Fungal and fungal-like diversity in marine sediments from the maritime Antarctic assessed using DNA metabarcoding

We assessed the fungal and fungal-like sequence diversity present in marine sediments obtained in the vicinity of the South Shetland Islands (Southern Ocean) using DNA metabarcoding through high-throughput sequencing (HTS). A total of 193,436 DNA reads were detected in sediment obtained from three locations: Walker Bay (Livingston Island) at 52 m depth (48,112 reads), Whalers Bay (Deception Island) at 151 m (104,704) and English Strait at 404 m (40,620). The DNA sequence reads were assigned to 133 distinct fungal amplicon sequence variants (ASVs) representing the phyla Ascomycota, Basidiomycota, Mortierellomycota, Chytridiomycota, Glomeromycota, Monoblepharomycota, Mucoromycota and Rozellomycota and the fungal-like Straminopila. Thelebolus balaustiformis, Pseudogymnoascus sp., Fungi sp. 1, Ciliophora sp., Agaricomycetes sp. and Chaetoceros sp. were the dominant assigned taxa. Thirty-eight fungal ASVs could only be assigned to higher taxonomic levels, and may represent taxa not currently included in the available databases or represent new taxa and/or new records for Antarctica. The total fungal community displayed high indices of diversity, richness and moderate to low dominance. However, diversity and taxa distribution varied across the three sampling sites. In Walker Bay, unidentified fungi were dominant in the sequence assemblage. Whalers Bay sediment was dominated by Antarctic endemic and cold-adapted taxa. Sediment from English Strait was dominated by Ciliophora sp. and Chaetoceros sp. These fungal assemblages were dominated by saprotrophic, plant and animal pathogenic and symbiotic taxa. The detection of an apparently rich and diverse fungal community in these marine sediments reinforces the need for further studies to characterize their richness, functional ecology and potential biotechnological applications

Biodiversity of yeasts from marine and terrestrial antarctic ecosystems and prospection of lipases.

O ambiente Antártico é caracterizado por condições ambientais restritivas de clima, hábitat e biogeografia. Neste sentido, o objetivo deste estudo foi avaliar a diversidade de leveduras em amostras marinhas e terrestres do ambiente Antártico, além da prospecção de lipases que atuem em processos que requerem baixas e moderadas temperaturas. De acordo com os resultados obtidos, 97 leveduras foram recuperadas por semeadura em meio de cultura PDA e YMA a partir de diferentes amostras coletadas na OPERANTAR XXVIII (verão 2010), das quais foram identificados 12 gêneros e 21 espécies. Destas, 45 apresentaram atividade de lipase e duas foram selecionadas para o ensaio de otimização da produção enzimática: C. laurentii L59 e L. scotti L117. A extração de lipase de L. scottii L117 foi avaliada por Sistema de Duas fases Aquosas e o sistema micelar utilizando TX-114 foi o mais eficiente para purificação parcial desta enzima. Por fim, a avaliação da diversidade das 405 leveduras isoladas de macroalgas e liquens (coletadas na OPERANTAR XXXII no verão de 2013) revelou a ocorrência de 24 taxons de leveduras a partir das amostras de macroalgas e 18 taxons a partir das amostras de liquens, sendo apenas cinco espécies comuns a estes dois substratos.The Antarctic continent is characterized by restrictive environmental conditions of climate, habitats and biogeography. In this sense, the objective of this study was to evaluate the diversity of yeasts in marine and terrestrial samples of the Antarctic environments, as well as the propspecting of lipases that can act in proceses that require low and moderate temperatures. According to the results, 97 yeasts were recovered from different samples collected in OPERANTAR XXVIII (summer 2010), representing 21 different species. Among them, 45 showed lipase activity and two were selected for the optimization of enzyme production: C. laurentii L59 and L. scotti L117. Extraction of L. scottii L117 lipase was assessed by Aqueous Two-Phase System and the micellar system TX-114 was the most efficient for partial enzyme purification. Finally, the diversity evaluation of 405 isolates from seaweeds and lichens (collected in XXXII OPERANTAR, summer of 2013) revealed the occurrence of 24 taxa from macroalgae samples and 18 taxa from lichens samples, being only five substrates common to these two substrata

Liquid-liquid Extraction Of Lipase Produced By Psychrotrophic Yeast Leucosporidium Scottii L117 Using Aqueous Two-phase Systems

