Biogas production through co-digestion of enzymatically pretreated corn bran and cow manure

Biogas production from wastes is an alternative that contributes positively to the environment and minimize the dependence on fossil energy sources. Additionally, the reuse of biomasses helps to reduce the waste production, but a pretreatment is required to use it in the anaerobic digestion. Here biogas was produced through co-digestion of enzymatically pretreated corn bran and cow manure. Firstly, it was selected the most hydrolysable waste (barley bagasse, sugar cane bagasse, elephant grass, thick orange pie, average orange pie, wheat bran, coffee grounds, orange peel, white sludge, vinasse, corn bran, soy bran, soy peel, cotton bran, cassava husk, cassava flour, banana peel, corn bran, sorghum stem, sorghum seed, total sorghum and wet distiller grain) by the crude extracts containing amylase (secreted by Aspergillus brasiliensis), xylanase (Aspergillus tamarii Kita) and cellulase (Trichoderma reesei, Novozymes®). Later on, different mixtures of these enzymes were studied using simplex-centroid designs. The most hydrolyzed waste by each enzyme individually (measured by reducing sugar using dinitrosalicylic acid, DNS) at 50°C, 120 rpm and 24 h were corn bran, banana peel and sorghum seed. Then, the simplex-centroid designs resulted in model equations and respective response surface contours. Amylase extract had a significant positive influence on corn bran hydrolysis by maximizing the reducing sugar yield when it was used individually (35g/L of reducing sugar). After it, the pretreated corn bran and a cow manure (1:2 g of volatile solids) were employed for biogas production in batch assays. It was found a biogas accumulation of 326 mL in the 12nd day of anaerobic codigestion, which were similar to the control (containing 35 g/L of glucose alone) and 53% higher than that found with corn bran without enzymatic pretreatment. In conclusion, it was observed that the crude extract optimized for amylase production affected significantly the corn bran hydrolyses and consequently the biogas production in a co-digestion with cow manure.CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico process 142139/2017-3)FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo process 2018/07522-6)FCT (Fundação para a Ciência e Tecnologia)info:eu-repo/semantics/publishedVersio

A qualidade das bases de dados como factor crucial em estudos ambientais: condições de referência e tipologia com base piscícola para rios portugueses

Com base num conjunto superior a 1000 amostragens piscícolas em rios portugueses (troços vadeáveis) realizadas por diversas entidades nacionais nos últimos 10 anos, seleccionou-se um grupo de 459 locais que cumprem critérios de normalização de amostragem, procedimentos no terreno, e acuidade nas identificações taxonómicas. Estes locais representam a diversidade de condições naturais e de impactos humanos em Portugal Continental. Para caracterizar a magnitude e tipo de perturbação humana em cada local foram aplicadas 15 variáveis de pressão – ao nível do troço, segmento e bacia – classificadas de acordo com o desvio às condições naturais (de 1 para ausência de desvio, até 5 para forte degradação). Este passo envolveu um longo período de trabalho e baseou-se na análise de várias fontes de informação geográfica e documental, em inquéritos e no conhecimento do terreno. Durante as fases de selecção de locais e construção das bases de dados ambientais e biológicas, toda a informação foi sujeita a procedimentos de controle de qualidade de dados. É proposta uma metodologia para a selecção de sítios de referência; desta forma, foram incluídos neste grupo os locais com pontuação 1 ou 2 em, pelo menos, 80% das variáveis, permitindo a classificação 3 em 20% (três variáveis), excepto para a variável “abundância de indivíduos exóticos”, que obrigatoriamente correspondeu à pontuação 1 ou 2. Uma abordagem estatística multivariada suportou os dois passos seguintes: a definição da tipologia com base nas ictiocomunidades e a alocação de todos os locais num tipo. Utilizando os sítios de referência, e com base na classificação de grupos funcionais piscícolas, foram estabelecidos 6 tipos (t.): t. salmonícola da região norte; t. transição salmonícola-ciprinícola da região norte; t. ciprinícola de pequena dimensão das regiões norte interior e sul; t. ciprinícola de média dimensão da região norte; t. ciprinícola de média dimensão da região sul e t. ciprinícola da região norte litoral. A análise discriminante múltipla (ADM) suportou a tipologia piscícola, alocando correctamente 71 a 93% dos locais para os seis grupos; temperatura média em Julho, área de drenagem, altitude, precipitação média anual e uma variável categórica de enquadramento geográfico foram as variáveis retidas pelo modelo final. A ADM evidenciou elevada robustez ao classificar a maioria dos locais de não referência num dos tipos da respectiva região geográfica e ao detectar variações longitudinais das comunidades piscícolas ao longo de vários rios

