Search results for: microbial metabolites

Authors: Zeenat Rupawalla, Nicole Robinson, Susanne Schmidt, Sijie Li, Selina Carruthers, Elodie Buisset, John Roles, Ben Hankamer, Juliane Wolf

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Agriculture has radically changed the global biogeochemical cycle of nitrogen (N). Fossil fuel-enabled synthetic N-fertiliser is a foundation of modern agriculture but applied to soil crops only use about half of it. To address N-pollution from cropping and the large carbon and energy footprint of N-fertiliser synthesis, new technologies delivering enhanced energy efficiency, decarbonisation, and a circular nutrient economy are needed. We characterised algae fertiliser (AF) as an alternative to synthetic N-fertiliser (SF) using empirical and modelling approaches. We cultivated microalgae in nutrient solution and modelled up-scaled production in nutrient-rich wastewater. Over four weeks, AF released 63.5% of N as ammonium and nitrate, and 25% of phosphorous (P) as phosphate to the growth substrate, while SF released 100% N and 20% P. To maximise crop N-use and minimise N-leaching, we explored AF and SF dose-response-curves with spinach in glasshouse conditions. AF-grown spinach produced 36% less biomass than SF-grown plants due to AF’s slower and linear N-release, while SF resulted in 5-times higher N-leaching loss than AF. Optimised blends of AF and SF boosted crop yield and minimised N-loss due to greater synchrony of N-release and crop uptake. Additional benefits of AF included greener leaves, lower leaf nitrate concentration, and higher microbial diversity and water holding capacity in the growth substrate. Life-cycle-analysis showed that replacing the most effective SF dosage with AF lowered the carbon footprint of fertiliser production from 2.02 g CO₂ (C-producing) to -4.62 g CO₂ (C-sequestering), with a further 12% reduction when AF is produced on wastewater. Embodied energy was lowest for AF-SF blends and could be reduced by 32% when cultivating algae on wastewater. We conclude that (i) microalgae offer a sustainable alternative to synthetic N-fertiliser in spinach production and potentially other crop systems, and (ii) microalgae biofertilisers support the circular nutrient economy and several sustainable development goals.

Keywords: bioeconomy, decarbonisation, energy footprint, microalgae

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Authors: Diana Puigserver Cuerda, Jofre Herrero Ferran, José M. Carmona Perez

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In the transition zone between aquifers and basal aquitards, DNAPL-pools of chlorinated solvents are more recalcitrant than at other depths in the aquifer. Although degradation of carbon tetrachloride (CT) and chloroform (CF) occurs in this zone, this is a slow process, which is why an adequate remediation strategy is necessary. The working hypothesis of this study is that the biostimulation of the transition zone of an aquifer contaminated by CT and CF can be an effective remediation strategy. This hypothesis has been tested in a site on an unconfined aquifer in which the major contaminants were CT and CF of industrial origin and where the hydrochemical background was rich in other compounds that can hinder natural attenuation of chloromethanes. Field studies and five laboratory microcosm experiments were carried out at the level of groundwater and sediments to identify: i) the degradation processes of CT and CF; ii) the structure of microbial communities; and iii) the microorganisms implicated on this degradation. For this, concentration of contaminants and co-contaminants (nitrate and sulfate), Compound Specific Isotope Analysis, molecular techniques (Denaturing Gradient Gel Electrophoresis) and clone library analysis were used. The main results were: i) degradation processes of CT and CF occurred in groundwater and in the lesser conductive sediments; ii) sulfate-reducing conditions in the transition zone were high and similar to those in the source of contamination; iii) two microorganisms (Azospira suillum and a bacterium of the Clostridiales order) were identified in the transition zone at the field and lab experiments that were compatible with the role of carrying out the reductive dechlorination of CT, CF and their degradation products (dichloromethane and chloromethane); iv) these two microorganisms were present at the high starting concentrations of the microcosm experiments (similar to those in the source of DNAPL) and continued being present until the last day of the lactate biostimulation; and v) the lactate biostimulation gave rise to the fastest and highest degradation rates and promoted the elimination of other electron acceptors (e.g. nitrate and sulfate). All these results are evidence that lactate biostimulation can be effective in remediating the source and plume, especially in the transition zone, and highlight the environmental relevance of the treatment of contaminated transition zones in industrial contexts similar to that studied.

Keywords: Azospira suillum, lactate biostimulation of carbon tetrachloride and chloroform, reductive dechlorination, transition zone between aquifer and aquitard

Procedia PDF Downloads 157

Authors: Andrés Quiroz, Emilio Hormazábal, Ana Mutis, Fernando Ortega, Loreto Méndez, Leonardo Parra

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Red clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), is the main insect pest associated to red clover, Trifolium pratense L. An average of 1.5 H. obscurus per plant can cause 5.5% reduction in forage yield in pastures of two to three years old. Moreover, insect attack can reach 70% to 100% of the plants. To our knowledge, there is no a chemical strategy for controlling this pest. Therefore alternative strategies for controlling H. obscurus are a high priority for red clover producers. One of this alternative is related to the study of secondary metabolites involved in intrinsic chemical defenses developed by plants, such as isoflavonoids. The isoflavonoids formononetin and daidzein have elicited an antifeedant and phagostimult effect on H. obscurus respectively. However, we do not know how is the dynamic variation of these isoflavonoids under field conditions. The main objective of this work was to evaluate the variation of the antifeedant isoflavonoids formononetin, the phagostimulant isoflavonoids daidzein, and their respective glycosides over time in different ecotypes of red clover. Fourteen red clover ecotypes (8 cultivars and 6 experimental lines), were collected at INIA-Carillanca (La Araucanía, Chile). These plants were established in October 2015 under irrigated conditions. The cultivars were distributed in a randomized complete block with three replicates. The whole plants were sampled in four times: 15th October 2016, 12th December 2016, 27th January 2017 and 16th March 2017 with sufficient amount of soil to avoid root damage. A polar fraction of isoflavonoid was obtained from 20 mg of lyophilized root tissue extracted with 2 mL of 80% MeOH for 16 h using an orbital shaker in the dark at room temperature. After, an aliquot of 1.4 mL of the supernatant was evaporated, and the residue was resuspended in 300 µL of 45% MeOH. The identification and quantification of isoflavonoid root extracts were performed by the injection of 20 µL into a Shimadzu HPLC equipped with a C-18 column. The sample was eluted with a mobile phase composed of AcOH: H₂O (1:9 v/v) as solvent A and CH₃CN as solvent B. The detection was performed at 260 nm. The results showed that the amount of aglycones was higher than the respective glycosides. This result is according to the biosynthetic pathway of flavonoids, where the formation of glycoside is further to the glycosides biosynthesis. The amount of formononetin was higher than daidzein. In roots, where H. obscurus spent the most part of its live cycle, the highest content of formononetin was found in G 27, Pawera, Sabtoron High, Redqueli-INIA and Superqueli-INIA cvs. (2.1, 1.8, 1.8, 1.6 and 1.0 mg g⁻¹ respectively); and the lowest amount of daidzein were found Superqueli-INIA (0.32 mg g⁻¹) and in the experimental line Sel Syn Int4 (0.24 mg g⁻¹). This ecotype showed a high content of formononetin (0.9 mg g⁻¹). This information, associated with cultural practices, could help farmers and breeders to reduce H. obscurus in grassland, selecting ecotypes with high content of formononetin and low amount of daidzein in the roots of red clover plants. Acknowledgements: FONDECYT 1141245 and 11130715.

