Avaliação do consumo alimentar de pacientes com síndrome do intestino irritável em acompanhamento ambulatorial / Evaluation of carbohydrate and fiber consumption in patients with irritable bowel syndrome in outpatient treatment

ABSTRACT BACKGROUND: Irritable bowel syndrome (IBS) is a chronic functional condition, which main symptoms of pain, discomfort and abdominal distension, constipation, diarrhea, altered fecal consistency and sensation of incomplete evacuation can be influenced by the presence of dietary fiber and fermentable carbohydrates (FODMAPs). This study aimed to assess the relationship between the quantity of fermentable carbohydrates (FODMAP) and fiber consumed by individuals diagnosed with IBS, and their classification according to the Rome III criteria. METHODS: A transversal study was carried out in the Intestinal Outpatient Clinic of the Gastroenterology Discipline of UNIFESP. The nutrients of interest for the study were: fiber, general carbohydrates and FODMAPs, with intake quantity measured in grams, analyzed through portions consumed. A nutrition log was used, along with a semi-quantitative questionnaire of consumption frequency. RESULTS: The sample included 63 adult patients; 21 with constipated IBS, 21 with diarrhea IBS, and 21 with mixed IBS. Carbohydrate intake was suboptimal in 55.6% of patients in all groups; excessive consumption was identified in 38.1% of the diarrhea group, 14.3% of the mixed group and 38.1% of the constipated group. Low consumption of carbohydrates was found in 28.6% of diarrhea patients and 47.6% of the mixed group. A mean intake of 23 g of fiber per day was identified, lower than recommended. CONCLUSION: The study identified a number of inadequacies in the consumption of different nutrients, excessive carbohydrate intake, especially FODMAPs, identified by the respondents as responsible for a worsening of their conditions. By contrast, other food groups such as meat, eggs and dairy were consumed by the sample population in insufficient quantities.

RESUMO CONTEXTO: A síndrome do intestino irritável é um distúrbio funcional crônico, no qual a dieta, principalmente o teor de fibra dietética e presença de carboidratos fermentativos (FODMAPs) podem influenciar nos principais sintomas: dores, desconforto e/ou distensão abdominal, constipação, diarreia, alteração na consistência das fezes, sensação de evacuação incompleta. OBJETIVO: Este estudo teve como objetivo avaliar as quantidades de carboidratos fermentativos (FODMAP) e fibras consumidas por indivíduos com o diagnóstico de síndrome do intestino irritável e relacionar com seu modelo da classificação, segundo os critérios Roma III. MÉTODOS: Estudo transversal, realizado no Ambulatório de Doenças Intestinais da Disciplina de Gastroenterologia/UNIFESP. Os nutrientes de interesse para o estudo foram: fibras, carboidratos em geral e FODMAPs, calculando-se suas quantidades em gramas, analisadas através das porções consumidas. Os instrumentos de pesquisa utilizados: ficha de acompanhamento nutricional e questionário de frequência alimentar semi-quantitativo. RESULTADOS: A amostra incluiu 63 pacientes adultos, com síndrome do intestino irritável constipado (21), diarreico (21) e misto (21). O consumo de carboidratos mostrou-se inadequado em 55,6% dos indivíduos em todos os grupos; os que tinham alto consumo (38,1%) pertenciam ao grupo diarreia, 14,3% ao misto e 38,1 % ao constipado. Baixo consumo deste nutriente foi 28,6% nos casos de diarreia e 47,6% do misto. Observamos uma ingestão média de fibras equivalente à 23 g/dia, nos três grupos, inferior ao recomendado. CONCLUSÃO: O estudo permitiu reconhecer várias inadequações no consumo dos diferentes grupos de alimentos, particularmente excesso de carboidratos, incluindo os classificados como FODMAPs, identificados pelos doentes como responsáveis pela piora das suas queixas. Em contrapartida, nutrientes fundamentais, como carnes, ovos, leite e derivados estiveram referidos em níveis abaixo do recomendado.

Intestinal fructose malabsorption is associated with increased lactulose fermentation in the intestinal lumen / Má absorção intestinal de frutose associa-se com maior fermentação de lactulose na luz intestinal

Abstract Objective: To study fructose malabsorption in children and adolescents with abdominal pain associated with functional gastrointestinal disorders. As an additional objective, the association between intestinal fructose malabsorption and food intake, including the estimated fructose consumption, weight, height, and lactulose fermentability were also studied. Methods: The study included 31 patients with abdominal pain (11 with functional dyspepsia, 10 with irritable bowel syndrome, and 10 with functional abdominal pain). The hydrogen breath test was used to investigate fructose malabsorption and lactulose fermentation in the intestinal lumen. Food consumption was assessed by food registry. Weight and height were measured. Results: Fructose malabsorption was characterized in 21 (67.7%) patients (nine with irritable bowel syndrome, seven with functional abdominal pain, and five with functional dyspepsia). Intolerance after fructose administration was observed in six (28.6%) of the 21 patients with fructose malabsorption. Fructose malabsorption was associated with higher (p < 0.05) hydrogen production after lactulose ingestion, higher (p < 0.05) energy and carbohydrate consumption, and higher (p < 0.05) body mass index z-score value for age. Median estimates of daily fructose intake by patients with and without fructose malabsorption were, respectively, 16.1 and 10.5 g/day (p = 0.087). Conclusion: Fructose malabsorption is associated with increased lactulose fermentability in the intestinal lumen. Body mass index was higher in patients with fructose malabsorption.

