Juliana Do Valle | Universidade Paranaense (original) (raw)
Papers by Juliana Do Valle
Journal of Microbiological Methods
Enzyme and Microbial Technology
The influence of green light on mycelium biomass growth and extracellular enzyme activities of ed... more The influence of green light on mycelium biomass growth and extracellular enzyme activities of edible mushrooms from the Pleurotus genus, which is popularly cultivated all over the world, were investigated. The mycelium of seven strains of five species of Pleurotus (P. citrinopileatus, P. djamor, P. eryngii, P. ostreatus, and P. pulmonarius) was grown in liquid medium at 28 °C in the dark or under green light (515-530 nm). The light source was light-emitting diodes (LED) with photon flux density adjusted to 20 μmol m-2 s-1 that was kept on throughout the cultivation period. After 12 days of growth, the mycelium was recovered and used for biomass determination and the cultivation medium was used to total cellulase, endoglucanase, xylanase, and laccase activities determination. Green light reduced the mycelial biomass growth of Pleurotus spp. but increased the cellulolytic and xylanolytic activities. The cellulolytic activity of most strains increased in the presence of green light with increases ranging from 1.5 times (P. ostreatus endoglucanase) to 8 times (P. citrinopileatus total cellulase and endoglucanase). Green light reduced laccase activity for most strains with the greatest reduction for P. eryngii (2.2 times lower). The specific enzymatic activity of cellulase and endoglucanase from P. citrinopileatus, increased by 31 times and 30 times, respectively, compared to the dark. Also, the specific laccase and xylanase activities of P. pulmonarius increased 4.4 times and 6.8 times, respectively, under green light. The use of light at particular wavelengths can be a viable strategy to increase the production of enzymes for different biotechnological applications and species of Pleurotus are particularly interesting for this purpose.
Semina: Ciências Agrárias, 2020
Bacteria from genus Bradyrhizobium can establish symbiosis with soybean and supply the plant nitr... more Bacteria from genus Bradyrhizobium can establish symbiosis with soybean and supply the plant nitrogen demands via biological nitrogen fixation (BNF). This study aimed to characterize genes related to BNF efficiency in B. japonicum strains contrasting in BNF efficiency. These gene sequences were previously identified in B. japonicum (strain S370) as probably related to the BNF efficiency in soybean using a DNA subtractive technique. These genes were amplified with primers based on B. japonicum USDA110 genome. The PCR products were digested with restriction endonucleases and the RFLP products were analyzed by horizontal electrophoresis. Among the four genes, only blr3208 and blr4511 amplified for most of the strains. Neither polymorphism of the restriction profile of blr3208 and blr4511 genes nor with endonuclease for PCR-RFLP was observed. The contrasting strains had blr3208 and blr4511 genes sequenced and the multiple alignment analysis of nucleotide sequences showed the presence of...
A especie Cochliomyia macellaria (Fabricius) caracteriza-se por ser exclusivamente saprofaga, pod... more A especie Cochliomyia macellaria (Fabricius) caracteriza-se por ser exclusivamente saprofaga, podendo causar miiases apenas em infestacoes secundarias de feridas. Apresenta ampla distribuicao geografica nas Americas, do sul do Canada ate a Argentina. Esta especie, por ser sinantropica, tem grande importância como vetor mecânico de microorganismos patogenicos. Na decada de 70, as Americas foram invadidas por especies do genero Chrysomya (Robineau-Desvoidy) cuja distribuicao geografica era confinada ao Velho Mundo. A introducao no continente Americano ocorreu atraves de intervencao humana. O estabelecimento de especies de Chrysomya no Novo Mundo afetou diretamente a fauna local de moscas varejeiras que exploram o mesmo recurso e mostram-se competidores eficientes. As especies nativas do Novo Mundo do genero Cochliomyia sao consideradas equivalentes ecologicos das especies de Chrysomya. A introducao e estabelecimento de especies deste genero na regiao Neotropical tem contribuido para alterar a distribuicao e frequencia de populacoes de C. macellaria no Brasil. O conhecimento basico da variabilidade genetica e evolucao intraespecifica e uma informacao necessaria para a compreensao da estrutura de populacao e eventos evolutivos recentes como introducoes e fluxo genico. Em insetos, o DNAmt tem mostrado ser um marcador genetico efetivo para obter informacoes sobre a variabilidade e estrutura genetica de populacoes. Neste trabalho, a analise de restricao do DNArnt e o calculo da divergencia de sequencia nucleotidica no DNArnt de 6 populacoes brasileiras de C. macellaria, foram conduzidos para examinar a variabilidade genetica e para fornecer bases para a compreensao da atual estrutura populacional de C. macellaria. Foi realizada tambem a analise cariotipica, para a obtencao de dados que relacionassem informacoes nucleares e citoplasmaticas de algumas populacoes. O uso de 13 endonucleases de restricao na analise do DNArnt de C. macellaria revelou que 3 enzimas (EcoRV, HindIII e PvuII) foram eficientes para detectar variabilidade. Baseado nos padroes de restricao obtidos para estas enzimas, foram caracterizados 11 haplotipos mitocondriais. Entre os haplotipos mitocondriais, os valores obtidos para divergencia de sequencia de nucleotideos (o) variaram de 0.002 a 0.009. Estes dados sugerem diferenciacao local do DNArnt de algumas populacoes. A analise cladistica dos haplotipos observados e da distribuicao geografica sugere que algumas populacoes de C. macellaria apresentam certa descontinuidade genetica com separacao espacial, dados que refletem barreiras ao fluxo genico. Por outro lado, algumas populacoes revelam que haplotipos intimamente relacionados nao apresentam localizacao geografica, o que pode refletir interacao dessas populacoes atraves de fluxo genico, migracao entre as diferentes localidades ou interferencia humana. A analise cariotipica de tres populacoes de C. macellaria (Ca, Cp e PU) revelou diferencas quanto a posicao dos centromeros e tamanho dos cromossomos sexuais, alem de detectar constricoes secundarias que podem ser utilizadas como marcadores interpopulacionais e interespecificos a nivel cromossomico. Apesar de ter sido detectada interacao via fluxo genico entre algumas das populacoes de C. macellaria e dos baixos valores de divergencia de sequencia, a alta variabilidade observada no DNArnt representa um marcador genetico molecular efetivo para a compreensao de eventos evolutivos recentes e no monitoramento de afunilamento e competicao entre as especies causadoras de miiases nativas e introduzidas da familia Calliphoridae Abstract
Research, Society and Development, 2020
Laccases are part of the family of ligninolytic enzymes and have played essential roles in severa... more Laccases are part of the family of ligninolytic enzymes and have played essential roles in several biological filamentous fungi processes, including fruiting body formation and lignin degradation. This study aimed to identify and characterize laccase genes in silico of several basidiomycete strains. The applied guaiacol oxidation test allowed the selection of seven out of 11 strains with ligninase activity, which were used for DNA extraction and amplification of the copper-binding region. A single amplicon of approximately 450 bp, was produced by all selected strains and they were further sequenced. Sequence analysis has suggested the presence of a new subdivision of the laccase genes. Clustering analysis confirmed the existence of two groups: cluster A with six strains and singleton B with U8-11 strain. The structural predictions of the U8-11 protein were dissimilar compared to other proteins described in our study due to the absence of the ALAVIN motif and, therefore, the U8-11 am...
