Resumo Durante o processo de produção do queijo, é necessária a presença de diversos microrganismos que contribuem na sua elaboração pela ação de seu metabolismo. Pouco é conhecido sobre os microrganismos presentes no queijo Colonial, além daqueles inseridos para iniciar o processo ou daqueles indesejados, potenciais causadores de infecção ou intoxicação. Este trabalho teve como objetivo principal identificar, através de MALDI-TOF e/ou sequenciamento de DNA, bactérias e leveduras isoladas de amostras coletadas nas principais etapas de produção de queijo Colonial, um tipo de queijo produzido na Região Sul do Brasil. Também foi verificada a capacidade lítica destes microrganismos a 5 °C e 30 °C. Foram analisados 58 isolados de bactérias, sendo identificadas 10 espécies de bactérias distribuídas nos gêneros Bacillus, Lactococcus, Paenibacillus, Staphylococcus, Citrobacter, Klebsiella e Raoutella. Dos 13 isolados de leveduras, foram identificadas três espécies: Candida pararugosa, Meyerozyma guilliermondii e Rhodotorula mucilaginosa. Em três isolados de leveduras, foi possível a identificação somente dos gêneros Candida sp. e Trichosporon sp. As espécies Lactococcus lactis (48%) e M. guilliermondii (46%) foram os isolados predominantes de bactéria e levedura, respectivamente. A maior atividade lítica dos microrganismos identificados foi na temperatura de 30 °C. Proporcionalmente, foi verificado maior número de isolados com atividade de lipase pelas leveduras e maior atividade proteolítica pelas bactérias. Menor atividade de caseínase e lipase foi observada a 5 °C, demonstrando a importância da refrigeração no controle da atividade microbiana. Este trabalho mostrou, através de cultivo, alguns dos microrganismos que fazem parte da microbiota do queijo Colonial e que vários deles possuem capacidade de hidrolisar vários compostos presentes no leite. Mostrou também que podem estar associados com a maturação do mesmo ou, em circunstâncias não controladas, que poderiam ser a causa de deterioração do produto.

Abstract Different types of microorganisms are important in cheese-making because of the contributions their metabolism offers during the process. Few microorganisms present in Colonial cheese are known, in addition to the ones that are introduced to kick-start the processes or the ones that are associated with infections or poisonings. This study aimed to identify, by MALDI-TOF and/or DNA sequencing, the bacteria and yeasts isolated from samples collected in the main stages of Colonial cheese production, i.e., a type of cheese produced in the southern region of Brazil. The lytic capacity of these microorganisms at 5 °C and 30 °C was also evaluated. The 58 bacterial strains were distributed in 10 species among the genera Bacillus, Citrobacter, Klebsiella, Lactococcus, Paenibacillus, Staphylococcus and Raoutella. From the 13 yeasts strains analyzed, three species were identified as following: Candida pararugosa; Meyerozyma guilliermondii; and Rhodotorula mucilaginosa. In three yeasts isolates it was possible to identify only the genus Candida sp. and Trichosporon sp. The species L. lactis (48%) and M. guilliermondii (46%) were, respectively, the predominant bacteria and yeasts species isolated. The highest microbial lytic activity observed was at 30 °C. Lipase activity on isolates was proportionally more observed with yeasts and proteolytic activity with bacteria. Lower caseinase and lipase activity was observed at 5 °C, demonstrating the importance of refrigeration in controlling microbial activity. This research highlighted the cultivation of some microorganisms that are part of the Colonial cheese microbiota as well as that several of them can hydrolyze various compounds present in milk and that could be associated with its maturation or, in uncontrolled circumstances, could be the cause of product deterioration.

