Diversity of Staphylococcus aureus clones in wild mammals in Aragon, Spain, with detection of MRSA ST130-mecC in wild rabbits.

AIMS: To determine the Staphylococcus aureus carriage rate in wild mammals in Aragon, northern Spain, to analyse their antimicrobial resistance phenotype/genotype and to characterize the recovered isolates. METHODS AND RESULTS: Nasal and rectal swabs of 103 mammals were collected in Aragón during the period 2012-2015. Antimicrobial susceptibility, the presence of antimicrobial resistance genes and virulence factors were investigated. Molecular characterization was carried out by spa, MLST, agr and SCCmec. Staphylococcus aureus were recovered from 23 animals (22%). Four of the 23 S. aureus were methicillin-resistant S. aureus (MRSA). Three MRSA were mecC-positive and were isolated from European rabbits and were typed as t843 (ascribed to CC130). The remaining MRSA was a mecA-carrying isolate from European hedgehog, typed as ST1-t386-SCCmecIVa-agrIII and it harboured the blaZ, erm(C), ant(6)-Ia and aph(3´)-IIIa resistance genes. A high diversity of spa-types was detected among the 19 methicillin-susceptible S. aureus isolates, which showed high susceptibility to the antimicrobials tested. The tst gene and different combinations of staphylococcal enterotoxins were found. CONCLUSIONS: Staphylococcus aureus were detected in nasal and rectal samples of wild mammals. Wild rabbits could be a reservoir of mecC-MRSA. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides information on the presence and characteristics of S. aureus from mammals in a defined geographic region in Spain.

Methyl aminolevulinate photodynamic therapy for onychomycosis: a multicentre, randomized, controlled clinical trial.

BACKGROUND: Onychomycosis is a common fungal nail infection that responds poorly to antifungals. OBJECTIVE: To investigate the efficacy and safety of methyl aminolevulinate (MAL) photodynamic therapy (PDT) in the treatment of onychomycosis. METHODS: A multicentre (3), randomized, placebo-controlled clinical trial compared the effects of three sessions of urea (40%) plus conventional MAL-PDT with urea (40%) plus placebo (red light) photodynamic therapy (pPDT) in onychomycosis patients. Efficacy, both clinical (onychomycosis severity index, OSI) and microbiological, was blindly evaluated after 36 weeks of follow-up. RESULTS: Forty patients were analysed in the trial. Twenty-two received MAL-PDT and 18 pPDT. A complete response (OSI = 0) was observed for four patients (18.18%) in the MAL-PDT group and one (5.56%) in the pPDT group (NTT 7.92, 95% CI: 2.98-9.69, P = 0.23). A decrease in OSI score of over 75% (OSI75) was achieved by 40.91% of the patients in the MAL-PDT group and 16.67% in the pPDT group (P = 0.096). Microbiological cure was achieved by seven patients (31.82%) in the MAL-PDT group and two (11.11%) in the pPDT group (P = 0.178). MAL-PDT resulted in better rates of clinical response [OSI >75%: 53.85% vs. 18.75% (P =0.048)] and microbiological cure [41.56% vs. 7.14% (P = 0.037)] in non-dystrophic vs. dystrophic onychomycosis patients. No significant side-effects were reported. The limitations of the study were the reduced sample size and the unexpected efficacy of the control treatment, which was attributed to the 40% urea pre-treatment. CONCLUSION: This study did not show significant differences between urea 40% + MAL-PDT and urea 40% + pPDT in the treatment of onychomycosis. However, some results suggest that this treatment may constitute an alternative for dermatophyte and non-dermatophyte mould onychomycosis in patients not eligible for systemic treatment, particularly in the absence of total nail dystrophy.

In vitro effect photodynamic therapy with differents photosensitizers on cariogenic microorganisms.

BACKGROUND: Antimicrobial photodynamic therapy has been proposed as an alternative to suppress subgingival species. This results from the balance among Streptococcus sanguis, Streptococcus mutans and Candida albicans in the dental biofilm. Not all the photosensitizers have the same photodynamic effect against the different microorganims. The objective of this study is to compare in vitro the photodynamic effect of methylene blue (MB), rose Bengal (RB) and curcumin (CUR) in combination with white light on the cariogenic microorganism S. mutans, S. sanguis and C. albicans. RESULTS: Photodynamic therapy with MB, RB and CUR inhibited 6 log 10 the growth of both bacteria but at different concentrations: 0.31-0.62 µg/ml and 0.62-1.25 µg/ml RB were needed to photoinactivate S. mutans and S. sanguis, respectively; 1.25-2.5 µg/ml MB for both species; whereas higher CUR concentrations (80-160 µg/ml and 160-320 µg/ml) were required to obtain the same reduction in S. mutans and S. sanguis viability respectively. The minimal fungicidal concentration of MB for 5 log10 CFU reduction (4.5 McFarland) was 80-160 µg/ml, whereas for RB it ranged between 320 and 640 µg/ml. For CUR, even the maximum studied concentration (1280 µg/ml) did not reach that inhibition. Incubation time had no effect in all experiments. CONCLUSIONS: Photodynamic therapy with RB, MB and CUR and white light is effective in killing S. mutans and S. sanguis strains, although MB and RB are more efficient than CUR. C. albicans required higher concentrations of all photosensitizers to obtain a fungicidal effect, being MB the most efficient and CUR ineffective.

