Search results for: xylose-fermenting yeast

Authors: Howaida I. Abd-Alla, Amal Z. Hassan, Maha Soltan, Atef G. Hanna, Mounir M. El-Safty

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Acokanthera oblongifolia (Apocynaceae) is used for treatment of several infection diseases and is a well-known cardiac glycoside-containing plant. The infusion of their leaves is gargled to treat tonsillitis and is used medicinally to treat snakebites. The total cardiac glycosides content in the leaves was determined by referring to gitoxigenin as a reference compound. Two triterpenes, lup-20(29)-en-3β-ol (1) and oleanolic acid (2); two cardenolides, acovenoside A (3) and acobioside A (4) were isolated from the ethyl acetate extract. Their structures were determined on the basis of spectral analysis. Major constituents isolated from this species were evaluated for cytotoxicity against normal lung cell line (Wi38) and antimicrobial activities against Gram-positive (two strains) and Gram-negative bacteria (four strains), yeast-like fungi (two strains) and fungi (five strains). The minimum inhibitory concentration (MIC) of the compounds was determined using broth microdilution method. Their viral inhibitory effects against avian influenza virus type A (AI-H5N1) and Newcastle disease virus (NDV) in specific pathogen free (SPF) embryonated chicken eggs (ECE), chicken embryo fibroblasts (CEF) and Vero cells were evaluated. The cardenolide (3) showed viral inhibitory effects against AI-H5N1 and NDV in SPF ECE. The two cardenolides isolated have shown potent cytotoxicity against Vero cells. Compound (3) showed potent anti-Gram-negative bacteria activity. These results suggested that acovenoside A might be promising for future antiviral and antimicrobial drug design.

Keywords: Acokanthera, AI-H5N1, Cardenolides, NDV, SPF-ECE, VERO, Wi38 , Microbe

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Authors: Ifeoluwa Omotehinse Oloruntoba

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Through comparative and causal models of translation, this paper examined the translation of ‘bread’ into the Yoruba language in three Yoruba versions of the Bible: Bibeli Yoruba Atoka (YBA), Bibeli Mimo ni Ede Yoruba Oni (BMY) and Bibeli Mimo (BM). In biblical times, bread was a very important delicacy that it was synonymous with food in general and in the Bible, bread sometimes refers to a type of food (a mixture of flour, water, and yeast that is baked) or food in general. However, this is not the case in the Yoruba culture. In fact, some decades ago, bread was not known in Nigeria and had no name in the Yoruba language until the 1900s when it was codified as burẹdi in Yoruba, a term borrowed from English and transliterated. Nevertheless, in Nigeria presently, bread is not a special food and it is not appreciated or consumed like in the West. This makes it difficult to translate bread in the Bible into Yoruba. From an investigation on the translation of this term, it was discovered that bread which has 330 occurrences in the English Bible translation (King James) has few occurrences in the three Yoruba Bible versions. In the first version (YBA) published in the 1880s, where bread is synonymous with food in general, it is mostly translated as oúnjẹ (food) or the verb jẹ (to eat), revealing that something is eaten but not indicating what it is. However, when the bread is a type of food, it is rendered as akara, a special delicacy of the Yoruba people made from beans flour. In the later version (BMY) published in the 1990s, bread as food, in general, is also mainly translated as oúnjẹ or the verb jẹ, but when it is a type of food, it is translated as akara with few occurrences of burẹdi. In the latest edition (BM), bread as food is either rendered as ounje or literally translated as burẹdi. Where it is a type of food in this version, it is mainly rendered as burẹdi with few occurrences of akara, indicating the assimilation of bread into the Yoruba culture. This result, although limited, shows that the Bible was translated into Yoruba to make it accessible to Yoruba speakers in their everyday language, hence the application of both domesticating and foreignising strategies. This research also emphasizes the role of the translator as an intermediary between two cultures.

Keywords: translation, Bible, Yoruba, cultural problems

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Authors: Nwafia Ifeyinwa Nkeiruka, Nwafia Walter Chukwuma

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Since the early 1980’s fungi have emerged as major causes of human diseases, especially among immunocompromised. The most commonly isolated yeast is Candida albicans and constitutes the 4th most common nosocomial BSI in humans. It is progressive and cumulative and become more complex over time.It can even lead to leaky gut syndrome that causes food and environmental allergies. It is worthy of note that all the available data on oral Candida risk factors in humans were documented essentially using data from studies conducted in other areas, hence there is need for comparative and complementary information from the South eastern part of Nigeria. Method: 200 subjects of all age groups of both sexes were randomly examined,by swabbing their palatine mucosa and dorsal tongue with sterile cotton wool,then cultured into Sabouraud dextrose agar plates supplemented with antibiotics and incubated aerobically at 37 degree for 48 hrs. Identification of Candida albicans was done by germ tubes tests, chlamydospores production on cornmeal agar supplemented with 1% Tween 80.Sugar and nitrogen assimilation test using API 20C Auxanogram and potassium nitrate agar. Results: Out of 30 samples that were positive for candida, 15 (50%) were candida albicans. Using the anova test (P < 0.05) this variation is significant (P = 0016). followed by C. dublinensis 3 (13%), C. tropicalis 3 (10%), C. pseudotropicalis 3 (10%), C, glabrata 2 (7%), C. parapsilosis 2 (7%) and lastly C. krusei 1 (3%).However, 53% of the patients were female while 47% were male. Among the HIV positive isolates.67% were HIV isolates not on drugs while 33% positives isolates were on drugs and the percentages of candida species in these patients were as follows C. albicans were 45% followed by C. glabrata and C.tropicalis which were 17% each, C.parapsilosis, C.dubliensis and C.pseudotropicalis were all 8% each. Conclusion: Oral Candidiasis is a marker of systemic diseases and in some cases, it may be the first clinical presentation. There is need for more intensive clinical and laboratory monitoring and possible early intervention to prevent the reoccurrence and resistance to treatment.