Keywords: daidzein, formononetin, isoflavonoid glycosides, trifolium pratense

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Authors: Shilpi, Indu Gaur, Neha Bhadauria, Susmita Goswami, Prabir K. Paul

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Cultivation and consumption of organically grown fruits and vegetables have increased by several folds. However, the presence of Human Enteric Pathogens on the surface of organically grown vegetables causing Gastro-intestinal diseases, are most likely due to contaminated water and fecal matter of farm animals. Human Enteric Pathogens are adapted to colonize the human gut, and also colonize plant surface. Microbes on plant surface communicate with each other to establish quorum sensing. The cross talk study is important because the enteric pathogens on phylloplane have been reported to mask the beneficial resident bacteria of plant. In the present study, HEPs and bacterial colonizers were identified using 16s rRNA sequencing. Microbial colonization patterns after interaction between Human Enteric Pathogens and natural bacterial residents on tomato phylloplane was studied. Tomato plants raised under aseptic conditions were inoculated with a mixture of Serratia fonticola and Klebsiella pneumoniae. The molecules involved in cross-talk between Human Enteric Pathogens and regular bacterial colonizers were isolated and identified using molecular techniques and HPLC. The colonization pattern was studied by leaf imprint method after 48 hours of incubation. The associated protein-protein interaction in the host cytoplasm was studied by use of crosslinkers. From treated leaves the crosstalk molecules and interaction proteins were separated on 1D SDS-PAGE and analyzed by MALDI-TOF-TOF analysis. The study is critical in understanding the molecular aspects of HEP’s adaption to phylloplane. The study revealed human enteric pathogens aggressively interact among themselves and resident bacteria. HEPs induced establishment of a signaling cascade through protein-protein interaction in the host cytoplasm. The study revealed that the adaptation of Human Enteric Pathogens on phylloplane of Solanum lycopersicum involves the establishment of complex molecular interaction between the microbe and the host including microbe-microbe interaction leading to an establishment of quorum sensing. The outcome will help in minimizing the HEP load on fresh farm produce, thereby curtailing incidences of food-borne diseases.

Keywords: crosslinkers, human enteric pathogens (HEPs), phylloplane, quorum sensing

Procedia PDF Downloads 251

Authors: Boce Zhang

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Strategic innovation of nanotechnology to promote food safety has drawn tremendous attentions among research groups, which includes the need for research support during the implementation of the Food Safety Modernization Act (FSMA) in the United States. There are urgent demands and knowledge gaps to the understanding of a) food-water-bacteria interface as for how pathogens persist and transmit during food processing and storage; b) minimum processing requirement needed to prevent pathogen cross-contamination in the food system. These knowledge gaps are of critical importance to the food industry. However, the lack of knowledge is largely hindered by the limitations of research tools. Our groups recently endeavored two novel engineering systems with biomimetics and microfluidics as a holistic approach to hazard analysis and risk mitigation, which provided unprecedented research opportunities to study pathogen behavior, in particular, contamination, and cross-contamination, at the critical food-water-pathogen interface. First, biomimetically-patterned surfaces (BPS) were developed to replicate the identical surface topography and chemistry of a natural food surface. We demonstrated that BPS is a superior research tool that empowers the study of a) how pathogens persist through sanitizer treatment, b) how to apply fluidic shear-force and surface tension to increase the vulnerability of the bacterial cells, by detaching them from a protected area, etc. Secondly, microfluidic devices were designed and fabricated to study the bactericidal kinetics in the sub-second time frame (0.1~1 second). The sub-second kinetics is critical because the cross-contamination process, which includes detachment, migration, and reattachment, can occur in a very short timeframe. With this microfluidic device, we were able to simulate and study these sub-second cross-contamination scenarios, and to further investigate the minimum sanitizer concentration needed to sufficiently prevent pathogen cross-contamination during the food processing. We anticipate that the findings from these studies will provide critical insight on bacterial behavior at the food-water-cell interface, and the kinetics of bacterial inactivation from a broad range of sanitizers and processing conditions, thus facilitating the development and implementation of science-based food safety regulations and practices to mitigate the food safety risks.

Keywords: biomimetic materials, microbial food safety, microfluidic device, nanotechnology

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

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Authors: Sandhya Parameswaran, Prajakta Dhuri

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Despite great improvements in health care, the world oral health report states that dental problems still persist, particularly among underprivileged groups in both developing and developed countries. Dental caries and periodontal diseases are identified as the most important oral health problems globally. Acidic foods and beverages can affect natural teeth, and chronic exposure often leads to the development of dental erosion, abrasion, and decay. In recent years, there has been an increased interest toward essential oils. These are secondary metabolites and possess antibacterial, antifungal and antioxidant properties. Essential oils are volatile and chemically unstable in the presence of air, light, moisture and high temperature. Hence many novel methods like a liposomal encapsulation of oils have been introduced to enhance the stability and bioavailability. This research paper focuses on two essential oils, clove and cinnamon oil. Clove oil was obtained from Syzygium aromaticum Linn using clavengers apparatus. It contains eugenol and β caryophyllene. Cinnamon oil, from the barks of Cinnamomum cassia, contains cinnamaldehyde, The objective of the current research was to develop a liposomal carrier system containing clove and cinnamon oil and study their synergistic activity against dental pathogens when formulated as a gel. Methodology: The essential oil were first tested for their antimicrobial activity against dental pathogens, Lactobacillus acidophillus (MTCC No. 10307, MRS broth) and Streptococcus Mutans (MTCC No .890, Brain Heart Infusion agar). The oils were analysed by UV spectroscopy for eugenol and cinnamaldehyde content. Standard eugenol was linear between 5ppm to 25ppm at 282nm and standard cinnamaldehde from 1ppm to 5pmm at 284nm. The concentration of eugenol in clove oil was found to be 62.65 % w/w, and that of cinnamaldehyde was found to be 5.15%s w/w. The oils were then formulated into liposomes. Liposomes were prepared by thin film hydration method using Phospholipid, Cholesterol, and other oils dissolved in a chloroform methanol (3:1) mixture. The organic solvent was evaporated in a rotary evaporator above lipid transition temperature. The film was hydrated with phosphate buffer (pH 5.5).The various batches of liposomes were characterized and compared for their size, loading rate, encapsulation efficiency and morphology. The prepared liposomes when evaluated for entrapment efficiency showed 65% entrapment for clove and 85% for cinnamon oil. They were also tested for their antimicrobial activity against dental pathogens and their synergistic activity studied. Based on the activity and the entrapment efficiency the amount of liposomes required to prepare 1gm of the gel was calculated. The gel was prepared using a simple ointment base and contained 0.56% of cinnamon and clove liposomes. A simultaneous method of analysis for eugenol and cinnamaldehyde.was then developed using HPLC. The prepared gels were then studied for their stability as per ICH guidelines. Conclusion: It was found that liposomes exhibited spherical shaped vesicles and protected the essential oil from degradation. Liposomes, therefore, constitute a suitable system for encapsulation of volatile, unstable essential oil constituents.