Autoregulacao colinergica e o nome dado a capacidade da acetilcolina de regular sua propria liber... more Autoregulacao colinergica e o nome dado a capacidade da acetilcolina de regular sua propria liberacao neuronal. A acetilcolina atua sobre receptores nicotinicos e muscarinicos presentes nos terminais nervosos motores. Tais receptores, em conjunto com outros nao colinergicos, permitem que os terminais colinergicos sejam regulados por agentes liberados proximos ou distantes dos nervos motores. O acionamento dos receptores nicotinicos e muscarinicos pre- sinapticos determinam, respectivamente, aumento e reducao da quantidade de acetilcolina liberada para a fenda sinaptica. Estudos farmacologicos sugerem que os receptores nicotinicos pre-sinapticos do terminal nervoso motor sejam do subtipo ganglionar Nn, por se comportarem de forma semelhante aqueles presentes nos gânglios autonomicos. Os muscarinicos, por outro lado, comportam-se farmacologicamente como receptores cardiacos do subtipo M2
Rennet is a natural complex of enzymes (aspartic endopeptidases) and is often used in the product... more Rennet is a natural complex of enzymes (aspartic endopeptidases) and is often used in the production of cheese. Those enzymes (proteases) are extracted from abomasums of young ruminant animals. The high cheese demand associated with the high curdle cost have been stimulated the searching for alternative protease sources. The objective of this study was to verify the time and temperature conditions for biomass, exopolysaccharide (EPS) and protease production by Pleurotus ostreatus in liquid cultivation. The mycelium was grown in Pontecorvo medium added with isolated soy protein (5%) and maintained at 22, 25 or 28oC, with samplings at 3, 6, 9, 14 or 17 days of cultivation. The biomass was separated by centrifugation, EPS were separated by ethanol precipitation and the proteolytic activity was determined by enzymatic extract action on alkaline soluble casein 1% (m/v). It was verified that time and temperature affect ( p ≤0.05) biomass, EPS and protease production. Most of P. ostreatus...
Evolução da curva de casos de COVID-19 nas cidades que pertencem à macrorregião noroeste do Estad... more Evolução da curva de casos de COVID-19 nas cidades que pertencem à macrorregião noroeste do Estado do Paraná, Brasil Evolution of COVID-19 case curve in cities belonging the northwest macro-region of the Paraná State, Brazil Evolución de la curva de casos de COVID-19 en ciudades pertenecientes a la macro
Os sais de lítio são usados no transtorno de humor, uma vez que este elemento químico atua no sis... more Os sais de lítio são usados no transtorno de humor, uma vez que este elemento químico atua no sistema nervoso central. Para aumentar a biodisponibilidade deste metal, estratégias são estudadas, como a bioacumulação em basidiomicetos. Desta forma, o objetivo deste trabalho foi realizar uma revisão bibliográfica sobre a capacidade de bioacumulação de lítio in vitro por basidiomicetos. Artigos científicos sobre biocumulação, lítio e basidiomicetos publicados nos anos de 1980 a 2020 foram pesquisados. Uma alternativa para o tratamento do transtorno de humor bipolar é o carbonato de lítio, porém sua faixa terapêutica é estreita, o que pode levar a intoxicação medicamentosa. A bioacumulação de lítio in vitro em basidiomicetos pode ajudar na redução dos efeitos colaterais e melhorar a biodisponibilidade do lítio. São escassos os estudos sobre bioacumulação de lítio em basidiomicetos, e a maioria utiliza LiCl como fonte de lítio, provavelmente pela sua alta solubilidade. O cultivo micelial ...
Basidiomicetos tem sido amplamente utilizados como produtores de enzimas, no entanto sao pouco ex... more Basidiomicetos tem sido amplamente utilizados como produtores de enzimas, no entanto sao pouco explorados quanto a sua capacidade de producao de proteases. Estes fungos sao reconhecidos pelas suas propriedades antitumorais, hipocolesterolemicas, antimutagenicas, antioxidantes entre outras. Assim, a associacao destas propriedades aos derivados do leite pode potencializar estes produtos como alimentos funcionais. Neste sentido, o objetivo deste trabalho foi selecionar basidiomicetos produtores de proteases, com potencial uso no processo de fabricacao de derivados do leite. Foram utilizadas 27 linhagens de fungos crescidas em meio minimo adicionado de 0,2% de caseina. A atividade proteolitica foi verificada pela formacao de halo pela adicao de uma solucao saturada de (NH4)2SO4. Concluiu-se que a producao de proteases nao apresenta relacao com o crescimento micelial. O melhor produtor de proteases e a linhagem Lentinula edodes U8/1, seguida por Pleurotus sp (U2/9, U6/10 e U2/11). Os bas...