Differential proteomic analysis by SWATH-MS unravels the most dominant mechanisms underlying yeast adaptation to non-optimal temperatures under anaerobic conditions

Elucidation of temperature tolerance mechanisms in yeast is essential for enhancing cellular robustness of strains, providing more economically and sustainable processes. We investigated the differential responses of three distinct Saccharomyces cerevisiae strains, an industrial wine strain, ADY5, a laboratory strain, CEN.PK113-7D and an industrial bioethanol strain, Ethanol Red, grown at sub- and supra-optimal temperatures under chemostat conditions. We employed anaerobic conditions, mimicking the industrial processes. The proteomic profile of these strains in all conditions was performed by sequential window acquisition of all theoretical spectra-mass spectrometry (SWATH-MS), allowing the quantification of 997 proteins, data available via ProteomeXchange (PXD016567). Our analysis demonstrated that temperature responses differ between the strains; however, we also found some common responsive proteins, revealing that the response to temperature involves general stress and specific mechanisms. Overall, sub-optimal temperature conditions involved a higher remodeling of the proteome. The proteomic data evidenced that the cold response involves strong repression of translation-related proteins as well as induction of amino acid metabolism, together with components related to protein folding and degradation while, the high temperature response mainly recruits amino acid metabolism. Our study provides a global and thorough insight into how growth temperature affects the yeast proteome, which can be a step forward in the comprehension and improvement of yeast thermotolerance.Financial support is acknowledged to Project ERA-IB “YeastTempTation” (ERA-IB-2-6/0001/2014), and FCT for the strategic funding of UIDB/04469/2020 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and BioTecNorte operation (NORTE-01-0145-FEDER-000004). Lallemand Ibéria, SA is acknowledged for the supply of yeast strains. Te proteomic analysis was carried out in the SCSIE University of Valencia Proteomics Unit, a member of the ISCIII ProteoRed Proteomics Platform. Authors thank to Luz Valero, researcher of the Proteomic unit, for her valuable support.info:eu-repo/semantics/publishedVersio

Harvest of endangered marine invertebrates in a priority area for conservation in Brazil

The harvesting of marine organisms for aquarium and ornamental art crafts is a very common and widespread activity, threatening a number of coral reef species. The goal of this manuscript is to report the intensive harvesting of two marine invertebrate species, the starfish Oreaster reticulatus and the gorgonian Phyllogorgia dilatata, in reef environments of a national priority area for the creation of a Marine Protected Area (MPA) in the Espírito Santo State, central coast of Brazil. These species were present on the 2003 Brazilian Red List of Endangered species as threatened of extinction, but P. dilatata was removed from the most recent 2014 Brazilian Red List, while O. reticulatus is currently considered vulnerable. Neither species have been assessed by IUCN. The harvest of these invertebrates was registered in 17% of the 51 days of monitoring (ten expeditions between March 2005 and February 2006), with catches of up to 24 starfishes and 1000 gorgonians in a single day. Starfishes used to be sold to local traders for US$0.50 each, and each gorgonian for US$ 0.15. This starfish is already in decline on a regional scale, and the actual harvesting rate of the gorgonian could lead to the extinction of the local population in about 17 years of exploitation. Due to the ecological importance of these two endangered species, their harvest threatens the entire local rocky reef ecosystem. Further ecological and fisheries studies on these species should be conducted, and the definition and creation of the MPA could help to manage all the unsustainable activities conducted and expected in the region

Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina.