Resumo Objetivo: Pesquisar a má absorção de frutose em crianças e adolescentes com dor abdominal associada com distúrbios funcionais gastrintestinais. Como objetivo adicional, estudou-se a relação entre a má absorção intestinal de frutose e a ingestão alimentar, inclusive a estimativa de consumo de frutose, o peso e a estatura dos pacientes e a capacidade de fermentação de lactulose. Métodos: Foram incluídos 31 pacientes com dor abdominal (11 com dispepsia funcional, 10 com síndrome do intestino irritável e 10 com dor abdominal funcional). O teste de hidrogênio no ar expirado foi usado para pesquisar a má absorção de frutose e a fermentação de lactulose na luz intestinal. O consumo alimentar foi avaliado por registro alimentar. Foram mensurados também o peso e a estatura dos pacientes. Resultados: Má absorção de frutose foi caracterizada em 21 (67,7%) pacientes (nove com síndrome do intestino irritável, sete com dor abdominal funcional e cinco com dispepsia funcional). Intolerância após administração de frutose foi observada em seis (28,6%) dos 21 pacientes com má absorção de frutose. Má absorção de frutose associou-se com maior produção de hidrogênio após ingestão de lactulose (p < 0,05), maior consumo de energia e carboidratos (p < 0,05) e maior valor de escore z de IMC para a idade (p < 0,05). As medianas da estimativa de ingestão diária de frutose pelos pacientes com e sem má absorção de frutose foram, respectivamente, 16,1 e 10,5 g/dia (p = 0,087). Conclusão: Má absorção de frutose associa-se com maior capacidade de fermentação de lactulose na luz intestinal. O índice de massa corporal foi maior nos pacientes com má absorção de frutose.

Produção de xilanases por fermentação em estado sólido de farelo de trigo utilizando consórcios fúngicos / Production of xylanases by solid state fermentation of wheat bran using fungal consortia

A fim de se agregar valor ao resíduo farelo de trigo gerado em indústrias do setor alimentício avaliou-se, no presente trabalho, o potencial deste subproduto como substrato para produção de enzima xilanase no cultivo em estado sólido, utilizando consórcios fúngicos bem como os fungos Aspergillus oryzae CCT nº 0975 (ATCC9362) eTrichoderma reesei CCT nº 2768 - QM 9414. Para tanto utilizou-se o farelo de trigo, não lavado e não autoclavado, como fonte de carbono e energia na fermentação em estado sólido pelo fungo Aspergillus oryzae que apresentou maior produção do percentual de proteína nas 72 horas de cultivo. Depois de realizado um Delineamento Composto Central Rotacional (DCCR) - planejamento fatorial 23,com três repetições no ponto central e seis pontos axiais - partiu-se para otimização dos fatores que foram considerados significativos no processo: umidade, pH e granulometria. Os fatores foram considerados significativos pela A NOVA com o nível de 95% de confiança e com o resultado otimizado de atividade enzimática de (1.84 ± 0.01) UI/mL utilizando pH 3,3, granulometria de 900,0 µm e umidade de 40%. O caldo enzimático obtido foi considerado eficiente na modificação de tipificação de farinhas de trigo pelo estudo dos parâmetros reológicos do falling number e alveografia sendo estável por cerca de 3 meses

This study aimed to find alternatives for wheat bran disposal destination generated in food industry sector,thus contributing to the reduction of the resultant impact of residue depo-sition in the environment. The poten-tial of the wheat bran as a substrate for xylanase production by solid-state fermentation using fungal con-sortiums as well as Aspergillus ory-zae (ATCC9362) and Trichoderma reesei (2768) was valued. The use of non-washed and non-autoclaved wheat bran as carbon and energy source in solid-state fermentation by A. oryzae fungus showed greater per-centage of produced protein after 72 h of cultivation. The use of a central composite rotatable design(CCRD), 23 factorial planning with three rep-etitions at the central point as well as six axial points, coupled with Sur-face Response Methodology (SRM) allowed to assay the influence of hu-midity, pH, and grain size (indepen-dent variables or factors) on the xy-lanase activity(dependent variable or response) as well as to optimize the best conditions for the enzyme production. The results showed that all factors and their combinations were significant at 95% confidence level. The optimized xylanase activi-ty was (1.84 ± 0.01) UI/mL, obtained at 40% humidity and pH 3.3 with a grain size of 900.0 µm. The produced broth was stable for 3 months and approximately had 50% of the initial xylanase activity at 4°C. SDS-PAGE assay showed that xylanase has 30 kDa molar mass. The obtained en-zymatic broth was efficient to modify wheat flours as shown by the falling number rheologic parameters and alveography assay

Enhancing inulinase yield by irradiation mutation associated with optimization of culture conditions

A new inulinase-producing strain was isolated from rhizosphere soils of Jerusalem artichoke collected from Shihezi (Xinjiang, China) using Jerusalem artichoke power (JAP) as sole carbon source. It was identified as an Aspergillus niger strain by analysis of 16S rRNA. To improve inulinase production, this fungus was subjected to mutagenesis induced by ⁶⁰Co γ-irradiation. A genetically stable mutant (designated E12) was obtained and it showed 2.7-fold higher inulinase activity (128 U/mL) than the parental strain in the supernatant of a submerged culture. Sequential methodology was used to optimize the inulinase production of stain E12. A screening trial was first performed using Plackett-Burman design and variables with statistically significant effects on inulinase bio-production were identified. These significant factors were further optimized by central composite design experiments and response surface methodology. Finally, it was found that the maximum inulinase production (185 U/mL) could be achieved under the optimized conditions namely pH 7.0, yeast extract concentration of 5.0 g/L, JAP concentration of 66.5 g/L, peptone concentration of 29.1 g/L, solution volume of 49.4 mL in 250-mL shake flasks, agitation speed of 180 rpm, and fermentation time of 60 h. The yield of inulinase under optimized culture conditions was approximately 1.4-fold of that obtained by using basal culture medium. These findings are of significance for the potential industrial application of the mutant E12.

Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO₂ concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking.