Brazilian Journal of Microbiology, 2019
White-rot basidiomycetes such as Lentinus crinitus produce laccases with potential use in dye bio... more White-rot basidiomycetes such as Lentinus crinitus produce laccases with potential use in dye biodegradation. However, high productivity and enzymes with specific properties are required in order to make viable laccase production. We aimed to produce laccase from Lentinus crinitus grown in sugarcane bagasse for dye decolorization. Solid state cultivation medium had sugarcane bagasse added with a nutrient solution of 10 g/L glucose, 1 g/L KH 2 PO 4 , 0.5 g/L MgSO 4 , 0.001 g/L FeSO 4 , 0.01 g/L ZnSO 4 , and 0.01 g/L MnSO 4. The addition of different nitrogen sources (peptone, urea, or peptone plus urea) and different nitrogen concentrations (0, 0.4, 0.6, 0.8, 1.0, and 1.2 g/L) were evaluated. Enzymatic extract was used in the decolorization of azo dyes, reactive blue 220 (RB220) and reactive black 5 (RB5), and anthraquinone dye, Remazol brilliant blue R (RBBR). The greatest laccase activity (4800 U/ g dry mass) occurred when the peptone and urea mixture was added to the solid state cultivation medium. When the nitrogen concentration was 1 g/L, the laccase activity increased to 6555 U/g dry mass. The laccase activity peak occurred on the 10th day, and the maximum decolorization within 24 h was observed with enzymatic extracts obtained on different cultivation days, i.e., 6th day for RB220, 10th day for RB5, and 9th day for RBBR. Manganese and lignin peroxidases were not produced when nitrogen was added to the cultivation medium. The crude enzymatic extract was more effective in the decolorization of azo dyes (RB220 and RB5), more than 90% of decolorization, than anthraquinone dye with 77% decolorization.
Chemosphere, 2019
Lentinus crinitus is an important basidiomycete consumed by ethnic groups from the Amazon, common... more Lentinus crinitus is an important basidiomycete consumed by ethnic groups from the Amazon, commonly found in decomposing trees with high lignolytic and antioxidant activities. Lithium is a mood stabilizer, antiepileptic, antipsychotic, and antidepressant used in clinical practice. This study aimed to evaluate L. crinitus mycelial biomass bioaccumulated with lithium in liquid cultivation medium. The malt extract medium was added from zero to 100 mg L-1 lithium from two lithium sources (Li 2 CO 3 and LiCl). The maximum mycelial biomass production was 7,218.89 mg L-1 in the culture medium added ACCEPTED MANUSCRIPT with 5 mg L-1 lithium from LiCl. The highest lithium concentration in the mycelial biomass was of 574.72 µg g-1 produced in the culture medium with 25 mg L-1 lithium from Li 2 CO 3. Pearson's correlation showed that Li 2 CO 3 reduces the mycelial biomass and increases lithium bioaccumulation. The maximum translocated lithium from cultivation medium to mycelial biomass was up to 19 or 28% with LiCl or Li 2 CO 3 , respectively. Therefore, although Li 2 CO 3 presents greater inhibition on the mycelial biomass production, it promoted greater lithium bioaccumulation in L. crinitus mycelial biomass and resulted in greater yield of lithium translocation. The equivalent daily dose of lithium for psychiatric treatment, without bioavailability studies, could be reached with 97.4 g lithium-enriched mycelial biomass and, based in the literature, for reduction of violence and criminality rates the amount could be reached with 0.24-0.58 mg. Thus, the development of lithium-enriched mycelial biomass could be an alternative functional food.
Brazilian Journal of Microbiology, 2019
This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelia... more This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelial viability of A. subrufescens after 5-year cryopreservation at − 70°C. Mycelia were grown in three types of whole or ground grains, with or without 5% glycerol addition in the substrate and/or in a cryotube. After 5 years of cryopreservation at − 70°C, every treatment was thawed and recovered in malt extract culture medium with 15 (solid culture medium) or 5 g L −1 (semisolid culture medium) of agar. The semisolid recovery culture medium increased the mycelial viability recovery capacity of A. subrufescens cryopreserved for 5 years in grains with glycerol only in the cryotube, and specifically with medium-hard wheat grain without glycerol addition at all. Agar-based substrates such as malt extract agar, agar-ground grain, or the one with glycerol addition to the substrate were not effective to keep the mycelial viability, regardless of the recovery culture medium consistency. Hard and medium-hard endosperm wheat grains or hard endosperm rye grains with addition of glycerol as cryoprotectant only to the cryotube were effective to cryopreserve the fungus for 5 years without cryoprotectant addition in the substrate.
Biocatalysis and Agricultural Biotechnology, 2018
Lentinus crinitus is a basidiomycete with the ability to produce laccase and other enzymes of bio... more Lentinus crinitus is a basidiomycete with the ability to produce laccase and other enzymes of biotechnological interest. Laccase production depends on cultivation conditions and can be induced by metallic, phenolic or aromatic compounds. This study aimed to evaluate the laccase production of L. crinitus cultivated in sugarcane vinasse and urea with addition of laccase inducers. L. crinitus was cultivated in sugarcane vinasse with urea and supplemented with copper sulfate (0, 50, 150, 250, 350 and 450 μM) and aromatic compounds (guaiacol, veratryl alcohol, or 2,5-xylidine) added alone or combined to the cultivation medium. Cooper (150 or 250 μM) increased laccase activity in 14.5 (49000 U/L) and 28.5% (55000 U/L), respectively. Guaiacol had no significant effect, but veratryl alcohol (~46000 U/L) and xylidine (~75000 U/L) increased laccase activity. Copper with other inducers had the most relevant effects on laccase production. Simultaneous guaiacol addition with 150 or 250 μM copper increased 108 and 151% laccase activity, respectively, when compared with only copper addition to vinasse-urea medium. Veratryl alcohol and 150 or 250 μM copper addition increased 100 and 138% laccase activity, respectively, when compared to only copper addition. Interaction analysis of the variables guaiacol or veratryl alcohol and copper (250 µM) added to cultivation medium demonstrated synergistic effects resulting in laccase activity increase. These results highlight the importance of L. crinitus as a laccase producer and emphasize the relevance of studies using a combination of inducers to increase laccase production.
