Determination of chitin content in fungal cell wall: an alternative flow cytometric method

The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted Glycosyl Phosphatidyl Inositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P < 0.001) than chs3Δ/chs3Δ and pga31Δ/Δ especially in the presence of caspofungin. Ca. parapsilosis, Ca. tropicalis, and Ca. albicans showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi.

Possible respiratory infection due to Aspergillus in workers from swineries and poultries

Recent epidemiologic studies clearly outline the link between fungal sensibilization and exarcebations of asthma, leading to increased morbidity and mortality. Amongst the filamentous fungi, Aspergillus scpecies have been strongly linked with exarcebations of asthma and other respiratory allergic diseases. Particles of approximately 1 to 4 pm are deposited in the lower respiratory tract. Therefore, conidia of A. fumigatus are small enough to traverse the terminal respiratory airways and reach the pulmonary alveoli, whereas the larger conidia of some other Aspergillus species, such as A. flavus and A. niger, tend to be deposited in the paranasal sinuses and upper airways. Exposute to environmental fungal spores has been associated with worsening asthma symptoms, lung function, hospital admissions and asthma-related deaths.

The efficiency of urine dipsticks for the diagnosis of urinary tract infection

Urinary tract infection (UTI) is one of the most prevalent pathologies in developed countries, particularly in women, characterized by the presence of bacterial growth in any part of the urinary system. Currently, urine culture is considered the gold standard method for the diagnosis of UTI. However, this method has several disadvantages including the time necessary for obtaining the results and the associated high costs. Therefore, it is important to evaluate new efficient and valuable methods for the diagnosis of these infections. Objectives: Presently, dipsticks are considered a possible valuable alternative to urine culture. This method has very low costs associated and the results can be obtained in few minutes. Here we aim to compare the sensibility, specificity, predictive value of a positive test and a negative test of both methods in order to determine the efficiency of the test strips method and also to characterize the microorganism more frequently isolated.