Keywords: cinnamon oil, clove oil, dental caries, liposomes

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Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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Authors: G. A. Gevorgyan, A. S. Mamyan, L. G. Stepanyan, L. R. Hambaryan

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Bacterio- and phytoplankton growth rates in 'Yerevanyan lich' reservoir and the Hrazdan river in Yerevan city, Armenia were investigated in April and June-August, 2015. Phytoplankton sampling and analysis were performed by the standard methods accepted in hydrobiological studies. The quantitative analysis of aerobic, coliform and E. coli bacteria is done by the 'RIDA COUNT' medium sheets (coated with ready-to-use culture medium). The investigations showed that the insufficient management of household discharges in Yerevan city caused the organic and fecal pollution of the Hrazdan river in this area which in turn resulted in an increase in bacterial count and increased sanitary and pathogenic risks to the environment and human health. During the investigation in April, the representatives of diatom algae prevailed quantitatively in the coastal area of 'Yerevanyan lich' reservoir, nevertheless, a significant change in the phytoplankton community in June occurred: due to green algae bloom in the reservoir, the quantitative parameters of phytoplankton increased significantly. This was probably conditioned by a seasonal increase in the water temperature in the conditions of the sufficient concentration of nutrients. However, a succession in phytoplankton groups during July-August occurred, and a dominant group (according to quantitative parameters) in the phytoplankton community was changed as follows: green algae-diatom algae-blue-green algae. Rapid increase in the quantitative parameters of diatom and blue-green algae in the reservoir may have been conditioned by increased organic matter level resulted from green algae bloom. Algal bloom in 'Yerevanyan lich' reservoir caused changes in phytoplankton community and an increase in bacterioplankton count not only in the reservoir but also in the Hrazdan river sites located in the downstream from the reservoir. Thus, the insufficient management of urban discharges and aquatic ecosystems in Yerevan city led to unfavorable changes in water quality and microbial and phytoplankton communities in “Yerevanyan lich” reservoir and the Hrazdan river which in turn caused increased sanitary and pathogenic risks to the environment and human health.

Keywords: algal bloom, bacterioplankton, phytoplankton, Hrazdan river, Yerevanyan lich reservoir

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Authors: Vuiswa Lucia Sethunya, Tonderayi Matambo, Diane Hildebrandt

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South African’s informal settlements produce tonnes of kitchen waste (KW) per year which is dumped into the landfill. These landfill sites are normally located in close proximity to the household of the poor communities; this is a problem in which the young children from those communities end up playing in these landfill sites which may result in some health hazards because of methane, carbon dioxide and sulphur gases which are produced. To reduce this large amount of organic materials being deposited into landfills and to provide a cleaner place for those within the community especially the children, an energy conversion process such as anaerobic digestion of the organic waste to produce biogas was implemented. In this study, the digestion of various kitchen waste was investigated in order to understand and develop a system that is suitable for household use to produce biogas for cooking. Three sets of waste of different nutritional compositions were digested as per acquired in the waste streams of a household at mesophilic temperature (35ᵒC). These sets of KW were co-digested with cow dung (CW) at different ratios to observe the microbial behaviour and the system’s stability in a laboratory scale system. The gas chromatography-flame ionization detector analyses have been performed to identify and quantify the presence of organic compounds in the liquid samples from co-digested and mono-digested food waste. Acetic acid, propionic acid, butyric acid and valeric acid are the fatty acids which were studied. Acetic acid (1.98 g/L), propionic acid (0.75 g/L) and butyric acid (2.16g/L) were the most prevailing fatty acids. The results obtained from organic acids analysis suggest that the KW can be an innovative substituent to animal manure for biogas production. The faster degradation period in which the microbes break down the organic compound to produce the fatty acids during the anaerobic process of KW also makes it a better feedstock during high energy demand periods. The C/N ratio analysis showed that from the three waste streams the first stream containing vegetables (55%), fruits (16%), meat (25%) and pap (4%) yielded more methane-based biogas of 317mL/g of volatile solids (VS) at C/N of 21.06. Generally, this shows that a household will require a heterogeneous composition of nutrient-based waste to be fed into the digester to acquire the best biogas yield to sustain a households cooking needs.

Keywords: anaerobic digestion, biogas, kitchen waste, household

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Authors: Olfat A. Mohamed

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Rhamnolipids biosurfactants have distinct properties given them importance in many industrial applications, especially their great new future applications in cosmetic and pharmaceutical industries. These applications have encouraged the search for diverse and renewable resources to control the cost of production. The experimental results were then applied to find a suitable mathematical model for obtaining the design criteria of the batch bioreactor. This research aims to produce Rhamnolipids from different oily wastewater sources such as petroleum crude oil (PO) and vegetable oil (VO) by using Pseudomonas aeruginosa ATCC 9027. Different concentrations of the PO and the VO are added to the media broth separately are in arrangement (0.5 1, 1.5, 2, 2.5 % v/v) and (2, 4, 6, 8 and 10%v/v). The effect of the initial concentration of oil residues and the addition of glycerol and palmitic acid was investigated as an inducer in the production of rhamnolipid and the surface tension of the broth. It was found that 2% of the waste (PO) and 6% of the waste (VO) was the best initial substrate concentration for the production of rhamnolipids (2.71, 5.01 g rhamnolipid/l) as arrangement. Addition of glycerol (10-20% v glycerol/v PO) to the 2% PO fermentation broth led to increase the rhamnolipid production (about 1.8-2 times fold). However, the addition of palmitic acid (5 and 10 g/l) to fermentation broth contained 6% VO rarely enhanced the production rate. The experimental data for 2% initially (PO) was used to estimate the various kinetic parameters. The following results were obtained, maximum rate or velocity of reaction (Vmax) = 0.06417 g/l.hr), yield of cell weight per unit weight of substrate utilized (Yx/s = 0.324 g Cx/g Cs) maximum specific growth rate (μmax = 0.05791 hr⁻¹), yield of rhamnolipid weight per unit weight of substrate utilized (Yp/s)=0.2571gCp/g Cs), maintenance coefficient (Ms =0.002419), Michaelis-Menten constant, (Km=6.1237 gmol/l), endogenous decay coefficient (Kd=0.002375 hr⁻¹). Predictive parameters and advanced mathematical models were applied to evaluate the time of the batch bioreactor. The results were as follows: 123.37, 129 and 139.3 hours in respect of microbial biomass, substrate and product concentration, respectively compared with experimental batch time of 120 hours in all cases. The expected mathematical models are compatible with the laboratory results and can, therefore, be considered as tools for expressing the actual system.

Keywords: batch bioreactor design, glycerol, kinetic parameters, petroleum crude oil, Pseudomonas aeruginosa, rhamnolipids biosurfactants, vegetable oil

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Authors: Kamile Nazan Turhan

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When work on edible packaging began, the hope was to provide a sustainable alternative to reduce plastic consumption. Edible packaging can take the form of a coating or film that provides better food protection. They have the potential to be a feasible alternative to traditional plastic food packaging as they can increase the shelf life of foods by reducing their respiration rate and water loss and protecting them from physical damage and microbial spoilage, preventing post-harvest loss. Edible films and coatings can extend the shelf life of the food product and improve food quality by regulating the transfer of moisture, oxygen, carbon dioxide, lipids, aroma, and taste compounds in food systems. The main advantage of using edible packaging is that it reduces the amount of plastic waste produced. Another interesting advantage is that some edible food packaging elements may have added vitamins, probiotics, and other nutrients, bioactive compounds that support technological and biological properties. Despite the use of many different biomaterials in the production of edible packaging, most of the problems experienced are similar. Among these problems are hydrophilicity, low water stability, high humidity sensitivity, poor moisture resistance, high water vapor permeability, poor mechanical properties and machinability, fragility, insolubility, low melting point, sensitivity to pH, temperature, ionic, electro reactions, brittle, and hard structure, low thermal stability, hardness, and hazy film are available. Edible films have the disadvantage of being difficult to apply to the food surface due to their weak barrier and mechanical properties. One of the most important problems is that the developed films have a brittle structure since biomaterials are generally hydrophilic. A plasticizer is added to increase the flexibility of the film, and as the plasticizer content increases, the film permeability also increases. The correct choice of plasticizer for a particular biopolymer allows optimization of the mechanical properties of the film with minimal increase in film permeability. The effect of the amount and type of plasticizer in achieving the desired mechanical properties with optimum permeability is constantly being investigated by researchers. However, it should be noted that in the studies conducted so far, plasticized biofilms cannot provide high mechanical strength or good flexibility compared to synthetic polymer materials. In this review, according to the results of the research in the literature, the disadvantages of implementing edible packaging will be revealed and possible solutions will be presented.