Biocatalysis and Agricultural Biotechnology, 2018
Panus strigellus is a native basidiomycete from Brazil with laccase production potential. Laccase... more Panus strigellus is a native basidiomycete from Brazil with laccase production potential. Laccases are involved in the biodegradation of dyes and their production can be stimulated by cultivation with agro industrial by-products. This study aimed to evaluate laccase production of P. strigellus cultivated with sugarcane bagasse on dye decolorization. Culture media had different concentrations of sugarcane bagasse (SB), sources of nitrogen (urea, yeast extract, ammonium sulfate and sodium nitrate), and concentrations of copper sulfate (0, 50, 100, 200, and 300 μM). Enzymatic extract from fungal cultivation was used in the decolorization of reactive blue 220 (RB220), malachite green (MG), and remazol brilliant blue R (RBBR). Laccase production was inversely proportional to the amount of SB in cultivation medium and higher titers of enzyme were obtained when 20 g/L SB (SB20) was used. Cultivation medium with SB20 and most nitrogen sources did not have a positive effect 2 on laccase production whereas the addition of urea or yeast-extract reduced laccase production. Most of the copper sulfate concentrations added to SB20 medium increased laccase production, and the maximum laccase production was 55342 U/L in the SB20 medium with 200 μM copper. The crude enzymatic extract was more effective in the decolorization of azo dye (RB220), with more than 90% of decolorization in 24 h, than anthraquinone dye (RBBR), with 60% decolorization, and triphenylmethane dye (MG), with 68% decolorization, both in 72 h.
Arquivos De Ciencias Veterinarias E Zoologia Da Unipar, May 6, 2013
Atividade antimicrobiana de óleos essenciais de condimentos frente a Staphylococcus spp isolados ... more Atividade antimicrobiana de óleos essenciais de condimentos frente a Staphylococcus spp isolados de mastite caprina.
Arquivos De Ciencias Veterinarias E Zoologia Da Unipar, May 9, 2013
A determinacao de biomassa micelial fungica crescida em substratos de cultivo solido particulado ... more A determinacao de biomassa micelial fungica crescida em substratos de cultivo solido particulado (SCSP) e ainda um desafio devido a dificuldade de separacao do micelio e o substrato. O objetivo deste trabalho foi avaliar a tecnica de microscopia de epifluorescencia para determinacao da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinacao da exatidao da metodologia P. ostreatus foi crescido em meio liquido de extrato de malte e; a biomassa micelial foi separada por centrifugacao, liofilizada e moida. Concentracoes conhecidas do po do micelio foram misturadas ao SCSP, composto de bagaco de cana de acucar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescencia. Para promover a variacao da biomassa micelial a ser determinada por microscopia de epifluorescencia, SCSP adicionado de diferentes concentracoes de ferro foram utilizados para o crescimento do fungo. Concluiu-se que a tecnica apresenta baixa precisao e exatidao, o que implica na necessidade de maiores estudos para aplicacao desta tecnica para a determinacao de biomassa micelial crescida em SCSP.
Current Microbiology, 2015
The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subr... more The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subrufescens (=A. blazei; =A. brasiliensis) obtained by different extraction methods were evaluated on Aspergillus nidulans conidia submitted to ultraviolet (UV) light. The aqueous extracts were obtained by three extraction methods: maceration, infusion, and decoction, at two different extraction times. The extracts of A. subrufescens did not present toxicity for A. nidulans conidia. A suspension of A. nidulans conidia was submitted to extracts before and after the exposure to UV light. All basidiocarp extracts, regardless of the extraction method or development stage, protected A. nidulans conidia against the damaging effects of the mutagenic agent. The antimutagenic and photoprotective activity was strengthened with extracts obtained by 168-h maceration, followed by 24-h maceration and 60-min infusion and, at last, by 30-min infusion. Although the extracts presented protector effect as well as recoverer effect to the action of UV light, the preventive effect was more evident. Differences in the biological activity in function of the different development stages were detected with greater antimutagenic and photoprotective activity for the opened basidiocarps. However, the extraction method is the most important factor to be considered when compared to the basidiocarp development stage to obtain better antimutagenic and photoprotective activity of A. subrufescens basidiocarps.
International Journal of Medicinal Mushrooms, 2015
Many alternative compounds have been tested to improve poultry performance but few of them have p... more Many alternative compounds have been tested to improve poultry performance but few of them have previously used mycelial-colonized substrate to partially replace standard diet in broiler chickens. The objective of this study was to evaluate broiler chicken production, health, and meat sensory characteristics, with partial replacement of the standard diet by Pleurotus ostreatus-colonized substrate. One hundred fifty 1-day-old male Cobb chicks were given standard diet partially replaced by 0, 5, 10, 100, or 200 g·kg-1 of P. ostreatus-colonized substrate and randomly distributed into five treatments. Each treatment had three replicates, with 10 birds per replicate, totaling 30 birds. The replacement of the standard diet by 10 g·kg-1 of colonized substrate increased (P≤0.05) chicken body mass up to 57% at 21 days, and up to 28% at 42 days. In general, partial replacement of standard diet by colonized substrate increased hematocrits and typical lymphocytes, and reduced low density lipoproteins. Also, it reduced chicken production period up to 21% and there is no meat taste alteration. The use of P. ostreatus-colonized substrate in chicken feeding is an alternative method to improve broiler chicken production.
