In vitro activities of antimicrobial agents against Candida species

doi:10.1016/S1079-2104(99)70293-3

Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology

Volume 87, Issue 1, January 1999, Pages 44-49

https://doi.org/10.1016/S1079-2104(99)70293-3 Get rights and content

Abstract

Objective. Antimicrobial mouthrinses may represent a valid alternative to topical antifungal agents. However, the action of antimicrobials could be affected by the different ingredients incorporated into mouthrinse products. The purpose of the present study was to investigate the in vitro antifungal and fungicidal activities of antimicrobials alone. Study Design. A broth macrodilution method was used to determine the minimum inhibitory concentration of 4 antimicrobial agents against Candida species. Minimum fungicidal concentration was also determined. Results. All antimicrobials showed antifungal activity against all tested organisms, but cetylpyridinium chloride received significantly lower minimum inhibitory concentrations (P < .005). Cetylpyridinium chloride also showed a greater fungicidal activity than chlorhexidine digluconate and hexetidine (P < .005), whereas sanguinarine chloride appeared to be less fungicidal against most of the isolates tested. Conclusions. These findings suggest that cetylpyridinium chloride may be used as a topical antifungal agent. Clinical trials are now required to assess its value in the management of oral candidosis.(Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1999;87:44-9)

Section snippets

Antimicrobial agents

CHX, CPC, HEX, and SNG were purchased from Sigma-Aldrich S.r.l. (Milano, Italy). Stock solutions were prepared just before their use at concentrations either 10 or 100 times the highest concentration to be tested (10× and 100× stocks, respectively).²⁸ The starting concentrations and the solvent in which the test compounds were dispersed were as follows: CHX and CPC, 20,000 μg/mL in sterile triple-distilled water (10× stocks); HEX and SNG, 200,000 and 781 μg/mL, respectively, in sterile 100%

RESULTS

All tested organisms produced detectable growth after 24 hours of incubation in the RPMI 1640 medium used. The inoculum sizes of the working suspensions fell within the range reported by the NCCLS (0.5 × 10³ to 2.5 × 10³ CFU/mL).²⁸ MIC and MFC data were normally distributed, as indicated by the KS test, and are expressed as means (± standard deviation) of duplicate determinations.

DISCUSSION

Antimicrobial agents, incorporated into mouthrinse, toothpaste, and gel formulations, have provided an improved chemical approach to prevention and treatment of gingivitis, periodontitis, and other oral infections.31, 32 Among these agents, CHX is established as the most effective plaque inhibitor to date, and oral products containing this agent have been recommended for a variety of uses in clinical dentistry.³³ CPC, HEX, and SNG have also been shown to possess good antibacterial activities,

References (48)

JB Epstein et al.
Risk factors for oropharyngeal candidiasis in patients who receive radiation therapy for malignant conditions of the head and neck
Oral Surg Oral Med Oral Pathol
(1993)
GN King et al.
Prevalence and risk factors associated with leukoplakia, hairy leukoplakia, erythematous candidiasis, and gingival hyperplasia in renal transplant recipients
Oral Surg Oral Med Oral Pathol
(1994)
L Bunetel et al.
Oral pathoses caused by Candida albicans during chemotherapy
Oral Surg Oral Med Oral Pathol Oral Radiol Endod
(1996)
JB Epstein et al.
Prophylaxis of candidiasis in patients with leukemia and bone marrow transplants
Oral Surg Oral Med Oral Pathol Oral Radiol Endod
(1996)
D. Greenspan
Treatment of oral candidiasis in HIV infection
Oral Surg Oral Med Oral Pathol
(1994)
JB Epstein et al.
Efficacy of chlorhexidine and nystatin rinses in prevention of oral complications in leukemia and bone marrow transplantation
Oral Surg Oral Med Oral Pathol
(1992)
AD Hamers et al.
Use of a microtiter plate assay to detect the rate of killing of adherent Candida albicans by antifungal agents
Oral Surg Oral Med Oral Pathol Oral Radiol Endod
(1996)
JM Thurmond et al.
Oral Candida albicans in bone marrow transplant patients given chlorhexidine rinses: Occurence and susceptibilities to the agent
Oral Surg Oral Med Oral Pathol
(1991)
SP Fisher-Hoch et al.
Opportunistic candidiasis: an epidemic of the 1980’s
Clin Infect Dis
(1995)
RD. Diamond
The growing problem of mycoses in patients infected with the human im. munodeficiency virus
Rev Infect Dis
(1991)

G Bodey et al.