Characterization of mannitol-fermenting methicillin-resistant staphylococci isolated from pigs in Nigeria

This study was conducted to determine the species distribution, antimicrobial resistance pheno- and genotypes and virulence traits of mannitol-positive methicillin-resistant staphylococci (MRS) isolated from pigs in Nsukka agricultural zone, Nigeria. Twenty mannitol-positive methicillin-resistant coagulase-negative staphylococcal (MRCoNS) strains harboring the mecA gene were detected among the 64 Staphylococcus isolates from 291 pigs. A total of 4 species were identified among the MRCoNS isolates, namely, Staphylococcus sciuri (10 strains), Staphylococcus lentus (6 strains), Staphylococcus cohnii (3 strains) and Staphylococcus haemolyticus (one strain). All MRCoNS isolates were multidrug-resistant. In addition to β-lactams, the strains were resistant to fusidic acid (85%), tetracycline (75%), streptomycin (65%), ciprofloxacin (65%), and trimethoprim/sulphamethoxazole (60%). In addition to the mecA and blaZ genes, other antimicrobial resistance genes detected were tet(K), tet(M), tet(L), erm(B), erm(C), aacA-aphD, aphA3, str, dfrK, dfrG, cat_pC221, and cat_pC223. Thirteen isolates were found to be ciprofloxacin-resistant, and all harbored a Ser84Leu mutation within the QRDR of the GyrA protein, with 3 isolates showing 2 extra substitutions, Ser98Ile and Arg100Lys (one strain) and Glu88Asp and Asp96Thr (2 strains). A phylogenetic tree of the QRDR nucleotide sequences in the gyrA gene revealed a high nucleotide diversity, with several major clusters not associated with the bacterial species. Our study highlights the possibility of transfer of mecA ...

Feasibility of biohydrogen production from industrial wastes using defined microbial co-culture

BACKGROUND: The development of clean or novel alternative energy has become a global trend that will shape the future of energy. In the present study, 3 microbial strains with different oxygen requirements, including Clostridium acetobutylicum ATCC 824, Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D, were used to construct a hydrogen production system that was composed of a mixed aerobic-facultative anaerobic-anaerobic consortium. The effects of metal ions, organic acids and carbohydrate substrates on this system were analyzed and compared using electrochemical and kinetic assays. It was then tested using small-scale experiments to evaluate its ability to convert starch in 5 L of organic wastewater into hydrogen. For the one-step biohydrogen production experiment, H1 medium (nutrient broth and potato dextrose broth) was mixed directly with GAM broth to generate H2 medium (H1 medium and GAM broth). Finally, Clostridium acetobutylicum ATCC 824, Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D of three species microbial co-culture to produce hydrogen under anaerobic conditions. For the two-step biohydrogen production experiment, the H1 medium, after cultured the microbial strains Enterobacter cloacae ATCC 13047 and Kluyveromyces marxianus 15D, was centrifuged to remove the microbial cells and then mixed with GAM broth (H2 medium). Afterward, the bacterial strain Clostridium acetobutylicum ATCC 824 was inoculated into the H2 medium to produce hydrogen by anaerobic fermentation. RESULTS: The experimental results demonstrated that the optimum conditions for the small-scale fermentative hydrogen production system were at pH 7.0, 35°C, a mixed medium, including H1 medium and H2 medium with 0.50 mol/L ferrous chloride, 0.50 mol/L magnesium sulfate, 0.50 mol/L potassium chloride, 1% w/v citric acid, 5% w/v fructose and 5% w/v glucose. The overall hydrogen production efficiency in the shake flask fermentation group was 33.7 mL/h-1.L-1, and those the two-step and the one-step processes of the small-scale fermentative hydrogen production system were 41.2 mLVh-1.L-1 and 35.1 mL/h-1.L-1, respectively. CONCLUSION: Therefore, the results indicate that the hydrogen production efficiency of the two-step process is higher than that of the one-step process.

Stress tolerant virulent strains of Cronobacter sakazakii from food

BACKGROUND: Cronobacter sakazakii is considered as an emerging foodborne pathogen. The aim of this study was to isolate and characterize virulent strains of Cronobacter sakazakii from food samples of Bangladesh. RESULT: Six (6) Cronobacter sakazakii was isolated and identified from 54 food samples on the basis of biochemical characteristics, sugar fermentation, SDS-PAGE of whole cell protein, plasmid profile and PCR of Cronobacter spp. specific genes (esak, gluA, zpx, ompA, ERIC, BOX-AIR) and sequencing. These strains were found to have moderately high antibiotic resistance against common antibiotics and some are ESBL producer. Most of the C. sakazakii isolates were capable of producing biofilm (strong biofilm producer), extracellular protease and siderophores, curli expression, haemolysin, haemagglutinin, mannose resistant haemagglutinin, had high cell surface hydrophobicity, significant resistance to human serum, can tolerate high concentration of salt, bile and DNase production. Most of them produced enterotoxins of different molecular weight. The isolates pose significant serological cross-reactivity with other gram negative pathogens such as serotypes of Salmonella spp., Shigella boydii, Shigella sonnei, Shigella flexneri and Vibrio cholerae. They had significant tolerance to high temperature, low pH, dryness and osmotic stress. CONCLUSION: Special attention should be given in ensuring hygiene in production and post-processing to prevent contamination of food with such stress-tolerant virulent Cronobacter sakazakii.

Utilização de sistemas poliméricos de duas fases aquosas (SPDFA) compostos por polietileno glicol/ácido poliacrílico (PEG/APA) para extração de ácido clavulânico / Utilization of aqueous two phase systems (ATPS) composed of polyethylene glycol/polyacrylic acid (PEG/APA) in the extraction of clavulanic acid