Journal of Microbiological Methods
Enzyme and Microbial Technology
The influence of green light on mycelium biomass growth and extracellular enzyme activities of ed... more The influence of green light on mycelium biomass growth and extracellular enzyme activities of edible mushrooms from the Pleurotus genus, which is popularly cultivated all over the world, were investigated. The mycelium of seven strains of five species of Pleurotus (P. citrinopileatus, P. djamor, P. eryngii, P. ostreatus, and P. pulmonarius) was grown in liquid medium at 28 °C in the dark or under green light (515-530 nm). The light source was light-emitting diodes (LED) with photon flux density adjusted to 20 μmol m-2 s-1 that was kept on throughout the cultivation period. After 12 days of growth, the mycelium was recovered and used for biomass determination and the cultivation medium was used to total cellulase, endoglucanase, xylanase, and laccase activities determination. Green light reduced the mycelial biomass growth of Pleurotus spp. but increased the cellulolytic and xylanolytic activities. The cellulolytic activity of most strains increased in the presence of green light with increases ranging from 1.5 times (P. ostreatus endoglucanase) to 8 times (P. citrinopileatus total cellulase and endoglucanase). Green light reduced laccase activity for most strains with the greatest reduction for P. eryngii (2.2 times lower). The specific enzymatic activity of cellulase and endoglucanase from P. citrinopileatus, increased by 31 times and 30 times, respectively, compared to the dark. Also, the specific laccase and xylanase activities of P. pulmonarius increased 4.4 times and 6.8 times, respectively, under green light. The use of light at particular wavelengths can be a viable strategy to increase the production of enzymes for different biotechnological applications and species of Pleurotus are particularly interesting for this purpose.
Semina: Ciências Agrárias, 2020
Bacteria from genus Bradyrhizobium can establish symbiosis with soybean and supply the plant nitr... more Bacteria from genus Bradyrhizobium can establish symbiosis with soybean and supply the plant nitrogen demands via biological nitrogen fixation (BNF). This study aimed to characterize genes related to BNF efficiency in B. japonicum strains contrasting in BNF efficiency. These gene sequences were previously identified in B. japonicum (strain S370) as probably related to the BNF efficiency in soybean using a DNA subtractive technique. These genes were amplified with primers based on B. japonicum USDA110 genome. The PCR products were digested with restriction endonucleases and the RFLP products were analyzed by horizontal electrophoresis. Among the four genes, only blr3208 and blr4511 amplified for most of the strains. Neither polymorphism of the restriction profile of blr3208 and blr4511 genes nor with endonuclease for PCR-RFLP was observed. The contrasting strains had blr3208 and blr4511 genes sequenced and the multiple alignment analysis of nucleotide sequences showed the presence of...
A especie Cochliomyia macellaria (Fabricius) caracteriza-se por ser exclusivamente saprofaga, pod... more A especie Cochliomyia macellaria (Fabricius) caracteriza-se por ser exclusivamente saprofaga, podendo causar miiases apenas em infestacoes secundarias de feridas. Apresenta ampla distribuicao geografica nas Americas, do sul do Canada ate a Argentina. Esta especie, por ser sinantropica, tem grande importância como vetor mecânico de microorganismos patogenicos. Na decada de 70, as Americas foram invadidas por especies do genero Chrysomya (Robineau-Desvoidy) cuja distribuicao geografica era confinada ao Velho Mundo. A introducao no continente Americano ocorreu atraves de intervencao humana. O estabelecimento de especies de Chrysomya no Novo Mundo afetou diretamente a fauna local de moscas varejeiras que exploram o mesmo recurso e mostram-se competidores eficientes. As especies nativas do Novo Mundo do genero Cochliomyia sao consideradas equivalentes ecologicos das especies de Chrysomya. A introducao e estabelecimento de especies deste genero na regiao Neotropical tem contribuido para alterar a distribuicao e frequencia de populacoes de C. macellaria no Brasil. O conhecimento basico da variabilidade genetica e evolucao intraespecifica e uma informacao necessaria para a compreensao da estrutura de populacao e eventos evolutivos recentes como introducoes e fluxo genico. Em insetos, o DNAmt tem mostrado ser um marcador genetico efetivo para obter informacoes sobre a variabilidade e estrutura genetica de populacoes. Neste trabalho, a analise de restricao do DNArnt e o calculo da divergencia de sequencia nucleotidica no DNArnt de 6 populacoes brasileiras de C. macellaria, foram conduzidos para examinar a variabilidade genetica e para fornecer bases para a compreensao da atual estrutura populacional de C. macellaria. Foi realizada tambem a analise cariotipica, para a obtencao de dados que relacionassem informacoes nucleares e citoplasmaticas de algumas populacoes. O uso de 13 endonucleases de restricao na analise do DNArnt de C. macellaria revelou que 3 enzimas (EcoRV, HindIII e PvuII) foram eficientes para detectar variabilidade. Baseado nos padroes de restricao obtidos para estas enzimas, foram caracterizados 11 haplotipos mitocondriais. Entre os haplotipos mitocondriais, os valores obtidos para divergencia de sequencia de nucleotideos (o) variaram de 0.002 a 0.009. Estes dados sugerem diferenciacao local do DNArnt de algumas populacoes. A analise cladistica dos haplotipos observados e da distribuicao geografica sugere que algumas populacoes de C. macellaria apresentam certa descontinuidade genetica com separacao espacial, dados que refletem barreiras ao fluxo genico. Por outro lado, algumas populacoes revelam que haplotipos intimamente relacionados nao apresentam localizacao geografica, o que pode refletir interacao dessas populacoes atraves de fluxo genico, migracao entre as diferentes localidades ou interferencia humana. A analise cariotipica de tres populacoes de C. macellaria (Ca, Cp e PU) revelou diferencas quanto a posicao dos centromeros e tamanho dos cromossomos sexuais, alem de detectar constricoes secundarias que podem ser utilizadas como marcadores interpopulacionais e interespecificos a nivel cromossomico. Apesar de ter sido detectada interacao via fluxo genico entre algumas das populacoes de C. macellaria e dos baixos valores de divergencia de sequencia, a alta variabilidade observada no DNArnt representa um marcador genetico molecular efetivo para a compreensao de eventos evolutivos recentes e no monitoramento de afunilamento e competicao entre as especies causadoras de miiases nativas e introduzidas da familia Calliphoridae Abstract
Research, Society and Development, 2020
Laccases are part of the family of ligninolytic enzymes and have played essential roles in severa... more Laccases are part of the family of ligninolytic enzymes and have played essential roles in several biological filamentous fungi processes, including fruiting body formation and lignin degradation. This study aimed to identify and characterize laccase genes in silico of several basidiomycete strains. The applied guaiacol oxidation test allowed the selection of seven out of 11 strains with ligninase activity, which were used for DNA extraction and amplification of the copper-binding region. A single amplicon of approximately 450 bp, was produced by all selected strains and they were further sequenced. Sequence analysis has suggested the presence of a new subdivision of the laccase genes. Clustering analysis confirmed the existence of two groups: cluster A with six strains and singleton B with U8-11 strain. The structural predictions of the U8-11 protein were dissimilar compared to other proteins described in our study due to the absence of the ALAVIN motif and, therefore, the U8-11 am...