Keywords: oral cavity, Candida species, oral Candidiasis, risk factors

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Authors: Sara D. Garduno-Diaz, Ramona Milcheva, Chanyu Xu

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Women’s reproductive stages (pre-pregnancy, pregnancy, and lactation) represent a time of higher micronutrient needs. With a healthy food selection as the first path of choice to cover these increased needs, tandem micronutrient supplementation is often required. Because pregnancy and lactation should be treated with care, all supplements consumed should be of quality ingredients and manufactured through controlled processes. This work describes the process followed for the development of plant-based multiple micronutrient supplements aimed at addressing the growing demand for natural ingredients of non-animal origin. A list of key nutrients for inclusion was prioritized, followed by the identification and selection of qualified raw ingredient providers. Nutrient absorption into the food matrix was carried out through natural processes. The outcome is a new line of products meeting the set criteria of being gluten and lactose-free, suitable for vegans/vegetarians, and without artificial conservatives. In addition, each product provides the consumer with 10 vitamins, 6 inorganic nutrients, 1 source of essential fatty acids, and 1 source of phytonutrients each (maca, moringa, and chlorella). Each raw material, as well as the final product, was submitted to microbiological control three-fold (in-house and external). The final micronutrient mix was then tested for human factor contamination, pesticides, total aerobic microbial count, total yeast count, and total mold count. The product was created with the aim of meeting product standards for the European Union, as well as specific requirements for the German market in the food and pharma fields. The results presented here reach the point of introduction of the newly developed product to the market, with acceptability and effectiveness results to be published at a later date.

Keywords: fertility, lactation, organic, pregnancy, vegetarian

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Authors: Rashida Rahmat Zohra, Afsheen Aman, Shah Ali Ul Qader

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Commercially important biopolymer, dextran, is enzymatically degraded into lower molecular weight fractions of vast industrial potential. Various organisms are associated with dextranase production, among which fungal, yeast and bacterial origins are used for commercial production. Dextranases are used to remove contaminating dextran in sugar processing industry and also used in oral care products for efficient removal of dental plaque. Among the hydrolytic products of dextran, isomaltooligosaccharides have prebiotic effect in humans and reduces the cariogenic effect of sucrose in oral cavity. Dextran derivatives produced by hydrolysis of high molecular polymer are also conjugated with other chemical and metallic compounds for usage in pharmaceutical, fine chemical industry, cosmetics, and food industry. Owing to the vast application of dextran and dextranases, current study focused on purification and analysis of kinetic parameters of dextranase from a newly isolated strain of Bacillus licheniformis KIBGE-IB25. Dextranase was purified up to 35.75 folds with specific activity of 1405 U/mg and molecular weight of 158 kDa. Analysis of kinetic parameters revealed that dextranase performs optimum cleavage of low molecular weight dextran (5000 Da, 0.5%) at 35ºC in 15 min at pH 4.5 with a Km and Vmax of 0.3738 mg/ml and 182.0 µmol/min, respectively. Thermal stability profiling of dextranase showed that it retained 80% activity up to 6 hours at 30-35ºC and remains 90% active at pH 4.5. In short, the dextranase reported here performs rapid cleavage of substrate at mild operational conditions which makes it an ideal candidate for dextran removal in sugar processing industry and for commercial production of low molecular weight oligosaccharides.

Keywords: Bacillus licheniformis, dextranase, gel permeation chromatograpy, enzyme purification, enzyme kinetics

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Authors: Manar Y. Abd El-Aziz, Alyaa E. Morgham, Amira A. El-Fallal, Heba Tolla E. Abo El Naga

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Different textile materials are usually used in upholstery. However, upholstery parts may become unhealthy when dust accrues and bacteria raise on the surface, which negatively affects the user's health. Also, leather and artificial leather were used in upholstery but, leather has a high cost and artificial leather has a potential chemical risk for users. Researchers have advanced vegie leather made from bacterial cellulose a symbiotic culture of bacteria and yeast (SCOBY). SCOBY remains a gelatinous, cellulose biofilm discovered floating at the air-liquid interface of the container. But this leather still needs some enhancement for its mechanical properties. This study aimed to prepare SCOBY, produce bamboo rib knitted fabrics with two different stitch densities, and cotton woven fabric then laminate these fabrics with the prepared SCOBY film to enhance the mechanical properties of the SCOBY leather at the same time; add anti-microbial function to the prepared fabrics. Laboratory tests were conducted on the produced samples, including tests for function properties; anti-microbial, thermal conductivity and light transparency. Physical properties; thickness and mass per unit. Mechanical properties; elongation, tensile strength, young modulus, and peel force. The results showed that the type of the fabric affected significantly SCOBY properties. According to the test results, the bamboo knitted fabric with higher stitch density laminated with SCOBY was chosen for its tensile strength and elongation as the upholstery of a bed model with antimicrobial properties and comfortability in the headrest design. Also, the single layer of SCOBY was chosen regarding light transparency and lower thermal conductivity for the creation of a lighting unit built into the bed headboard.

Keywords: anti-microbial, bamboo, rib, SCOPY, upholstery

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Authors: N. Noori, P. Taherkhani, A. Akhondzadeh Basti, H. Gandomi, M. Alimohammadi

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Research on natural antimicrobial agents, especially of plant origin, highly noticed in recent years and evaluation of antimicrobial effects of native plants such as Bunium persicum Boiss. is especially important. In the present study, sensory characteristics and microbiological properties of Gouda cheese affected by different concentrations of Bunium persicum Boiss. essential oil were investigated. Extraction of the essential oil was performed by hydro distillation. The oil was analyzed by GC using flame ionization (FID) and GC/ MS for detection. The antimicrobial effects were determined against various microbial groups (aerobic mesophilic bacteria, enterococci, mesophilic lactobacilli, enterobacteriaceae, lactococcus and yeasts). Microbial groups were counted during ripening period using plate count on specific culture media. Organoleptic evaluation including teture, flavor, odor, color and total acceptability were determined at the end of aging. According to results, the essential oil yield was 4/1 % ( W/ W). Twenty- six compounds were identified in the oil that concluded 99.7 % of the total oil. The major components of Bunium persicum Boiss. essential oil were γ- terpinene- 7- al (26.9 %) and cuminaldehyde (23.3 %). Generally, the increase of Black Cumin essential oil concentration led to reduction in microbial counts in different groups. The maximum antimicrobial effect was seen in yeast that reduced by 2 log compared to the control group at EO concentration of 4µl/ ml at day 90.The minimum reduction was observed in enterobacteriaceae that showed only 0.75 log decreese compared to the control at the same concentration of EO. Addition of EO improved organoleptic properties of Gouda cheese especially in the case of flavor and odor characteristic. However, no significant differences were observed in texture and color between treatment and control groups. Bunium persicum Boiss. essential oil could be used as preservative material and flavoring agent in some kinds of food such as cheese and also could be provided consumers health.