Fungal infections in cancer patients: an international autopsy survey

Eur J Clin Microbiol Infect Dis

(1992)

CV. Paya

Fungal infections in solid-organ transplantation

Clin Infect Dis

(1993)

MA. Pfaller

Nosocomial candidiasis: emerging species, reservoirs, and modes of transmission

Clin Infect Dis

(1996)

J. Nikoskelainen

Oral infections related to radiation and immunosuppressive therapy

J Clin Periodontol

(1990)

EC-Clearinghouse on Oral Problems related to HIV Infection and WHO Collaborating Centre on Oral Manifestations of the Human Immunodeficiency Virus

Classification and diagnostic criteria for oral lesions in HIV infection

J Oral Pathol Med

(1993)

A Heimdhal et al.

Oral yeast infections in immunocompromised and seriously diseased patients

Acta Odontol Scand

(1990)

Working Party of the British Society for Antimicrobial Chemotherapy

Chemoprophylaxis for candidosis and aspergillosis in neutropenia and transplantation: a review and recommendations

J Antimicrob Chemother

(1993)

E Budtz-Jörgensen et al.

Antifungal therapy in the oral cavity

Periodontol 2000

(1996)

JH Rex et al.

Resistance of Candida species to fluconazole

Antimicrob Agents Chemother

(1995)

EM Johnson et al.

Emergence of azole drug resistance in Candida species from HIV-infected patients receiving prolonged fluconazole therapy for oral candidosis

J Antimicrob Chemother

(1995)

RA Monteil et al.

In vitro antifungal resistance of oral Candida albicans strains in non-AIDS patients

Oral Microbiol Immunol

(1997)

F Barchiesi et al.

Emergence of oropharyngeal candidiasis caused by non-albicans species of Candida in HIV-infected patients

Eur J Epidemiol

(1993)

D Sullivan et al.

Oligonucleotide fingerprinting of isolates of Candida species other than C. albicans and of atypical Candida species from human immunodeficiency virus-positive and AIDS patients

J Clin Microbiol

(1993)

YU Samaranayake et al.

Candida krusei: biology, epidemiology, pathogenicity and clinical manifestations of an emerging pathogen

J Med Microbiol

(1994)

Cited by (34)