Keywords: edible packaging, biyomaterial, disadvantages, production

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Authors: Nafees Ahmed, Nur Izyani Kamaruzman, Saralla Nathan, Mohd Ezharul Hoque Chowdhury, Anuar Zaini Md Zain, Iekhsan Othman, Sharifah Binti Syed Hassan

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Meat quality is always subject to consumer scrutiny when purchasing from retail markets on mislabeling as fresh meat. Various physiological and biochemical changes influence the quality of meat. As a major component of muscle tissue, proteins play a major role in muscle foods. In meat industry, freezing is the most common form of storage of meat products. Repeated cycles of freezing and thawing are common in restaurants, kitchen, and retail outlets and can also occur during transportation or storage. Temperature fluctuation is responsible for physical, chemical, and biochemical changes. Repeated cycles of ‘freeze-thaw’ degrade the quality of meat by stimulating the lipid oxidation and surface discoloration. The shelf life of meat is usually determined by its appearance, texture, color, flavor, microbial activity, and nutritive value and is influenced by frozen storage and subsequent thawing. The main deterioration of frozen meat during storage is due to protein. Due to the large price differences between fresh and frozen–thawed meat, it is of great interest to consumer to know whether a meat product is truly fresh or not. Researchers have mainly focused on the reduction of moisture loss due to freezing and thawing cycles of meat. The water holding capacity (WHC) of muscle proteins and reduced water content are key quality parameters of meat that ultimately changes color and texture. However, there has been limited progress towards understanding the actual mechanisms behind the meat quality changes under the freeze–thaw cycles. Furthermore, effect of freeze-thaw process on integrity of proteins is ignored. In this paper, we have studied the effect of ‘freeze-thawing’ on physicochemical changes of chicken meat protein. We have assessed the quality of meat by pH, spectroscopic measurements, Western Blot. Our results showed that increase in freeze-thaw cycles causes changes in pH. Measurements of absorbance (UV-visible and IR) indicated the degradation of proteins. The expression of various proteins (CREB, AKT, MAPK, GAPDH, and phosphorylated forms) were performed using Western Blot. These results indicated the repeated cycles of freeze-thaw is responsible for deterioration of protein, thus causing decrease in nutritious value of meat. It damges the use of these products in Islamic Sharia.

Keywords: chicken meat, freeze-thaw, halal, protein, western blot

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Authors: D. Desai, J.Dixon, N. Jain, M. Datta

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Microbial infections of scalp consist of symptomatic appearances associated with seborrhoeic dermatitis, folliculitis, furuncles, carbuncles and ringworm. The main causative organisms in these scalp-based infections are bacteria like S. aureus, P. aeruginosa and a fungus M. Furfur. Allopathic treatment of these infections is available and efficient, but occasionally, topical applications have been found to cause side effects. India is known as the botanical garden of the world and considered as the epicentre for utilization of traditional drugs. Many treatments based on herb extracts are commonly used in India. It has been observed treatment with ethnomedicines requires a higher dosage and greater time period. Additionally, repeated applications are required to obtain the full efficacy of the treatment. An attempt has been made to imbibe the traditional knowledge with nanotechnology to generate a proficient therapeutic against scalp infections. We have imbibed metallic nanoparticles with extracts from traditional medicines and propose to formulate an antimicrobial hair massager. Four commonly used herbs for treatment against scalp disorders like Zingiber officinale (ginger), Allium sativum (garlic), Azadirachta indica (neem) leaves and Citrus limon (lemon) peel was taken. 30 gms of dried homogenized powder was obtained and processed for obtaining the aqueous and ethanolic extract by soxhlet apparatus. The extract was dried and reconstituted to obtain working solution of 1mg/ml. Phytochemical analysis for the obtained extract was done. Synthesis of nanoparticles was mediated by incubating 1mM silver nitrate with extracts of various herbs to obtain silver nanoparticles. The formation of the silver nanoparticles (AgNPs) was monitored using UV-Vis spectroscopy. The AgNPs thus obtained were centrifuged and dried. The AgNPs thus formed were characterized by X Ray Diffraction, scanning electron microscopy and transmission electron microscopy. The size of the AgNPs varied from 10-20 nm and was spherical in shape. P. aeruginosa was plated on nutrient agar and comparative antibacterial activity was tested. Comparative antimicrobial potential was calculated for the extracts and the corresponding nanoconstructs. It was found AgNPs were more efficient than their aqueous and ethanolic counterparts except in the ase of C. limon. Statistical analysis was performed to validate the results obtained.

Keywords: ethnomedicine, nanoconstructs, scalp infections, Zingiber officinale

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Authors: Andrés Durán, Brian Castellanos, Marta Quicazán, Carlos Zuluaga-Domínguez

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Bee-pollen is an apicultural-derived food product, with a growing appreciation among consumers given the remarkable nutritional and functional composition, in particular, protein (24%), dietary fiber (15%), phenols (15 – 20 GAE/g) and carotenoids (600 – 900 µg/g). These properties are given by the geographical and climatic characteristics of the region where it is collected. There are several countries recognized by their pollen production, e.g. China, United States, Japan, Spain, among others. Beekeepers use traps in the entrance of the hive where bee-pollen is collected. After the removal of foreign particles and drying, this product is ready to be marketed. However, in countries located along the equator, the absence of seasons and a constant tropical climate throughout the year favors a more rapid spoilage condition for foods with elevated water activity. The climatic conditions also trigger the proliferation of microorganisms and insects. This, added to the factor that beekeepers usually do not have adequate processing systems for bee-pollen, leads to deficiencies in the quality and safety of the product. In contrast, the Andean region of South America, lying on equator, typically has a high production of bee-pollen of up to 36 kg/year/hive, being four times higher than in countries with marked seasons. This region is also located in altitudes superior to 2500 meters above sea level, having extremes sun ultraviolet radiation all year long. As a mechanism of defense of radiation, plants produce more secondary metabolites acting as antioxidant agents, hence, plant products such as bee-pollen contain remarkable more phenolics and carotenoids than collected in other places. Considering this, the improvement of bee-pollen processing facilities by technical modifications and the implementation of an integrated cleaning and drying system for the product in an apiary in the area was proposed. The beehives were modified through the installation of alternative bee-pollen traps to avoid sources of contamination. The processing facility was modified according to considerations of Good Manufacturing Practices, implementing the combined use of a cabin dryer with temperature control and forced airflow and a greenhouse-type solar drying system. Additionally, for the separation of impurities, a cyclone type system was implemented, complementary to a screening equipment. With these modifications, a decrease in the content of impurities and the microbiological load of bee-pollen was seen from the first stages, principally with a reduction of the presence of molds and yeasts and in the number of foreign animal origin impurities. The use of the greenhouse solar dryer integrated to the cabin dryer allowed the processing of larger quantities of product with shorter waiting times in storage, reaching a moisture content of about 6% and a water activity lower than 0.6, being appropriate for the conservation of bee-pollen. Additionally, the contents of functional or nutritional compounds were not affected, even observing an increase of up to 25% in phenols content and a non-significant decrease in carotenoids content and antioxidant activity.