Keywords: Bunium persicum Boiss. essential oil, Microbiological properties, sensory evaluation, gouda cheese

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Authors: P. Mandree, S. O. Ramchuran, F. O'Brien, L. Sethunya, S. Khumalo

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South Africa is identified as one of the five emerging waste management markets, globally. The waste sector in South Africa influences the areas of energy, water and food at an economic and social level. Recently, South African industries have focused on waste valorization and diversification of the current product offerings in an attempt to reduce industrial waste, target a zero waste-to-landfill initiative and recover energy. South Africa has a number of waste streams including industrial and agricultural biomass, municipal waste and marine waste. Large volumes of agricultural and forestry residues, in particular, are generated which provides significant opportunity for production of bio-based fuels and chemicals. This could directly impact development of a rural economy. One of the largest agricultural industries is the sugar industry, which contributes significantly to the country’s economy and job creation. However, the sugar industry is facing challenges due to fluctuations in sugar prices, increasing competition with low-cost global sugar producers, increasing energy and agricultural input costs, lower consumption and aging facilities. This study is aimed at technology development for the production of various bio-based chemicals using feedstock from the sugar refining process. Various indigenous bacteria and yeast species were assessed for the potential to produce platform chemicals in flask studies and at 30 L fermentation scale. Quantitative analysis of targeted bio-based chemicals was performed using either gas chromatography or high pressure liquid chromatography to assess production yields and techno-economics in order to compare performance to current commercial benchmark processes. The study also creates a decision platform for the research direction that is required for strain development using Industrial Synthetic Biology.

Keywords: bio-based chemicals, biorefinery, industrial synthetic biology, waste valorization

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Authors: Lorenzo Cocola, Massimo Fedel, Dragiša Savić, Bojana Danilović, Luca Poletto

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An instrument for the detection and evaluation of gaseous carbon dioxide in the headspace of closed containers has been developed in the context of Packsensor Italian-Serbian joint project. The device is based on Tunable Diode Laser Absorption Spectroscopy (TDLAS) with a Wavelength Modulation Spectroscopy (WMS) technique in order to accomplish a non-invasive measurement inside closed containers of fermented dairy products (yogurts and fermented cheese in cups and bottles). The purpose of this instrument is the continuous monitoring of carbon dioxide concentration during incubation and storage of products over a time span of the whole shelf life of the product, in the presence of different microorganisms. The instrument’s optical front end has been designed to be integrated in a thermally stabilized incubator. An embedded computer provides processing of spectral artifacts and storage of an arbitrary set of calibration data allowing a properly calibrated measurement on many samples (cups and bottles) of different shapes and sizes commonly found in the retail distribution. A calibration protocol has been developed in order to be able to calibrate the instrument on the field also on containers which are notoriously difficult to seal properly. This calibration protocol is described and evaluated against reference measurements obtained through an industry standard (sampling) carbon dioxide metering technique. Some sets of validation test measurements on different containers are reported. Two test recordings of carbon dioxide concentration evolution are shown as an example of instrument operation. The first demonstrates the ability to monitor a rapid yeast growth in a contaminated sample through the increase of headspace carbon dioxide. Another experiment shows the dissolution transient with a non-saturated liquid medium in presence of a carbon dioxide rich headspace atmosphere.

Keywords: TDLAS, carbon dioxide, cups, headspace, measurement

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Authors: Aurora Gitto, Philipp Proksch

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The presence of various microorganisms (bacteria, fungi) in surface water and groundwater represents an important issue for human health worldwide. The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) has emerged as a promising and reliable tool for bacteria identification in clinical diagnostic microbiology and environmental strains thanks to an ionization technique that uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation. The study aims first to conceptualise and set up library information and create a comprehensive database of MALDI-TOF-MS spectra from environmental water samples. The samples were analysed over a year (2021-2022) using membrane filtration methodology (0.45 μm and 0.22 μm) and then isolated on R2A agar for a period of 5 days and Yeast extract agar growing at 22 °C up to 4 days and 37 °C for 48 hours. The undetected organisms by MALDI-TOF-MS were analysed by PCR and then sequenced. The information obtained by the sequencing was further implemented in the MALDI-TOF-MS library. Among the culturable bacteria, the results show how the incubator temperature affects the growth of some genera instead of others, as demonstrated by Pseudomonas sp., which grows at 22 °C, compared to Bacillus sp., which is abundant at 37 °C. The bacteria community shows a variation in composition also between the media used, as demonstrated with R2A agar which has been defined by a higher presence of organisms not detected compared to YEA. Interesting is the variability of the Genus over one year of sampling and how the seasonality impacts the bacteria community; in fact, in some sampling locations, we observed how the composition changed, moving from winter to spring and summer. In conclusion, the bacteria community in groundwater and river bank filtration represents important information that needs to be added to the library to simplify future water quality analysis but mainly to prevent potential risks to human health.

Keywords: water quality, MALDI-TOF-MS, sequencing, library

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Authors: Mervat Kassem, Nourhan Fanaki, F. Dabbous, Hamida Abou-Shleib, Y. R. Abdel-Fattah

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With increasing environmental awareness and emphasis on a sustainable society in harmony with the global environment, biosurfactants are gaining prominence and have already taken over for a number of important industrial uses. They are produced by living organisms, for examples Pseudomonas aeruginosa which produces rhamnolipids, Candida (formerly Torulopsis) bombicola, which produces high yields of sophorolipids from vegetable oils and sugars and Bacillus subtilis which produces a lipopeptide called surfactin. The main goal of this work was to optimize biosurfactants production by an environmental Gram positive isolate for large scale production with maximum yield and low cost. After molecular characterization, phylogenetic tree was constructed where it was found to be B. subtilis, which close matches to B. subtilis subsp. subtilis strain CICC 10260. For optimizing its biosurfactants production, sequential statistical design using Plackett-Burman and response surface methodology, was applied where 11 variables were screened. When analyzing the regression coefficients for the 11 variables, pH, glucose, glycerol, yeast extract, ammonium chloride and ammonium nitrate were found to have a positive effect on the biosurfactants production. Ammonium nitrate, pH and glucose were further studied as significant independent variables for Box-Behnken design and their optimal levels were estimated and were found to be 7.328 pH value, 3 g% glucose and 0.21g % ammonium nitrate yielding high biosurfactants concentration that reduced the surface tension of the culture medium from 72 to 18.16 mN/m. Next, kinetics of cell growth and biosurfactants production by the tested B. subtilis isolate, in bioreactor was compared with that of shake flask where the maximum growth and specific growth (µ) in the bioreactor was higher by about 25 and 53%, respectively, than in shake flask experiment, while the biosurfactants production kinetics was almost the same in both shake flask and bioreactor experiments.