A viabilidade da produção em escala industrial de produtos biotecnológicos de interesse comercial e terapêutico, como os fármacos, depende significativamente das técnicas de separação e purificação utilizadas. A aplicação do sistema de duas fases aquosas (SDFA) é proposta como alternativa para a purificação, pois permite a separação e análise de biomoléculas, de modo que estas não percam sua atividade ou propriedades desejadas. Esta técnica é interessante para a purificação em larga escala, pois permite partição seletiva, com potencial de obtenção de altos rendimentos, além de apresentar boa relação custo-benefício. O presente trabalho estudou a purificação por extração líquido-líquido do ácido clavulânico em SDFA utilizando um novo sistema polimérico aquoso, formado pelos polímeros polietileno glicol (PEG) e ácido poliacrílico (APA). Foram estudadas diferentes composições do sistema polimérico aquoso PEG/APA, empregando diferentes massas molares e concentrações para o PEG e utilizando a massa molar 8000g/mol para o APA. Com base nas informações obtidas o melhor ponto de extração para o ácido clavulânico na presença de Na2SO4 foi definido como MPEG=400 g/mol, CPEG=17,5% (m/m) e CNaPA=22,5% (m/m) com K= 19,14, ηT=91,21%, BM=101,69 e R=0,45. Enquanto que na presença de NaCl, o melhor ponto encontrado foi: MPEG=400 g/mol, CPEG=35% (m/m) e CNaPA=10% (m/m) com K=11,96 ηT=80,04%, BM=90,18 e R=0,66. No trabalho será avaliada, também, a influência da temperatura, pH e força iônica nesse sistema. Estabeleceram-se os melhores parâmetros de separação do ácido clavulânico presente em meio fermentado produzido por Streptomyces clavuligerus utilizando a metodologia de fermentação extrativa com SDFA PEG/APA. O efeito do ácido clavulânico no diagrama de fases do sistema PEG-APA, bem como sua partição na forma pura e na presença de homogeneizado celular, foi estudado principalmente através da determinação do coeficiente de partição e recuperação do respectivo fármaco

The viability of industrial scale production of commercial and therapeutical biotechnological products, such as drugs, is significantly dependent on the separation and purification techniques applied. The use of two-aqueous phase systems (ATPS) is proposed as an alternative to purification because it allows the separation and analysis of biomolecules, so that they do not lose their activities or desired properties. This technique is interesting for large scale purification because it allows selective partition with high potential yield and good cost/benefit ratio. The present work studied the purification of clavulanic acid (CA) by liquid-liquid extraction in ATPS applying a new aqueous polymeric system composed of two polymers, namely polyethylene-glicol (PEG) and sodium polyacrylate (NaPA). Different compositions of the aqueous polymeric system (PEG/PAA) were utilized, employing different PEG molar masses (MPEG) and concentrations (CPEG) and a molar mass of PAA of 8000 g/mol. In the light of the results obtained, the best conditions for clavulanic acid extraction, in the presence of Na2SO4, were MPEG = 400 g/mol, CPEG = 17.5% (m/m) and CNaPA = 22.5% (m/m), which allowed obtaining a partition coefficient (K) of 19.14, a yield in the top phase (ηT) of 91.21%, a mass balance (MB) of 101.69 and a volume ratio (R) of 0.45. On the other hand, in the presence of NaCl, the best results (K = 11.96, ηT = 80.04%, MB = 90.18 and R = 0.66) were found at: MPEG = 400 g/mol, CPEG = 35% m/m and CNaPA = 10% m/m. The effect of clavulanic acid in the PEG-PAA system phase diagram and its partition either in its pure form or in the cell homogenate were studied mainly through both the determination of the partition coefficient and the recovery of the drug selected for this study

Efeito da suplementação de cobre e selênio na dieta de novilhos Brangus sobre o desempenho e fermentação ruminal / The effects of copper and selenium supplementation in the diet of Brangus steers on performance and rumen fermentation

Vinte e oito bovinos Brangus foram usados para determinar o efeito da suplementação de cobre e selênio no desempenho e na fermentação ruminal. Os animais foram divididos em: 1) C(Controle) - sem a suplementação de cobre e selênio; 2) Se - 2mg Se/kg de matéria seca na forma de selenito de sódio; 3) Cu - 40mg Cu/kg de matéria seca na forma de sulfato de cobre; 4) Se/Cu - 2mg Se/kg de matéria seca na forma de selenito de sódio e 40mg Cu/kg de matéria seca na forma de sulfato de cobre. As pesagens dos animais foram feitas em intervalos de 28 dias, após jejum completo de 18 horas. Foram colhidas amostras de líquido ruminal para análises de ácidos graxos voláteis e pH. O ganho de peso diário aumentou com a suplementação de Se (P<0,05). A ingestão de matéria seca não foi alterada pelos tratamentos (P>0,05). Os animais submetidos à suplementação com Cu apresentaram menor pH ruminal quando comparado com a suplementação Se/Cu (P<0,05). Os animais suplementados com Se/Cu apresentaram maior proporção de ácido acético quando comparado com o controle (P<0,05). Para o ácido propiônico e butírico, não houve diferenças significativas (P>0,05) entre os tratamentos. Apesar do pouco efeito na fermentação ruminal, as suplementações de selênio, cobre e selênio/cobre proporcionaram uma melhor eficiência alimentar.

Twenty-eight Brangus cattle were used to determine the effect of copper and selenium supplementation on performance and rumen fermentation. The animals were divided in: 1) (Control) - without supplementation of copper and selenium; 2) Se - 2mg Se/kg of dry matter as sodium selenite; 3) Cu - 40mg Cu/kg of dry matter as copper sulphate; 4) Se/Cu - 2mg Se/kg of dry matter as sodium selenite and 40mg Cu/kg of dry matter as copper sulphate. Animal weighing was performed in 28 day intervals, after 18 hours of fasting. Samples of rumen liquid were collected for analysis of volatile fatty acid and pH. The daily weight gain increased with selenium supplementation (P<0.05). The dry matter intake was not altered by treatments (P>0.05). The animals that received copper supplementation had lower pH rumen compared with selenium/copper supplementation (P<0.05). The animals supplemented with selenium/copper had a high proportion of acetic acid when compared to control (P<0.05). For propionic and butyric acid, there was no difference (P>0.05) among treatments. Despite the little effect on rumen fermentation, the supplementations of selenium, copper and selenium/copper provided better feed efficiency.