Keywords: beekeeping, drying, food processing, food safety

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Authors: Mariyah Poonawala, Selvan Ravindran, Anuradha Vaidya

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Despite much progress in understanding the regulatory factors and cytokines that support the maturation of the various cell lineages of the hematopoietic system, factors that govern the self-renewal and proliferation of hematopoietic stem cells (HSCs) is still a grey area of research. Hematopoietic stem cell transplantation (HSCT) has evolved over the years and gained tremendous importance in the treatment of both malignant and non-malignant diseases. However, factors such as graft rejection and multiple organ failure have challenged HSCT from time to time, underscoring the urgent need for development of milder processes for successful hematopoietic transplantation. An emerging concept in the field of stem cell biology states that the interactions between the bone-marrow micro-environment and the hematopoietic stem and progenitor cells is essential for regulation, maintenance, commitment and proliferation of stem cells. Understanding the role of mesenchymal stromal cells in modulating the functionality of HSCs is, therefore, an important area of research. Trans-resveratrol has been extensively studied for its various properties to combat and prevent cancer, diabetes and cardiovascular diseases etc. The aim of the present study was to understand the effect of trans-resveratrol on HSCs using single and co-culture systems. We have used KG1a cells since it is a well accepted hematopoietic stem cell model system. Our preliminary experiments showed that low concentrations of trans-resveratrol stimulated the HSCs to undergo proliferation whereas high concentrations of trans-resveratrol did not stimulate the cells to proliferate. We used a murine fibroblast cell line, M210B4, as a stromal feeder layer. On culturing the KG1a cells with M210B4 cells, we observed that the stimulatory as well as inhibitory effects of trans-resveratrol at low and high concentrations respectively, were enhanced. Our further experiments showed that low concentration of trans-resveratrol reduced the generation of reactive oxygen species (ROS) and nitric oxide (NO) whereas high concentrations increased the oxidative stress in KG1a cells. We speculated that perhaps the oxidative stress was imposing inhibitory effects at high concentration and the same was confirmed by performing an apoptotic assay. Furthermore, cell cycle analysis and growth kinetic experiments provided evidence that low concentration of trans-resveratrol reduced the doubling time of the cells. Our hypothesis is that perhaps at low concentration of trans-resveratrol the cells get pushed into the G0/G1 phase and re-enter the cell cycle resulting in their proliferation, whereas at high concentration the cells are perhaps arrested at G2/M phase or at cytokinesis and therefore undergo apoptosis. Liquid Chromatography-Quantitative-Time of Flight–Mass Spectroscopy (LC-Q-TOF MS) analyses indicated the presence of trans-resveratrol and its metabolite(s) in the supernatant of the co-cultured cells incubated with high concentration of trans-resveratrol. We conjecture that perhaps the metabolites of trans-resveratrol are responsible for the apoptosis observed at the high concentration. Our findings may shed light on the unsolved problems in the in vitro expansion of stem cells and may have implications in the ex vivo manipulation of HSCs for therapeutic purposes.

Keywords: co-culture system, hematopoietic micro-environment, KG1a cell line, M210B4 cell line, trans-resveratrol

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Authors: Abubakar Bello Usman, Alhassan Muhammad Gani, Kolo Ibrahim

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The use of botanical raw materials to produce pharmaceuticals, herbal remedies, teas, spirits, cosmetics, sweets, dietary supplements, special industrial compounds and crude materials constitute an important global resource in terms of healthcare and economy. In Nigeria and other developing countries, the indigenous knowledge on the uses of plants lies with the older generation and the traditional healers. However, these custodians are decreasing in number due to death and other unforeseen occurrences. An Ethno-botanical survey was carried out to obtain information on the ethno medical values of wide range of plants used by the people of Gombe State, North-Eastern Nigeria, in the practice of healing and cure of typhoid (enteric) fever. Oral interviews were conducted so as to consider those with low literacy level who are involved in the practice of traditional medicine and thirty four (34) informants availed themselves for the interview and were consulted. All relevant information obtained from the respondents was recorded. A recent and valid nomenclature, along with local names, family names, part of the plant(s) used, methods of preparation and administration and fifty four (54) plant species belonging to 27 families as well as 7 unidentified species that are commonly used by the people of the state in ethnomedical treatment of the ailment were tabulated. Those interviewed included traditional practitioners, local herb sellers, traditional rulers, hunters, farmers and patients. Specific questions were asked and information supplied by informants was promptly documented. Results showed that the people of Gombe State are knowledgeable on herbal medicine in the treatment of diseases and ailments. Furthermore, the aqueous leaf extracts of Senna siamea, the plant species with the highest PPK (percentage of people who have knowledge about the use of a species for treating typhoid fever) in this ethnobotanical survey, was tested for its activity against clinical isolates of Salmonella typhi using the agar well diffusion method. The aqueous extracts showed some activity (zones of inhibition 11, 9, 7.5, 3.5, 1.3 mm) at 2000, 1800, 1600, 1400, 1200 µg/ml concentrations respectively. Preliminary phytochemical studies of the aqueous leaf extracts of the plant revealed the presence of secondary metabolites such as alkaloids, saponins, tannins, flavonoids and cardiac glycosides. Though a large number of traditionally used plants for the treatment of enteric fever were identified, further scientific validation of the traditional claims of anti-typhoid properties is imperative. This would establish their candidature for any possible future research for active principles and the possible development of new cheaper and more effective anti-typhoid drugs, as well as in the conservation of this rich diversity of medicinal plants.

Keywords: antimicrobial activities, ethnobotany, gombe state, north-eastern Nigeria, phytochemical screening, senna siamea, typhoid fever

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Authors: José M. Carmona, Diana Puigserver, Jofre Herrero

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Chlorinated solvents belong to the group of nonaqueous phase liquids (DNAPL) and have been involved in many contamination episodes. They are carcinogenic and recalcitrant pollutants that may be found in granular aquifers as: i) pools accumulated on low hydraulic conductivity layers; ii) immobile residual phase retained at the pore-scale by capillary forces; iii) dissolved phase in groundwater; iv) sorbed by particulate organic matter; and v) stored into the matrix of low hydraulic conductivity layers where they penetrated by molecular diffusion. The transition zone between granular aquifers and basal aquitards constitute the lowermost part of the aquifer and presents numerous fine-grained interbedded layers that give rise to significant textural contrasts. These layers condition the transport and fate of contaminants and lead to differences from the rest of the aquifer, given that: i) hydraulic conductivity of these layers is lower; ii) DNAPL tends to accumulate on them; iii) groundwater flow is slower in the transition zone and consequently pool dissolution is much slower; iv) sorbed concentrations are higher in the fine-grained layers because of their higher content in organic matter; v) a significant mass of pollutant penetrates into the matrix of these layers; and vi) this contaminant mass back-diffuses after remediation and the aquifer becomes contaminated again. Thus, contamination sources of chlorinated solvents are extremely more recalcitrant in transition zones, which has far-reaching implications for the environment. The aim of this study is to analyze the spatial and temporal differences in the evolution of biogeochemical processes in the transition zone and in the rest of the aquifer. For this, an unconfined aquifer with a transition zone in the lower part was selected at Vilafant (NE Spain). This aquifer was contaminated by perchloroethylene (PCE) in the 80’s. Distribution of PCE and other chloroethenes in groundwater and porewater was analyzed in: a) conventional piezometers along the plume and in two multilevel wells at the source of contamination; and b) porewater of fine grained materials from cores recovered when drilled the two multilevel wells. Currently, the highest concentrations continue to be recorded in the source area in the transition zone. By contrast, the lowest concentrations in this area correspond to the central part of the aquifer, where flow velocities are higher and a greater washing of the residual phase initially retained has occurred. The major findings of the study were: i) PCE metabolites were detected in the transition zone, where conditions were more reducing than in the rest of the aquifer; ii) however, reductive dechlorination was partial since only the formation of cis-dicholoroethylene (DCE) was reached; iii) In the central part of the aquifer, where conditions were predominantly oxidizing, the presence of nitrate significantly hindered the reductive declination of PCE. The remediation strategies to be implemented should be directed to enhance dissolution of the source, especially in the transition zone, where it is more recalcitrant. For example, by combining chemical and bioremediation methods, already tested at the laboratory scale with groundwater and sediments of this site.