Usefulness of Photodynamic Therapy in the Management of Onychomycosis.

Onychomycosis, or fungal infection of the nails, is one of the most prevalent fungal diseases in the general population. Treatment is of limited effectiveness, tedious, and must be administered for long periods. Furthermore, systemic antifungal agents are associated with adverse effects. Photodynamic therapy (PDT) may prove to be a viable alternative in the treatment of superficial skin infections, including onychomycosis. We review articles relating to the usefulness of PDT in onychomycosis in both in vitro and in vivo settings and discuss the potential and limitations of various photosensitizing agents. In vivo, methylene blue and 5-aminolevulinic acid have led to cure rates in 80% and 43% of cases, respectively, at 12 months. Finally, based on data in the literature and our own experience, we propose a protocol of 3 PDT sessions, separated by an interval of 1 or 2 weeks, using methyl aminolevulinate 16% as a photosensitizing agent and red light (λ=630 nm, 37 J.cm(-2)). Each session is preceded by the topical application of urea 40% over several days. Clinical trials are needed to optimize PDT protocols and to identify those patients who will benefit most from this treatment.

Photodynamic fungicidal efficacy of hypericin and dimethyl methylene blue against azole-resistant Candida albicans strains.

Antimicrobial photodynamic therapy (aPDT) is an emerging alternative to treat infections based on the use of photosensitisers (PSs) and visible light. To investigate the fungicidal effect of PDT against azole-resistant Candida albicans strains using two PSs with a different mechanism of action, hypericin (HYP) and 1,9-dimethyl methylene blue (DMMB), comparing their efficacy and the reactive oxygen species (ROS) species involved in their cytotoxicity. Azole-resistant and the azole-susceptible C. albicans strains were used. Solutions of 0.5 and 4 McFarland inoculum of each Candida strain were treated with different concentrations of each PS, and exposed to two light-emitting diode light fluences (18 and 37 J cm⁻²). Mechanistic insight was gained using several ROS quenchers. The minimal fungicidal concentration of HYP for ≥3 log10 CFU reduction (0.5 McFarland) was 0.62 µmol l⁻¹ for most strains, whereas for DMMB it ranged between 1.25 and 2.5 µmol l⁻¹. Increasing the fluence to 37 J cm⁻² allowed to reduce the DMMB concentration. Higher concentrations of both PSs were required to reach a 6 log10 reduction (4 McFarland). H2O2 was the main phototoxic species involved in the fungicidal effect of HYP-aPDT whereas ¹O2 was more important for DMMB-based treatments. aPDT with either HYP or DMMB is effective in killing of C. albicans strains independent of their azole resistance pattern. HYP was more efficient at low fungal concentration and DMMB at higher concentrations.

Detection and characterization of a ST6 clone of vanB2-Enterococcus faecalis from three different hospitals in Spain.

Thirteen vancomycin-resistant and teicoplanin-susceptible Enterococcus faecalis isolates were recovered from unrelated patients in three Spanish hospitals from November 2009 to December 2010. All isolates carried the vanB2 gene, showed indistinguishable or closely-related PFGE patterns and were ascribed to the sequence type ST6 (included into the high-risk clonal-complex CC2). They showed a multiresistance phenotype (erythromycin, tetracycline, ciprofloxacin and high-level-resistance to streptomycin, gentamicin and kanamycin) and harboured the aac(6')-aph(2"), ant(6)-Ia, and tet(M)+/-tet(L) genes. All isolates produced gelatinase and harboured the gelE gene, but not the esp or hyl genes. The inclusion of the vanB2 gene into the Tn5382 transposon was demonstrated in one isolate. Clonal dissemination of vanB2-containing the E. faecalis strain is demonstrated.

Tinea capitis caused by Trichophyton soudanense and Microsporum audouinii in an adult: a case report.