Cinética e parâmetros de fermentação ruminal in vitro de silagens de parte aérea e raízes de mandioca / Kinetic parameters and in vitro rumen fermentation of cassava foliage and roots silages

Foram avaliados os efeitos do emurchecimento e da inclusão de raízes 0, 15, 30 e 45% da matéria natural sobre a cinética e os parâmetros da fermentação ruminal de silagens da parte aérea da mandioca. A produção acumulada de gases (PAG) às 96 horas de incubação foi influenciada significativamente (P<0,05) pela interação emurchecimento versus inclusão de raízes. Para a PAG, houve elevação linear de 1,88mL a cada unidade percentual de raiz adicionada às silagens não emurchecidas e efeito quadrático nas silagens emurchecidas, com produção máxima de 226mL de gases, com a inclusão de 30,5% de raízes. A degradabilidade verdadeira in vitro da MS (DVIMS) das silagens apresentou significância (P<0,05) para o emurchecimento e para a interação inclusão de raízes versus tempo de incubação. O emurchecimento reduziu, aproximadamente, 2% da degradabilidade da silagem, enquanto a inclusão de raízes propiciou elevação linear dessa variável, que atingiu o valor máximo às 24 horas de incubação.

The effects of wilting and roots inclusion (0, 15, 30 and 45% fresh matter) on rumen fermentation kinetics and parameters of cassava foliage silages were evaluated. Total gas production (TGP) after 96 hours of incubation was significantly influenced (P <0.05) by the interaction between wilting and roots inclusion. TGP was linearly increased by 1.88 mL for each percent unit of root added to the wilted silage, whereas, for wilted ones, a quadratic effect was observed, with a maximum gas cumulative output of 226 mL for 30.5% roots addition. The true DM degradability of silages in vitro (IVTDMD) showed significance (P<0.05) to wilting and interaction between root inclusion and incubation time. Wilting reduced silage degradability in approximately 2%, while root inclusion led to linear increases in this variable, which reached its peak at 24h of incubation.

Interações entre bactérias láticas produtoras de bacteriocinas e a microbiota autóctone de charque / Interactions between bacteriocin-producing lactic acid bacteria and the autochthonous microbiota from charqui

O charque é um produto cárneo tipicamente brasileiro, salgado e seco ao sol, ainda produzido de maneira artesanal. Durante sua produção há uma etapa de fermentação, realizada pela microbiota naturalmente presente na matéria-prima, o que dificulta a padronização do produto, e pode influenciar negativamente em suas características sensoriais e qualidade microbiológica. O controle da etapa de fermentação do charque seria uma alternativa para minimizar este problema e, neste contexto, as bactérias láticas produtoras de bacteriocinas se enquadram de forma interessante. A microbiota autóctone de charque inclui principalmente bactérias láticas e micro-organismos halofílicos e halotolerantes, sendo assim, este produto apresenta potencial como fonte para o isolamento de novas bactérias láticas produtoras de bacteriocinas. Assim, este trabalho teve por objetivo isolar e identificar culturas de bactérias láticas produtoras de bacteriocinas naturalmente presentes no charque, caracterizar parcialmente as bacteriocinas produzidas por essas culturas, avaliar seu potencial de aplicação neste produto para a melhoria de sua qualidade microbiológica e avaliar seu efeito na ecologia microbiana do charque, nas diferentes etapas de sua fabricação. Através da técnica de tripla camada em ágar foi isolada uma cepa de Lactococcus lactis subsp. lactis apresentando o gene codificador para nisina Z e com capacidade de inibir, in vitro, micro-organismos medianamente e altamente halotolerantes isolados de charque, além de outros micro-organismos deteriorantes e patogênicos importantes em alimentos, como Lactobacillus spp., Listeria monocytogenes e Staphylococcus aureus. A bacteriocina produzida pela cepa isolada neste estudo também possui características interessantes para sua aplicação na bioconservação de alimentos, como resistencia ao calor, presença de agente químicos e altos teores de NaCl, além de não ser afetada pelo pH. A aplicação dessa cepa em charque modelo resultou na redução de até 2 ciclos log na população de micro-organismos halotolerantes, indicando apresentar um potencial de aplicação como agente de bioconservação do produto. Os ensaios de avaliação da ecologia microbiana, empregando DGGE, indicaram que a fermentação natural do charque ocorreu com a participação de bactérias láticas dos gêneros Lactobacillus, Streptococcus, Lactococcus e de micro-organismos halotolerantes do gênero Staphylococcus. Além disso, os estudos referentes à dinâmica populacional demonstraram que a adição da cepa bacteriocinogênica ao charque não influenciou, de forma qualitativa, as populações presentes no produto

Charqui is a Brazilian traditional meat product, salted and sun-dried, still manufactured without control of the fermentation step, which is performed by the indigenous microbiota. This fact interferes on the standardization of the product and can negatively affect the sensorial properties and microbiological quality. The application of a known microbiota would be an alternative to minimize this problem and the bacteriocin-producing lactic acid bacteria can can fit in this purpose. The charqui indigenous microbiota mainly includes lactic acid bactéria and halophilic and halotolerant microorganisms, therefore, this product presents a potencial as a source for the isolation of new bacteriocin-producing lactic acid bacteria. The aim of the present work was to isolate and identify bacteriocin-producing lactic acid bacteria from charqui, characterize the bacteriocins produced by the isolated culture, evaluate its potential as biopreservative in charqui and its influence on the microbial populations during the manufacture of the product. A bacteriocinogenic Lactococcus lactis subsp. lactis strain was isolated from charqui through the triple-layer agar technique. This strain produces a nisin-like bacteriocin capable to inhibit in vitro medium and highly halotolerant bacteria isolated from charqui and other food-borne pathogenic and spoilage microorganisms. The application of this strain for charqui manufacturing caused a reduction of up to 2 log in the halotolerant bacteria population, evidencing its potential application for charqui biopreservation. Studies in the populational dynamics using DGGE indicated that the presence of the bacteriocinogenic strain did not affect the microbial populations in the product

Influencia de las regiones de estabilidad del biorreactor en la producción eficiente de alcohol carburante / Influence of bioreactor stability regions in the efficient production of fuel ethanol