Keywords: chlorinated solvents, chloroethenes, DNAPL, partial reductive dechlorination, PCE, transition zone to basal aquitard

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Authors: Marcin Dębowski, Marcin Zieliński, Mirosław Krzemieniewski

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As methane fermentation is a complex series of successive biochemical transformations, its subsequent stages are determined, to a various extent, by physical and chemical factors. A specific state of equilibrium is being settled in the functioning fermentation system between environmental conditions and the rate of biochemical reactions and products of successive transformations. In the case of physical factors that influence the effectiveness of methane fermentation transformations, the key significance is ascribed to temperature and intensity of biomass agitation. Among the chemical factors, significant are pH value, type, and availability of the culture medium (to put it simply: the C/N ratio) as well as the presence of toxic substances. One of the important elements which influence the effectiveness of methane fermentation is the pre-treatment of organic substrates and the mode in which the organic matter is made available to anaerobes. Out of all known and described methods for organic substrate pre-treatment before methane fermentation process, the ultrasound disintegration is one of the most interesting technologies. Investigations undertaken on the ultrasound field and the use of installations operating on the existing systems result principally from very wide and universal technological possibilities offered by the sonication process. This physical factor may induce deep physicochemical changes in ultrasonicated substrates that are highly beneficial from the viewpoint of methane fermentation processes. In this case, special role is ascribed to disintegration of biomass that is further subjected to methane fermentation. Once cell walls are damaged, cytoplasm and cellular enzymes are released. The released substances – either in dissolved or colloidal form – are immediately available to anaerobic bacteria for biodegradation. To ensure the maximal release of organic matter from dead biomass cells, disintegration processes are aimed to achieve particle size below 50 μm. It has been demonstrated in many research works and in systems operating in the technical scale that immediately after substrate supersonication the content of organic matter (characterized by COD, BOD5 and TOC indices) was increasing in the dissolved phase of sedimentation water. This phenomenon points to the immediate sonolysis of solid substances contained in the biomass and to the release of cell material, and consequently to the intensification of the hydrolytic phase of fermentation. It results in a significant reduction of fermentation time and increased effectiveness of production of gaseous metabolites of anaerobic bacteria. Because disintegration of Virginia fanpetals biomass via ultrasounds applied in order to intensify its conversion is a novel technique, it is often underestimated by exploiters of agri-biogas works. It has, however, many advantages that have a direct impact on its technological and economical superiority over thus far applied methods of biomass conversion. As for now, ultrasound disintegrators for biomass conversion are not produced on the mass-scale, but by specialized groups in scientific or R&D centers. Therefore, their quality and effectiveness are to a large extent determined by their manufacturers’ knowledge and skills in the fields of acoustics and electronic engineering.

Keywords: ultrasound disintegration, biomass, methane fermentation, biogas, Virginia fanpetals

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Authors: Zeynep Yilmazer Hitit, Patrick C. Hallenbeck

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Fuel reserve requirements due to depletion of fossil fuels have increased interest in biohydrogen since the 1990’s. In fermentative hydrogen production, pure, mixed, and co-cultures can be used to produce hydrogen. Several previous studies have evaluated hydrogen production by pure cultures of Clostridium butyricum or Enterobacter aerogenes. Evaluating hydrogen production by co-culture of these microorganisms is an interestıng approach since E. aerogenes is a facultative microorganism with resistance to oxygen in contrast to the strict anaerobe C. butyricum, and therefore has the ability to maintain anaerobic conditions. It was found that using co-cultures of facultative E. aerogenes (as a reducing agent and H2 producer) and the obligate anaerobe C. butyricum for producing hydrogen increases the yield of hydrogen by about 50% compared to C. butyricum by itself. Also, using different types of microorganisms for hydrogen production eliminates the need to use expensive reducing agents. C. butyricum strain pre-cultured anaerobically at 37 0C for 15h by inoculating 100 mL of GP medium (pH 6.8) consisting of 1% glucose, 2% polypeptone, 0.2% KH2PO4, 0.05% yeast extract, 0.05% MgSO4. 7H2O and E. aerogenes strain was pre-cultured aerobically at 30 0C, 150 rpm for 9 h by inoculating 100 mL of TGY medium (pH 6.8), consisting of 0.1% glucose, 0.5% tryptone, 0.1% K2HPO4, 0.5% yeast extract. All duplicate batch experiments were conducted in 100 mL bottles with different inoculum ratios of Clostridium butyricum and Enterobater aerogenes (C:E) using 5x diluted rich media (GP) consisting of 2 g/L glucose, 4g/L polypeptone, 0.4 g/L KH2PO4, 0.1 g/L yeast extract, 0.1 MgSO4.7H2O. The range of inoculum ratio of C. butyricum to E. aerogenes were 2:1,4:1,8:1, 1:2,1:4, 1:8, 1:0, 0:1. Using glucose as a carbon source aided in the observation of microbial behavior as well as making the effect of inoculum ratio more evident. Nearly all the glucose in the medium was used to produce hydrogen, except at a 1:0 ratio of inoculum (i.e. containing only C. butyricum). Low glucose consumption leads to a higher hydrogen yield due to cumulative hydrogen production and consumption of glucose, but not as much as C:E, 8:1. The lowest hydrogen yield was achieved in 1:8 inoculum ratio of C:E, 71.9 mL, 1.007±0.01 mol H2/mol glucose and the highest cumulative hydrogen, hydrogen yield and dry cell weight were achieved in 8:1 inoculum ratio of C:E, 117.4 mL, 2.035±0.082 mol H2/mol glucose, 0.4 g/L respectively. In this study effect of inoculum ratio on dark fermentative biohydrogen production using C. butyricum and E. aerogenes was investigated. The maximum hydrogen yield of 2.035mol H2/mol glucose was obtained using 2g/L glucose, an initial pH of 6 and an inoculum ratio of C. butyricum to E. aerogenes of 8:1. Results showed that inoculum ratio is an important parameter on hydrogen production due to competition between the two microorganisms in using substrate for growth and production of by-products. The results presented here could be of great significance for further waste management studies using co-culture hydrogen production.

