Many countries have seen a two-wave pattern in reported cases of coronavirus disease-19 during the 2020 pandemic, with a first wave during spring followed by the current second wave in late summer and autumn. Empirical data show that the characteristics of the effects of the virus do vary between the two periods. Differences in age range and severity of the disease have been reported, although the comparative characteristics of the two waves still remain largely unknown. Those characteristics are compared in this study using data from two equal periods of 3 and a half months. The first period, between 15th March and 30th June, corresponding to the entire first wave, and the second, between 1st July and 15th October, corresponding to part of the second wave, still present at the time of writing this article. Two hundred and four patients were hospitalized during the first period, and 264 during the second period. Patients in the second wave were younger and the duration of hospitalization and case fatality rate were lower than those in the first wave. In the second wave, there were more children, and pregnant and post-partum women. The most frequent signs and symptoms in both waves were fever, dyspnea, pneumonia, and cough, and the most relevant comorbidities were cardiovascular diseases, type 2 diabetes mellitus, and chronic neurological diseases. Patients from the second wave more frequently presented renal and gastrointestinal symptoms, were more often treated with non-invasive mechanical ventilation and corticoids, and less often with invasive mechanical ventilation, conventional oxygen therapy and anticoagulants. Several differences in mortality risk factors were also observed. These results might help to understand the characteristics of the second wave and the behaviour and danger of SARS-CoV-2 in the Mediterranean area and in Western Europe. Further studies are needed to confirm our findings.

We provide an overview of opportunistic fungal infections caused by Acremonium (Cephalosporium) species and discuss the classification of these species as well as the diagnosis and treatment of acremonium infections. We used a microdilution broth method to compare in vitro susceptibilities and minimum inhibitory concentrations and minimum fungicidal concentrations of amphotericin B, miconazole, itraconazole, 5-fluorocytosine, fluconazole, and ketoconazole for 33 clinical and environmental isolates of Acremonium. In general, the isolates tested displayed little susceptibility to the antifungals tested. Fluconazole and 5-fluorocytosine were ineffective in all cases. The efficacy of the remaining drugs was dependent on the strain. Amphotericin B showed the best results.

The MICs of amphotericin B, miconazole, ketoconazole, flucytosine, itraconazole and fluconazole for 19 isolates of Fusarium oxysporum, 16 Fusarium solani, seven Fusarium verticilliodes, four Fusarium proliferatum, four Fusarium dimerum, three Fusarium equiseti, and one each of the following species: Fusarium graminearum, Fusarium chlamydosporum, Fusarium semitectum, Fusarium avenaceum and Fusarium subglutinans were determined by a broth microdilution method. Thirty-eight of these isolates were of clinical origin and 20 from environmental sources. In general, Fusarium spp. strains showed resistance to all the antifungals tested. However, the most active agent was amphotericin B. Fluconazole and flucytosine were not active against any of the isolates tested. A correlation study of in-vitro testing with in-vivo outcome of amphotericin B of the cases of disseminated fusarium infections published is reported.

Although rarely, Arcobacter spp. have been associated with diarrhoea and bacteraemia. We report a persistent case in a healthy 26-year-old Spanish male of bloody diarrhoea, which was attributed to Campylobacter but in fact was caused by Arcobacter cryaerophilus , as determined by sequencing of the rpoB gene. The isolate was re-identified by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) and genotyped for five putative virulence genes and for seven genes included in the Arcobacter multilocus sequence typing database. The low score obtained by MALDI-TOF indicates the need to complement the database with more isolates. Only the ciaB gene, which encodes for an invasin, was detected. Despite the isolate belonging to a new sequence type, three of the alleles ( glnA , pgm and tkt ) had been found previously in isolates from faeces of patients with diarrhoea. This study, together with the reviewed literature, indicates that Arcobacter can produce bacteraemia and that the isolation from patients with diarrhoea range from 0.11% to 1.25%. This study also demonstrates that Arcobacter species are confused with Campylobacter spp., as previously suggested. This is one of the factors that leads to underestimation of their incidence together with the use of inappropriate detection and identification methods.