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Aqueous two-phase systems (ATPS) have been used in biomolecules separation and as an efficient alternative to traditional purification systems for lipases extraction. Here, we investigated the partitioning and recovery of lipase derived from Leucosporidium scottii L117 using ATPS and aqueous two-phase micellar systems (ATPMS). Thus, we evaluated three ATPS: (i) polyethylene glycol (PEG)/phosphate salts and (ii) PEG/polyacrylic acid (NaPA) in different molecular weights (1500, 4000 and 8000 g/mol). (iii) Triton X-114 (TX-114)/McIlvaine buffer pH 7.0 in different conditions (2.0% (w/w) of TX-114 at 25.0 and 28.0 degrees C). The PEG/phosphate and PEG/NaPA systems resulted in a great loss of enzymatic activity; thus these systems do not represent viable alternatives for these lipase extraction. The micellar systems yielded the best results for lipase extraction with enzyme activity balances ranging between 84.7% and 113.05%. After optimizing the micellar system by experimental design of the partition coefficient of lipase increased by 10.3-fold (0.75-7.76). Lipase preferentially partitioned into the micelle-rich phase with K-Lip = 7.76, % RECBot = 93.85% and PF = 1.2 at 25.03 degrees C, 5.1 pH and 10.38% TX-114 and K-Lip, = 4.77, % RECBot = 73.53% and PF = 1.97 at 28.00 degrees C, 4.5 pH and 8.0% TX-114, indicating that the ATPMS represents an alternative to purification/extraction of lipase L scottii L117. A crude lipase extract was also evaluated to define the optimum pH and temperature. Lipase reached optimal activity at 40 degrees C, and remained stable in pH values ranging from pH 3.0 to 8.0 and temperatures from 20.0 to 45.0 degrees C, with relative residual lipase activity above 80% after 30 min of incubation. (C) 2015 Elsevier B.V. All rights reserved.156215225Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [2010/08352-5, 2010/17033-0, 2013/19486-0

Liquid–liquid extraction of lipase produced by psychrotrophic yeast leucosporidium scottii L117 using aqueous two-phase systems

Aqueous two-phase systems (ATPS) have been used in biomolecules separation and as an efficient alternative to traditional purification systems for lipases extraction. Here, we investigated the partitioning and recovery of lipase derived from Leucosporidium scottii L117 using ATPS and aqueous two-phase micellar systems (ATPMS). Thus, we evaluated three ATPS: (i) polyethylene glycol (PEG)/phosphate salts and (ii) PEG/polyacrylic acid (NaPA) in different molecular weights (1500, 4000 and 8000 g/mol). (iii) Triton X-114 (TX-114)/McIlvaine buffer pH 7.0 in different conditions (2.0% (w/w) of TX-114 at 25.0 and 28.0 °C). The PEG/phosphate and PEG/NaPA systems resulted in a great loss of enzymatic activity; thus these systems do not represent viable alternatives for these lipase extraction. The micellar systems yielded the best results for lipase extraction with enzyme activity balances ranging between 84.7% and 113.05%. After optimizing the micellar system by experimental design of the partition coefficient of lipase increased by 10.3-fold (0.75–7.76). Lipase preferentially partitioned into the micelle-rich phase with KLip = 7.76, %RECBot = 93.85% and PF = 1.2 at 25.03 °C, 5.1 pH and 10.38% TX-114 and KLip = 4.77, %RECBot = 73.53% and PF = 1.97 at 28.00 °C, 4.5 pH and 8.0% TX-114, indicating that the ATPMS represents an alternative to purification/extraction of lipase L. scottii L117. A crude lipase extract was also evaluated to define the optimum pH and temperature. Lipase reached optimal activity at 40 °C, and remained stable in pH values ranging from pH 3.0 to 8.0 and temperatures from 20.0 to 45.0 °C, with relative residual lipase activity above 80% after 30 min of incubation156Part 2215225CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPnão temnão tem2010/08352-5; 2010/17033-0; 2013/19486-

Honey and bee pollen produced by Meliponini (Apidae) in Alagoas, Brazil: multivariate analysis of physicochemical and antioxidant profiles