This report presents a rare case of tinea capitis caused by Trichophyton soudanense and Microsporum audouinii in a 31-year-old woman from Senegal. The patient showed atrophic skin lesions causing cicatricial alopecia, scarring being caused by two aetiological agents uncommon in Spain.

Infection and colonisation due to Scedosporium in Northern Spain. An in vitro antifungal susceptibility and molecular epidemiology study of 60 isolates.

Since the latest taxonomical changes in the genus Scedosporium by Gilgado et al. in 2010, no species-specific studies on epidemiology and antifungal susceptibility patterns (AFSP) have so far been published. This study aimed to provide qualitative epidemiological data of Scedosporium spp. isolated from cystic fibrosis (CF) patients and immunocompromised patients from Northern Spain. Isolates were identified by using amplified fragment length polymorphism (AFLP), and species-specific AFSP were generated for all currently available antifungal compounds. AFLP was a useful tool for identification to species-level and for the discrimination of inter- and intra-patient isolates. Scedosporium prolificans represents the most prevalent species in the respiratory tract of CF patients and immunocompromised patients in Northern-Spain, followed by Pseudallescheria boydii, P. apiosperma, and P. ellipsoidea. CF patients were exclusively colonised with either P. boydii or S. prolificans. Patients were colonised over years exclusively with isolates affiliated to one species, but some patients were colonised with multiple strains with different AFSP. The sum of those co-colonising strains in one patient, may appear in vitro and in vivo as a multi-resistant S. prolificans isolate, as strains are morphologically identical and might therefore be regarded as only one strain. A majority of Scedosporium strains (with exception of S. prolificans) were found susceptible for voriconazole and micafungin.

Successful treatment of chromoblastomycosis of 36 years duration caused by Fonsecaea monophora.

We report a case of chromoblastomycosis in a 67-year-old female farmer, which involved a large (20 x 30 cm) cicatricial erythematous plaque on the inner side of her right thigh. The lesion was initially a small nodule which gradually extended over 36 years. Direct microscopic examination revealed a granulomatous lesion with muriform cells surrounded by giant cells. The mould recovered in cultures was dark olivaceous and identified as Fonsecaea monophora by ribosomal internal transcribe spacer (ITS) sequence data. The lesion was successfully cured after 4 months treatment with itraconazole, but there was a relapse.

Cavitary pneumonia caused by Nocardia otitidiscaviarum.

We report a case of cavitary pneumonia caused by N. otitidiscaviarum in a man with diabetes mellitus and thrombocytopenia treated with systemic corticosteroid. Taxonomic identification involved phenotypic testing and molecular identification that was carried out by DNA sequencing of the 16SrRNA gene.

First Spanish case of nocardiosis caused by Nocardia takedensis.

Nocardia takedensis is a recently described species isolated from soil. The first clinical isolate in Japan has recently been reported. This report describes the first clinical isolate of N. takedensis in Spain from a respiratory specimen.

A prospective international Aspergillus terreus survey: an EFISG, ISHAM and ECMM joint study.

OBJECTIVES: A prospective international multicentre surveillance study was conducted to investigate the prevalence and amphotericin B susceptibility of Aspergillus terreus species complex infections. METHODS: A total of 370 cases from 21 countries were evaluated. RESULTS: The overall prevalence of A. terreus species complex among the investigated patients with mould-positive cultures was 5.2% (370/7116). Amphotericin B MICs ranged from 0.125 to 32 mg/L, (median 8 mg/L). CONCLUSIONS: Aspergillus terreus species complex infections cause a wide spectrum of aspergillosis and the majority of cryptic species display high amphotericin B MICs.

[Mycological view of dermatophytes in humans.]. / Perspectiva micológica de los dermatofitos en el ser humano.

Dermatophytes are a group of closely related fungi that have the capacity to invade keratinized tissue (skin, hair, and nails) of humans and animals to produce an infection, dermatophytosis, commonly referred to as ringworm. Dermatophytoses are common of world wide: in the United States, Microsporum audouinii and Microsporum canis, once the major agents of tinea capitis, have been superseded by Trichophyton tonsurans. Since the 1950s, T. tonsuranshas advanced from Mexico and the Caribbean and is now the prevalent cause of tinea capitisin North America. M. canisis the prevalent agent of tinea capitis in many regions of the world, including Spain, at this moment. This could be related to close association of humans with their pets. M. canis is more prevalent in urban areas and Trichophyton mentagrophytes in rural ones. The superficial dermatophyte infections of the skin do not represent a single disease, their clinical appearance is dependent largely on the region of the body affected. There are more antifungal preparations available today than at any other time in medical history. Oral antifungals are indicated or required to treat hyperkeratotic areas such as nails, palms, soles and tinea capitis, patients with disabling or extensive disease, patients intolerant to or who have failed topical therapy, tose with chronic infection, those with granulomatous lesions and patients immunosuppressed by disease or by therapy. A successfull eradication of the fungi is now possible with relatively short treatment regimens.