Colombia ha desarrollado un proyecto ambicioso en la producción de alcohol carburante, y actualmente es uno de los países líderes de América Latina en la producción de biocombustibles. No obstante, algunas de sus plantas industriales productoras de etanol presentan problemas durante el arranque y la operación de los fermentadores, que afectan adversamente la productividad del proceso global. Estudios teóricos y experimentales demuestran que el comportamiento dinámico del proceso depende de los fenómenos de estabilidad que exhibe el sistema. En este trabajo se muestra la influencia de las regiones de estabilidad sobre la eficiencia del proceso tomando como criterios de evaluación índices de impacto económico y ambiental. Para esto, se realizó un análisis de estabilidad donde se tomó como parámetro de operación principal la velocidad de dilución. Se empleó un programa propio desarrollado sobre Matlab® para generar diagramas de bifurcación y de fase. El algoritmo de reducción de residuos (WAR) desarrollado por la Agencia de Protección Ambiental de los Estados Unidos, y el software SuperPro Designer®, fueron utilizados respectivamente para hacer la evaluación ambiental y económica del proceso. Como resultado, se evidenció que los procesos continuos de fermentación se pueden llevar a cabo en diferentes regiones de estabilidad, y que las variables de respuesta, tales como el rendimiento, dependen de la región de operación seleccionada. Finalmente, se llegó a la conclusión de que la evaluación de la estabilidad de los procesos de fermentación debe incluirse en las etapas de diseño de bioprocesos para garantizar su buen desempeño.

Colombia has developed an ambitious project to produce fuel ethanol. Nowadays, Colombia is one of the leading countries in Latin America for the biofuels production. However, some of its industrial plants producing ethanol present problems during the startup and the operation of fermenters, which adversely affects the productivity of the global process. Theoretical and experimental studies show that the process’ dynamic behavior depends on the stability phenomenon exhibited by the system. This work shows the influence of the stability regions on the process’ efficiency. For this, economic and environmental index were used. A stability analysis was carried out. The dilution rate was selected as the main operating parameter. An own program developed on Matlab® was used to generate bifurcation diagrams and phase diagrams. The Waste Reduction Algorithm (WAR) developed by the Environmental Protection Agency of the United States, and the SuperPro Designer® software were used for estimating the environmental and economic assessment of the process. As a result, it was shown that continuous fermentation processes can be carried out in different stability regions. Likewise, it was shown that the response variables, such as the yield, depend on the selected operating region. Finally, it was concluded that the evaluation of fermentation processes’ stability must be included in the stages for designing of bioprocess in order to ensure a good performance.

Utilización de Candida guilliermondii aislada del corozo chiquito (Bactris guineensis) en la producción de xilitol / Utilization of Candida guilliermondii isolated of corozo chiquito (Bactris guineensis) in the production of xylitol

La levadura Candida guilliermondii es objeto de estudio debido a su capacidad de producir xilitol aprovechando compuestos hemicelulósicos ricos en xilosa, dado esto, la cepa Candida guilliermondii aislada del fruto del corozo chiquito (Bactris guineensis) fue usada en este estudio con el fin de evaluar su capacidad para producir xilitol sobre un sustrato hidrolizado de cascarilla de arroz. El objetivo de este trabajo fue determinar los parámetros fermentativos como producción de xilitol, productividad volumétrica (Qp) y rendimiento de sustrato en producto (Yp/s) durante la fermentación con la cepa nativa Candida guilliermondii. Se emplearon 200 ml de medio de cultivo hidrolizado de cascarilla de arroz, el cual contenía una concentración de xilosa de 27,5 g/L. La fermentación se llevó a cabo bajo las siguientes condiciones: temperatura 30 ºC, pH del medio 5,8, agitación 120 rpm e inóculo adaptado de 3 g/L. Los resultados mostraron que después de 120 horas de fermentación se obtuvieron 2,6 g/L de xilitol con productividad volumétrica (Qp) de 0,02 g/L-h y rendimiento de sustrato en producto (Yp/s) de 0,13 g/g. De esta manera, la cepa nativa Candida guilliermondii, aislada del fruto de Corozo chiquito (Bactris guineensis), produjo xilitol bajo condiciones específicas de fermentación.

The yeast Candida guilliermondii has been studied due to its ability to produce xylitol in xylose-rich hemicellulosic compounds, Candida guilliermondii strain isolated from the fruit of Corozo chiquito (Bactris guineensis) was used in this study to assess their ability to xylitol production on these substrates. The aim of this study was to determine the fermentation parameters such as xylitol production, volumetric productivity (Qp) and yield of xylitol production (Yp/s) during fermentation with the native strain Candida guilliermondii. Was used 200 ml of culture medium rice husk hydrolysate, which contained a xylose concentration of 27.5 g/L. The fermentation was carried out under the following conditions: temperature 30 ºC, pH of 5.8, agitation 120 rpm and adapted inoculum of 3 g/L. The results showed that after 120 hours of fermentation 2.6 g / L of xylitol was achieved with volumetric productivity (Qp) 0.02 g/L-h and 0.13 g/g yield of xylitol production (Yp/s). The native strain Candida guilliermondii, isolated from the fruit of Corozo chiquito (Bactris guineensis) produced xylitol fermentation under specific conditions.

Leveduras no rúmen de caprinos e bovinos de corte criados em pastagem tropicais / Yeasts in the rumen of beef steers and goats raised on tropical pasture

Yeasts suplemented in the rumen have been produced benefic interations in the digestion and in the health of the ruminants. This study aimed to quantify, to isolate and, to identify aerobic fungi and yeasts naturally present in the rumen of goats and cattle raised on tropical pastures of the North of Minas Gerais State, Brazil. Samples of 15mL of ruminal juice from 18 hibrid goats and 31 crossbred Nellore steers were used. The physico-chemical characteristics of the samples were evaluated and mycological culture, quantification, and identification of the aerobic fungi were performed. The results indicated the absence of yeasts in the ruminal fluid of steers. However, theses microorganisms were cultured from ruminal juice for all evaluated goats, at an average concentration of 3.2 x 10VCFU/mL. The species Pichia membranifaciens was the most frequently identified yeast, suggesting its participation in the ruminal microbiot of theses small ruminants.