Keywords: biohydrogen, Clostridium butyricum, dark fermentation, Enterobacter aerogenes, inoculum ratio in biohydrogen production

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Authors: N. Pérez-Rodríguez, D. García-Bernet, A. Torrado-Agrasar, J. M. Cruz, A. B. Moldes, J. M. Domínguez

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World economy is based on non-renewable, fossil fuels such as petroleum and natural gas, which entails its rapid depletion and environmental problems. In EU countries, the objective is that at least 20% of the total energy supplies in 2020 should be derived from renewable resources. Biogas, a product of anaerobic degradation of organic substrates, represents an attractive green alternative for meeting partial energy needs. Nowadays, trend to circular economy model involves efficiently use of residues by its transformation from waste to a new resource. In this sense, characteristics of agricultural residues (that are available in plenty, renewable, as well as eco-friendly) propitiate their valorisation as substrates for biogas production. Corn cob is a by-product obtained from maize processing representing 18 % of total maize mass. Corn cob importance lies in the high production of this cereal (more than 1 x 109 tons in 2014). Due to its lignocellulosic nature, corn cob contains three main polymers: cellulose, hemicellulose and lignin. Crystalline, highly ordered structures of cellulose and lignin hinders microbial attack and subsequent biogas production. For the optimal lignocellulose utilization and to enhance gas production in anaerobic digestion, materials are usually submitted to different pretreatment technologies. In the present work, enzymatic hydrolysis, ultrasounds and combination of both technologies were assayed as pretreatments of corn cob for biogas production. Enzymatic hydrolysis pretreatment was started by adding 0.044 U of Ultraflo® L feruloyl esterase per gram of dry corncob. Hydrolyses were carried out in 50 mM sodium-phosphate buffer pH 6.0 with a solid:liquid proportion of 1:10 (w/v), at 150 rpm, 40 ºC and darkness for 3 hours. Ultrasounds pretreatment was performed subjecting corn cob, in 50 mM sodium-phosphate buffer pH 6.0 with a solid: liquid proportion of 1:10 (w/v), at a power of 750W for 1 minute. In order to observe the effect of the combination of both pretreatments, some samples were initially sonicated and then they were enzymatically hydrolysed. In terms of methane production, anaerobic digestion of the corn cob pretreated by enzymatic hydrolysis was positive achieving 290 L CH4 kg MV-1 (compared with 267 L CH4 kg MV-1 obtained with untreated corn cob). Although the use of ultrasound as the only pretreatment resulted detrimentally (since gas production decreased to 244 L CH4 kg MV-1 after 44 days of anaerobic digestion), its combination with enzymatic hydrolysis was beneficial, reaching the highest value (300.9 L CH4 kg MV-1). Consequently, the combination of both pretreatments improved biogas production from corn cob.

Keywords: biogas, corn cob, enzymatic hydrolysis, ultrasound

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Authors: Shalini TV, Gangadharan GG, Sriranjini S Jaideep, ASN Seshasayee, Awadhesh Pandit

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Introduction: Prakriti (body-mind constitution of an individual) is a conventional, customized and unique understanding of which is essential for the personalized medicine described in Ayurveda, Indian System of Medicine. Based on the Doshas( functional, bio humoral unit in the body), individuals are categorized into three major Prakriti- Vata, Pitta, and Kapha. The human gut microbiome hosts plenty of highly diverse and metabolically active microorganisms, mainly dominated by the bacteria, which are known to influence the physiology of an individual. Few researches have shown the correlation between the Prakriti and the biochemical parameters. In this study, an attempt was made to explore any correlation between the Prakriti (phenotype of an individual) with the Genetic makeup of the gut microbiome in healthy individuals. Materials and methods: 270 multi-ethnic, healthy volunteers of both sex with the age group between 18 to 40 years, with no history of antibiotics in the last 6 months were recruited into three groups of Vata, Pitta, and Kapha. The Prakriti of the individual was determined using Ayusoft, a software designed by CDAC, Pune, India. The volunteers were subjected to initial screening for the assessment of their height, weight, Body Mass Index, Vital signs and Blood investigations to ensure they are healthy. The stool and saliva samples of the recruited volunteers were collected as per the standard operating procedure developed, and the bacterial DNA was isolated using Qiagen kits. The extracted DNA was subjected to 16s rRNA sequencing using the Illumina kits. The sequencing libraries are targeting the variable V3 and V4 regions of the 16s rRNA gene. Paired sequencing was done on the MiSeq system and data were analyzed using the CLC Genomics workbench 11. Results: The 16s rRNA sequencing of the V3 and V4 regions showed a diverse pattern in both the oral and stool microbial DNA. The study did not reveal any specific pattern of bacterial flora amongst the Prakriti. All the p-values were more than the effective alpha values for all OTUs in both the buccal cavity and stool samples. Therefore, there was no observed significant enrichment of an OTU in the patient samples from either the buccal cavity or stool samples. Conclusion: In healthy volunteers of multi-ethnicity, due to the influence of the various factors, the correlation between the Prakriti and the gut microbiome was not seen.

Keywords: gut microbiome, ayurveda Prakriti, sequencing, multi-ethnic urban population

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Authors: Bhupendra Kumar Poudel, Sadhana Amatya, Tirtha Maiya Shrestha, Bharatmani Pokhrel, Mohan Prasad Amatya

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Background: H. salicifolia is a dense, branched, multipurpose, deciduous, nitrogen fixing, thorny willow-like small to moderate tree, restricted to the Himalaya. Among the two species of Nepal (Hippophae salicifolia and H. tibetana), it has been traditionally used as food additive, anticancer (bark), and treating toothache, tooth inflammation (anti-inflammatory) and radiation injury; while people of Western Nepal have largely undermined its veiled treasure by using it for fuel, wood and soil stabilization only. Therefore, the main objective of this study was to explore biological properties (analgesic, antidiabetic, cytotoxic and anti-inflammatory properties of this plant. Methodology: The transverse section of leaves and stems were viewed under microscope. Extracts obtained from soxhlation subjected to tests for phytochemical and biological studies. Rats (used to study antidiabetic and anti-inflammatory properties) and mice (used to study analgesic, CNS depressant, muscle relaxant and locomotor properties) were assumed to be normally distributed; then ANOVA and post hoc tukey test was used to find significance. The data obtained were analyzed by SPSS 17 and Excel 2007. Results and Conclusion: Pharmacognostical analysis revealed the presence of long stellate trichomes, double layered vascular bundle 5-6 in number and double layered compact sclerenchyma. The preliminary phytochemical screening of the extracts was found to exhibit the positive reaction tests for glycoside, steroid, tannin, flavonoid, saponin, coumarin and reducing sugar. The brine shrimp lethality bioassay tested in 1000, 100 and 10 ppm revealed cytotoxic activity inherent in methanol, water, chloroform and ethyl acetate extracts with LC50 (μg/ml) values of 61.42, 99.77, 292.72 and 277.84 respectively. The cytotoxic activity may be due to presence of tannins in the constituents. Antimicrobial screening of the extracts by cup diffusion method using Staphylococcus aereus, Escherichia coli and Pseudomonas aeruginosa against standard antibiotics (oxacillin, gentamycin and amikacin respectively) portrayed no activity against the microorganisms tested. The methanol extract of the stems and leaves showed various pharmacological properties: and antidiabetic, anti-inflammatory, analgesic [chemical writhing method], CNS depressant, muscle relaxant and locomotor activities in a dose-dependent fashion, indicating the possibility of the presence of different constituents in the stems and leaves responsible for these biological activities. All the effects when analyzed by post hoc tukey test were found to be significant at 95% confidence level. The antidiabetic activity was presumed to be due to flavonoids present in extract. Therefore, it can be concluded that this plant’s secondary metabolites possessed strong antidiabetic, anti-inflammatory and cytotoxic activity which could be isolated for further investigation.

