The aim of this work was the partial purification and subsequent evaluation of chitinase expression during the various growth phases of Paracoccidioides brasiliensis. Initially, PbCTS1r was expressed as a recombinant protein and displayed enzymatic activity against 4-MU-[N-acetylglucosamine (GlcNAc)]3 and 4-MU-(GlcNAc)2. Two proteins, 45 kDa and 39 kDa in size, were partially purified from P. brasiliensis yeast crude extract using cation-exchange chromatography coupled with HPLC and were characterised as PbCTS1 and PbCTS2, respectively. Anti-PbCTS1r antibody recognised two proteins in the crude extracts of yeast and the transitional stage between mycelial and yeast phases. In crude extracts of mycelium, only the 45 kDa protein was detected. However, quantitative real-time polymerase chain reaction led to the detection of small quantities of Pbcts2 transcript in the mycelial phase. In the yeast cell wall extract, only the 39 kDa protein was detected. Moreover, both proteins were secreted by the yeast parasitic phase, suggesting that these proteins participate in the modulation of the fungal environment. Phylogenetic analysis of the predicted PbCTS1 and PbCTS2 proteins indicated that they code for distinct chitinases in P. brasiliensis. During evolution, P. brasiliensis could have acquired the paralogues Pbcts1 and Pbcts2 for growth and survival in diverse environments in both saprophytic and parasitic phases.

Neste trabalho foi investigado o índice de HMF (5-hidroximetilfurfural) em méis comercializados em Porto Alegre - RS, utilizando Cromatografia Capilar Eletrocinética Micelar. O HMF, produto da condensação da frutose, é um indicador da qualidade e conservação do mel. Foram analisadas 11 marcas de méis comercializados na cidade de Porto Alegre. O composto estudado esteve presente em todas as amostras, em um intervalo de concentração de 0,191 a 6,206 mg.kg-1. Para quantificar o HMF presente nos méis, utilizou-se a técnica de adição de padrão. A taxa de recuperação foi de 98% e o limite de detecção foi de 0,025 mg.kg-1. O limite permitido de HMF em méis, segundo a legislação brasileira, é de 60 mg.kg-1.

In this work, the occurrence of HMF (5-hydroxymethylfurfural) in honey marketed in Porto Alegre - RS was investigated using Micellar Electrokinetic Capillary Chromatography. The HMF, which is the product of the fructose condensation, is an indicator of honey quality and conservation. Eleven types of honey commercialized in Porto Alegre were analyzed, and all of them contained HMF in a range from 0.191 to 6.206 mg.kg-1. In order to quantify the HMF present in the samples, the technique of standard addition was employed. The recovery was 98% and the detection limit was 0.025 mg.kg-1. The allowed limit of HMF in honey, according to the Brazilian Legislation, is 60 mg.kg-1.

O presente estudo teve por objetivo determinar os padrões moleculares e de resistência aos antimicrobianos de isolados de E. coli provenientes do trato urinário de suínos no Sul do Brasil. Os fatores estudados dividiram os patotipos ETEC, STEC e UPEC. Trinta e quatro (38,63%) isolados avaliados apresentavam um ou mais dos fatores de virulência pesquisados. A freqüência dos genes de virulência detectados foram: pap (10,97%), hlyA (10,97%), iha (9,75%), lt (8,53%), sta (7,31%) sfa (6,09%), f4 (4,87%), f5 (4,87%), stb (4,87%), f6 (1,21%) e f41 (1,21%). Os isolados foram resistentes à penicilina (95,12%), lincomicina (93,9%), eritromicina (92,68%), tetraciclina (90,24%), amoxacilina (82,92%), ampicilina (74,39%), josamicina (79,26%), norfloxacina (58,53%), enrofloxacina (57,31%), gentamicina (39,02%), neomicina (37,8%), apramicina (30,48%), colistina (30,48%) e cefalexina (6,09%). Trinta e dois (39,02%) isolados de E. coli continham plasmídeos.

The present study determined the molecular and resistance patterns of E. coli isolates from urinary tract of swine in Southern of Brazil. Molecular characterization of urinary vesicle samples was performed by PCR detection of virulence factors from ETEC, STEC and UPEC. From a total of 82 E. coli isolates, 34 (38.63%) harbored one or more virulence factors. The frequency of virulence factors genes detected by PCR were: pap (10.97%), hlyA (10.97%), iha (9.75%), lt (8.53%), sta (7.31%) sfa (6.09%), f4 (4.87%), f5 (4.87%), stb (4.87%), f6 (1.21%) and f41 (1.21%). Isolates were resistant to penicillin (95.12%), lincomycin (93.9%), erythromycin (92.68%), tetracycline (90.24%), amoxicillin (82.92%), ampicillin (74.39%), josamycin (79.26%), norfloxacin (58.53%), enrofloxacin (57.31%), gentamicin (39.02%), neomycin (37.8%), apramycin (30.48%), colistine (30.48%) and cefalexin (6.09%). A number of 32 (39.02%) E. coli isolates harbored plasmids.

A patulina é uma micotoxina produzida por diversas espécies de fungos dos gêneros Penicillium e Aspergillus, encontrada em diversas espécies frutíferas e, notadamente, em maçãs e produtos derivados da maçã. A patulina tem um amplo espectro de toxicidade, incluindo carcinogenicidade e teratogenicidade em animais. Devido a sua estabilidade, níveis consideráveis de patulina permanecem nos produtos derivados da maçã, mesmo após as etapas do processamento. Este artigo revisa os diferentes métodos analíticos usados para a determinação de patulina e métodos empregados para reduzir os níveis de patulina nos produtos alimentícios.

Patulin is a mycotoxin produced by several fungal species of the genera Penicillium and Aspergillus, found on several fruit species and, remarkably, in apples and apple products. Patulin has a broad spectrum of toxicity, including carcinogenicity and teratogenicity in animals. Due to the stability of the molecule, considerable amounts of patulin still remain in apple products after processing. This paper reviews different analytical methods for patulin determination and methods to reduce levels of patulin in apple products as well.

Comparou-se a suscetibilidade de 69 amostras de C. neoformans isoladas de pacientes com SIDA com 13 amostras de C. neoformans isoladas do meio ambiente, frente a quatro agentes antifúngicos. Com base na metodologia preconizada pelo NCCLS (M27-A) as concentrações inibitórias mínimas (CIMs) obtidas para a anfotericina B, itraconazol e cetoconazol foram muito semelhantes nos dois grupos estudados. Todavia, frente ao fluconazol, os isolados clínicos evidenciaram menor sensibilidade do que os provenientes do meio ambiente. Alguns aspectos envolvendo a importância e dificuldades dos testes de suscetibilidade com Cryptococcus neoformans são também discutidos pelos autores.

The purpose of the present study was to compare the susceptibility to four antifungal agents of 69 Cryptococcus neoformans strains isolated from AIDS patients with that of 13 C. neoformans strains isolated from the environment. Based on the NCCLS M27-A methodology the Minimal Inhibitory Concentrations (MICs) obtained for amphotericin B, itraconazole and ketoconazole were very similar for clinical and environmental isolates. Clinical isolates were less susceptible to fluconazole than environmental isolates. The significance of these findings and aspects concerning the importance, role and difficulties of C. neoformans susceptibility testing are also discussed.