Avaliação dos modelos logístico bicompartimental e de Gompertz na estimativa da dinâmica de fermentação ruminal in vitro do farelo e da torta de babaçu (Orbignya martiana) / Avaliation of bicompartimental logistic and Gompertz mathematical models to estimate gas production from babassu (Orbignya martiana) meal and pie using the semi-automated in vitro technique

Utilizaram-se dois modelos matemáticos para avaliar a produção de gases do farelo e da torta de babaçu, pela técnica in vitro semiautomática de produção de gases. Foram utilizados o modelo logístico e o de Gompertz. Os parâmetros de validação usados foram o quadrado médio do erro (QME), o coeficiente de determinação (R²), o desvio médio absoluto dos resíduos (DMA) e a análise gráfica dos resíduos. O modelo logístico bicompartimental apresentou menores valores (P<0,05) para o QME e o DMA em relação ao de Gompertz, e não houve diferença (P>0,05) quanto ao R². Os gráficos de dispersão mostraram semelhanças nos ajustes dos dois modelos. Na análise gráfica dos resíduos, os dois modelos descreveram bem cinética de produção de gases da matéria seca. No entanto, o modelo logístico apresentou melhor valor de QME. Para avaliação da cinética de fermentação ruminal do farelo e da torta de babaçu pela técnica in vitro semiautomática de produção de gases, recomenda-se adotar o modelo logístico.

Two mathematical models were used to evaluate gas production from the meal and pie of babassu using the semi-automated gas production technique. The logistic and Gompertz models were used and the validation parameters for both models were the residual mean square (RMS), the coefficient of determination (R 2 ), the absolute average residual (AAR), and the graphical analysis of residues. The logistic model showed lower RMS (P<0.05) and AAR in comparison to Gompertz model. Both Gompertz and logistic models showed similar R 2 . The dispersion graphics showed similarity for both models and graphics analyses demonstrated that both models describe well the kinetic of gas production of dry matter. However, the bicompartimental logistic model showed the best RMS value. Logistic models are recommended to describe the kinetic of gas production from babassu foods using the semi-automated in vitro technique.

Ethanol production from sweet sorghum juice under very high gravity conditions: batch, repeated-batch and scale up fermentation

Batch ethanol fermentations from sweet sorghum juice by Saccharomyces cerevisiae NP 01 were carried out in a 500 ml air-locked Erlenmeyer flask under very high gravity (VHG) and static conditions. The maximum ethanol production efficiency was obtained when 9 g l-1 of yeast extract was supplemented to the juice. The ethanol concentration (P), productivity (Qp) and yield (Yp/s) were 120.24 +/- 1.35 g l-1, 3.01 +/- 0.08 g l-1 h-1 and 0.49 +/- 0.01, respectively. Scale up ethanol fermentation in a 5-litre bioreactor at an agitation rate of 100 rev min-1 revealed that P, Qp and Yp/s were 139.51 +/- 0.11 g l-1, 3.49 +/- 0.00 g l-1 h-1 and 0.49 +/- 0.01, respectively, whereas lower P (119.53 +/- 0.20 g l-1) and Qp (2.13 +/- 0.01 g l-1 h-1) were obtained in a 50-litre bioreactor. In the repeated-batch fermentation in the 5-litre bioreactor with fill and drain volume of 50 percent of the working volume, lower P and Qp were observed in the subsequent batches. P in batch 2 to 8 ranged from 103.37 +/- 0.28 to 109.53 +/- 1.06 g l-1.

Production of a novel halophilic dextranase from a honey isolate, bacillus subtilis NRC-B233 under solid-state fermentation

A GRAM-POSITIVE, sporulating halophilic bacteria, designated NRC-B233, was isolated from the honey produced in Saudi Arabia. It was identified by the 16-23S intergenic region as Bacillus subtilis NRC-B233. Screening of the wastes and agro-products for dextranase production under solid state fermentation showed that corn flour was the best substrate [61.323 U/g]. The optimum conditions for dextranase productions were 37°C, pH 9, 32 hr incubation period, and 200% moisture content. The most favorable nitrogen and carbon sources for enzyme production were 2% peptone and 5% starch [1076.768, 1553.364 U/g]. respectively. A unique character of this isolate is its ability to continuously produce dextranase in the absence and presence of NaCl 5-20 g/l. The addition of 0.175 Mm CrCl[3] increased the dextranase production about 4.5 fold. The enzyme has been partially purified about 112-fold from crude extract by only two purification steps involving ultra-filtration. The purified dextranase showed its maximum activity at pH 9.2 and 70°C. It retained fill activity [100%] at 75°C for one hour. Dextranase activity increased about four fold in the presence of 10% NaCl. On the other hand, CaCl[2] [0.050M], EDTA [0.100M], and KCI [0.100M] had great influence in enzyme activity. The enzyme showed variable degradation effects on different types of dextran and its derivatives. These results suggest that the dextranase secreted by Bacillus subtilis NRC-B233 is industrially important from the perspectives of its activity at across pH range [5.0-100], its thermo-activity in addition to its halophilic character and its ability to degrade different types of alpha-1,4 and alpha-1,6 glycosidic linkages

Fermentação ruminal de ovinos alimentados com alto concentrado e grãos de girassol ou gordura protegida / Ruminal fermentation of sheep fed high-concentrated diet containing sunflower grains or protected fat

Seis ovinos machos, não castrados, da raça Santa Inês, com média de peso de 30kg, fistulados no rúmen, foram distribuídos em delineamento de quadrado-latino duplo (3x3). Três períodos e três dietas, uma controle, sem inclusão de fonte de lipídio, e duas com inclusão de grãos de girassol ou gordura protegida, foram testados quanto aos parâmetros ruminais. Foram verificadas diferenças (P<0,05) entre as dietas quanto à concentração ruminal de amônia (18mg/dL), mas não houve efeito sobre o pH (6,1), a produção total de ácidos graxos de cadeia curta (98mM), a proporção de acetato (66,4 por cento), de propionato (20 por cento) e de butirato (13 por cento) e sobre a razão acetato:propionato (3,2:1). As bactérias sólido-aderidas isoladas do conteúdo ruminal dos animais recebendo a dieta-controle apresentaram maior teor de nitrogênio (10,7 por cento) que as das dietas com gordura protegida (9,8 por cento) ou com grãos de girassol (9,1 por cento). A produção de nitrogênio pelas bactérias sólido-aderidas da dieta-controle (170mg/g) não diferiu da dieta com grãos de girassol (153mg/kg) ou com gordura protegida (160mg/kg). A inclusão de grãos de girassol ou gordura protegida na dieta com alto concentrado para ovinos propiciou ambiente adequado para fermentação ruminal.