Resumo: A artrite-encefalite caprina (CAE) é diagnosticada rotineiramente pela técnica de imunodifusão em gel de agarose (IDGA), que é considerada pouco sensível. Objetivou-se com este estudo padronizar testes de Elisa-i e Western Blot (WB) para diagnóstico precoce de anticorpos em caprinos contra CAEV e comparar os resultados obtidos nesses testes com a prova de IDGA. Para a padronização dos testes Elisa-i e WB, utilizaram-se diferentes concentrações e diluições de antígeno, soros e conjugado. No Elisa-i, adotaram-se microplacas rígidas com 96 poços, sendo a combinação de concentração de 0,5µg/poço de antígeno e diluições de 1:100 de soro e 1:1500 de conjugado a que apresentou melhor resultado. No WB foram utilizadas membranas de nitrocelulose, definindo-se as diluições de 1:50 de soro e 1:15000 de conjugado. Para avaliar o desempenho das técnicas, 222 amostras de soro caprino foram testadas e os dados obtidos foram comparados com o IDGA. A sensibilidade e a especificidade do Elisa-i/IDGA, WB/IDGA e WB/Elisa-i foram de 70% e 91%, 100% e 72,6%, 84,6% e 76,5%, concomitantemente. O índice Kappa desses testes foi de 0,35, 0,2 e 0,36, respectivamente. As técnicas de Elisa-i e WB apresentaram-se mais sensíveis que a IDGA, podendo ser utilizadas como ferramentas para o diagnóstico precoce da CAE. [Standardization of indirect Elisa and Western Blot for diagnosis of Caprine Arthritis-Encephalitis]. Abstract: Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE

Wharton's jelly cells from sheep umbilical cord maintained with different culture media are permissive to in vitro infection by Small Ruminant Lentiviruses.

Abstract: This study aimed to isolate cells from the Wharton's jelly of umbilical cord (WJUC) of sheep collected during natural parturition using different culture media, in addition to reporting for the first time the permissiveness of these cells to in vitro infection by small ruminant lentiviruses. Ten umbilical cords were collected from healthy sheep. Each cord explants were grown in different media consisting of MEM, low glucose DMEM, M199, and RPMI-1640. The permissiveness of infection of sheep cells from WJUC was tested with CAEV-Cork and MVV-K1514 strains, inoculating 0.1 MOI of each viral strain. Four supernatants from each strain were obtained from WJUC sheep cell cultures infected in different media. The results demonstrated the presence of cytopathic effect after the in vitro infection by CAEV-Cork and MVV-K1514 with all of the tested culture media. Nested-PCR detected proviral DNA in all supernatants. Supernatants containing CAEV-Cork viruses had TCID50/ml titres of 105.5 in MEM, 104.0 in low glucose DMEM, 105.0 in M199, and 105.7 in RPMI-1640. Supernatants containing the MVV-K1514 virus had TCID50/ml titres of 104.3 in MEM, 103.5 in low-glucose DMEM, 104.7 in M199, and 103.5 in RPMI-1640. Sheep cells from WJUC are permissive to in vitro infection by small ruminant lentivirus. [Células da geleia de Wharton do cordão umbilical ovino mantidas em diferentes meios de cultivo são permissivas à infecção in vitro por lentivírus de pequenos ruminantes]. Resumo: O objetivo deste estudo foi isolar células da geleia de Wharton do cordão umbilical (GWCU) ovino coletado por ocasião do parto natural, utilizando-se diferentes meios de cultivo, além de relatar, pela primeira vez, sua permissividade à infecção in vitro por lentivírus de pequenos ruminantes (LVPRs). Dez cordões umbilicais foram coletados de ovelhas hígidas e soronegativas para LVPRs pelo teste de imunodifusão em gel de agarose (IDGA). De cada cordão, explantes foram cultivados em quatro meios distintos que consistiram em MEM, DMEM baixa glicose, meio 199 e RPMI-1640, todos acrescidos de 10% de soro fetal bovino em estufa com atmosfera úmida e 5% de CO2 a 37ºC. A permissividade de infecção das células GWCU ovino foi testada frente às cepas CAEV-Cork e MVV-K1514, inoculando-se 0,1 MOI de cada cepa viral e corando as monocamadas com May Grunwald Giemsa para visualização do efeito citopático. Foram obtidos quatro sobrenadantes CAEV-Cork e quatro MVV-K1514, provenientes do cultivo de células GWCU ovino infectadas por 21 dias em meios distintos, dos quais foram realizadas titulação em membrana sinovial caprina e extração do DNA pró-viral para realização de nested-PCR e eletroforese em gel de agarose a 2%. Os resultados demonstraram a presença de efeito citopático na infecção in vitro tanto por CAEV-Cork como por MVV-K1514 em todos os meios de cultivo, sendo visualizados sincícios e lise celular em microscópio invertido. A nested-PCR detectou o DNA pró-viral tanto do CAEV-Cork como do MVV-K1514 em todos os sobrenadantes. Os sobrenadantes contendo o vírus CAEV-Cork apresentaram títulos em TCID50/mL de 105,5 em MEM, 104,0 em DMEM baixa glicose, 105,0 em meio 199 e 105,7 em RPMI-1640. Os sobrenadantes contendo o vírus MVV-K1514 apresentaram título em TCID50/mL de 104,3 em MEM, 103,5 em DMEM baixa glicose, 104,7 em meio 199 e 103,5 em RPMI-1640. Células GWCU ovino são permissivas à infecção in vitro pelos lentivírus de pequenos ruminantes CAEV-Cork e MVV-K1514.[ALICE ANDRIOLI PINHEIRO]