Autoregulacao colinergica e o nome dado a capacidade da acetilcolina de regular sua propria liber... more Autoregulacao colinergica e o nome dado a capacidade da acetilcolina de regular sua propria liberacao neuronal. A acetilcolina atua sobre receptores nicotinicos e muscarinicos presentes nos terminais nervosos motores. Tais receptores, em conjunto com outros nao colinergicos, permitem que os terminais colinergicos sejam regulados por agentes liberados proximos ou distantes dos nervos motores. O acionamento dos receptores nicotinicos e muscarinicos pre- sinapticos determinam, respectivamente, aumento e reducao da quantidade de acetilcolina liberada para a fenda sinaptica. Estudos farmacologicos sugerem que os receptores nicotinicos pre-sinapticos do terminal nervoso motor sejam do subtipo ganglionar Nn, por se comportarem de forma semelhante aqueles presentes nos gânglios autonomicos. Os muscarinicos, por outro lado, comportam-se farmacologicamente como receptores cardiacos do subtipo M2
Rennet is a natural complex of enzymes (aspartic endopeptidases) and is often used in the product... more Rennet is a natural complex of enzymes (aspartic endopeptidases) and is often used in the production of cheese. Those enzymes (proteases) are extracted from abomasums of young ruminant animals. The high cheese demand associated with the high curdle cost have been stimulated the searching for alternative protease sources. The objective of this study was to verify the time and temperature conditions for biomass, exopolysaccharide (EPS) and protease production by Pleurotus ostreatus in liquid cultivation. The mycelium was grown in Pontecorvo medium added with isolated soy protein (5%) and maintained at 22, 25 or 28oC, with samplings at 3, 6, 9, 14 or 17 days of cultivation. The biomass was separated by centrifugation, EPS were separated by ethanol precipitation and the proteolytic activity was determined by enzymatic extract action on alkaline soluble casein 1% (m/v). It was verified that time and temperature affect ( p ≤0.05) biomass, EPS and protease production. Most of P. ostreatus...
Evolução da curva de casos de COVID-19 nas cidades que pertencem à macrorregião noroeste do Estad... more Evolução da curva de casos de COVID-19 nas cidades que pertencem à macrorregião noroeste do Estado do Paraná, Brasil Evolution of COVID-19 case curve in cities belonging the northwest macro-region of the Paraná State, Brazil Evolución de la curva de casos de COVID-19 en ciudades pertenecientes a la macro
Os sais de lítio são usados no transtorno de humor, uma vez que este elemento químico atua no sis... more Os sais de lítio são usados no transtorno de humor, uma vez que este elemento químico atua no sistema nervoso central. Para aumentar a biodisponibilidade deste metal, estratégias são estudadas, como a bioacumulação em basidiomicetos. Desta forma, o objetivo deste trabalho foi realizar uma revisão bibliográfica sobre a capacidade de bioacumulação de lítio in vitro por basidiomicetos. Artigos científicos sobre biocumulação, lítio e basidiomicetos publicados nos anos de 1980 a 2020 foram pesquisados. Uma alternativa para o tratamento do transtorno de humor bipolar é o carbonato de lítio, porém sua faixa terapêutica é estreita, o que pode levar a intoxicação medicamentosa. A bioacumulação de lítio in vitro em basidiomicetos pode ajudar na redução dos efeitos colaterais e melhorar a biodisponibilidade do lítio. São escassos os estudos sobre bioacumulação de lítio em basidiomicetos, e a maioria utiliza LiCl como fonte de lítio, provavelmente pela sua alta solubilidade. O cultivo micelial ...
Basidiomicetos tem sido amplamente utilizados como produtores de enzimas, no entanto sao pouco ex... more Basidiomicetos tem sido amplamente utilizados como produtores de enzimas, no entanto sao pouco explorados quanto a sua capacidade de producao de proteases. Estes fungos sao reconhecidos pelas suas propriedades antitumorais, hipocolesterolemicas, antimutagenicas, antioxidantes entre outras. Assim, a associacao destas propriedades aos derivados do leite pode potencializar estes produtos como alimentos funcionais. Neste sentido, o objetivo deste trabalho foi selecionar basidiomicetos produtores de proteases, com potencial uso no processo de fabricacao de derivados do leite. Foram utilizadas 27 linhagens de fungos crescidas em meio minimo adicionado de 0,2% de caseina. A atividade proteolitica foi verificada pela formacao de halo pela adicao de uma solucao saturada de (NH4)2SO4. Concluiu-se que a producao de proteases nao apresenta relacao com o crescimento micelial. O melhor produtor de proteases e a linhagem Lentinula edodes U8/1, seguida por Pleurotus sp (U2/9, U6/10 e U2/11). Os bas...