Six not castrated Santa Inês breed male sheep, weighing 30kg, fistulated in the rumen, were randomly alloted in a double Latin-Square design (3x3). Three periods and three diets, a control without addition of the lipid source, and two with addition of sunflower seeds or protected fat, were tested on the ruminal fermentation. Differences (P<0.05) between diets for ruminal ammonia concentration (18mg/dL) were observed. However, effects were not detected on pH (6.1); the total production of short-chain fatty acids (98mM); proportions of acetate (66.4 percent), propionate (20 percent), and butyrate (13 percent); and acetate:propionate ratio (3.2:1). The solid-adherent bacteria isolated from the rumen content of animals fed control diet had higher nitrogen content (10.7 percent) than those fed protected fat diet (9.8 percent) or sunflower seeds diet (9.1 percent). The production of nitrogen by bacteria attached to solid-control diet (170mg/kg) did not differ from sunflower seeds (153mg/kg) or protected fat diets (160mg/kg). The inclusion of sunflower seeds or protected fat in diets with high concentrate for sheep provided suitable environment for rumen fermentation.

Remoção de endotoxina presente em meio fermentado contendo biomoléculas utilizando sistemas micelares de duas fases aquosas / Endotoxin removal from fermentation broth containing biomolecules using two-phase micellar system

Foi investigada a utilização de Sistema Micelar de Duas Fases Aquosas (SMDFA) para remoção de lipolissacarídeos (LPS) de preparações contendo proteínas recombinantes de interesse farmacêutico, como a proteína verde fluorescente (GFPuv). Os SMDFA são constituídos por soluções de tensoativos contendo micelas e oferecem ambientes hidrofóbico e hidrofílico, que possibilitam seletividade na partição de biomoléculas de acordo com sua hidrofobicidade, permitindo a remoção de LPS contaminante. Neste trabalho, foi realizada a implementação do método para a quantificação de LPS em amostras contaminadas e a obtenção de LPS e GFPuv puros a partir de cultivo de E. coli recombinante. Além disso, foi estudada a influência do Triton X-114 na metodologia de quantificação de LPS, e a adição de MgSO4, CaCl2, KI e (NH4)2SO4 na partição de GFPuv e LPS puros em SMDFA. E ainda, realizou-se um planejamento experimental (22) para avaliar os maiores KGFPuv e porcentoRECGFPuv. O homogeneizado celular de E. coli foi testado nas melhores condições obtidas com o planejamento experimental. E finalmente, o processo por cromatografia de afinidade por íons metálicos (IMAC) foi empregado para investigar a adsorção de LPS em matriz IDA-Ca2+. Conforme os resultados obtidos, o TX-114 causou elevada interferência no método cinético cromogênico, em função da similaridade desta molécula com os LPS. Os LPS apresentaram partição preferencial para a fase concentrada em micelas, com altos valores de remoção, por centoREMLPS>98,0 por cento. Ao contrário, a GFPuv foi recuperada preferencialmente na fase diluída, na qual existe maior volume disponível, resultando em valores de KGFPuv>1. A adição de sais ocasionou diminuição nos valores KGFPuv, provavelmente por causa da carga negativa que GFPuv adquiriu nas condições avaliadas. Os resultados do planejamento experimental mostraram que a melhor condição de partição obtida foi na região do ponto central, 4,0 por cento (p/p) a 60,0°C, com KGFPuv>10. O processo por IMAC apresentou as maiores...

The Aqueous Two-Phase Micellar System (ATPMS) was investigated for endotoxin (LPS) removal from preparations containing recombinant proteins of pharmaceutical interest, such as the green fluorescent protein (GFPuv). These systems usually consist of micellar surfactants solutions and offer both hydrophobic and hydrophilic environments, providing selectivity to the biomolecules partitioning according to its hydrophobicity. In this work, the implementation of the method for LPS quantification in contaminated samples was accomplished, as well as the obtaining of pure LPS and GFPuv from recombinant E. coli. Furthermore, the influence of Triton X-114 in the methodology for LPS quantification was studied, as the addition of MgSO4, CaCl2, KI, and (NH4)2SO4 into the partition of pure GFPuv and LPS in ATPMS. In addition, a statistical design (22) was carried out to evaluate the highest KGFPuv and percentRECGFPuv. The E. coli cell lysate was tested under optimum conditions obtained with the statistical design. And, finally, the process by ionmetal affinity chromatography (IMAC) was used to investigate the adsorption of LPS in IDA-Ca2+ matrix. The results showed that the TX-114 caused high interference in the kinetic chromogenic method, according to the similarity of this molecule to LPS. The LPS showed preferential partitioning to the micellerich phase, with high values of removal, percentREMLPS>98.0 percent. In the other hand, the GFPuv was preferentially recovered in the micelle-poor phase, in which there is greater volume available resulting in values of KGFPuv>1. The addition of salts caused a reduction in the values KGFPuv, probably because of the negative charge that the GFPuv acquired at the conditions evaluated. The results of the statistical design showed that the best partitioning condition obtained was in the central point region, 4.0 percent (wt/wt) at 60.0°C, with KGFPuv>10. The process by IMAC showed the highest adsorption of LPS-IDA-Ca+2 capacities at the conditions of lower pH...