Polyphasic approach including MALDI-TOF MS/MS analysis for identification and characterisation of Fusarium verticillioides in Brazilian corn kernels

Fusarium verticillioides is considered one of the most important global sources of fumonisin contamination in food and feed. Corn is one of the main commodities produced in the Northeastern Region of Brazil. The present study investigated potential mycotoxigenic fungal strains belonging to the F. verticillioides species isolated from corn kernels in 3 different Regions of the Brazilian State of Pernambuco. A polyphasic approach including classical taxonomy, molecular biology, MALDI-TOF MS and MALDI-TOF MS/MS for the identification and characterisation of the F. verticillioides strains was used. Sixty F. verticillioides strains were isolated and successfully identified by classical morphology, proteomic profiles of MALDI-TOF MS, and by molecular biology using the species-specific primers VERT-1 and VERT-2. FUM1 gene was further detected for all the 60 F. verticillioides by using the primers VERTF-1 and VERTF-2 and through the amplification profiles of the ISSR regions using the primers (GTG)5 and (GACA)4. Results obtained from molecular analysis shown a low genetic variability among these isolates from the different geographical regions. All of the 60 F. verticillioides isolates assessed by MALDI-TOF MS/MS presented ion peaks with the molecular mass of the fumonisin B1 (721.83 g/mol) and B2 (705.83 g/mol)Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior-CAPES/Brazil for the Financial Suppor

Cellulolytic ability of Penicillium strains isolated from soil of the Brazilian Atlantic forest

Penicillium spp. are capable of degrading plant wastes by producing large amounts of enzymes such as cellulases. These form a complex capable of acting on cellulosic materials and producing sugars with industrial interest (e.g., ethanol production). Cellulases are also used for (a) pulp and paper industry (b) in the textile industry. The aim of this study was to evaluate the cellulolytic capability of 17 strains of Penicillium isolated from soil of the Brazilian Atlantic Forest and conserved under mineral oil at the URM Culture Collection. All strains were re-grown from mineral oil and re-identifiied. Each strain was grown in synthetic medium with carboxymethylcellulose as the carbon source and incubated for 5 days at 28°C. Strains were subjected to heat shock for 16h at 50°C. Thereafter, onto each colony was added 5 ml of Congo red stain solution in Tris-HCl. After 30 min this solution was removed and the cultures were washed and submerged under 0.1 M NaCl aqueous solution for 5 min. Finally, an enzymatic index was calculated from the ratio of the diameter of the halo around each colony to the diameter of the colony. All of the 17 strains tested showed a halo of cellulose degradation, indicating enzyme production. The enzymatic ratios varied between 0.2 (Penicillium brevicompactum URM5994) and 3.3 (Penicillium glabrum URM6009). Thus, Penicillium glabrum URM6009 is evaluated as a high producer of cellulase. It was selected for quantitative production of this enzyme and additional studies are taking place in order to verify potential industrial application for clarification of fruit juices

Genotyping of two Neisseria gonorrhoeae fluroquinolone-resistant strains in the Brazilian Amazon Region

We report two ciprofloxacin and ofloxacin-resistant Neisseria gonorrhoeae strains that were isolated from the urethral discharge of male patients at the sexually transmitted diseases outpatient clinic of the Alfredo da Matta Foundation (Manaus, state of Amazonas, Brazil). The gonococci displayed minimal inhibitory concentrations (> 32.00 µg/mL) and three mutations in the quinolone resistance-determining region (S91F and D95G in GyrA and S87R in ParC). Both isolates were genotyped using N. gonorrhoeae multi-antigen sequence typing and the analysis showed that the ST225 which represented an emerging widespread multi-resistant clone that has also been associated with reduced susceptibility to ceftriaxone. We recommend continued surveillance of this pathogen to assess the efficacy of anti-gonococcal antibiotics in Brazil

DISTRIBUIÇÃO DA MEIOFAUNA EM DOIS HABITATS ESTUARINOS DA APA DO RIO MAMANGUAPE (NE BRASIL)

