RESUMO: O objetivo deste estudo foi genotipar isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único (SNP) de sequências de rDNA. Anteriormente, um multiplex-PCR baseado em SNP foi capaz de distinguir P. insidiosum em três diferentes clados. Dessa forma, utilizamos este método para avaliar isolados clínicos de P. insidiosum da América do Sul (n=32), cepas padrão da Costa Rica (n=4), Tailândia (n=3), Japão (n=1) e Índia (n=1), uma cepa padrão de Pythium aphanidermatum e isolados ambientais brasileiros de Pythium torulosum; Pythium rhizo-oryzae e Pythium pachycaule voucher (n=3). Os isolados analisados foram alocados aos clados: I (americanos), II (isolados da Índia, Japão e Tailândia), e III (um isolado tailandês). P. aphanidermatum, P.torulosum, P.rhizo-oryzae e P.pachycaule voucher não foram amplificados. Pela primeira vez, um isolado de P. insidiosum do Uruguai foi incluído em análises moleculares. Através da multiplex-PCR baseada em SNP, foi possível realizar a identificação e genotipagem dos isolados sul-americanos de P. insidiosum, demonstrando características genéticas semelhantes entre esses isolados.

ABSTRACT: We aimed to genotype the South American clinical isolates of Pythium insidiosum using the single nucleotide polymorphisms (SNP) of the ribosomal DNA sequences (rDNA). Previously, an SNP-based multiplex-PCR was able to distinguish three different clades of P. insidiosum isolates. Thus, we used this assay to evaluate South American clinical isolates of P. insidiosum (n=32), standard strains from Costa Rica (n=4), Thailand (n=3), Japan (n=1), and India (n=1), a standard strain of Pythium aphanidermatum, and Brazilian environmental isolates of Pythium torulosum, Pythium rhizo-oryzae and Pythium pachycaule voucher (n=3). It was possible to allocate each American P. insidiosum isolate to clade I, the isolates of India, Japan, and Thailand to clade II, and the Thai isolate to clade III. P. aphanidermatum, P.torulosum, P.rhizo-oryzae and P.pachycaule voucher isolates were not amplified. For the first time, a P. insidiosum isolate from Uruguay, South America, was included in molecular analyzes. By SNP-based multiplex-PCR, it was possible to perform the identification and genotyping of the South American isolates of P. insidiosum, demonstrating similar genetic characteristics of these isolates.

RESUMO: O presente trabalho objetivou a extração do óleo da polpa de macaúba utilizando extração por solvente a baixa pressão, bem como a caracterização dos extratos e farelos obtidos, utilizando diferentes solventes (n-hexano, acetato de etila e isopropanol). Os resultados reportam maiores rendimentos em óleo, 27,43%, na utilização do isopropanol (P<0,05), sendo que os maiores teores em β-caroteno no extrato, 348,30mg 100g-1, foram obtidos com este solvente. Na composição dos extratos, predominam os ácidos graxos monoinsaturados e relata-se que o tipo de solvente não apresentou influência significativa (P>0,05) na composição, no entanto, na quantificação dos compostos livres de glicerol, o isopropanol apresentou os maiores teores, 104,15mg 100g-1, desses compostos (P<0,05). Os farelos apresentaram maiores teores de cinzas, proteína e fibras, quando comparados à polpa. A remoção do óleo favorece a capacidade de retenção de água, no entanto, não tem influência nas demais características tecnológicas avaliadas.

ABSTRACT: This study aimed to extract the oil from macauba pulp using a low-pressure solvent extraction, as well as characterisation of the extracts and defatted meal obtained using different solvents (n-hexane, ethyl acetate and isopropanol). Results reported higher yields in oil, 27.43%, using isopropanol (P<0.05), wherein the highest levels of β-carotene in the extract, 348.30mg 100g-1, were obtained with this solvent. In the composition of the extracts, monounsaturated fatty acids were predominantly found, and it was reported that the type of solvent had no significant influence (P>0.05) on the composition; however, in the quantification of free glycerol compounds, isopropanol showed higher levels, 104.15mg 100g-1, of these compounds (P<0.05). Meals had higher ash, protein and fibre content when compared to pulp. Thus, oil removal can be stated to promote water retention capacity; however, it has no influence on the other technological characteristics evaluated.

The most promising microorganisms for use as starter cultures are those isolated from the native microbiota of traditional fermented products. The aim of this study was to evaluate the use of lactic acid bacteria selected from the native microbiota of sausages produced by spontaneous fermentation as starter cultures for the production of sausage. Strains of Lactobacillus plantarum 503 and 341 have the potential for use as starter cultures in the manufacture of Italian sausage type. The population of lactic acid bacteria in sausages was >8 log CFU.g-1 during fermentation, which caused the pH to decrease to <4.5. This decrease in pH and the water activity of < 0.90 of sausages ensures the safety and preservation of this product. Sausages produced with these lactic cultures fulfill the requirements for microbiological quality and composition of Italian sausage type. Our results suggest the possibility of using these starter cultures for the production of sausages with peculiar characteristics that contribute to the identity of the product.

O estudo da ecologia de salames fermentados faz-se necessário para entender as mudanças físicas e químicas que ocorrem durante a fermentação e maturação. O objetivo deste trabalho foi determinar as características microbiológicas de salames produzidos por fermentação espontânea. Foram analisadas cinquenta amostras de salames de diferentes pequenas indústrias em relação à seguinte microbiota: bactérias aeróbias mesófilas; Micrococcaceae; bolores e leveduras; bactérias lácticas; coliformes totais e fecais; Staphylococcus coagulase positiva; e Salmonella. Na grande maioria das amostras (72%), a presença de bactérias lácticas foi superior a 10(6) ufc.g-1, sendo que as amostras com as maiores contagens tiveram quantidades acima de 10(8) ufc.g-1. A concentração de Micrococcaceae, na maioria das amostras, ficou entre 10(5) e 10(7) ufc.g-1. Em relação à concentração de bolores e leveduras, houve uma grande variação entre as amostras, com quantidades próximas a 10² ufc.g-1 até 10(6) ufc.g-1. Pelos dados obtidos, concluiu-se que, das amostras analisadas, 24% estão em desacordo com as normas vigentes no Brasil, pois apresentam quantidades superiores do máximo permitido para coliformes a 45 ºC ou Staphylococcus coagulase positiva.

The study of the ecology of fermented sausage is fundamental to understand the physical and chemical changes that happen during fermentation and maturation. The aim of the present study was to determine the microbiological characteristics of sausages produced by spontaneous fermentation. Fifty samples of sausages produced in the South of Brazil by different small manufacturers were analyzed for the following microbiota: aerobic mesophilic bacteria; Micrococcaceae; mold and yeast; lactic acid bacteria; total and fecal coliforms; coagulase-positive Staphylococcus, and Salmonella. In most samples (72%), the count of lactic bacteria was higher than 6 log10 cfu.g-1, and the samples with the highest counts were above 8 log10 cfu.g-1. The counts of Micrococcaceae in most samples were between 5 log10 and 7 log10 cfu.g-1. With respect to the presence of molds and yeasts, there was a significant variation among the samples with counts ranging from 2 log10 cfu.g-1 and 6 log10 cfu.g-1. From the data obtained, it was possible to conclude that 24% of the analyzed samples did not comply with the current law in Brazil since the levels of fecal coliforms or coagulase-positive Staphylococcus exceeded the maximum limit allowed.