Antifungal prescribing pattern and attitude towards the treatment of oral candidiasis among dentists in Jordan
2015, International Dental Journal
Aim: The aim of this study was to evaluate the attitude of Jordanian dentists towards the treatment of oral candidiasis and their current antifungal prescribing habits, shedding more light on the possible influence of their socio-professional factors on the pattern of prescribing and practice. Methods: A structured validated questionnaire was developed and tested; it was then emailed to a random sample of 600 Jordanian dental practitioners during the period of this cross-sectional survey. The questionnaire recorded practitioners’ personal details and their attitude and prescribing of antifungal therapy for oral candidiasis. Statistical significance was based on probability values of <0.05 and was measured using the chi-square and Fisher’s exact tests. Multiple logistic regression analysis was used to analyse the influence of respondents’ socio-professional factors on their attitude towards oral candidiasis. Results: Of the 423 questionnaires returned, only 330 were included. The attitude of respondents was significantly influenced by their experience [odds ratio (OR) = 0.14; P < 0.001] and workplace (OR = 4.70; P < 0.001). Nystatin was the most commonly prescribed antifungal agent (78.2%), followed by miconazole (62.4%), which was prescribed for topical use. Systemic antifungals were prescribed by 21.2% of respondents, with a significant (P < 0.05) association with the country in which their qualification was obtained. Conclusion: The attitude towards the treatment of oral candidiasis is much better among the least-experienced dentists working in private practice. Nystatin and miconazole are the most popular choices of antifungal agents among Jordanian dentists.
Chlorhexidine is a highly effective topical broad-spectrum agent against Candida spp.
2013, International Journal of Antimicrobial Agents
Citation Excerpt :
It was effective against all Candida spp. tested at concentrations detected in saliva when using standard dosing regimens. These findings are in line with previous limited data [28]. Susceptibility to CHX was compared with that of FLZ for a wide range of Candida spp.
The objective of this study was to compare the in vitro antifungal activities of chlorhexidine (CHX) and fluconazole (FLZ) against Candida isolates comprising eight different species associated with oral candidosis. A broth microdilution method as described in Clinical and Laboratory Standards Institute (CLSI) protocol M27-A3 was used to determine susceptibility. A total of 79 clinical isolates and reference strains belonging to eight different Candida spp. was tested. The minimum inhibitory concentration (MIC) was the lowest drug concentration that reduced growth by 50% for FLZ at 48 h and by 80% for CHX at 24 h and 48 h. The geometric mean MIC (and MIC range) at 48 h for CHX was 3.03 mg/L (0.78–6.25 mg/L) and for FLZ was 19.12 mg/L (≤0.125–256 mg/L). Of the 79 isolates, 14 (18%) were resistant to FLZ (MIC ≥ 64 mg/L). All isolates were effectively inhibited by ≤6.25 mg/L CHX, and Candida CHX MICs are below the CHX levels found in saliva following normal dosing. No cross-resistance between CHX and FLZ was detected (r_s = 0.039, P = 0.733). CLSI M27-A3 methodology proved to provide reproducible results with clear endpoints for CHX. In conclusion, the findings showed that CHX has excellent broad-spectrum antifungal activity in vitro. It was effective at concentrations detected in saliva when using standard dosing regimens. Moreover, no cross-resistance was detected between CHX and FLZ, even among Candida spp. highly resistant to FLZ.
Assessment of in vitro activity and stability of antifungal suspensions for mouthrinses: To a reappraisal of empiric practices?
2012, Pathologie Biologie
L’instauration d’une prophylaxie efficace des candidoses buccales par traitement local est primordiale chez les patients immunodéprimés. Le but de l’étude est d’évaluer l’activité et la stabilité de suspensions antifongiques utilisées pour réaliser des bains de bouche. Les suspensions testées comprennent un solvant (eau stérile, eau de source ou bicarbonate de sodium) associé à l’amphotericine B (Fungizone^®) ou la nystatine (Mycostatine^®). Deux autres mélanges sont évalués : Mycostatine^®-bicarbonate et Mycostatine^®-Hextril^®-bicarbonate ainsi que les deux antifongiques purs. L’activité in vitro est testée sur cinq espèces de Candida (C. albicans, C. glabrata, C. krusei, C. parapsilosis, C. tropicalis) après mise en contact durant cinq minutes des levures avec la suspension à tester. Une étude de stabilité galénique est conduite pendant trois jours. Les mélanges associant un polyène et du bicarbonate de sodium n’ont aucune activité antifongique sur C. albicans, les autres mélanges présentant un taux d’inhibition de pousse des levures compris entre 35 et 68 %. Les résultats d’activité des mélanges à base d’Hextril^® ne sont pas interprétables, la présence d’alcool dans sa composition pouvant biaiser les résultats. Les mélanges à base d’eau de source sont à proscrire car une contamination microbiologique apparaît au bout de 48 heures. Le mélange Mycostatine^®-Hextril^®-bicarbonate n’est pas stable pendant trois jours. Tous ces bains de bouche, peu efficaces sauf chez C. glabrata, doivent être évités. La Mycostatine^® pure présente une très bonne activité antifongique sauf chez C. krusei et son utilisation peut être recommandée.
Establishment of an effective prophylaxis against oral candidiasis by local treatment is essential for immunocompromised patients. The aim of the study is to assess effectiveness and stability of antifungal suspensions for mouthrinses. The assessed suspensions are compounded by one solvent among sterile water, spring water or sodium bicarbonate associated with amphotericin B (Fungizone^®) or nystatine (Mycostatine^®). Two others mixes are assessed: Mycostatine^®-bicarbonate and Mycostatine^®-Hextril^®-bicarbonate as well as the two straight antifungal. In vitro activity is tested on five Candida species (C. albicans, C. glabrata, C. krusei, C. parapsilosis, C. tropicalis) after a five minutes contact between yeasts and the assessed suspension. A galenic study is realized during 3 days. Mixes associating a polyene with sodium bicarbonate have no effectiveness on Candida albicans, others mixes shows intermediate effectiveness (the percentage of yeast growth inhibition lies between 35% and 68%). Effectiveness results of Hextril^®-based mixes are not explainable because of alcohol in its composition. Spring water-based mixes must be evicted due to microbiologic contaminations after 48 hours. Mycostatine^®-Hextril^®-bicarbonate mix is not stable during 3 days. All those mouthrinses, poorly effective, excepted on C. glabrata, should be avoided. Straight Mycostatine^® shows a good antifungal effectiveness excepted on C. krusei and its use should be recommended.
