Estelle Cateau

In vitro activity of terpenes against Candida biofilms

The antibiofilm activity of 10 terpenes was tested in vitro against three Candida species by 24-h treatment of biofilms aged 1-5 days. Treatment of 24-h-old Candida albicans biofilms with carvacrol, geraniol or thymol (0.06%) resulted in >80% inhibition. Carvacrol (0.03%) inhibition was > or =75% independent of the age of the C. albicans biofilm. Carvacrol (0.125%) inhibition was >75% against Candida glabrata and Candida parapsilosis biofilms. Geraniol (> or =0.125%) and thymol (0.06% or 0.125%) inhibition was >75% against C. parapsilosis biofilms whatever their age. This study demonstrates the antibiofilm activity of terpenes and points out the exceptional efficiency of carvacrol, geraniol and thymol, which could represent candidates in the treatment of candidiasis associated with medical devices.

In vitro efficacies of caspofungin or micafungin catheter lock solutions on Candida albicans biofilm growth

OBJECTIVES Caspofungin and micafungin belong to the echinocandins; the mechanism of action of echinocandins is based on the inhibition of (1,3)-beta-D-glucan synthase. The aim of this study was to investigate in vitro the optimal antifungal lock treatment details against a Candida albicans biofilm. METHODS An in vitro model of a C. albicans (ATCC 3153 or ATCC 66396) biofilm associated with 100% silicone catheters was used. The effectiveness of the antifungal treatment was assayed against biofilms aged 12 h or 5 days, after exposure to caspofungin (2 mg/L) or micafungin (5 mg/L) for 12 h. The durability of the reduction in the biofilm metabolic activity was investigated (1-3 days after echinocandin treatment). The efficacy of caspofungin and micafungin was determined by evaluating a significant decrease (P < 0.0001) in the metabolic activity of biofilm yeasts. RESULTS The results showed that the tested antifungal agents used as lock solution significantly (P < 0.0001) reduced the metabolic activity of C. albicans, whatever the biofilm maturation stage (12 h or 5 days old biofilms). The reduction in the metabolic activity of biofilm yeasts was maintained, even after 48 h. CONCLUSIONS These data suggest that caspofungin (2 mg/L) and micafungin (5 mg/L) could represent good candidates for the reduction or control of fungal biofilms associated with silicone medical devices, as part of an antifungal lock. They were able to induce a significant and persistent reduction in the yeast metabolic activity of intermediate and mature biofilms, 12 h and 5 days old, respectively, when used as catheter lock solutions.

Possible role of azole and echinocandin lock solutions in the control of Candida biofilms associated with silicone

Until now, management of candidiasis related to implanted devices has remained problematic. The aim of this study was to investigate antifungal lock strategies against Candida albicans and Candida glabrata biofilms in vitro. Three antifungal agents were used against eight C. albicans and six C. glabrata clinical strains isolated from infected catheters. Caspofungin and micafungin, both echinocandins, as well as the azole posaconazole were tested. An in vitro model of Candida biofilm on 100% silicone catheters was used. Efficacy of the antifungal lock was tested against biofilms aged 12h and 5 days following exposure to caspofungin (5mg/L and 25mg/L), micafungin (5mg/L and 15 mg/L) and posaconazole (10mg/L) for 12h. Persistence of antibiofilm activity was investigated 1-3 days following drug elimination. Antifungal lock was considered effective in the event of a significant decrease (P<0.001) in the metabolic activity of the biofilm yeast. The results showed that micafungin had significant inhibitory effectiveness against young and mature C. albicans and C. glabrata biofilms. Moreover, this activity appeared to persist for up to 3 days. Caspofungin displayed similar activity against all C. albicans biofilms, but the activity was less persistent for C. glabrata biofilms. Posaconazole was less effective against C. albicans biofilms, but its activity was sustained. Echinocandin lock therapy could significantly enhance the management of candidiasis in patients with indwelling catheters by combating biofilms and enabling device maintenance in situ.

Free-living amoebae: what part do they play in healthcare-associated infections?

Free-living amoebae (FLA) are ubiquitous protozoa that do not require a host organism for survival. They are found in natural environments such as water or soil, and man-made environments including tap water or swimming pools, where they may interact with other micro-organisms, including bacteria, fungi and viruses. FLA can harbour micro-organisms including those found in hospital water systems, offering them protection against hostile conditions, providing a vehicle of dissemination, and enabling them to prepare for subsequent survival in macrophages. The interaction between Legionella pneumophila and FLA has been studied extensively; subsequent investigations have shown that FLA may serve as a reservoir for other bacteria including mycobacteria, Pseudomonas aeruginosa, Acinetobacter baumannii, or even fungi and viruses. Amoebae found in hospital water systems can serve as a reservoir of potential pathogens and thus be indirectly related to healthcare-associated infections.