Keywords: biosurfactants, B. subtilis, molecular identification, phylogenetic trees, Plackett-Burman design, Box-Behnken design, 16S rRNA

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Authors: Priya Tomar, Pallavi Mittal

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As industrialization is inevitable and progress with rapid acceleration, the need for innovative ways to get rid of waste has increased. Recent advancement in bioresource technology paves novel ideas for recycling of factory waste that has been polluting the agro-industry, soil and water bodies. Paper industries in India are in a considerable number, where molasses and impure alcohol are still being used as raw materials for manufacturing of paper. Paper mills based on nonconventional agro residues are being encouraged due to increased demand of paper and acute shortage of forest-based raw materials. The colouring body present in the wastewater from pulp and paper mill is organic in nature and is comprised of wood extractives, tannin, resins, synthetic dyes, lignin and its degradation products formed by the action of chlorine on lignin which imparts an offensive colour to the water. These mills use different chemical process for paper manufacturing due to which lignified chemicals are released into the environment. Therefore, the chemical oxygen demand (COD) of the emanating stream is quite high. This paper presents some new techniques that were developed for the efficiency of bioremediation on paper industry. A short introduction to paper industry and a variety of presently available methods of bioremediation on paper industry and different strategies are also discussed here. For solving the above problem, two bacterial strains (Pseudomonas aeruginosa and Bacillus subtilis) and their consortia (Pseudomonas aeruginosa and Bacillus subtilis) were utilized for the pulp and paper mill effluent. Pseudomonas aeruginosa and Bacillus subtilis named as T–1, T–2, T–3, T–4, T–5, T–6, for the decolourisation of paper industry effluent. The results indicated that a maximum colour reduction is (60.5%) achieved by Pseudomonas aeruginosa and COD reduction is (88.8%) achieved by Bacillus subtilis, maximum pH changes is (4.23) achieved by Pseudomonas aeruginosa, TSS reduction is (2.09 %) achieved by Bacillus subtilis, and TDS reduction is (0.95 %) achieved by Bacillus subtilis. When the wastewater was supplemented with carbon (glucose) and nitrogen (yeast extract) source and data revealed the efficiency of Bacillus subtilis, having more with glucose than Pseudomonas aeruginosa.

Keywords: bioremediation, paper and pulp mill effluent, treated effluent, lignin

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Authors: Snehal D. Ganjave, Hardik Dodia, Avinash V. Sunder, Swati Madhu, Pramod P. Wangikar

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Batch cultivation of recombinant bacteria in shake flasks results in low cell density due to nutrient depletion. Previous protocols on high cell density cultivation in shake flasks have relied mainly on controlled release mechanisms and extended cultivation protocols. In the present work, we report an optimized fed-batch process for high cell density cultivation of recombinant E. coli BL21(DE3) for protein production. A cybernetic model-based, multi-objective optimization strategy was implemented to obtain the optimum operating variables to achieve maximum biomass and minimized substrate feed rate. A syringe pump was used to feed a mixture of glycerol and yeast extract into the shake flask. Preliminary experiments were conducted with online monitoring of dissolved oxygen (DO) and offline measurements of biomass and glycerol to estimate the model parameters. Multi-objective optimization was performed to obtain the pareto front surface. The selected optimized recipe was tested for a range of proteins that show different extent soluble expression in E. coli. These included eYFP and LkADH, which are largely expressed in soluble fractions, CbFDH and GcanADH , which are partially soluble, and human PDGF, which forms inclusion bodies. The biomass concentrations achieved in 24 h were in the range 19.9-21.5 g/L, while the model predicted value was 19.44 g/L. The process was successfully reproduced in a standard laboratory shake flask without online monitoring of DO and pH. The optimized fed-batch process showed significant improvement in both the biomass and protein production of the tested recombinant proteins compared to batch cultivation. The proposed process will have significant implications in the routine cultivation of E. coli for various applications.

Keywords: cybernetic model, E. coli, high cell density cultivation, multi-objective optimization

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Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

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Authors: Muhammad Dildar Gogi, Muhammad Jalal Arif, Muhammad Junaid Nisar, Mubashir Iqbal, Waleed Afzal Naveed, Muhammad Ahsan Khan, Ahmad Nawaz, Muhammad Sufian, Muhammad Arshad, Amna Jalal

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Fruit flies of Bactrocera genus cause heavy losses in fruits and vegetables globally and insecticide-application for their control creates issues of ecological backlash, environmental pollution, and food safety. There is need to explore alternatives and food-baits application is considered safe for the environment and effective for fruit fly management. Present experiment was carried out to assess the attractancy of five phagostimulant-Mixtures (PHS-Mix) prepared by mixing banana-squash, mulberry, protein-hydrolysate and molasses with some phagostimulant-lure sources including beef extract, fish extract, yeast, starch, rose oil, casein and cedar oil in five different ratios i.e., PHS-Mix-1 (1 part of all ingredients), PHS-Mix-2 (1 part of banana with 0.75 parts of all other ingredients), PHS-Mix-3 (1 part of banana with 0.5 parts of all other ingredients), PHS-Mix-4 (1 part of banana with 0.25 parts of all other ingredients) and PHS-Mix-5 (1 part of banana with 0.125 parts of all other ingredients). These were evaluated in comparison with a standard (GF-120). PHS-Mix-4 demonstrated 40.5±1.3-46.2±1.6% AI for satiated flies (class-II i.e., moderately attractive) and 59.5±2.0-68.6±3.0% AI for starved flies (class-III i.e., highly attractive) for both B. dorsalis and B. zonata in olfactometric study while the same exhibited 51.2±0.53% AI (class-III i.e., highly attractive) for B. zonata and 45.4±0.89% AI (class-II i.e., moderately attractive) for B. dorsalis in field study. PHS-Mix-1 proved non-attractive (class-I) and moderately attractive (class-II) phagostimulant in olfactometer and field studies, respectively. PHS-Mix-2 exhibited moderate attractiveness for starved lots in olfactometer and field-lot in field studies. PHS-Mix-5 proved non-attractive to starved and satiated lots of B. zonata and B. dorsalis females in olfactometer and field studies. Overall PHS-Mix-4 proved better phagostimulant-mixture followed by PHS-Mix-3 which was categorized as class-II (moderately attractive) phagostimulant for starved and satiated lots of female flies of both species in olfactometer and field studies; hence these can be exploited for fruit fly management.

