Avaliação de diferentes métodos de cozimento e de cisalhamento na determinação da maciez de lombos suínos de diferentes qualidades / Evaluation of different cooking and shear methods to assess tenderness of pork loins from different qualities

Objetivou-se avaliar a maciez de lombos suínos (M. Longissimus thoracis) de diferentes classes de qualidade (RFN, PSE e RSE) por dois métodos de cozimento (micro-ondas e grelha) e dois protocolos (SSF e WBsSF) de determinação de força de cisalhamento (FC). As classes de qualidade de carne suína não difeririam (P>0,05) quanto a FC, independentemente do método de cozimento ou protocolo de cisalhamento usado. Maior repetibilidade da FC foi observada quando do cozimento em micro-ondas (SSF = 0,77; WBsSF = 0,75), tendo alta correlação (r = 0,72) entre os dois protocolos. Concluiu-se que que as classes de qualidade da carne suína não diferem quanto à maciez instrumental e que o cozimento em micro-ondas associado ao protocolo SSF é altamente promissor para avaliação da maciez instrumental da carne suína.

A new isolate of thermophilic and organic solvent tolerant bacteria for lipase production using basal medium of palm kernel cake 80-87

@#Aims: This research focused on the selection of potential strains especially bacteria that can grow effectively in palm kernel cake (PKC) and produce high amount of thermostable and solvent tolerant (TS-OST) lipase. The work involved the exploration of renewable PKC as potential fermentation medium for discovery to novel TS-OST lipase that would have excellent tolerance and activity in presence of organic solvents with high temperatures for industrial applications.Methodology and results: Using palm kernel cake (PKC) as source of thermophilic bacteria, 53 bacterial strains were found survived at temperature 65 °C. However, after subcultured several times, only 17 strains were found as pure thermophilic strains. Preliminary screening both qualitative and quantitative was performed to all 17 potential thermophilic bacterial strains and showed that only 11 purified thermophilic strains are lipase producer. Strain PKC-P1 produced highest enzyme activity (11.13 U/g), followed by PKC-P13 and PKC-C9. The lowest enzymeactivity was lipase produced byPKC-C10 (0.76U/g). Strain PKC-P1 has been classified as Gram negative bacteria and identified as Bacillus smithiistrain PKC_P1.Conclusion, significance and impact of study: PKC as a by-product of oil palm industry consistsof many nutrients that can give benefits towards industry and can be utilized in order to produce enzymes like lipases. From these results, it could be concluded that this lipase stable at temperature 65 °C and pH 7 and may be a potential candidate to be used in a variety of biotechnological applications. This finding revealed that a bacterial strain obtained from oil-rich environment which is PKC through isolation process has potential as a source of more economical enzyme to be applied in biotechnology industr

Production of α-galactosidase from Aspergillus foetidus MTCC 6322 by solid state fermentation and its application in soymilk hydrolysis.

The production of α-galactosidase from the wild fungal strain Aspergillus foetidus MTCC 6322 using solid state fermentation (SSF), its characterization, and its efficacy in the hydrolysis of soymilk using response surface methodology were studied. The optimum conditions for production of α-galactosidase by SSF were: wheat bran (10 g), moisture content (64%), inoculum volume (1.0 mL; 6 × 107 spores/mL) with a yield of 4.1 × 103 units per gram dry substrate (U/gds) at 96 h. The enzyme showed optimum activity at pH 6.0, temperature 40°C, pH stability between 5.0-8.0, and temperature stability between 30-40°C. The enzyme was stable in the presence of trypsin, lipase, and collagenase and it showed susceptibility of the substrates such as raffinose, melibiose, guar gum and soymilk to hydrolysis in varying degrees. The optimized conditions for soymilk hydrolysis were: soymilk (10 mL) from defatted soybean meal (1.5%), α-galactosidase (0.15 UmL-1) at 30°C, pH 6.0 and duration of 1 h.

