Exploring the dynamics of mixed-species biofilms involving Candida spp. and bacteria in cystic fibrosis.

Cystic fibrosis (CF) is an inherited disease that results from mutations in the gene responsible for the cystic fibrosis transmembrane conductance regulator (CFTR). The airways become clogged with thick, viscous mucus that traps microbes in respiratory tracts, facilitating colonization, inflammation and infection. CF is recognized as a biofilm-associated disease, it is commonly polymicrobial and can develop in biofilms. This review discusses Candida spp. and both Gram-positive and Gram-negative bacterial biofilms that affect the airways and cause pulmonary infections in the CF context, with a particular focus on mixed-species biofilms. In addition, the review explores the intricate interactions between fungal and bacterial species within these biofilms and elucidates the underlying molecular mechanisms that govern their dynamics. Moreover, the review addresses the multifaceted issue of antimicrobial resistance in the context of CF-associated biofilms. By synthesizing current knowledge and research findings, this review aims to provide insights into the pathogenesis of CF-related infections and identify potential therapeutic approaches to manage and combat these complex biofilm-mediated infections.

Evaluation of mixed biofilm production by Candida spp. and Staphylococcus aureus strains co-isolated from cystic fibrosis patients in northwest Algeria.

Cystic fibrosis patients' lungs are chronically colonized by multiple microbial species capable of forming biofilms. This study aimed to characterize the polymicrobial biofilm formed by Candida spp. and S. aureus, co-isolated from sputum samples of cystic fibrosis patients regarding microbial density, metabolic activity, and structure. 67 samples from 28 patients were collected with a 96% alteration rate. 34% showed alterations by both Candida spp. and Gram-positive bacteria, predominantly Candida spp. and S. aureus in 77% of cases, accounting for 6 associations. Biofilm biomass was quantified using the crystal violet assay, and metabolic activity was assessed using the MTT reduction assay. Scanning electron microscopy analyzed the C. tropicalis/S. aureus24 biofilm architecture. Candida spp. isolates demonstrated the ability to form mixed biofilms with S. aureus. The C. tropicalis/S. aureus24 association exhibited the highest production of biofilm and metabolic activity, along with the C. albicans17/C. rugosa/S. aureus7 in both single and mixed biofilms.

Initial characteristics of cystic fibrosis in Algeria: Description of 34 pediatric cases.

BACKGROUND: Cystic fibrosis (CF) is a rare disease in Algeria, and its prognosis is poor in developing countries. The clinical and demographic knowledge of Algerian pediatric patients diagnosed with CF is incomplete due to the nonexistence of a national medical registry. Hence, the present study is the first Algerian multicentre study on CF. METHODS: This retrospective study was conducted in western Algeria. Over 1 year, the study included all pediatric patients with a confirmed diagnosis of CF in the pediatric hospital of Oran. Patient characteristics, clinical manifestations, and the prescribed treatment were reported. RESULTS: Thirty-four children (16 boys and 18 girls) participated in this study. Only 15 were diagnosed before the age of 6 months. The sweat chloride test was positive in all patients. Respiratory manifestations were found in all patients, chronic diarrhoea in 29 of them, and growth retardation in 10. Moreover, 25 (73.5%) had low to low intermediate socioeconomic levels. After diagnosis, respiratory complications marked the evolution of the 34 patients, with bronchial congestion observed in 33 of them, while 10 (29.4%) patients presented severe bronchopneumonia and 4 (11.8%) were affected by asthma. Consequently,  three (8.8%) died at an average age of 9 years mainly because of respiratory failure. CONCLUSION: The prognosis of CF is poor in Algeria compared to other developed countries due to the longer diagnostic delay and limited therapeutic alternatives. This representative subset of Algerian pediatric patients with CF will serve as a reference for future studies on CF in Algeria.

In vitro evaluation of biofilm formation by Candida parapsilosis and Enterobacter cloacae. Scanning electron microscopy and efficacy of antimicrobial combinations study.

Fungal-bacterial infections are being increasingly recognized in clinical settings, and the interaction between these species in polymicrobial biofilms often lead to infections that are highly resistant to treatment. In this in vitro study, we analyzed the formation of mixed biofilms using clinically isolated Candida parapsilosis and Enterobacter cloacae. Additionally, we assessed the potential of conventional antimicrobials, both alone and in combination, for treating polymicrobial biofilms built by these human pathogens. Our results demonstrate that C. parapsilosis and E. cloacae are capable of forming mixed biofilms, as confirmed by scanning electron microscopy. Interestingly, we found that colistin alone or in combination with antifungal drugs was highly effective reducing up to 80% of the total biomass of polymicrobial biofilms.

Influence of ascorbic acid and α-tocopherol on the autoxidation and in vitro antifungal activity of amphotericin B.