As assembleias infaunais são conhecidas por terem um papel importante em termos do uso da energia em ecossistemas estuarinos através de processos de decomposição. Estes organismos têm diferentes funções ecológicas, tais como ciclagem de nutrientes, degradação de poluentes, a dispersão, o processo de produção secundária, aeração e bioturbação, desempenhando um papel chave na cadeia alimentar dos ecossistemas de mangue. O objetivo deste estudo foi comparar as assembleias infaunais de dois ambientes estuarinos com diferentes tipos de sedimentos. As amostras foram coletadas em quatro estações ao longo do estuário, considerando dois tipos de substrato (areia e lama). Um total de 102 indivíduos pertencentes a nove táxons foram registrados. Foram observadas diferenças significativas na densidade dentro e entre os substratos amostrados, observando-se o maior número médio de indivíduos por cm³ e diversidade no fundo lamacento. Maior riqueza e equitabilidade foram encontradas na areia. Assim, as informações sobre as assembleias infaunais nos dois tipos de substratos estudados , contribui para a descrição biológica da área de estudo , tornando-se estudos adicionais necessários que se concentram em fatores físico- químicos, tamanho de partículas e matéria orgânica , uma vez que esses fatores têm sido fortemente correlacionada com a distribuição de organismos bentônicos

Background choice and immobility as context dependent tadpole responses to perceived predation risk

The association of immobility and camouflage is widespread as a defensive mechanism in prey from varied taxa. However, many experiments assessing the reaction of prey to predator cues are conducted under artificial laboratory conditions. In a previous experiment we observed the tadpoles of Ololygon machadoi (Hylidae) to respond to predator visual and/or chemical cues by choosing backgrounds that improve their disruptive properties, but detected no associated reduction of movement. Here we experimentally demonstrate this response in the species’ natural habitat, on backgrounds where the tadpoles are likely to achieve their best camouflage. We also tested whether previous experiences could influence both background choice and immobility in O. machadoi tadpoles. These novel experimental results suggest that a defensive behavior—i.e., reduction of movement—in these tadpoles is more strongly expressed under the natural conditions where they evolved, compared to laboratory conditions where prey and predator were brought into closer contact. Besides, previous experiences are likely to play an important role in expressed defensive responses

Identification of radiologic and clinicopathologic variables associated with tumor regression pattern and distribution of cancer cells after short-course radiotherapy and consolidation chemotherapy in patients with rectal cancer

BackgroundKnowledge of the pattern of regression and distribution of residual tumor cells may assist in the selection of candidates for rectum-sparing strategies.ObjectiveTo investigate and identify factors associated with tumor regression pattern and distribution of residual tumor cells.MethodsWe conducted a prospective study of patients with T3/T4 N0/N+ adenocarcinoma of the middle and lower third of the rectum (≤10 cm) treated with radiotherapy (5×5 Gy) followed by 6 cycles of CAPOX chemotherapy. The pattern of tumor regression was classified as fragmented or solid. Microscopic intramural spread was measured. We used a model of distribution of residual tumor cells not yet applied to rectal cancer, defined as follows: type I (luminal), type II (invasive front), type III (concentric), and type IV (random).ResultsForty patients were included with a median age of 66 years; 23 (57.5%) were men. A fragmented pattern was identified in 18 patients (45.0%), and a solid pattern in 22 (55.0%). Microscopic intramural spread was identified in 25 patients (62.5%), extending from 1 to 18 mm (median, 4 mm). There were 14 cases (35.0%) of microscopic intramural spread ≥10 mm. All cases of fragmented regression pattern, except one, showed microscopic intramural spread. Within the fragmented pattern, microscopic intramural spread was 4–8 mm in 4 cases and ≥10 mm in the remaining cases. All cases of microscopic intramural spread ≥ 10 mm were within the fragmented pattern. Regarding the distribution pattern of residual tumor cells, 11 cases (31.5%) were classified as type I, 14 (40.0%) as type II, 10 (28.5%) as type III, and none as type IV. Carcinoembryonic antigen levels >5 ng/mL, downsizing <50%, residual mucosal abnormality >20 mm, and anatomopathologic lymph node involvement were significantly associated with the occurrence of fragmentation (P<0.05). Having received all 6 cycles of CAPOX chemotherapy and absence of microscopic intramural spread were significantly associated with the type I distribution pattern (P<0.05).ConclusionThe occurrence of a fragmented regression pattern is common, as is the presence of microscopic intramural spread. We could identify radiologic and clinicopathologic factors associated with the pattern of tumor regression and a type I distribution pattern