“Comparación de técnicas de laboratorio para el diagnóstico de Giardia intestinalis” / “Comparison of Laboratory Techniques For the Diagnosis of Giardia Intestinalis”

Evaluar la sensibilidad y especificidad de tres técnicas de laboratorio para el diagnóstico de Giardia intestinalis. Materiales y Métodos: 31 muestras de heces provenientes de niños en edad preescolar, se procesaron a través del examen microscópico con SSF-lugol, método de concentración de Ritchie y método inmunológico “Giardia-Strip”. La Técnica de Ritchie fue considerada como “Gold Standard”. Resultados: Se identificaron quistes de Giardia intestinalis en 6 muestras (19,35%) a través de la técnica de Ritchie. La técnica del examen al fresco identificó en 5 de ellas quistes del protozoario (16,1%), mostrando una sensibilidad de 83% y especificidad del 100%. La técnica Giardia-Strip identificó quistes en 4 muestras (12,9%) con sensibilidad de 66,66% y especificidad del 100%. El valor predictivo positivo para el método de “Giardia-Strip” fue 14% y valor predictivo negativo de 93%. El examen al fresco mostró valor predictivo positivo del 100% y valor predictivo negativo de 96%. Conclusiones: El concentrado de Ritchie y el examen al fresco mostraron mayor sensibilidad y especificidad en el diagnóstico de Giardia intestinalis, cuando las muestras contienen solo quistes. Sin embargo, el método Giardia-Strip, demostró mayor rapidez en la obtención de los resultados.

Objective: To evaluate the sensitivity and specificity of three laboratory techniques for the diagnosis of Giardia intestinalis. Materials and Methods: 31 fecal samples from preschoolers were submitted to microscopic examination with SSF-Lugol, the Ritchie concentration method and the “Giardia-Strip” immunological method. The Ritchie technique was considered the “gold standard.“ Results: G. intestinalis cysts were identified in 6 samples (19.35%) using the Ritchie technique. The fresh test identified 5 cases of protozoan cysts (16.1%), showing a sensitivity of 83 % and specificity of 100 %. The Giardia-Strip identified cysts in 4 samples (12.9%) with 66.66 % sensitivity and 100% specificity. The positive predictive value for the “Giardia Strip” method was 14% and the negative predictive value was 93%. The fresh test showed a positive predictive value of 100% and a negative predictive value of 96 %. Conclusions: Ritchie concentrate and the fresh test showed higher sensitivity and specificity in the diagnosis of Giardia intestinalis, when samples contain only cysts. However, the Giardia-Strip method proved faster in obtaining results.

Evaluación de las enzimas celulolíticas producidas por hongosnativos mediante fermentación en estado sólido (SSF) utilizando residuos de cosecha de caña de azúcar / Evaluation of cellulolytic enzymes produced by native fungi through solid state fermentation (SSF) using sugarcane harvesting residues

Los residuos agrícolas de cosecha de caña de azúcar (RAC), se constituyen en una materia prima alternativa para la producción de etanol carburante, dado su contenido de celulosa próximo al 40%. El aprovechamiento de la celulosa depende de la aplicación de tratamientos fisicoquímicos o bioquímicos, que permitan la liberación de la glucosa y su posterior utilización en procesos fermentativos. La hidrólisis enzimática de estos residuos requiere un complejo celulolítico producido por microorganismos, comprendido por tres actividades enzimáticas: Endoglucanasas, Exoglucanasas y β-Glucosidasas. En el presente estudio, se evaluaron las enzimas celulolíticas producidas por dos hongos nativos del género Aspergillus spp., CH 2016 y CH 2001, mediante procesos de fermentación en estado sólido utilizando como sustrato RAC pre-tratados con organosolventes (deslignificado) y sin este pre-tratamiento. La cepa CH 2016 presentó la mayor actividad endoglucanasa 11,0773 U/mL en el sustrato sin pre-tratar el día siete de fermentación; esta misma cepa, en el sustrato deslignificado presentó la mayor actividad exoglucanasa (0,042 U/mL) y celulasa total (0,287 UPF/mL) en el día cinco de fermentación. La cepa CH 2001 presentó la mayor actividad β-glucosidasa (0,1778 U/mL) en el sustrato sin pre-tratar el día cinco de fermentación. Se observó que las variables sustrato y tiempo de fermentación, inciden en la expresión de las enzimas celulolíticas obteniendo en este trabajo extractos enzimáticos que pueden llevar a cabo una acción hidrolítica sinérgica sobre la celulosa.