BACKGROUND AND PURPOSE: Amphotericin B (AmB) is the standard treatment for systemic fungal infections; however, the formation of reactive oxygen species reduces the efficacy and stability of this molecule. The present study aimed to evaluate the effect of the combination of AmB with ascorbic acid and α-tocopherol on its autoxidation and antifungal activity. MATERIALS AND METHODS: The antifungal activity against Candida albicans was evaluated by the viable cell counting method and checking their morphological changes with a scanning electron microscope. Monomer state of AmB was assessed by scanning the UV absorbance in the range of 300-450 nm and the lipid peroxidation was measured using quantification of thiobarbituric acid reactive-substances (TBARS). RESULTS: Based on the findings, the addition of ascorbic acid (3×102 µg/mL) and α-tocopherol (16 µg/mL) to the reaction medium of AmB increased its antifungal  activity while maintaining its molecular stability. Moreover, the level of TBARS formed in the reaction medium of AmB was significantly reduced after combination  with ascorbic acid and α-tocopherol. CONCLUSION: Given their availability, their anti-free radical activity, and their low toxicity, the incorporation of ascorbic acid and α-tocopherol into the reaction medium of AmB seems to be a promising approach to obtain an effective antifungal formulation.

Erratum: Touil, H.F.Z. et al. Optimum Inhibition of Amphotericin-B-Resistant Candida albicans Strain in Single- and Mixed-Species Biofilms by Candida and Non-Candida Terpenoids. Biomolecules 2020, 10, 342.

The authors wish to make the following correction to this paper [1] [...].

Optimum Inhibition of Amphotericin-B-Resistant Candida albicans Strain in Single- and Mixed-Species Biofilms by Candida and Non-Candida Terpenoids.

Candida albicans is one of the most common human fungal pathogens and represents the most important cause of opportunistic mycoses worldwide. Surgical devices including catheters are easily contaminated with C. albicans via its formation of drug-resistant biofilms. In this study, amphotericin-B-resistant C. albicans strains were isolated from surgical devices at an intensive care center. The objective of this study was to develop optimized effective inhibitory treatment of resistant C. albicans by terpenoids, known to be produced naturally as protective signals. Endogenously produced farnesol by C. albicans yeast and plant terpenoids, carvacrol, and cuminaldehyde were tested separately or in combination on amphotericin-B-resistant C. albicans in either single- or mixed-infections. The results showed that farnesol did not inhibit hyphae formation when associated with bacteria. Carvacrol and cuminaldehyde showed variable inhibitory effects on C. albicans yeast compared to hyphae formation. A combination of farnesol with carvacrol showed synergistic inhibitory activities not only on C. albicans yeast and hyphae, but also on biofilms formed from single- and mixed-species and at reduced doses. The combined terpenoids also showed biofilm-penetration capability. The aforementioned terpenoid combination will not only be useful in the treatment of different resistant Candida forms, but also in the safe prevention of biofilm formation.

Cosmetics Preservation: A Review on Present Strategies.

Cosmetics, like any product containing water and organic/inorganic compounds, require preservation against microbial contamination to guarantee consumer's safety and to increase their shelf-life. The microbiological safety has as main goal of consumer protection against potentially pathogenic microorganisms, together with the product's preservation resulting from biological and physicochemical deterioration. This is ensured by chemical, physical, or physicochemical strategies. The most common strategy is based on the application of antimicrobial agents, either by using synthetic or natural compounds, or even multifunctional ingredients. Current validation of a preservation system follow the application of good manufacturing practices (GMPs), the control of the raw material, and the verification of the preservative effect by suitable methodologies, including the challenge test. Among the preservatives described in the positive lists of regulations, there are parabens, isothiasolinone, organic acids, formaldehyde releasers, triclosan, and chlorhexidine. These chemical agents have different mechanisms of antimicrobial action, depending on their chemical structure and functional group's reactivity. Preservatives act on several cell targets; however, they might present toxic effects to the consumer. Indeed, their use at high concentrations is more effective from the preservation viewpoint being, however, toxic for the consumer, whereas at low concentrations microbial resistance can develop.

Risk assessment for the spread of Candida sp. in dental chair unit waterlines using molecular techniques.

AIMS: The aim of this study was to evaluate the presence of yeasts in dental chair unit waterlines (DCUWLs) and to test their ability to form biofilms. MATERIALS AND METHODS: Eighteen dental waterlines were analysed by culture in liquid Sabouraud in order to allow the quantification and the purification of isolated yeasts from their internal surfaces. All isolates were identified by standard laboratory procedures, including CHROMagar Candida medium for orientation, commercial yeast identification system Api Candida, MALDI-TOF MS and DNA sequencing. To evaluate their kinetics of antifungal susceptibility during different phases of biofilm formation, these yeasts were subjected to three antifungal agents. RESULTS: From the 18 DCUWLs studied, 10 were altered (55.56%). Eleven strains of Candida sp. [Candida albicans (2), Candida guilliermondii (5) and Candida glabrata (4)] and two species of non-Candida; Rhodotorula spp. (1) and Trichosporon spp. (2) were identified. The majority of yeasts in planktonic form were susceptible to amphotericin B, caspofungin and voriconazole, except C. albicans was resistant to voriconazole. In the biofilm form, caspofungin was the most effective antifungal agent for all isolated strains. For the other antifungal agents, sessile cells were resistant. CONCLUSION: Several types of yeasts were identified; the most frequently isolated genus was Candida. The majority of these yeasts had the ability to form biofilms and resisted antifungal agents used in this study.