In vitro antifungal effect of mouth rinses containing chlorhexidine and thymol
2011, Journal of Dental Sciences
Citation Excerpt :
This in vitro study to assess the antifungal activities of chlorhexidine- and thymol-containing mouth rinses was carried out by means of MIC and time-kill assays. The results compared favorably with investigations by Giuliana et al.2,12 However, the concentration of chlorhexidine required for growth inhibition of C. albicans was less than that required for C. tropicalis. The 0.2% chlorhexidine-containing mouthwash was able to kill all strains of C. albicans in a shorter time compared to the thymol-containing mouthwash.
In this in vitro study, we assessed the antifungal effect of mouth rinses containing chlorhexidine and thymol.
The fungistatic activities of chlorhexidine- and thymol-containing mouth rinses were assessed by means of the minimum inhibitory concentration (MIC) and the fungicidal activity was determined by a time-kill assay.
The chlorhexidine-containing mouthwash was able to kill all strains of Candida albicans and Candida tropicalis in shorter times compared to the thymol-containing mouthwash. Hexidine showed an MIC of 1:32 for both Candida species, whereas Listerine respectively showed MICs of 1:8 and 1:16 for C. albicans and C. tropicalis.
Antimicrobial agents used in the study had good in vitro activity against the two Candida species. Mouth rinses containing chlorhexidine showed superior antifungal and fungicidal activities compared to the thymol-containing mouth rinse. Both antimicrobial agents may be suggested for use as topical antifungal agents.
A randomized, double-blind clinical study to assess the antimicrobial effects of a cetylpyridinium chloride mouth rinse on dental plaque bacteria
2009, Clinical Therapeutics
Background: Studies with cetylpyridinium chloride (CPC) mouth rinses that range from 1 use to 6 months of use have documented the clinical efficacy of these formulations on supragingival plaque and gingivitis.
Objective: The objective of the present study was to compare the effects of a commercially available mouth rinse containing 0.05% CPC versus a fluoride mouth rinse on the anaerobic bacteria found in dental plaque. Antimicrobial effects on the organisms of the supragingival plaque, a natural biofilm, were determined after 1 use and after 14 days of use of each mouth rinse.
Methods: After enrollment, adult subjects from China completed a 1-week washout period and provided baseline samples of supragingival plaque for analysis of anaerobic bacteria. Subjects were randomly assigned to receive a commercially available mouth rinse formulated with 0.05% CPC or a fluoride mouth rinse. Subjects were assigned to each group according to a computer-generated randomization sequence. They were instructed to rinse with 20 mL of either the CPC or the fluoride mouth rinse for 30 seconds. Microbiologic analyses of dental plaque samples were conducted 12 hours after the first use of assigned mouth rinse. Subjects were instructed to continue twice-daily rinsing with their assigned mouth rinse for the next 14 days in addition to brushing their teeth with a commercial fluoride toothpaste. Dental plaque samples for microbiologic analyses were collected on day 15; this was done 12 hours after the final use of the assigned mouth rinses. A dentist conducted oral examinations before each sample collection to evaluate hard and soft tissue health over the course of the study.
Results: The study included 117 adults (62 females, mean age, 28.70 years; 55 males, mean age, 30.41 years). Subjects rinsing with the CPC mouthwash (n = 58; mean age, 29.41 years) reported significant reductions in anaerobic bacteria versus those issued the fluoride rinse (n = 59; mean age, 29.61 years) 12 hours after 1 use and 12 hours after 14 days of use (P < 0.001). The mean percent reduction in anaerobic bacteria between the CPC mouth rinse and the fluoride mouth rinse was 29.98% after 1 use and 57.90% after 14 days of use. All enrolled subjects completed the study without any adverse events.
Conclusion: Use of the CPC mouth rinse was associated with significant reductions in the anaerobic bacteria of supragingival plaque compared with fluoride mouth rinse use in these adult subjects.
Sensitivity of Candida albicans biofilm cells grown on denture acrylic to antifungal proteins and chlorhexidine
2009, Archives of Oral Biology
Citation Excerpt :
Chlorhexidine is a cationic chlorophenyl bisbiguanide that binds to negatively charged surfaces. It has a broad spectrum of antimicrobial activity24 including C. albicans.25–27 Chlorhexidine is more effective than nystatin or amphotericin B in killing adherent C. albicans cells.27,28
Candida albicans cells form biofilms on polymeric surfaces of dentures and other prostheses introduced into the oral cavity. Many biofilm microorganisms exhibit resistance to antimicrobial agents; C. albicans cells may also develop resistance to naturally occurring antifungal peptides in human saliva including histatins (Hsts) and defensins (hBDs). Therefore, we evaluated Hst 5 activity on C. albicans biofilm cells compared to planktonic cells and measured whether surface treatment of denture acrylic with Hst 5, hBD-3, or chlorhexidine gluconate could inhibit in vitro biofilm development.
Acrylic disks were preconditioned with 500 μl saliva for 30 min, and inoculated with C. albicans cells (10⁶ cells/ml) for 1 h, at 37 °C. Non-adherent cells were removed by washing and disks and were incubated in YPD growth medium for 24, 48, and 72 h at 37 °C. Candidacidal assays were performed on 48-h-biofilms and on planktonically grown cells using Hst 5 (15.5, 31.25, and 62 μM). Cell adhesion was compared on disks pre-coated with 0.12% chlorhexidine gluconate, 50 μM Hst 5, or 0.6 μM hBD-3 after 24, 48, and 72 h growth.
No significant difference was observed in sensitivity to Hst 5 of biofilm cells compared to planktonic cells (p > 0.05). Pre-coating disks with hBD-3 did not inhibit biofilm development; however, Hst 5 significantly inhibited biofilm development at 72 h, while 0.12% chlorhexidine significantly inhibited biofilm development at all time intervals (p < 0.05).
C. albicans biofilm cells grown on denture acrylic are sensitive to killing by Hst 5. Surface coating acrylic with chlorhexidine or Hst 5 effectively inhibits biofilm growth and has potential therapeutic application.