Keywords: attractive index, field conditions, olfactometer, Tephritid flies

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Authors: Dong-Gi Lee, Suyeon Cha, Yong-Sun Bahn

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Sterol lipid is essential for cell membrane structure in eukaryotic cells. In mammalian cells, sterol regulatory element binding proteins (SREBPs) act as principal regulators of cellular cholesterol which is essential for proper cell membrane fluidity and structure. SREBP and sterol regulation are related to levels of cellular oxygen because it is a major substrate for sterol synthesis. Upon cellular sterol and oxygen levels are depleted, SREBP is translocated to the Golgi where it undergoes proteolytic cleavage of N terminus, then it travels to the nucleus to play a role as transcription factor. In yeast cells, synthesis of ergosterol is also highly oxygen consumptive, and Sre1 is a transcription factor known to play a central role in adaptation to growth under low oxygen condition and sterol homeostasis in Cryptococcus neoformans. In this study, we observed phenotypes in other strains of Cryptococcus species by constructing hob1Δ and sre1Δ mutants to confirm whether the functions of both genes are conserved in most serotypes. As a result, hob1Δ showed no noticeable phenotype under treatment of antifungal drugs and most environmental stresses in R265 (C. gattii) and XL280 (C. neoformans), suggesting that Hob1 is related to sterol regulation only in H99 (serotype A). On the other hand, the function of Sre1 was found to be conserved in most serotypes. Furthermore, mating experiment of hob1Δ or sre1Δ showed dramatic defects in serotype A (H99) and D (XL280). It revealed that Hob1 and Sre1 related to mating ability in Cryptococcus species, especially cell fusion efficiency. In conclusion, HOB1 and SRE1 play crucial role in regulating sterol-homeostasis and differentiation in C. neoformans, moreover, Hob1 is specific gene in Cryptococcus neoformans. It suggests that Hob1 is considered as potent factor-targeted new safety antifungal drug.

Keywords: cryptococcus neoformans, Hob1, Sre1, sterol regulatory element binding proteins

Procedia PDF Downloads 229

Authors: K. Premakumar, R. G. Lakmali, S. M. A. C. U. Senarathna

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In the present world, production and consumption of fruit and vegetable beverages have increased owing to the healthy life style of the people. Therefore, a study was conducted to develop composite vegetable squash by incorporating nutritional, medicinal and organoleptic properties of tomato, pumpkin and ginger. Considering the finding of several preliminary studies, five formulations in different combinations tomato pumpkin were taken and their physico-chemical parameters such as pH, TSS, titrable acidity, ascorbic acid content and total sugar and organoleptic parameters such as colour, aroma, taste, nature, overall acceptability were analyzed. Then the best sample was improved by using 1 % ginger (50% tomato+ 50% pumpkin+ 1% ginger). Best three formulations were selected for storage studied. The formulations were stored at 30 °C room temperature and 70-75% of RH for 12 weeks. Physicochemical parameters , organoleptic and microbial activity (total plate count, yeast and mold, E-coil) were analyzed during storage periods and protein content, fat content, ash were also analysed%.The study on the comparison of physico-chemical and sensory qualities of stored Squashes was done up to 12 weeks storage periods. The nutritional analysis of freshly prepared tomato pumpkin vegetable squash formulations showed increasing trend in titratable acidity, pH, total sugar, non -reducing sugar, total soluble solids and decreasing trend in ascorbic acid and reducing sugar with storage periods. The results of chemical analysis showed that, there were the significant different difference (p < 0.05) between tested formulations. Also, sensory analysis also showed that there were significant differences (p < 0.05) for organoleptic character characters between squash formulations. The highest overall acceptability was observed in formulation with 50% tomato+ 50% pumpkin+1% ginger and all the all the formulations were microbiologically safe for consumption. Based on the result of physico-chemical characteristics, sensory attributes and microbial test, the Composite Vegetable squash with 50% tomato+50% pumpkin+1% ginger was selected as best formulation and could be stored for 12 weeks without any significant changes in quality characteristics.

Keywords: nutritional analysis, formulations, sensory attributes, squash

Procedia PDF Downloads 174

Authors: Wisdom Robert Duruji

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Local farmers and fruit processors in developing countries of West Africa use different ripening agents to accelerate the ripening process of plantain and banana. This study reports on the influence of different ripening agents on the shelf-life and microbial load of organic and inorganic plantain (Musa paradisiaca) and banana (Musa sapientum) during ripening process and the health implication for food security in Nigeria. The experiment consisted of four treatments, namely: Calcium carbide, Irvingia gabonensis fruits, Newbouldia laevis leaves and a control, where no ripening agent was applied to the fingers of plantain and banana. The unripe and ripened plantain and banana were subjected to microbial analysis by isolating their micro flora (Bacteria, Yeast and Mould) using pour plate method. Microbes present in the samples were enumerated, characterized and classified to genera and species. The result indicated that the microbial load of inorganic plantain from (Urban day) open market in Ile-Ife increased from 8.00 for unripe to 12.11 cfu/g for ripened; and the microbial load of organic plantain from Obafemi Awolowo University Teaching and Research Farm (OAUTRF) increased from 6.00 for unripe to 11.60 cfu/g for ripened. Also, the microbial load of inorganic banana from (Urban day) open market in Ile-Ife increased from 8.00 for unripe to 11.50 cfu/g for ripened; while the microbial load of organic banana from OAUTRF increased from 6.50 for unripe to 9.40 cfu/g for ripened. The microbial effects of the ripening agents increased from 10.00 for control to 16.00 cfu/g for treated (ripened) organic and inorganic plantain; while that of organic and inorganic banana increased from 7.50 for control to 14.50 cfu/g for ripened. Visual observation for the presence of fungal colonies and deterioration rates were monitored till seven days after the plantain and banana fingers have fully ripened. Inorganic plantain and banana from (Urban day) open market in Ile-Ife are more contaminated than organic plantain and banana fingers from OAUTRF. The ripening accelerators reduced the shelf life, increased senescence, and microbial load of plantain and banana. This study concluded that organic Agriculture is better and microbial friendlier than inorganic farming.