Multicenter survey of in vitro antifungal resistance in yeasts of medical importance isolated from Spanish patients.

Twelve Spanish laboratories collected 325 yeast clinical isolates during a 30 day's period, among them 224 Candida albicans, 30 Candida glabrata, and 27 Candida parapsilosis. In vitro antifungal susceptibility to amphotericin B, ketoconazole, fluconazole and itraconazole was determined by an agar diffusion test (Neo-Sensitabs, Rosco, Denmark). All the isolates tested were susceptible in vitroto amphotericin B and nearly all (97.2%) to itraconazole. In vitrosusceptibility to fluconazole and ketoconazole was high (90.2% and 91.4% of isolates, respectively) but showed variations depending on the species tested. Resistance to fluconazole and ketoconazole was low in C. albicans (4% and 3%, respectively), but 30% of Candida guilliermondii and 36% of C. glabrata isolates were resistant to fluconazole. Ketoconazole resistance was observed in 40% of C. glabrata, and 17% of Candida tropicalis. Resistance to antifungal drugs is very low in Spain and it is related to non-C. albicans isolates.

[In vitro activity of fluconazole, voriconazole and posaconazole against Candida spp]. / Actividad in vitro de fluconazol, voriconazol y posaconazol frente a Candida spp.

Voriconazole and posaconazole are two new triazoles with broad-spectrum antifungal activity. The in vitro activity of these agents was compared with that of fluconazole against 113 clinical isolates of Candida spp. exhibiting a wide range of MICs for fluconazole (< or =0.125 to >64 mg/l). They included 22 C. albicans isolates, 22 C. glabrata, 20 C. tropicalis, 15 C. dubliniensis, 13 C. parapsilosis, 13 C. krusei, 4 C. lusitaniae, 3 C. guilliermondii and 1 C. famata. MICs were determined by broth microdilution test performed according to the National Committee for Clinical Laboratory Standards' M27-A document. Voriconazole and posaconazole were quite active against all Candida spp. isolates, with MIC(90) of 0.25 and 0.5 mg/l, respectively. Fluconazole MIC(50) and MIC(90) were 1 and 32 mg/l. Voriconazole and posaconazole showed in vitro activity against fluconazole-susceptible and -resistant Candida isolates. Higher voriconazole and posaconazole MICs were observed in isolates exhibiting higher fluconazole MICs (>/=16 mg/l) than in those of fluconazole-susceptible isolates (MIC(90) for voriconazole and posaconazole in the fluconazole-resistant isolates were 0.5 mg/l versus 0.06 mg/l for voriconazole and 0.25 mg/l for posaconazole in the fluconazole-susceptible strains). C. dubliniensis was the most susceptible species, and voriconazole the most active antifungal agent against all Candida spp. tested.

[Comparative study of Salmonella and Campylobacter infections in Huesca. 1996-1999]. / Estudio comparado de infección por Salmonella y Campylobacter en Huesca. 1996-1999.

BACKGROUND: The Salmonella and Campylobacter genera are the main bacterial agents causing enteritis in humans in the developed countries. The purpose of this paper is to study the descriptive characteristics of the cases of infection by Salmonella and by Campylobacter and to describe their trend and seasonal variance in the province of Huesca for the 1996-1999 period. METHOD: The sources of information were the Hospital Microbiological Laboratories (HML's) which conduct coproculture analyses of both outpatients and inpatients. The municipality is taken as the reference unit for plotting the space distribution. For the study of the trend, a multiplicative determinist model is used for grouping the data into four-week periods. RESULTS: A total of 781 cases of infection by Salmonella and 654 cases of infection by Campylobacter have been included, respectively showing mean annual rates of 95 and 79.5 x 10(5) inhabitants. The enteritis by Campylobacter occurs mainly in children under five years of age (73.4%), as compared to the 35.5% of the cases of salmonellosis. The situation is the reverse in the case of the hospital admission percentages, hospitalization due to salmonellosis hence being nearly five times greater. Both of these genera are of a clear-cut seasonal nature, showing a marked peak in the month of August. CONCLUSIONS: A growing trend in the number of positive isolations for both of these organisms and a clear seasonality in the summertime have been found.