Abstract This study evidenced the physicochemical composition and antioxidant activity (AA) of honeys (n = 31) and pollen (n = 25) of stingless bees species from Alagoas, Brazil. Fifteen parameters were studied under the light of a multivariate analysis. A dendrogram with three groups of honeys was stablished for the different bees, being the group III formed exclusively by samples from genus Melipona. Plebeia sp. and Tetragona clavipes Fabricius (1804) produced honeys with higher pH/acidity, electrical conductivity, phenolic total and AA. Even different bees from the same meliponary, as M. asilvai and T. clavipes, produced honeys with different AA and chemical profiles, possibly due their different nectar preference. The multivariate analysis of the bee pollen samples also showed three principal components responsible for 74.52% of their variability, clustering nine groups strongly influenced by total phenolics and AA. Honey and pollen produced by T. clavipes had the highest phenolic content and AA. Therefore, the chemical characteristics and AA of the Meliponini honey and bee pollen showed here can give support to the market to influence their incorporation in the human diet as sources of potential functional foods

Yeasts From Macroalgae And Lichens That Inhabit The South Shetland Islands, Antarctica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Antarctic terrestrial ecosystems are largely dominated by lichens, while shallow coastal environments are mainly covered by macroalgae. The aim of this study was to isolate and to evaluate the diversity of yeasts in different species of macroalgae and lichens collected in South Shetland Islands, Antarctica. A total of 405 yeasts were recovered (205 from macroalgae and 200 from lichens). The yeast community from macroalgae was most diversity than the yeast community from lichen. The dominance index was similar for both substrates. A total of 24 taxa from macroalgae and 18 from lichens were identified, and only 5 were common to both substrates. Metschnikowia australis, Mrakia sp., Rhodotorula glacialis and Glaciozyma litorale were the most abundant yeasts in macroalgae and Cryptococcus victoriae, Rhodotorula laryngis, Rhodotorula arctica, Trichosporon sp. 1 and Mrakia sp. were the most abundant in lichens. Based on molecular and phylogenetic analyses, four yeast from macroalgae and six from lichens were considered potential new species. This is the first study to report the yeast communities from the Antarctic macroalgae Himantothallus grandifolius and lichen Ramalina terebrata. Results suggest that Antarctic phyco and lichensphere represent a huge substrate for cold-adapted yeasts and enhanced the knowledge of the microbiota from extreme environments.85874885Brazilian Antarctic Program (PROANTAR)INCT CRIOSFERACoordination for Higher Level Graduate Improvement (CAPES/Brazil)State of Sao Paulo Research Foundation (FAPESP/Brazil) [2010/17033-0, 2013/19486-0]CAPESFAPESP [2010/08352-5]National Council for Scientific and Technological Development (CNPq) [304103/2013-6]Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Physicochemical composition and antioxidant potential of bee pollen from different botanical sources in Alagoas, Brazil

ABSTRACT Bee pollen results from the mixture of pollen and floral nectar with the salivary substances of bees and has increasingly been used as a food with therapeutic properties. In this study, 30 samples of bee pollen from Apis mellifera apiaries in three mesoregions of Alagoas (Brazil) - Mata Atlântica, South Coast and Caatinga, were collected during the dry season of 2008/09 and analysed. Mata Atlântica and Caatinga had good production of bee pollen. In the same season, Mata Atlântica contained a higher diversity of pollen types for feeding bees with a predominance of herbaceous pollen (63%), whereas the Caatinga samples contained monofloral pollen. Physicochemical data were analysed with the nonparametric Kruskal-Wallis statistical test. The Caatinga samples were analysed to determine their contents of total phenolic compounds (25.85 ± 10.80 mg gallic acid eq/g) and flavonoids (45.62 ± 32.19 mg quercetin eq/g) and their antioxidant activity (for instance, 70.62 ± 4.50% in the DPPH test), which were possibly affected by the environmental conditions of this biome

Composition and antioxidant activity of honey from Africanized and stingless bees in Alagoas (Brazil): a multivariate analysis

The purpose of this study was to evaluate the antioxidant activity of honey from different entomological sources which were harvested in the dry season of 2008-2009 from distinct mesoregions of the State of Alagoas in the North East of Brazil. Honey produced by five different species of bees, even from the same region and season, showed a statistically significant difference (p <0.05) in the content of phenols, flavonoids and antioxidants, with higher levels of these compounds found in honey produced by Plebeia spp. and A. mellifera. Honey from stingless bees was quite different from that of A. mellifera, especially from the Plebeia spp. A dendrogram of the five species of bees showed the formation of 3 groups, one being formed by Apis mellifera, one by the genus Melipona (M. subnitida, M. quadrifasciata and M. scutellaris) and another formed by Plebeia spp.Banco do Nordeste SA BNBBanco do Nordeste SA- BNBFundacao de Amparo a Pesquisa de Alagoas FAPEALFundacao de Amparo a Pesquisa de Alagoas - FAPEA