Keywords: organic agriculture, food security, Musaceae, calcium carbide, Irvingia gabonensis, Newbouldia laevis

Procedia PDF Downloads 486

Authors: Dibakar Chandra Deka, Anamika Kalita Deka

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Fermented alcoholic beverage production has its own stand among the tribal communities of North-East India. This biological oxidation method is followed by Ahom, Dimasa, Nishi, Miri, Bodo, Rabha tribes of this region. Bodo tribes among them not only prepare fermented alcoholic beverage but also store it for various time periods like 3 months, 6 months, 9 months, 12 months and 15 months etc. They prepare alcoholic beverage Jou (rice beer) following the fermentation of Oryza sativa with traditional yeast culture Amao. Saccharomyces cerevisiae is the main domain strain present in Amao. Dongphangrakep (Scoparia dulcis), Mwkhna (Clerodendrum viscosum), Thalir (Musa balbisina) and Khantal Bilai (Ananas cosmos) are the main plants used for Amao preparation. The stored Jou is known as Joufinai. They store the fermented mixture (rice and Amao) in anaerobic conditions for the preparation of Joufinai. We observed a successive increase in alcohol content from 3 months of storage period with 11.79 ± 0.010 (%, v/v) to 15.48 ± 0.070 (%, v/v) at 15 months of storage by a simple, reproducible and solution based colorimetric method. A positive linear correlation was also observed between pH and ethanol content with storage having correlation coefficient 0.981. Here, we optimised the detection of change in constituents of Joufinai during storage using reverse phase HPLC method. We found acetone, ethanol, acetic acid, glycerol as main constituents present in Joufinai. A very good correlation was observed from 3 months to 15 months of storage periods with its constituents. Increase in glycerol content was also detected with storage periods and hence Joufinai can be use as a precursor of above stated compounds. We also observed antioxidant activities increase from 0.056 ±2.80 mg/mL for 3 months old to 0.078± 5.33 mg/mL (in ascorbic acid equivalents) for 15 month old beverage by DPPH radical scavenging method. Therefore, we aimed for scientific validation of storage procedure used by Bodos in Joufinai production and to convert the Bodos’ traditional alcoholic beverage to a commercial commodity through our study.

Keywords: Amao, correlation, beverage, joufinai

Procedia PDF Downloads 288

Authors: O. A. Adebiyi, A. O. Oni, A. O. K. Adeshehinwa, I. O. Adejumo

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In forty nine (49) days, twenty four (24) growing pigs (Landrace x Large white) with an average weight of 17 ±2.1kg were allocated to four experimental treatments T1 (dry mash without probiotics), T2 (wet feed without probiotics), T3 (dry mash + Saccaromyces cereviseae probiotics) and T4 (wet feed + Saccaromyces cereviseae probiotics) which were replicated three times with two pigs per replicate in a completely randomised design. The basal feed (dry feed) was formulated to meet the nutritional requirement of the animal with crude protein of 18.00% and metabolisable energy of 2784.00kcal/kgME. Growth pattern, faecal microbial load and behavioural activities (eating, drinking, physical pen interaction and frequency of visiting the drinking troughs) were accessed. Pigs fed dry mash without probiotics (T1) had the highest daily feed intake among the experimental animals (1.10kg) while pigs on supplemented diets (T3 and T4) had an average daily feed intake of 0.95kg. However, the feed conversion ratio was significantly (p < 0.05) affected with pigs on T3 having least value of 6.26 compared those on T4 (wet feed + Saccaromyces cereviseae) with means of 7.41. Total organism counts varied significantly (p < 0.05) with pigs on T1, T2, T3 and T4 with mean values of 179.50 x106cfu; 132.00 x 106cfu; 32.00 x 106cfu and 64.50 x 106cfu respectively. Coliform count was also significantly (p < 0.05) different among the treatments with corresponding values of 117.50 x 106cfu; 49.00 x 106cfu, 8.00 x 106cfu for pigs in T1, T2 and T4 respectively. The faecal Saccaromyces cereviseae was significantly lower in pigs fed supplemented diets compared to their counterparts on unsupplemented diets. This could be due to the inability of yeast organisms to be voided easily through feaces. The pigs in T1 spent the most time eating (7.88%) while their counterparts on T3 spent the least time eating. The corresponding physical pen interaction times expressed in percentage of a day for pigs in T1, T2, T3 and T4 are 6.22%, 5.92%, 4.04% and 4.80% respectively. These behavioural responses exhibited by these pigs (T3) showed that little amount of dry feed supplemented with probiotics is needed for better performance. The water intake increases as a result of the dryness of the feed with consequent decrease in pen interaction and more time was spent resting than engaging in other possible vice-habit like fighting or tail biting. Pigs fed dry feed (T3) which was supplemented with Saccaromyces cereviseae probiotics had a better overall performance, least faecal microbial load than wet fed pigs either supplemented with Saccaromyces cereviseae or non-supplemented.

Keywords: behaviour, feed forms, feed utilization, growth, microbial

Procedia PDF Downloads 316

Authors: Dorota Zielińska, Monika Trząskowska, Anna Łepecka, Katarzyna Neffe-Skocińska, Beata Bilska, Marzena Tomaszewska, Danuta Kołożyn-Krajewska

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Minimum durability date (MDD) labeled food is known to have a long shelf life. A properly stored or transported food retains its physical, chemical, microbiological, and sensory properties up to MDD. The aim of the study was to assess the sensory quality and microbiological safety of selected thermally processed products,i.e., mayonnaise, jam, and canned tuna within and after MDD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the Enterobacteriaceae count and the total yeast and mold (TYMC) count on the last day of MDD and after 1 and 3 months of storage, after the MDD expired. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of MDD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of differentiators (quality features), and overall quality were defined and determined. It was found that during three months storage of tested food products, after the MDD expired, the microbiological quality slightly decreased, however, regardless of the tested sample, TVC was at the level of <3 log cfu/g, similarly, the Enterobacretiaceae, what indicates the good microbiological quality of the tested foods. The TYMC increased during storage but did not exceed 2 logs cfu/g of product. Salmonella and Listeria monocytogenes were not found in any of the tested food samples. The sensory quality of mayonnaise negatively changed during storage. After three months from the expiry of MDD, a decrease in the "fat" and "egg" taste and aroma intensity, as well as the "density" were found. The "sour" taste intensity of blueberry jam after three months of storage was slightly higher, compared to the jam tested on the last day of MDD, without affecting the overall quality. In the case of tuna samples, an increase in the "fishy" taste and aroma intensity was observed during storage, and the overall quality did not change. Tested thermally processed products (mayonnaise, jam, and canned tuna) were characterized by good microbiological and sensory quality on the last day of MDD, as well as after three months of storage under conditions recommended by the producer. These findings indicate the possibility of reducing food losses by extending or completely abolishing the MDD of selected thermal processed food products.