The MICs and minimum fungicidal concentrations (MFCs) of amphotericin B, miconazole, itraconazole, ketoconazole, fluconazole, and flucytosine for 52 isolates of Paecilomyces species were evaluated by the broth microdilution method, largely based on the recommendations of the National Committee for Clinical Laboratory Standards (document M27-A). The fungal isolates tested included 16P. variotii, 11 P. lilacinus, 9 P. marquandii, 6 P. fumosoroseus, 4 P. javanicus, and 2 P. viridis isolates and 1 isolate of each of the following species: P. carneus, P. farinosus, P. fulvus, and P. niveus. The MFCs and the MICs at which 90% of isolates were inhibited (MIC90s) for the six antifungal agents were remarkably high; the MIC50s indicated that amphotericin B, miconazole, itraconazole, and ketoconazole had good activities, while fluconazole and flucytosine demonstrated poor efficacy. The ranges of the MICs were generally wider and lower than those of the MFCs. There were significant susceptibility differences among the species. All species with the exception of P. variotii were highly resistant to fluconazole and flucytosine; P. variotii was susceptible to flucytosine. Amphotericin B and the rest of the azoles showed good activity against P. variotii, while all the antifungal agents assayed showed low efficacy against P. lilacinus.

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An evaluation of broth dilution antifungal susceptibility tests was performed by determining both the microand macrodilution MICs of amphotericin B, fluconazole, ketoconazole, 5-fluorocytosine, miconazole, and itraconazole against representative species of opportunistic hyphomycetes (Fusarium spp. and Cladosporium [Cladophialophora] spp.) and ascomycetes (Chaetomium spp.). A total of 78 strains were tested, the majority of them twice and some three times on different days. Both methods were performed according to the recommendations of the National Committee for Clinical Laboratory Standards (Document M27-P), with the exception of the temperature of incubation, which was 25؇C in our case. A spectrophotometric method for inoculum preparation, RPMI 1640 medium buffered with morpholinepropanesulfonic acid (pH 7.0), and an additive drug dilution procedure were used. The MICs obtained by the two methods were read after 48, 72, and 96 h of incubation for Fusarium spp. and after 72, 96, and 120 h for the remaining isolates. The kappa test was used to calculate the degree of agreement. Considering the three fungal groups together, a good agreement between the results of both tests was observed with almost all the drugs at the different incubation times. There were no cases of poor agreement. The highest level (kappa index ‫؍‬ 1) was observed with ketoconazole at the second-day reading. These results support the further evaluation of the broth microdilution test as an alternative to the reference broth macrodilution susceptibility test.

Infections caused by dermatophytes are probably the most common communicable fungal diseases affecting humans. Although a wide variety of both topically and systemically administered compounds with activities against these fungi are available, some of these infections are still difficult to resolve completely and remissions and relapses are often observed (3, 9). Remissions and relapses are more likely due to the inability of the antifungal drug to penetrate the site of infection rather than to the intrinsic resistance of the fungus. In recent years there has been growing interest in the development of a reference method for in vitro antifungal susceptibility testing of dermatophytes. The studies that have been described in the literature (1,8,11,12,14) are based on slight modifications of the broth macro-and microdilution techniques for molds recommended by the NCCLS (document M38-P) (13). However, these methods may not be the most practical procedures for use in the routine clinical laboratory, mainly due to the need for the subjective determination of endpoints.The addition of an oxidation-reduction colorimetric indicator, such as Alamar Blue, which changes from blue to red in the presence of metabolically active growing organisms, has been shown to facilitate the reading of MIC endpoints (4, 7) and could be an alternative to broth macro-and microdilution techniques for molds for use in a general laboratory. The Sensititre YeastOne Colorimetric Antifungal panel (Trek Diagnostic Systems Ltd., East Grinstead, United Kingdom) is a commercial microdilution plate that is already available and that contains dried serial dilutions of five antifungal agents (amphotericin B, flucytosine, fluconazole, itraconazole, and ketoconazole) in a diluent with Alamar Blue. The aim of this study was to compare the in vitro activities of four antifungals both by tests with the Sensititre YeastOne panel and by an adaptation of the NCCLS broth microdilution method, performed independently.Sensititre method. The Sensititre YeastOne test panels were provided by IZASA S.A. (Barcelona, Spain). A total of 49 clinical isolates belonging to six of the most common species of dermatophytes were tested (Table 1). Paecilomyces variotii ATCC 36257 was included as a reference strain. The fungi were subcultured on potato dextrose agar, and stock inoculum suspensions were prepared according to the recommendations of the NCCLS (13). This suspension was then adjusted with a spectrophotometer to 65 to 70% transmittance for dermatophytes and to 74 to 76% transmittance for P. variotii at a wavelength of 530 nm. The working suspension was made by dilution of the suspensions 1:100 in RPMI 1640 to produce the final test concentration of the inoculum. Aliquots of 100 l of the diluted suspension were inoculated into the wells with antifungals and the growth control well (containing only diluent and colorimetric indicator) with a multichannel pipette. The concentrations of the amphotericin B, itraconazole, and ketoconazole dilutions ranged from 0.008 to 16 g...