Children’s coping styles and trauma symptoms after an explosion disaster

Background: The negative impact of trauma on children and adolescents is well documented. However, few studies have investigated the relationship between coping and trauma and distress symptoms after man-made disasters, especially those not related to war. Objective: This study investigated the relationship between children’s coping styles and their self-reported levels of trauma and distress symptoms after an explosion disaster in a residential area. Method: Participants were recruited through the local public school that served the affected residential area. A total of 333 children and adolescents from grades 3 through 10 participated in the study 16 months after the explosion. All participants filled out questionnaires to assess their trauma and distress symptoms as well as their coping strategies. The adolescents answered additional questions about pre-, peri-, and post-traumatic factors and filled out questionnaires about their trauma and distress symptoms, including aspects of somatization and negative affectivity. Results: The following variables were associated with a higher degree of trauma symptoms for children in grades 6 through 10 and explained 39% to 48% of the unique variance in these symptoms: female gender; the experience of traumatic events pre-disaster; the destruction of property or danger to life occurring during the disaster; the experience of traumatic events post-disaster; and the use of self-blame, emotion regulation, wishful thinking, and cognitive restructuring. For the younger children, pre-, peri-, and post-disaster factors were not measured. However, female gender and the use of self-blame as a coping strategy explained 26% of the variance in trauma symptoms. Conclusions: This study generally supports the findings of the limited literature addressing coping skills after man-made disasters. However, contrary to previous findings in community samples after episodes of terrorism, adaptive coping strategies such as cognitive restructuring were found to influence the variance of trauma and distress symptoms

Frequência de anticorpos contra o vírus da língua azul em ovinos do Estado do Ceará, Brasil.

