.	 Clinical	 microbiology	
reviews.	(2010);	23(2),	253–273.		
35. Abdalwahd	 N.,	 Al-Saigh	 R.J.,	 Al-Humadi	
H.W.	 Assessment	 of	 antifungal	 drugs’	 activity	
against	 some	 Candida	 albicans	 isolates	 in	 the	

presence	 or	 absence	 of	 human	 albumin:	 a	 study	
employing	 an	 in	 vitro	 pharmacokinetics	 /	
pharmacodynamics	model.	 Rev.	 Clin.	 Pharmacol.	
Pharmacokinet.	 Int.	 Ed.	 38(Sup2):	 39-42	 (2024).	
https://doi.org/10.61873/SEXH5182		
36. Bhavan,	 P.,	 Rajkumar,	 R,	 Subramanian,	
Radhakrishnan,	 Seenivasan,	 Dr.	 Chandrasekar	 &	
Kannan,	 Soundarapandian.	 Culture	 and	
Identification	 of	 Candida	 Albicans	 from	 Vaginal	
Ulcer	 and	 Separation	 of	 Enolase	 on	 SDS-PAGE.	
International	 Journal	 of	 Biology.	 (2010);	 Vol	 2.	
10.5539/ijb.v2n1p84.	
37. Abbas,	Nada	Fadhil.	Comparison	between	
Conventional	 Methods	 and	 Molecular	 Diagnosis	
for	 Candida	 albicans	 and	 Candida	 dubliniensis	
isolated	 from	Cancer	 Patients	 Infected	with	Oral	
Candidiasis.	Medical	Journal	of	Babylon	21(Suppl	
2):p	 S276-S281,	 November	 2024.	 |	 DOI:	
10.4103/MJBL.MJBL_1079_23	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
5	

Table	1.	Culture	positive	and	culture	negative	for	Staphylococcus	aureus	and	Candida	albicans.	
Type	of	Microorganisms	 Gender	 Culture	Positive	 Culture	Negative	 Total	
Staphylococcus	aureus	 Male	 16	 32	 4	 8	 40	Female	 16	 4	
Candida	albicans	 Male	 13	 27	 7	 13	 40	Female	 14	 6	
Total	 	 59	 21	 80	

	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	
	



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
6	

	
Figure	1.	The	bar	graph	illustrates	the	inhibition	zones	(mm)	for	Staphylococcus	aureus	isolates	at	three-time	intervals	(T0	(A),	T1	(B),	T2	
(C)),	when	exposed	to	six	different	antibiotics:	Amoxicillin,	Vancomycin,	Ampicillin,	Erythromycin,	Azithromycin,	and	Clindamycin.		Statistical	
comparisons	between	groups	are	indicated	with	asterisks	(*	for	p	<	0.05,	**	for	p	<	0.01,	***	for	p	<	0.001)	and	"ns"	for	non-signiXicant	differ-
ences.		



  Impact of Fixed OrthodonEc Appliances on Staphylococcus aureus and Candida albicans 

Vol 13, No 1 (2025)    DOI 10.5195/d3000.2025.975 

	http://dentistry3000.pitt.edu	
7	

	
Figure	2.	The	bar	graph	illustrates	the	inhibition	zones	(mm)	for	Candida	albicans	isolates	at	three-time	intervals	(T0	(A),	T1	(B),	T2	(C)),	when	
exposed	to	six	antifungal	agents:		Clotrimazole,	Fluconazole,	Nystatin,	5-Flucytosine,	Caspofungin,	and	Amphotericin	B.	Statistical	com-
parisons	between	groups	are	annotated	with	***	for	p	<	0.001	and	“ns”	for	non-signiXicant	differences.