View all citing articles on Scopus

^☆: Reprint requests: Giuseppe Pizzo, DDS,Via Nairobi, 31,90129 Palermo,Italy

^☆☆: 1079-2104/99/$8.00 + 0 7/13/94071

View full text

In vitro activities of antimicrobial agents against Candida species☆,☆☆

Abstract

Section snippets

Antimicrobial agents

RESULTS

DISCUSSION

Oral Surg Oral Med Oral Pathol

Oral Surg Oral Med Oral Pathol

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Oral Surg Oral Med Oral Pathol

Oral Surg Oral Med Oral Pathol

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Oral Surg Oral Med Oral Pathol

Opportunistic candidiasis: an epidemic of the 1980’s

Clin Infect Dis

The growing problem of mycoses in patients infected with the human im. munodeficiency virus

Rev Infect Dis

Fungal infections in cancer patients: an international autopsy survey

Eur J Clin Microbiol Infect Dis

Fungal infections in solid-organ transplantation

Clin Infect Dis

Nosocomial candidiasis: emerging species, reservoirs, and modes of transmission

Clin Infect Dis

Oral infections related to radiation and immunosuppressive therapy

J Clin Periodontol

Classification and diagnostic criteria for oral lesions in HIV infection

J Oral Pathol Med

Oral yeast infections in immunocompromised and seriously diseased patients

Acta Odontol Scand

Chemoprophylaxis for candidosis and aspergillosis in neutropenia and transplantation: a review and recommendations

J Antimicrob Chemother

Antifungal therapy in the oral cavity

Periodontol 2000

Resistance of Candida species to fluconazole

Antimicrob Agents Chemother

Emergence of azole drug resistance in Candida species from HIV-infected patients receiving prolonged fluconazole therapy for oral candidosis

J Antimicrob Chemother

In vitro antifungal resistance of oral Candida albicans strains in non-AIDS patients

Oral Microbiol Immunol

Emergence of oropharyngeal candidiasis caused by non-albicans species of Candida in HIV-infected patients

Eur J Epidemiol

Oligonucleotide fingerprinting of isolates of Candida species other than C. albicans and of atypical Candida species from human immunodeficiency virus-positive and AIDS patients

J Clin Microbiol

Candida krusei: biology, epidemiology, pathogenicity and clinical manifestations of an emerging pathogen

J Med Microbiol