Chemical composition, antibacterial, antifungal and antioxidant activities of Algerian Eryngium tricuspidatum L. essential oil.

This study describes the chemical composition and the antibacterial, antifungal and antioxidant activities of the essential oil extracted from aerial parts of the Algerian Eryngium tricuspidatum L., obtained by hydrodistillation and analysed by using the combination of gas chromatography (GC) and GC/mass spectrometry. A total of 63 compounds were identified accounting for 93.1% of the total oil. Chemical composition of oil was characterised by a high proportion of oxygenated sesquiterpenes (49.6%) among which α-bisabolol (32.6%) was the predominant compound. The sesquiterpene hydrocarbons represent the second major fraction (31.9%) with α-curcumene (6.5%) being the predominant one. Antibacterial and antifungal activities of the oil were tested using the micro-well determination of minimum inhibitory concentration (MIC) assay against eleven bacteria and two Candida species. It was found that the aerial parts of E. tricuspidatum exhibited interesting antibacterial and anticandidal activities (MIC = 9 µg/mL against several strains of bacteria and MIC = 4.6 µg/mL against Candida albicans). The antioxidant effect of this oil was evaluated using the 2,2-diphenyl-l-1-picrylhydrazil (DPPH) and ferric reducing antioxidant power (FRAP) assays. Results revealed significant activities (DPPH method: IC50 = 510 µg/mL; FRAP assay: reducing power of oil increases from 0.0188 at 5 µg/mL to 0.5016 at 1000 µg/mL).

Assessment of the types of catheter infectivity caused by Candida species and their biofilm formation. First study in an intensive care unit in Algeria.

Nosocomial candidiasis remains a potential risk in intensive care units (ICUs), wherein Candida albicans is most responsible for its occurrence. Equally, non-C. albicans species, especially C. glabrata, are also involved. These infections are frequently associated with biofilms that contaminate medical devices, such as catheters. These biofilms constitute a significant clinical problem, and cause therapeutic failures, because they can escape the immune response and considerably decrease sensitivity to antifungal therapy. The diagnosis of catheter-related candidiasis is difficult; however, the differentiation between an infection of the catheter (or other medical implant) and a simple contamination is essential to start an antifungal treatment. Among the methods used for this type of study is the Brun-Buisson method, but this method only examines the infectivity of catheters caused by bacteria. For this reason, we wanted to adapt this method to the yeast cells of Candida spp. To assess the various types of infectivity of catheters (contamination, colonization, or infection) and their corresponding rates, as well as the responsible yeast species, we conducted our study, between February 2011 and January 2012, in the ICU at the University Hospital Center of Sidi Bel Abbes, Algeria; during this study, we took photographic images of the tongue of one patient and of that patient's implanted orobronchial catheter. In addition, catheters contaminated by C. albicans biofilms were observed by scanning electron microscopy.

Antifungal activity of the Algerian Lawsonia inermis (henna).

CONTEXT: Lawsonia inermis Linn. (Lythraceae) or henna has been used since the earliest times as a medicine, preservative, and cosmetic. It has long been recommended in traditional medicine as an astringent, purgative, and abortifacient. OBJECTIVE: Lawsone and six extracts of L. inermis plant, used by Algerian traditional healers to treat infectious diseases, were screened for their antifungal activity against filamentous fungi. MATERIALS AND METHODS: Water and five organic extracts - DMSO, ethanol, chloroform, ethyl acetate, and di-ethyl ether - of L. inermis leaves, collected in the area of Adrar (Algeria), were prepared by soaking 25 g of powdered plant in 100 mL of solvent. The extracts were screened for antifungal activity using the poisoned food technique against five filamentous fungi. RESULTS: Results demonstrated that the best yield (8.03%) was obtained with the ethanol extract. The commercial lawsone showed potentially interesting MICs against the strains Fusarium oxysporum (12 µg/mL) and Aspergillus flavus (50 µg/mL). The ethanol extract showed the only interesting MIC (230 µg/mL of crude extract) against the strain F. oxysporum compared with other extracts. DISCUSSION AND CONCLUSION: These results suggest that the Algerian L. inermis plant has antifungal activity that can be related to the presence of lawsone in the leaves plant. The results can be exploited largely in research of new antifungal drugs.