Protótipo acessível de um filtro com materiais reutilizáveis para captação de água da chuva em calhas residenciais e industriais / Affordable prototype of a filter with reusable materials for rainwater harvesting in residential and industrial gutters

O objetivo desse projeto é a elaboração da prototipagem de um filtro com materiais financeiramente acessíveis, captando água da chuva através de calhas no telhado. A água captada e tratada pelo filtro, poderá ser utilizada para diversos fins, salientando que o método de filtragem proposto não contempla a capacidade de ingestão da água filtrada. O insumo terá um objetivo de ser empregado nos ambientes industriais e residenciais. O filtro se apresenta como uma proposta inicial de sustentabilidade simples e eficiente, com aspectos de utilização do insumo tratado, por exemplo: lavagem de quintal, carro, moto, louça, roupas e irrigação de hortas entre outras funcionalidades, apresentando como vantagem o custo benefício atrelado ao padrão sustentável, viável e econômico ao usuário. Compreendendo a escassez de água nas diversas regiões, o protótipo torna-se uma excelente opção para sustentabilidade da água, logo o serviço doméstico economizará água e os processos industriais utilizarão da água coletada pelo filtro empregando no processo fabril, não necessitando de utilizar a água principalmente potável. A região municipal onde ocorreu o desenvolvimento da pesquisa, a precipitação atmosférica municipal apresentou valores relevantes no âmbito municipal, regional, estadual e nacional. Outro fator relevante que contribui para a continuidade das pesquisas e aperfeiçoamento do protótipo é que o município, situa-se na região do Vale do Paraíba, localizado no estado de São Paulo, consistindo em um cenário fértil e relevante de indústrias regionais, nacionais, internacionais e multinacionais, concluindo assim a eficácia para os habitantes e indústrias. Os resultados obtidos foram satisfatórios, a capacidade volumétrica do filtro foi de aproximadamente vinte litros de água e o valor monetário para o desenvolvimento do filtro e instalação na residência ou indústria, no ano de 2018 que foi desenvolvido, não ultrapassou o valor de cem reais

Predicting the Proteins of Angomonas deanei, Strigomonas culicis and Their Respective Endosymbionts Reveals New Aspects of the Trypanosomatidae Family