Resumo: O objetivo deste trabalho foi avaliar a ocorrência de ovinos soropositivos para o vírus da língua azul (VLA) no Estado do Ceará, Brasil, e analisar as proteínas imunogênicas das cepas virais circulantes nesses rebanhos. O teste de imunodifusão em gel de agarose (IDGA) foi utilizado para pesquisar 271 amostras de soro oriundas de 16 rebanhos. Os resultados demonstraram que 27,3% (74/271) das amostras analisadas apresentaram anticorpos contra o agente e 68,8% (11/16) das propriedades tiveram animais positivos. O immunoblotting (IB) foi utilizado para analisar as proteínas imunogênicas do VLA a partir dos soros de animais positivos no IDGA. Os soros demonstraram forte reação contra a proteína viral VP2. Para o VLA, das sete proteínas estruturais, a VP2 é a principal a estimular a resposta imune protetora. Concluiu-se que a soropositividade para a língua azul (LA) nos rebanhos ovinos estudados no Ceará é alta, apesar dos animais não apresentarem sinais clínicos, indicativo de que o vírus ocorra de forma endêmica. Além disso, a resistência à doença apresentada pelos animais pode estar relacionada com a forte reação imunológica desses à proteína VP2. Sendo assim, outros estudos são necessários para melhor esclarecer a situação epidemiológica da LA no país, através da identificação dos vetores e sorotipos virais circulantes nas diferentes regiões. [Antibodies against the bluetongue virus in sheep flocks of Ceará State, Brazil.]. Abstract: The objective of this work was to verify the occurrence of sheep serologically positive for bluetongue virus (BTV) in the state of Ceará, Brazil, and analyze immunogenic proteins of circulating viral strains in these flocks. The agar gel immunodifusion test (AGID) was used to examine 271 serum samples from 16 herds. The results demonstrated that 27.3% (74/271) of the analyzed samples presented antibodies for the agent, and that 68.8% (11/16) of the properties presented positive animals. Immunoblotting (IB) was used to analyze the immunogenic proteins of BTV derived from AGID positive sera. Sera showed strong reaction against viral protein VP2. Of the seven BTV structural proteins, VP2 is the major protein to elicit protective immune responses. It was concluded that bluetongue (BT) seropositivity in sheep flocks studied in Ceará is high, despite that the animal's do not show clinical signs, indicating that it occurs in an endemic form. The animals? resistance to the disease may be related to the strong immune response to the protein VP2. Therefore, further studies are needed to better clarify the epidemiological situation of BT in Brazilian sheep flocks, through the identification of viral vectors and serotypes circulating in different regions

Comparison in the Trichoderma longibrachiatum xyloglucanase production using tamarind (Tamarindus indica) and jatobá (Hymenaea courbaril) seeds: factorial design and immobilization on ionic supports

in the control of the stretching and expansion of the plant cell wall. There are five types of enzymes known to be capable of cleaving the linear chain of xyloglucan, the most famous of them being the xyloglucanase (XEG). The immobilization can be used to solve problems related to stability, besides the economic benefits brought by the possibility of repeated use and recovery, decreasing the costs of production. Therefore, this study aims the optimization of the production of a xyloglucanase from Trichoderma longibrachiatum, with the aid of factorial design, using tamarind (Tamarindus indica) and jatobá (Hymenaea courbaril) seeds as carbon source; and the immobilization of the enzyme on ionic supports, such as MANAE (monoamino-N-aminoethyl), DEAE (diethylaminoethyl)-cellulose, CM (carboxymethyl)-cellulose and PEI (polyethyleneimine). High concentrations of carbon source in the culture medium, especially tamarind seeds, were the most favorable conditions for the greater activity of the xyloglucanase from T. longibrachiatum. The scaling up from Erlenmeyer flasks to the bioreactor was an essential strategy to increase the content of secreted enzyme. Regarding the biochemical characterization of the crude extract, the optimal temperature was 50-55 °C and the optimal pH 5.0. Regarding the stabilities to pH and to temperature, the enzyme was not stable for prolonged periods, which was crucial for the performing of immobilization on ionic resins (CM-cellulose, DEAE-cellulose, MANAE, and PEI), being the first time described in literature the immobilization of a xyloglucanase on these supports.We thank the Fundação de Amparo à Pesquisa do estado de São Paulo (process 2018/07522-6; 2014/50884-5), and Conselho Nacional de Dsenvolvimento Científico (process 301963/2017-7; 465319/2014-9).info:eu-repo/semantics/publishedVersio