Brazilian Journal of Microbiology, 2019
White-rot basidiomycetes such as Lentinus crinitus produce laccases with potential use in dye bio... more White-rot basidiomycetes such as Lentinus crinitus produce laccases with potential use in dye biodegradation. However, high productivity and enzymes with specific properties are required in order to make viable laccase production. We aimed to produce laccase from Lentinus crinitus grown in sugarcane bagasse for dye decolorization. Solid state cultivation medium had sugarcane bagasse added with a nutrient solution of 10 g/L glucose, 1 g/L KH 2 PO 4 , 0.5 g/L MgSO 4 , 0.001 g/L FeSO 4 , 0.01 g/L ZnSO 4 , and 0.01 g/L MnSO 4. The addition of different nitrogen sources (peptone, urea, or peptone plus urea) and different nitrogen concentrations (0, 0.4, 0.6, 0.8, 1.0, and 1.2 g/L) were evaluated. Enzymatic extract was used in the decolorization of azo dyes, reactive blue 220 (RB220) and reactive black 5 (RB5), and anthraquinone dye, Remazol brilliant blue R (RBBR). The greatest laccase activity (4800 U/ g dry mass) occurred when the peptone and urea mixture was added to the solid state cultivation medium. When the nitrogen concentration was 1 g/L, the laccase activity increased to 6555 U/g dry mass. The laccase activity peak occurred on the 10th day, and the maximum decolorization within 24 h was observed with enzymatic extracts obtained on different cultivation days, i.e., 6th day for RB220, 10th day for RB5, and 9th day for RBBR. Manganese and lignin peroxidases were not produced when nitrogen was added to the cultivation medium. The crude enzymatic extract was more effective in the decolorization of azo dyes (RB220 and RB5), more than 90% of decolorization, than anthraquinone dye with 77% decolorization.
Chemosphere, 2019
Lentinus crinitus is an important basidiomycete consumed by ethnic groups from the Amazon, common... more Lentinus crinitus is an important basidiomycete consumed by ethnic groups from the Amazon, commonly found in decomposing trees with high lignolytic and antioxidant activities. Lithium is a mood stabilizer, antiepileptic, antipsychotic, and antidepressant used in clinical practice. This study aimed to evaluate L. crinitus mycelial biomass bioaccumulated with lithium in liquid cultivation medium. The malt extract medium was added from zero to 100 mg L-1 lithium from two lithium sources (Li 2 CO 3 and LiCl). The maximum mycelial biomass production was 7,218.89 mg L-1 in the culture medium added ACCEPTED MANUSCRIPT with 5 mg L-1 lithium from LiCl. The highest lithium concentration in the mycelial biomass was of 574.72 µg g-1 produced in the culture medium with 25 mg L-1 lithium from Li 2 CO 3. Pearson's correlation showed that Li 2 CO 3 reduces the mycelial biomass and increases lithium bioaccumulation. The maximum translocated lithium from cultivation medium to mycelial biomass was up to 19 or 28% with LiCl or Li 2 CO 3 , respectively. Therefore, although Li 2 CO 3 presents greater inhibition on the mycelial biomass production, it promoted greater lithium bioaccumulation in L. crinitus mycelial biomass and resulted in greater yield of lithium translocation. The equivalent daily dose of lithium for psychiatric treatment, without bioavailability studies, could be reached with 97.4 g lithium-enriched mycelial biomass and, based in the literature, for reduction of violence and criminality rates the amount could be reached with 0.24-0.58 mg. Thus, the development of lithium-enriched mycelial biomass could be an alternative functional food.
Brazilian Journal of Microbiology, 2019
This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelia... more This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelial viability of A. subrufescens after 5-year cryopreservation at − 70°C. Mycelia were grown in three types of whole or ground grains, with or without 5% glycerol addition in the substrate and/or in a cryotube. After 5 years of cryopreservation at − 70°C, every treatment was thawed and recovered in malt extract culture medium with 15 (solid culture medium) or 5 g L −1 (semisolid culture medium) of agar. The semisolid recovery culture medium increased the mycelial viability recovery capacity of A. subrufescens cryopreserved for 5 years in grains with glycerol only in the cryotube, and specifically with medium-hard wheat grain without glycerol addition at all. Agar-based substrates such as malt extract agar, agar-ground grain, or the one with glycerol addition to the substrate were not effective to keep the mycelial viability, regardless of the recovery culture medium consistency. Hard and medium-hard endosperm wheat grains or hard endosperm rye grains with addition of glycerol as cryoprotectant only to the cryotube were effective to cryopreserve the fungus for 5 years without cryoprotectant addition in the substrate.
Biocatalysis and Agricultural Biotechnology, 2018
Lentinus crinitus is a basidiomycete with the ability to produce laccase and other enzymes of bio... more Lentinus crinitus is a basidiomycete with the ability to produce laccase and other enzymes of biotechnological interest. Laccase production depends on cultivation conditions and can be induced by metallic, phenolic or aromatic compounds. This study aimed to evaluate the laccase production of L. crinitus cultivated in sugarcane vinasse and urea with addition of laccase inducers. L. crinitus was cultivated in sugarcane vinasse with urea and supplemented with copper sulfate (0, 50, 150, 250, 350 and 450 μM) and aromatic compounds (guaiacol, veratryl alcohol, or 2,5-xylidine) added alone or combined to the cultivation medium. Cooper (150 or 250 μM) increased laccase activity in 14.5 (49000 U/L) and 28.5% (55000 U/L), respectively. Guaiacol had no significant effect, but veratryl alcohol (~46000 U/L) and xylidine (~75000 U/L) increased laccase activity. Copper with other inducers had the most relevant effects on laccase production. Simultaneous guaiacol addition with 150 or 250 μM copper increased 108 and 151% laccase activity, respectively, when compared with only copper addition to vinasse-urea medium. Veratryl alcohol and 150 or 250 μM copper addition increased 100 and 138% laccase activity, respectively, when compared to only copper addition. Interaction analysis of the variables guaiacol or veratryl alcohol and copper (250 µM) added to cultivation medium demonstrated synergistic effects resulting in laccase activity increase. These results highlight the importance of L. crinitus as a laccase producer and emphasize the relevance of studies using a combination of inducers to increase laccase production.