Endosymbiont-bearing trypanosomatids have been considered excellent models for the study of cell evolution because the host protozoan co-evolves with an intracellular bacterium in a mutualistic relationship. Such protozoa inhabit a single invertebrate host during their entire life cycle and exhibit special characteristics that group them in a particular phylogenetic cluster of the Trypanosomatidae family, thus classified as monoxenics. in an effort to better understand such symbiotic association, we used DNA pyrosequencing and a reference-guided assembly to generate reads that predicted 16,960 and 12,162 open reading frames (ORFs) in two symbiont-bearing trypanosomatids, Angomonas deanei (previously named as Crithidia deanei) and Strigomonas culicis (first known as Blastocrithidia culicis), respectively. Identification of each ORF was based primarily on TriTrypDB using tblastn, and each ORF was confirmed by employing getorf from EMBOSS and Newbler 2.6 when necessary. the monoxenic organisms revealed conserved housekeeping functions when compared to other trypanosomatids, especially compared with Leishmania major. However, major differences were found in ORFs corresponding to the cytoskeleton, the kinetoplast, and the paraflagellar structure. the monoxenic organisms also contain a large number of genes for cytosolic calpain-like and surface gp63 metalloproteases and a reduced number of compartmentalized cysteine proteases in comparison to other TriTryp organisms, reflecting adaptations to the presence of the symbiont. the assembled bacterial endosymbiont sequences exhibit a high A+T content with a total of 787 and 769 ORFs for the Angomonas deanei and Strigomonas culicis endosymbionts, respectively, and indicate that these organisms hold a common ancestor related to the Alcaligenaceae family. Importantly, both symbionts contain enzymes that complement essential host cell biosynthetic pathways, such as those for amino acid, lipid and purine/pyrimidine metabolism. These findings increase our understanding of the intricate symbiotic relationship between the bacterium and the trypanosomatid host and provide clues to better understand eukaryotic cell evolution.Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)ERC AdG SISYPHEUniv Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Lab Ultraestrutura Celular Hertha Meyer, BR-21941 Rio de Janeiro, BrazilUniv Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Lab Metab Macromol Firmino Torres de Castro, BR-21941 Rio de Janeiro, BrazilLab Bioinformat, Lab Nacl Computacao Cient, Rio de Janeiro, BrazilINRIA Grenoble Rhone Alpes, BAMBOO Team, Villeurbanne, FranceUniv Lyon 1, CNRS, UMR5558, Lab Biometrie & Biol Evolut, F-69622 Villeurbanne, FranceUniv Estadual Campinas, Inst Biol, Dept Genet Evolucao & Bioagentes, São Paulo, BrazilUniv São Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Dept Ciencias Farmaceut, São Paulo, BrazilLab Nacl Ciencia & Tecnol Bioetano, São Paulo, BrazilUniv Fed Minas Gerais, Inst Ciencias Biol, Dept Bioquim & Imunol, Belo Horizonte, MG, BrazilUniv Fed Goias, Inst Ciencias Biol, Mol Biol Lab, Goiania, Go, BrazilFundacao Oswaldo Cruz, Inst Carlos Chagas, Lab Biol Mol Tripanossomatideos, Curitiba, Parana, BrazilFundacao Oswaldo Cruz, Inst Carlos Chagas, Lab Genom Func, Curitiba, Parana, BrazilUniv Estadual Campinas, Ctr Pluridisciplinar Pesquisas Quim Biol & Agr, São Paulo, BrazilUniv Fed Minas Gerais, Inst Ciencias Biol, Dept Parasitol, Belo Horizonte, MG, BrazilUniv Fed Santa Catarina, Dept Microbiol Imunol & Parasitol, Ctr Ciencias Biol, Lab Protozool & Bioinformat, Florianopolis, SC, BrazilUniv Fed Vicosa, Dept Bioquim & Biol Mol, Ctr Ciencias Biol & Saude, Vicosa, MG, BrazilInst Butantan, Lab Especial Ciclo Celular, São Paulo, BrazilUniv São Paulo, Dept Biol, Fac Filosofia Ciencias & Letras Ribeirao Preto, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilWeb of Scienc

Ocorrência de leveduras na cavidade oral e traqueia de aves de rapina

Objetivou-se com esta pesquisa isolar e identificar leveduras da cavidade oral e traqueia de aves de rapina. Para este estudo, utilizaram-se 17 aves de rapina (três carcarás (Caracara plancus), 11 gaviões-carijós (Rupornis magnirostris), 1 coruja-das-torres (Tyto furcata) e duas corujas-orelhuda (Rhinoptynx clamator). As amostras foram colhidas da cavidade oral e traqueia com auxílio de swabs estéreis. Foram cultivadas 34 amostras em ágar Sabouraud dextrose com cloranfenicol (100 mg/L) e incubadas em aerobiose à temperatura ambiente por um período mínimo de sete dias e máximo de 15 dias, sendo observadas diariamente. Foram isolados 14 (93,3%) amostras de leveduras da cavidade oral e apenas uma (6,7%) da traqueia. Dos 15 isolados, 14 (93,3%) foram obtidos de amostras de gavião-carijós (Rupornis magnirostris) e uma (6,67%) em carcará (Caracara plancus). As espécies identificadas foram: Candida albicans (13,3%), C. parapsilosis (20%), C. tropicallis (26,7%), C. magnoliae (6,78%), Candida sp. (13,3%) e Trichosporon cutaneum (20%). O isolamento e identificação de leveduras em aves de rapina é epidemiologicamente importante para uma melhor compreensão dos processos patológicos na cavidade oral e na traqueia das espécies estudadas, uma vez que seu conhecimento permite adoção de medidas para a prevenção de doenças causadas por esses agentes oportunistas