Antimicrobial and anti-inflammatory activities of Apis mellifera honey on the Helicobacter pylori infection of Wistar rats gastric mucosa

Abstract Considering that Helicobacter pylori, a bacterium able to colonize the upper gastrointestinal tract and cause mucosal injury, not always can be effectively eradicated by the traditional approaches, there is an interest in alternative therapies until a vaccine be available. Honey is a food supplement with high carbohydrate content and antioxidant activity, as well as broad antimicrobial spectrum. After analyzing the physicochemical and in vitro antimicrobial properties of an Apis mellifera honey from the Atlantic forest of Alagoas / Brazil, the purpose of the present work was evaluate its in vivo effects against Helicobacter pylori in the gastric mucosa of Wistar rats. First, it was verified the success of inoculation/infection of the pathogen in the gastric mucosa of the rats, through the subsequent removal of their stomachs for histological analysis (hematoxylin and eosin stain and Giemsa stain). Then, four groups of animals were treated with sterilized distilled deionized water, the Apis mellifera honey, a combination of omeprazole, amoxicillin and clarithromycin, and an association of such medicines and honey (1:1). Except the control, all treatments were effective in combating infection, however, honey reduced the inflammatory process, whilst the antibiotics increase the number of eosinophils

Socialização de conceitos de microbiologia no ensino fundamental através de atividades práticas

As abordagens tradicionais de ensino tendem a ser menos estimulantes e de maior dificuldade de compreensão, principalmente ao tratar-se de assuntos mais distantes da realidade dos alunos e, de certa forma, abstratos, como alguns conceitos de Microbiologia. Tal realidade reforça a importância do estudo e aplicação de novas formas de ensino no ambiente escolar. Nesse sentido, o objetivo deste artigo é relatar a experiência de ações de extensão realizadas com o tema “Dia do Cientista”, nas quais os métodos utilizados foram baseados no protagonismo dos alunos, através da participação ativa em experimentos, discussões e visitações. Ao longo da execução das ações, observou-se o despertar do interesse em assuntos até então desconhecidos e distantes da realidade dos estudantes do Ensino Fundamental I (4.º e 5.º ano), que puderam reconhecer o que a Microbiologia representa no seu cotidiano, a partir da execução de atividades práticas, como: I) Detecção de microrganismos em diferentes espaços da escola, como salas de aula e dos professores, banheiros e pátio; II) “Carimbo de mãos”; III) Utilização de fermento biológico para o crescimento microbiano e IV) Visita à Universidade Federal de Alagoas - Campus Arapiraca, onde puderam conhecer laboratórios de ensino prático do tema. Dessa forma, percebeu-se que estratégias com abordagens ativas e práticas auxiliaram numa maior interação dos alunos com os temas propostos, além de aguçar a curiosidade, estimular o aprendizado e permitir maior crescimento pessoal por parte dos escolares, bem como de todos os participantes do projeto. Palavras-chave: Extensão comunitária; Ensino; Aprendizagem   Socialization of microbiology concepts in elementary school through practical activities Abstract: Traditional teaching approaches tend to be less stimulating and more difficult to understand, especially when dealing with subjects that are more distant from the reality of students and, in a way, abstract, such as some topics of microbiology. This reality reinforces the importance of studying and applying new forms of teaching in the school environment. In this sense, the objective of this article is to report the experience of extension actions carried out with the theme "Scientist's Day", in which the methods used were based on the protagonism of the students, through active participation in experiments, discussions, and visits. Throughout the actions, it was possible to notice an awakening of interest in subjects previously unknown and distant from the reality of the students at elementary school (4th and 5th), who were able to recognize what microbiology represents in their daily lives, from the execution of practical activities such as I) Detection of microorganisms in different locations of the school, such as classrooms and teachers’ room, bathrooms and courtyard; II) "Hand Stamp"; III) Use of biological yeast for microbial growth and IV) Visit to the Federal University of Alagoas - Campus Arapiraca (Arapiraca, Brazil), where they got to know the laboratories. Therefore, it was noticed that strategies with active and practical approaches made the interaction of students with the proposed themes easier, besides to whet curiosity, stimulate learning and allow greater personal growth by the schoolers, as well as all the participants of the project. Keywords: Community extension; Teaching; Learnin