Sugarcane harvesting residues are considered as a raw material for fuel ethanol production due its high content of cellulose, around 40% DS. The use of cellulose depends of the application of physicochemical or biochemical treatments that allow the release of glucose and its subsequent uses in fermentation processes. The enzymatic hydrolysis of these residues requires a cellulolytic complex produced by microorganisms, including three enzymatic activities: Endoglucanases, β-Glucosidases and Exoglucanases. In the present study, cellulolytic enzymes produced by two native fungi Aspergillus spp., CH 2016 and CH 2001 was assessment, through of solid-state fermentation processes using as raw substrate RAC and pre-treated with organosolvents (delignified). Strain CH 2016 had the highest endoglucanase activity 11.0773 U/mL in the raw substrate on day seven of fermentation, the same strain, in the delignified substrate showed the highest activity exoglucanasa (0.042 U/mL ) and total cellulase (0.287 UPF/mL) on day five of fermentation. Strain CH 2001 got the highest β-glucosidase activity (0.1778 U/mL) in the substrate without pre-treatment on day 5 of fermentation. It was observed that the variables as substrate and fermentation time affected the expression of cellulolytic enzymes.

Characterization of Solid State Fermentation Culture Conditions for Growth and Mananase Production by Aspergillus niger USM F4 on Rice Husk in Tray System.

Aim: The study evaluated various fermentation conditions for the production of mannanase. Place and Duration of Study: Industrial Biotechnology Research Laboratory (IBRL), School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia between May 2009 and September 2010. Methodology: Solid substrate fermentation was carried out in a shallow aluminum tray system (16 cm x 16 cm x 5 cm) for maximum mannanase production by Aspergillus niger USM F4 using rice husk as a substrate. Results: The maximum mannanase activity of 119.91 U/g substrate was achieved on the 6 days of cultivation when the optimized physical parameters were used (substrate thickness of 1.6 cm or equivalent to 80 g of 0.75 mm rice husk, moisture content to substrate ratio of 1:1 (w/v), cultivation temperature at room temperature (28±2ºC), inoculum size of 6x106 spores/ml and in static condition (no mixing during the fermentation process). The results showed an increment of about 30.79% of mannanase activity after the optimization (119.91 U/g substrate) compared to before optimization (91.68 U/g substrate). Conclusion: The results obtained from this study revealed that rice husk can be used as a substrate for mannanase production in solid state fermentation process.

A biodegradation study of forest biomass by Aspergillus niger F7: correlation between enzymatic activity, hydrolytic percentage and biodegradation index

Aspergillus niger F7 isolated from soil was found to be the potent producer of cellulase and xylanase. The residue of forest species Toona ciliata, Celtris australis, Cedrus deodara and Pinus roxburghii was selected as substrate for biodegradation study due to its easy availability and wide use in industry. It was subjected to alkali (sodium hydroxide) treatment for enhancing its degradation. Biodegradation of forest waste by hydrolytic enzymes (cellulase and xylanase) secreted by A. niger under solid state fermentation (SSF) was explored. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Highest extracellular enzyme activity of 2201±23.91 U/g by A. niger was shown in pretreated C. australis wood resulting in 6.72±0.20 percent hydrolysis and 6.99±0.23 biodegradation index (BI). The lowest BI of 1.40±0.08 was observed in untreated saw dust of C. deodara having the least enzyme activity of 238±1.36 U/g of dry matter. Biodegradation of forest biomass under SSF was increased many folds when moistening agent i.e. tap water had been replaced with modified basal salt media (BSM). In BSM mediated degradation of forest waste with A. niger, extracellular enzyme activity was increased up to 4089±67.11 U/g of dry matter in turn resulting in higher BI of 15.4±0.41 and percent hydrolysis of 19.38±0.81 in pretreated C. australis wood. A. niger exhibited higher enzyme activity on pretreated biomass when moistened with modified BSM in this study. Statistically a positive correlation has been drawn between these three factors i.e. enzyme activity, BI and percent hydrolysis of forest biomass thus proving their direct relationship with each other.