Keywords: food wastes, food quality and safety, mayonnaise, jam, tuna

Procedia PDF Downloads 98

Authors: Raviv Harris, Maggie Levy

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Natural product-based pesticides may serve as an alternative to the traditional synthetic pesticides, which have a potentially damaging effect, both to human health and for the environment. Along with plants, microorganisms are a prospective source of such biological pesticides. A unique and active strain of P. aphidis (designated isolate L12, Israel 2004), an epiphytic and non-pathogenic basidiomycete yeast, was isolated in our lab from strawberry leaves. P. aphidis L12 secretions were found to inhibit broad range of plant pathogens. This work demonstrates that metabolites isolated from the biocontrol agent P. aphidis (isolate L12) can inhibit varied fungal and bacterial phytopathogens. Biologically active metabolites were extracted from P. aphidis biomass, using the organic solvent ethyl acetate. The antimicrobial activity of the extract was demonstrated, both in vitro and in planta. Using disk diffusion assays, the following inhibition zones were obtained: 43cm² for Pseudomonas syringae pv. tomato, 28.5cm² for Xanthomonas campestris pv. vesicatoria, 59cm² for Clavibacter michiganensis subsp. michiganensis, 34cm² for Erwinia amylovora and 34cm² for Agrobacterium tumefaciens. Additionally, strong inhibitory activity of the extract against fungi mycelial growth was established, with IC₅₀ values of 606µg ml⁻¹ for Botrytis cinerea, 221µg ml⁻¹ for Pythium spp., 519µg ml⁻¹ for Rhizoctonia solani, 455µg ml⁻¹ for Sclerotinia sclerotiorum, 2270µg ml⁻¹ for Fusarium oxysporum f. sp. lycopersici, and 2038µg ml⁻¹ for Alternaria alternata. The results of the in planta experiments demonstrated a dose-dependent reduction in disease infection. Significant inhibition of B. cinerea lesions on tomato plants was obtained when a spore suspension of this pathogen was treated with extract concentrations higher than 4.2mg ml⁻¹. Concentration of 7mg ml⁻¹ caused a reduction of over 95% in the lesion size of B. cinerea on tomato plants. The strong antimicrobial activity demonstrated both in vitro and in planta against varied phytopathogens, may indicate that the extracted antimicrobial metabolites have potential to serve as natural pesticides in the field.

Keywords: antimicrobial, B. cinerea, metabolites, natural pesticides, P. aphidis

Procedia PDF Downloads 207

Authors: Patarasuda Chaisupa, R. Clay Wright

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The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.

Keywords: synthetic biology, bioengineering, molecular biology, biotechnology

Procedia PDF Downloads 59

Authors: Dorith Tavor, Adi Wolfson

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In the past two decades a variety of green solvents have been proposed, including water, ionic liquids, fluorous solvents, and supercritical fluids. However, their implementation in industrial processes is still limited due to their tedious and non-sustainable synthesis, lack of experimental data and familiarity, as well as operational restrictions and high cost. Several years ago we presented, for the first time, the use of glycerol-based solvents as alternative sustainable reaction mediums in both catalytic and non-catalytic organic synthesis. Glycerol is the main by-product from the conversion of oils and fats in oleochemical production. Moreover, in the past decade, its price has substantially decreased due to an increase in supply from the production and use of fatty acid derivatives in the food, cosmetics, and drugs industries and in biofuel synthesis, i.e., biodiesel. The renewable origin, beneficial physicochemical properties and reusability of glycerol-based solvents, enabled improved product yield and selectivity as well as easy product separation and catalyst recycling. Furthermore, their high boiling point and polarity make them perfect candidates for non-conventional heating and mixing techniques such as ultrasound- and microwave-assisted reactions. Finally, in some reactions, such as catalytic transfer-hydrogenation or transesterification, they can also be used simultaneously as both solvent and reactant. In our ongoing efforts to design a viable protocol that will facilitate the acceptance of glycerol and its derivatives as sustainable solvents, pure glycerol and glycerol triacetate (triacetin) as well as various glycerol-triacetin mixtures were tested as sustainable solvents in several representative organic reactions, such as nucleophilic substitution of benzyl chloride to benzyl acetate, Suzuki-Miyaura cross-coupling of iodobenzene and phenylboronic acid, baker’s yeast reduction of ketones, and transfer hydrogenation of olefins. It was found that reaction performance was affected by the glycerol to triacetin ratio, as the solubility of the substrates in the solvent determined product yield. Thereby, employing optimal glycerol to triacetin ratio resulted in maximum product yield. In addition, using glycerol-based solvents enabled easy and successful separation of the products and recycling of the catalysts.