Acanthamoeba sp. promotes the survival and growth of Acinetobacter baumanii

Acinetobacter baumanii, which may be found in water, is an important emerging hospital-acquired pathogen. Free-living amoebae can be recovered from the same water networks, and it has been shown that these protozoa may support the growth of other bacteria. In this paper, we have studied potential relationships between A. baumanii and Acanthamoeba species. Two strains of A. baumanii isolated from hospital water were co-cultivated with the trophozoites or supernatants of two free-living amoebae strains: Acanthamoeba castellanii or Acanthamoeba culbertsoni. Firstly, the presence of the amoebae or their supernatants induced a major increase in A. baumanii growth, compared with controls. Secondly, A. baumanii affected only the viability of A. culbertsonii, with no effect on A. castellanii. Electron microscopy observations of the cultures investigating the bacterial location in the protozoa showed persistence of the bacteria within cyst wall even after 60 days of incubation. In our study, the survival and growth of A. baumanii could be favored by Acanthamoeba strains. Special attention should consequently be paid to the presence of free-living amoebae in hospital water systems, which can promote A. baumanii persistence.

Stenotrophomonas maltophilia and Vermamoeba vermiformis relationships: bacterial multiplication and protection in amoebal-derived structures.

Stenotrophomonas maltophilia, a bacteria involved in healthcare-associated infections, can be found in hospital water systems. Other microorganisms, such as Free Living amoebae (FLA), are also at times recovered in the same environment. Amongst these protozoa, many authors have reported the presence of Vermamoeba vermiformis. We show here that this amoeba enhances S. maltophilia growth and harbors the bacteria in amoebal-derived structures after 28 days in harsh conditions. These results highlight the fact that particular attention should be paid to the presence of FLA in hospital water systems, because of their potential implication in survival and growth of pathogenic bacterial species.

Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identification of Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review

Microscopy is the reference standard for routine laboratory diagnosis in faecal parasitology but there is growing interest in alternative methods to overcome the limitations of microscopic examination, which is time-consuming and highly dependent on an operators skills and expertise. Compared with microscopy, DNA detection by PCR is simple and can offer a better turnaround time. However, PCR performances remain difficult to assess as most studies have been conducted on a limited number of positive clinical samples and used in-house PCR methods. Our aim was to evaluate a new multiplex PCR assay (G-DiaParaTrio; Diagenode Diagnostics), targeting Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica. To minimize the turnaround time, PCR was coupled with automated DNA extraction (QiaSymphony; Qiagen). The PCR assay was evaluated using a reference panel of 185 samples established by routine microscopic examination using a standardized protocol including Ziehl-Neelsen staining and adhesin detection by ELISA (E. histolytica II; TechLab). This panel, collected from 12 French parasitology laboratories, included 135 positive samples for G. intestinalis (n = 38), C. parvum/C. hominis (n = 26), E. histolytica (n = 5), 21 other gastrointestinal parasites, together with 50 negative samples. In all, the G-DiaParaTrio multiplex PCR assay identified 38 G. intestinalis, 25 C. parvum/C. hominis and five E. histolytica leading to sensitivity/specificity of 92%/100%, 96%/100% and 100%/100% for G. intestinalis, C. parvum/C. hominis and E. histolytica, respectively. This new multiplex PCR assay offers fast and reliable results, similar to microscopy-driven diagnosis for the detection of these gastrointestinal protozoa, allowing its implementation in routine clinical practice.

Relationships between free living amoebae and Exophiala dermatitidis: a preliminary study

Free living amoebae can play a role as reservoirs for pathogens isolated from hospital water. We have investigated the potential interactions between two protozoa (Acanthamoeba castellanii and Hartmanella vermiformis) that may be recovered from hospital water tips and Exophiala dermatitidis, a black yeast often recovered from water sources. We showed that the presence of trophozoites or supernatants of culture of H. vermiformis increased fungal growth, whereas the same phenomenon was observed only with the supernatant of A. castellanii cultures. These preliminary results highlight the fact that the recovering of free-living amoebae in hospital water taps could lead to the development of fungal nosocomial pathogens.

Hartmanella vermiformis can be permissive for Pseudomonas aeruginosa

Aims:  The amoebae of the genus Hartmanella are frequently recovered from hospital water taps, whereas Pseudomonas aeruginosa is often implicated in nosocomial infections. Previous works suggested that free living amoebae can act as vehicles of bacterial transmission. The present work investigates the relationships between a strain of Hartmanella vermiformis and three strains of P. aeruginosa: a reference strain, a strain from a patient and an environmental strain.

Enterobius vermicularis in the kidney: an unusual location.

A woman was admitted to hospital with abdominal pain. A large kidney stone was recovered and a nephrectomy was performed. Histology revealed the unusual presence of multiple Enterobius vermicularis ova. However, no other parasitic element was recovered on further investigations.