Xylanase production by Aspergillus awamori under solid state fermentation conditions on tomato pomace

In this work, tomato pomace, a waste abundantly available in the Mediterranean and other temperate climates agro-food industries, has been used as raw material for the production of some hydrolytic enzymes, including xylanase, exo-polygalacturonase (exo-PG), cellulase (CMCase) and ¥á-amylase. The principal step of the process is the solid state fermentation (SSF) of this residue by Aspergillus awamori. In several laboratory experiments, maximum xylanase and exo-PG activities were measured during the first days of culture, reaching values around 100 and 80 IU/gds (international units of enzyme activity per gram of dried solid), respectively. For CMCase and ¥á-amylase production remained almost constant along fermentation, with average values of 19 and 21.5 IU/gds, respectively. Experiments carried out in a plate-type bioreactor at lab scale showed a clear positive effect of aeration on xylanase and CMCase, while the opposite was observed for exo-PG and ¥á-amylase. In general, xylanase was the enzyme produced in higher levels, thus the optimum conditions for the determination of the enzyme activity was characterized. The xylanase activity shows an optimum pH of 5 and an optimum temperature of 50 ¨¬C. The enzyme is activated by Mg2+, but strongly inhibited by Hg2+ and Cu2+. The enzymatic activity remains quite high if the extract is preserved in a range of pH from 3 to 10 and a temperature between 30 ¨¬C to 40 ¨¬C.

Extra-cellular isoamylase production by Rhizopus oryzae in solid-state fermentation of agro wastes

Extra-cellular isoamylase was produced by Rhizopus oryzae PR7 in solid-state fermentations of various agro wastes, among which millet, oat, tapioca, and arum (Colocasia esculenta) showed promising results. The highest amount of enzyme production was obtained after 72 h of growth at 28°C. The optimum pH for enzyme production was - 8.0. Among the various additives tested, enzyme production increased with ions such as Ca2+, Mg2+ and also with cysteine, GSH, and DTT. The enzyme synthesis was reduced in the presence of thiol inhibitors like Cu2+ and pCMB. The surfactants like Tween-40, Tween-80 and Triton X-100 helped in enhancing the enzyme activity. The production could be further increased by using the combinations of substrates. The ability to produce high amount of isoamylase within a relatively very short period and the capability of degrading wastes could make the strain suitable for commercial production of the enzyme.

Tannase production by Penicillium atramentosum KM under SSF and its applications in wine clarification and tea cream solubilization

Tannin acyl hydrolase commonly known as tannase is an industrially important enzyme having a wide range of applications, so there is always a scope for novel tannase with better characteristics. A newly isolated tannase-yielding fungal strain identified as Penicillium atramentosum KM was used for tannase production under solid-state fermentation (SSF) using different agro residues like amla (Phyllanthus emblica), ber (Zyzyphus mauritiana), jamun (Syzygium cumini), Jamoa (Eugenia cuspidate) and keekar (Acacia nilotica) leaves. Among these substrates, maximal extracellular tannase production i.e. 170.75 U/gds and 165.56 U/gds was obtained with jamun and keekar leaves respectively at 28ºC after 96 h. A substrate to distilled water ratio of 1:2 (w/v) was found to be the best for tannase production. Supplementation of sodium nitrate (NaNO3) as nitrogen source had enhanced tannase production both in jamun and keekar leaves. Applications of the enzyme were studied in wine clarification and tea cream solubilization. It resulted in 38.05 percent reduction of tannic acid content in case of jamun wine, 43.59 percent reduction in case of grape wine and 74 percent reduction in the tea extract after 3 h at 35ºC.

Evaluation of Potential Fungal Species for the in situ Simultaneous Saccharification and Fermentation (SSF) of Cellulosic Material

Three fungal species were evaluated for their abilities to saccharify pure cellulose. The three species chosen represented three major wood-rot molds; brown rot (Gloeophyllum trabeum), white rot (Phanerochaete chrysosporium) and soft rot (Trichoderma reesei). After solid state fermentation of the fungi on the filter paper for four days, the saccharified cellulose was then fermented to ethanol by using Saccharomyces cerevisiae. The efficiency of the fungal species in saccharifying the filter paper was compared against a low dose (25 FPU/g cellulose) of a commercial cellulase. Total sugar, cellobiose and glucose were monitored during the fermentation period, along with ethanol, acetic acid and lactic acid. Results indicated that the most efficient fungal species in saccharifying the filter paper was T. reesei with 5.13 g/100 g filter paper of ethanol being produced at days 5, followed by P. chrysosporium at 1.79 g/100 g filter paper. No ethanol was detected for the filter paper treated with G. trabeum throughout the five day fermentation stage. Acetic acid was only produced in the sample treated with T. reesei and the commercial enzyme, with concentration 0.95 and 2.57 g/100 g filter paper, respectively at day 5. Lactic acid production was not detected for all the fungal treated filter paper after day 5. Our study indicated that there is potential in utilizing in situ enzymatic saccharification of biomass by using T. reesei and P. chrysosporium that may lead to an economical simultaneous saccharification and fermentation process for the production of fuel ethanol.