Keywords: glycerol, green chemistry, sustainability, catalysis

Procedia PDF Downloads 597

Authors: Natalia Moreno-Castellanos, Amaia Rodriguez, Gema Frühbeck

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Introduction: In adipocytes, the cytoskeleton undergoes important expression and distribution in adipocytes rearrangements during adipogenesis and in obesity. Indeed, a role for these proteins in the regulation of adipocyte differentiation and response to insulin has been demonstrated. Recently, septins have been considered as new components of the cytoskeletal network that interact with other cytoskeletal elements (actin and tubulin) profoundly modifying their dynamics. However, these proteins have not been characterized as yet in adipose tissue. In this work, were examined the cellular, molecular and functional features of a member of this family, septin 11 (SEPT11), in adipocytes and evaluated the impact of obesity on the expression of this protein in human adipose tissue. Methods: Adipose gene and protein expression levels of SEPT11 were analysed in human samples. SEPT11 distribution was evaluated by immunocytochemistry, electronic microscopy, and subcellular fractionation techniques. GST-pull down, immunoprecipitation and a Yeast-Two Hybrid (Y2H) screening were used to identify the SEPT11 interactome. Gene silencing was employed to assess the role of SEPT11 in the regulation of insulin signaling and lipid metabolism in adipocytes. Results: SEPT11 is expressed in human adipocytes, and its levels increased in both omental and subcutaneous adipose tissue in obesity, with SEPT11 mRNA content positively correlating with parameters of insulin resistance in subcutaneous fat. In non-stimulated adipocytes, SEPT11 immunoreactivity showed a ring-like distribution at the cell surface and associated to caveolae. Biochemical analyses showed that SEPT11 interacted with the main component of caveolae, caveolin-1 (CAV1) as well as with the fatty acid-binding protein, FABP5. Notably, the three proteins redistributed and co-localized at the surface of lipid droplets upon exposure of adipocytes to oleate. In this line, SEPT11 silencing in 3T3-L1 adipocytes impaired insulin signaling and decreased insulin-induced lipogenesis. Conclusions: Those findings demonstrate that SEPT11 is a novel component of the adipocyte cytoskeleton that plays an important role in the regulation of lipid traffic, metabolism and can thus represent a potential biomarker of insulin resistance in obesity in adipocytes through its interaction with both CAV1 and FABP5.

Keywords: caveolae, lipid metabolism, obesity, septins

Procedia PDF Downloads 173

Authors: Iman Farasat, Howard M. Salis

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The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.

Keywords: biophysical model, CRISPR, Cas9, genome editing

Procedia PDF Downloads 377

Authors: Mohammed Sabbah, Asmaa Al-Asmar, Doaa Abu-Hani, Fuad Al-Rimawi

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Plant extracts are an increasingly popular natural product with a wide range of potential applications in food, industrial, and health care industries. They are rich in polyphenolic compounds and flavonoids, which have been demonstrated to possess a variety of beneficial properties, including antimicrobial and antioxidant activity. Plant extracts have been found to possess antimicrobial activity against a variety of foodborne pathogens and can be used as a natural preservative to extend the shelf life of food products. They have also strong antioxidant activity, which can reduce the formation of free radicals and oxidation of food components. Recently there is an increase interest in bio-based polymers to be used as innovative “bioplastics” for industrial exploitation e.g. packaging materials for food products. Additionally, incorporation of active compounds (e.g. antioxidants and antimicrobials) in bio-polymer materials is of particular interest since such active polymers can be used as active packaging materials (with antimicrobial and antioxidant activity). In this work, different plant extracts have been characterized for their phenolic compounds, flavonoids content, antioxidant activity (both as free radical scavenging ability and reducing ability), and antimicrobial activity against gram positive and negative bacteria (Escherichia coli; Staphylococcus aureus, and Pseudomonas aeruginosa) as well as antifungal activities (against yeast, mold and Botrytis cinera/a plant pathogen). Results showed that many extracts are rich with polyphenolic compounds and flavonoids and have strong antioxidant activities, and rich with phytochemicals (e.g. rutin, quercetin, oleuropein, tyrosol and hydroxytyrosol). Some extracts showed antibacterial activity against both gram positive and negative bacteria as well as antifungal activities and can work, therefore, as preservatives for food or pharmaceutical industries. As an application, two extracts were used as additive to pectin-based packaging film, and results showed that the addition of these extracts significantly improve their functionality as antimicrobial and antioxidant activity. These biomaterials, therefore can be used in food packaging materials to extend the shelf life of food products.

Keywords: plant extracts, antioxidants, flavonoids, bioplastic, edible biofilm, packaging materials

Procedia PDF Downloads 45

Authors: Bengi Hakguder Taze, Sevcan Unluturk

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Şalak apricot (Prunus armeniaca L., cv. Şalak) is a specific variety grown in Igdir, Turkey. The fruit has distinctive properties distinguish it from other cultivars, such as its unique size, color, taste and higher water content. Drying is the widely used method for preservation of apricots. However, fresh consumption is preferred for Şalak apricot instead of drying due to its low dry matter content. Higher amounts of water in the structure and climacteric nature make the fruit sensitive against rapid quality loss during storage. Hence, alternative processing methods need to be introduced to extend the shelf life of the fresh produce. Mild heat (MH) treatment is of great interest as it can reduce the microbial load and inhibit enzymatic activities. Therefore, the aim of this study was to evaluate the impact of mild heat treatment on the natural microflora found on Şalak apricot surfaces and some physical quality parameters of the fruit, such as color and firmness. For this purpose, apricot samples were treated at different temperatures between 40 and 60 ℃ for different periods ranging between 10 to 60 min using a temperature controlled water bath. Natural flora on the fruit surfaces was examined using standard plating technique both before and after the treatment. Moreover, any changes in color and firmness of the fruit samples were also monitored. It was found that control samples were initially containing 7.5 ± 0.32 log CFU/g of total aerobic plate count (TAPC), 5.8±0.31 log CFU/g of yeast and mold count (YMC), and 5.17 ± 0.22 log CFU/g of coliforms. The highest log reductions in TAPC and YMC were observed as 3.87-log and 5.8-log after the treatments at 60 ℃ and 50 ℃, respectively. Nevertheless, the fruit lost its characteristic aroma at temperatures above 50 ℃. Furthermore, great color changes (ΔE ˃ 6) were observed and firmness of the apricot samples was reduced at these conditions. On the other hand, MH treatment at 41 ℃ for 10 min resulted in 1.6-log and 0.91-log reductions in TAPC and YMC, respectively, with slightly noticeable changes in color (ΔE ˂ 3). In conclusion, application of temperatures higher than 50 ℃ caused undesirable changes in physical quality of Şalak apricots. Although higher microbial reductions were achieved at those temperatures, temperatures between 40 and 50°C should be further investigated considering the fruit quality parameters. Another strategy may be the use of high temperatures for short time periods not exceeding 1-5 min. Besides all, MH treatment with UV-C light irradiation can be also considered as a hurdle strategy for better inactivation results.

Keywords: color, firmness, mild heat, natural flora, physical quality, şalak apricot

Procedia PDF Downloads 111

Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

Procedia PDF Downloads 142