Abstract The formation of microbial biofilms in materials used in the industrial production of dairy may lead to deterioration of these foods. Yarrowia lipolytica biofilms are widely found in dairy products and can modify the final characteristics of these products. Thus, this study investigated the effectiveness of hygienization by detergents and sodium hypochlorite on the formation of Y. lipolytica biofilms in different utensils usually employed during industrial cheese production, like polypropylene, hoses, and nylon/polyethylene. The utensils were sanitized using solutions of mild and alkaline detergents, and sodium hypochlorite, according to the cheese industry Standard Operation Procedure. Results showed that in all coupons there was biofilm formation with Y. lipolytica isolates. The contact angle measurements were favored to promote the adhesion of the biofilm in the evaluated surfaces. Even after treatment with sanitizers, a significant survival rate of planktonic cells was observed in all coupons tested. These results indicate that Y. lipolytica biofilms show a significant ability to adhere to polypropylene, presenting an important impact on the quality of colonial cheese.

ABSTRACT The indiscriminate use of pesticides on grape crops is harmful for consumers´ healthin “in natura” consumption and in the ingestion of wine and grape juice. During winemaking, a rapid and efficient fermentation stage is critical to avoid proliferation of contaminating microorganisms and to guarantee the product´s quality. Polymerase chain reaction (PCR) has the advantage of detecting these contaminants in the early stages of fermentation. However,this enzymatic reaction may also be susceptible to specific problems, reducing its efficiency. Agricultural practices, such as fungicide treatments, may be a source of PCR inhibiting factors and may also interfere in the normal course of fermentation.The action of the pesticides captan and folpet on PCR and on yeast metabolism was evaluated, once these phthalimide compounds are widely employed in Brazilian vineyards. DNA amplification was only observed at 75 and 37.5 µg/mL of captan concentrations, whereas with folpet, amplification was observed only in the two lowest concentrations tested (42.2 and 21.1µg/mL).Besides the strong inhibition on Taq polymerase activity, phthalimides also inhibited yeast metabolism at all concentrations analyzed.Grape must containing captan and folpet residues could not be transformed into wine due to stuck fermentation caused by the inhibition of yeast metabolism. Non-compliance with the waiting period for phthalimide fungicides may result in financial liabilities to the viticulture sector.The use of yeasts with high fungicide sensitivity should be selected for must fermentation as a strategy for sustainable wine production and to assure that products comply with health and food safety standards.

As infecções causadas por Dipodascus capitatus são raras e de difícil tratamento. Aqui se relata um caso em paciente com leucemia mielocítica aguda. O isolamento fúngico ocorreu a partir de hemocultura e a identificação fenotípica baseou-se em métodos micológicos clássicos; a identificação genotípica foi realizada através do sequenciamento da região D1/D2 do 26 rDNA. Os testes de suscetibilidade foram realizados através do Etest® e microdiluição em caldo. A antifungicoterapia foi ineficaz, registrando-se óbito da paciente no 17° dia após o diagnóstico. Os autores comparam o caso com relatos similares e discutem a emergência destas infecções bem como suas dificuldades diagnósticas e terapêuticas.

The infections caused by Dipodascus capitatus are rare, and the treatment is difficult. We reported a case of a patient with acute myeloid leukemia. The fungus was first isolated from hemocultures, and the phenotypic identification was based on mycological methods. The genotyping was carried out by sequencing the region D1/D2 from 26 rDNA. The susceptibility tests were assayed by Etest® and by the microdilution technique. None of the antifungal treatments employed were effective. The patient died on day 17 after the mycological diagnosis. The authors discussed the emergence of such infections as well as the difficulty regarding the diagnosis and treatment.

Leveduras em pelotas fecais de pequenos mamíferos provenientes de fragmentos de Mata Atlântica tiveram contagem média acima de 106 UFC/g. A maioria das 55 espécies isoladas foi de ascomicetos fermentativos, com as mais freqüentes sendo Debaryomyces hansenii, Pichia membranifaciens e Issatchenkia orientalis, enquanto Rhodotorula mucilaginosa foi a levedura mais freqüente de afinidade basidiomicética.

Yeasts had mean counts of above 106 CFU/g in the fecal pellets of small mammals from tropical forest fragments. Most of the 55 species isolated were fermentative ascomycetes, with the most frequent being Debaryomyces hansenii, Pichia membranifaciens and Issatchenkia orientalis, whereas Rhodotorula mucilaginosa was the most frequent yeast of basidiomycetous affinity.