Production of á-Galactosidase by Aspergillus oryzae through solid-state fermentation and its application in soymilk Galactooligosaccharide hydrolysis

á-Galactosidase was produced by Aspergillus oryzae on red gram plant waste-wheat bran based media in solid-state fermentation (SSF). Optimum temperature for á-galactosidase production was 35 0C and upto 4 cm of bed height of substrate had no inhibitory effect on enzyme production. Hydrolysis of galactooligosaccharides in soymilk was carried out by á-galactosidase. Optimum temperature and pH for the hydrolysis of raffinose and stachyose of soymilk were 55(0)C and 5.2-6.2, respectively. The enzymatic treatment for 3 h completely removed the raffinose oligosaccharides in soymilk. Crude extract also showed considerable amount of invertase activity.

Producción de eaanol a partir de almidón de yuc utilizando la estrategia de prroceso sacarificación- fermentación simultáneas (ssf) / Ethanol prodctioon from cassva starch using the process strategy simultaneoous saccharification-fermentation

La tendencia mundial en el manejo de los combustibles, en especial los biocombustibles como el etanol, ha llevado a explorar nuevas metodologías de proceso para optimizar su producción; por tal razón se aborda en esta investigación el proceso sacarificación fermentación simultáneas, se evalúa la influencia de la concentración de azúcares reductores y la dosificación de la enzima Spirizyme fuel® sobre la productividad y concentración final de etanol, bajo el proceso SSF (sacarificación -fermentación simultáneas), partiendo del licuado de almidón de yuca como sustrato. El proceso SSF se compara con un control con características de sacarificación-fermentación independientes (SHF), proceso convencional. Sólo el factor concentración inicial de sustrato presenta efecto sobre la productividad de etanol. Las cinéticas de proceso, frente a las del control, presentan reducciones de tiempo de 47 y 33% para los niveles de sustrato evaluados. Los niveles de productividad son mayores en un 33% para el nivel de 150 g/l de AR (azúcares reductores) y se mantiene constante para 200 g/l. La glucosa en la estrategia SSF, conforme se produce se transforma en etanol, no permitiendo alcanzar concentraciones superiores a 100 g/l, lo que se traduce en que no se presentan inhibiciones por sustrato. La concentración de etanol no afecta la reacción de la enzima en el proceso de sacarificación. El proceso SSF demuestra su viabilidad técnica en la producción de alcohol, al reducir los tiempos y necesidades de energía en la producción de alcohol carburante a partir de almidón de yuca.

The world trend on fuel management, in special biofuels like ethanol, have gone to explorer new methodologies of process to optimize its production by this reason in this research is about simultaneous sacarification fermentation process and evaluate initial concentration of reducing sugar, and enzyme dosing of Spirizyme fuel® are evaluated on productivity and final concentration of ethanol, under SSF (Simultaneous Saccharification and Fermentation) process, from the product of the licuefaction process of cassava starch as substrate. The SSF process is evaluated against SHF (Independent Saccharification and Fermentation) process as control. Only the factor, initial concentration of substrate presents effect over ethanol productivity. The kinetic of SSF process, in opposite to the SHF process, presents time diminution of the global process around 47 y 33% to substrate levels of 150 and 200 g/l respectively. The productivity values are most at a 33% to 150 g/l of reducing sugar, and they keep constant to 200 g/l reducing sugar. The glucose in SSF strategy, at the time it is producing, it is transformed to ethanol, does not allowing to reach superior concentration to 100 g/l of reducing sugar, this implicates there is not substrate inhibition. The ethanol concentration doesn't affect the enzymatic process of sacharification. The SSF process demonstrates his technical viability on the ethanol production, to reduce time an energy requirements on the ethanol production from cassava flour.