Biocatalysis and Agricultural Biotechnology, 2018
Panus strigellus is a native basidiomycete from Brazil with laccase production potential. Laccase... more Panus strigellus is a native basidiomycete from Brazil with laccase production potential. Laccases are involved in the biodegradation of dyes and their production can be stimulated by cultivation with agro industrial by-products. This study aimed to evaluate laccase production of P. strigellus cultivated with sugarcane bagasse on dye decolorization. Culture media had different concentrations of sugarcane bagasse (SB), sources of nitrogen (urea, yeast extract, ammonium sulfate and sodium nitrate), and concentrations of copper sulfate (0, 50, 100, 200, and 300 μM). Enzymatic extract from fungal cultivation was used in the decolorization of reactive blue 220 (RB220), malachite green (MG), and remazol brilliant blue R (RBBR). Laccase production was inversely proportional to the amount of SB in cultivation medium and higher titers of enzyme were obtained when 20 g/L SB (SB20) was used. Cultivation medium with SB20 and most nitrogen sources did not have a positive effect 2 on laccase production whereas the addition of urea or yeast-extract reduced laccase production. Most of the copper sulfate concentrations added to SB20 medium increased laccase production, and the maximum laccase production was 55342 U/L in the SB20 medium with 200 μM copper. The crude enzymatic extract was more effective in the decolorization of azo dye (RB220), with more than 90% of decolorization in 24 h, than anthraquinone dye (RBBR), with 60% decolorization, and triphenylmethane dye (MG), with 68% decolorization, both in 72 h.
Arquivos De Ciencias Veterinarias E Zoologia Da Unipar, May 6, 2013
Atividade antimicrobiana de óleos essenciais de condimentos frente a Staphylococcus spp isolados ... more Atividade antimicrobiana de óleos essenciais de condimentos frente a Staphylococcus spp isolados de mastite caprina.
Arquivos De Ciencias Veterinarias E Zoologia Da Unipar, May 9, 2013
A determinacao de biomassa micelial fungica crescida em substratos de cultivo solido particulado ... more A determinacao de biomassa micelial fungica crescida em substratos de cultivo solido particulado (SCSP) e ainda um desafio devido a dificuldade de separacao do micelio e o substrato. O objetivo deste trabalho foi avaliar a tecnica de microscopia de epifluorescencia para determinacao da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinacao da exatidao da metodologia P. ostreatus foi crescido em meio liquido de extrato de malte e; a biomassa micelial foi separada por centrifugacao, liofilizada e moida. Concentracoes conhecidas do po do micelio foram misturadas ao SCSP, composto de bagaco de cana de acucar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescencia. Para promover a variacao da biomassa micelial a ser determinada por microscopia de epifluorescencia, SCSP adicionado de diferentes concentracoes de ferro foram utilizados para o crescimento do fungo. Concluiu-se que a tecnica apresenta baixa precisao e exatidao, o que implica na necessidade de maiores estudos para aplicacao desta tecnica para a determinacao de biomassa micelial crescida em SCSP.
Current Microbiology, 2015
The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subr... more The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subrufescens (=A. blazei; =A. brasiliensis) obtained by different extraction methods were evaluated on Aspergillus nidulans conidia submitted to ultraviolet (UV) light. The aqueous extracts were obtained by three extraction methods: maceration, infusion, and decoction, at two different extraction times. The extracts of A. subrufescens did not present toxicity for A. nidulans conidia. A suspension of A. nidulans conidia was submitted to extracts before and after the exposure to UV light. All basidiocarp extracts, regardless of the extraction method or development stage, protected A. nidulans conidia against the damaging effects of the mutagenic agent. The antimutagenic and photoprotective activity was strengthened with extracts obtained by 168-h maceration, followed by 24-h maceration and 60-min infusion and, at last, by 30-min infusion. Although the extracts presented protector effect as well as recoverer effect to the action of UV light, the preventive effect was more evident. Differences in the biological activity in function of the different development stages were detected with greater antimutagenic and photoprotective activity for the opened basidiocarps. However, the extraction method is the most important factor to be considered when compared to the basidiocarp development stage to obtain better antimutagenic and photoprotective activity of A. subrufescens basidiocarps.
International Journal of Medicinal Mushrooms, 2015
Many alternative compounds have been tested to improve poultry performance but few of them have p... more Many alternative compounds have been tested to improve poultry performance but few of them have previously used mycelial-colonized substrate to partially replace standard diet in broiler chickens. The objective of this study was to evaluate broiler chicken production, health, and meat sensory characteristics, with partial replacement of the standard diet by Pleurotus ostreatus-colonized substrate. One hundred fifty 1-day-old male Cobb chicks were given standard diet partially replaced by 0, 5, 10, 100, or 200 g·kg-1 of P. ostreatus-colonized substrate and randomly distributed into five treatments. Each treatment had three replicates, with 10 birds per replicate, totaling 30 birds. The replacement of the standard diet by 10 g·kg-1 of colonized substrate increased (P≤0.05) chicken body mass up to 57% at 21 days, and up to 28% at 42 days. In general, partial replacement of standard diet by colonized substrate increased hematocrits and typical lymphocytes, and reduced low density lipoproteins. Also, it reduced chicken production period up to 21% and there is no meat taste alteration. The use of P. ostreatus-colonized substrate in chicken feeding is an alternative method to improve broiler chicken production.