Keywords: Hippophae salicifolia, constituents, antidiabetic, inflammatory, brine shrimp

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Authors: Peter C. St. John, Michael F. Crowley, Yannick J. Bomble

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Production of chemicals from engineered organisms in a batch culture typically involves a trade-off between productivity, yield, and titer. However, strategies for strain design typically involve designing mutations to achieve the highest yield possible while maintaining growth viability. Such approaches tend to follow the principle of designing static networks with minimum metabolic functionality to achieve desired yields. While these methods are computationally tractable, optimum productivity is likely achieved by a dynamic strategy, in which intracellular fluxes change their distribution over time. One can use multi-stage fermentations to increase either productivity or yield. Such strategies would range from simple manipulations (aerobic growth phase, anaerobic production phase), to more complex genetic toggle switches. Additionally, some computational methods can also be developed to aid in optimizing two-stage fermentation systems. One can assume an initial control strategy (i.e., a single reaction target) in maximizing productivity - but it is unclear how close this productivity would come to a global optimum. The calculation of maximum theoretical yield in metabolic engineering can help guide strain and pathway selection for static strain design efforts. Here, we present a method for the calculation of a maximum theoretical productivity of a batch culture system. This method follows the traditional assumptions of dynamic flux balance analysis: that internal metabolite fluxes are governed by a pseudo-steady state and external metabolite fluxes are represented by dynamic system including Michealis-Menten or hill-type regulation. The productivity optimization is achieved via dynamic programming, and accounts explicitly for an arbitrary number of fermentation stages and flux variable changes. We have applied our method to succinate production in two common microbial hosts: E. coli and A. succinogenes. The method can be further extended to calculate the complete productivity versus yield Pareto surface. Our results demonstrate that nearly optimal yields and productivities can indeed be achieved with only two discrete flux stages.

Keywords: A. succinogenes, E. coli, metabolic engineering, metabolite fluxes, multi-stage fermentations, succinate

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Authors: Kamran Jawed, Anu Jose Mattam, Zia Fatma, Saima Wajid, Malik Z. Abdin, Syed Shams Yazdani

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Worldwide demand of natural and sustainable fuels and chemicals have encouraged researchers to develop microbial platform for synthesis of short chain fatty acids as they are useful precursors to replace petroleum-based fuels and chemicals. In this study, we evaluated the role of fatty acid synthesis and β-oxidation cycle of Escherichia coli to produce butyric acid, a 4-carbon short chain fatty acid, with the help of three thioesterases, i.e., TesAT from Anaerococcus tetradius, TesBF from Bryantella formatexigens and TesBT from Bacteroides thetaiotaomicron. We found that E. coli strain transformed with gene for TesBT and grown in presence of 8 g/L glucose produced maximum butyric acid titer at 1.46 g/L, followed by that of TesBF at 0.85 g/L and TesAT at 0.12 g/L, indicating that these thioesterases were efficiently converting short chain fatty acyl-ACP intermediate of fatty acid synthesis pathway into the corresponding acid. The titer of butyric acid varied significantly depending upon the plasmid copy number and strain genotype. Deletion of genes for fatty acyl-CoA synthetase and acyl-CoA dehydrogenase, which are involved in initiating the fatty acid degradation cycle, and overexpression of FadR, which is a dual transcriptional regulator and exerts negative control over fatty acid degradation pathway, reduced up to 30% of butyric acid titer. This observation suggested that β-oxidation pathway is working synergistically with fatty acid synthesis pathway in production of butyric acid. Moreover, accelerating the fatty acid elongation cycle by overexpressing acetyl-CoA carboxyltransferase (Acc) and 3-hydroxy-acyl-ACP dehydratase (FabZ) or by deleting FabR, the transcription suppressor of elongation, did not improve the butyric acid titer, rather favored the long chain fatty acid production. Finally, a balance between cell growth and butyric acid production was achieved with the use of phosphorous limited growth medium and 14.3 g/L butyric acid, and 17.5 g/L total free fatty acids (FFAs) titer was achieved during fed-batch cultivation. We have engineered an E. coli strain which utilizes the intermediate of both fatty acid synthesis and degradation pathway, i.e. butyryl-ACP and -CoA, to produce butyric acid from glucose. The strategy used in this study resulted in highest reported titers of butyric acid and FFAs in engineered E. coli.

Keywords: butenoic acid, butyric acid, Escherichia coli, fed-batch fermentation, short chain fatty acids, thioesterase

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Authors: Mahboobe Daneshvar

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Introduction: Clinical studies on the effect of systemic conditions on periodontal diseases have shown that some systemic deficiencies may provide grounds for the onset of periodontal diseases. One of these systemic problems is osteoporosis, which may be a risk factor for the onset and exacerbation of periodontitis. This study tends to evaluate periodontal indices in osteoporotic menopausal women and compare them with healthy controls. Materials and Methods: In this case-control study, participants included 45-75-year-old menopausal women referred to rheumatology wards of the Khatamolanbia Clinic and Shahid Sadoughi Hospital in Yazd; Their bone density was determined by DEXA-scan and by imaging the femoral-lumbar bone. Thirty patients with osteoporosis and 30 subjects with normal BMD were selected. Then, informed consent was obtained for participation in the study. During the clinical examinations, tooth loss (TL), plaque index (PI), gingival recession, pocket probing depth (PPD), clinical attachment loss (CAL), and tooth mobility (TM) were measured to evaluate the periodontal status. These clinical examinations were performed to determine the periodontal status by catheter, mirror and probe. Results: During the evaluation, there was no significant difference in PPD, PI, TM, gingival recession, and CAL between case and control groups (P-value>0.05); that is, osteoporosis has no effect on the above factors. These periodontal factors are almost the same in both healthy and patient groups. In the case of missing teeth, the following results were obtained: the mean of missing teeth was 22.173% of the total teeth in the case group and 18.583% of the total teeth in the control group. In the study of the missing teeth in the case and control groups, there was a significant relationship between case and control groups (P-value = 0.025). Conclusion: In fact, since periodontal disease is multifactorial and microbial plaque is the main cause, osteoporosis is considered a predisposing factor in exacerbation or persistence of periodontal disease. In patients with osteoporosis, usually pathological fractures, hormonal changes, and aging lead to reduced physical activity and affect oral health, which leads to the manifestation of periodontal disease. But this disease increases tooth loss by changing the shape and structure of bone trabeculae and weakening them. Osteoporosis does not seem to be a deterministic factor in the incidence of periodontal disease, since it affects bone quality rather than bone quantity.

Keywords: plaque index, Osteoporosis, tooth mobility, periodontal packet

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Authors: Fatemeh Elmi, Zahra Etemadifar

Abstract:

Improvement of soil mechanical properties is crucial before its use in construction, as the low mechanical strength and unstable structure of soil in many parts of the world can lead to the destruction of engineering infrastructure, resulting in financial and human losses. Although, conventional methods, such as chemical injection, are often utilized to enhance soil strength and stiffness, they are generally expensive, require heavy machinery, and cause significant environmental effects due to chemical usage, and also disrupt urban infrastructure. Moreover, they are not suitable for treating large volume of soil. Recently, an alternative method to improve various soil properties, including strength, hardness, and permeability, has received much attention: the application of biological methods. One of the most widely used is biocementation, which is based on the microbial precipitation of calcium carbonte crystalls using ureolytic bacteria However, there are still limitations to its large-scale use that need to be resolved before it can be commercialized. These issues have not received enough attention in prior research. One limitation of MICP (microbially induced calcium carbonate precipitation) is that microorganisms cannot operate effectively in harsh and variable environments, unlike the controlled conditions of a laboratory. Another limitation of applying this technique on a large scale is the high cost of producing a substantial amount of bacterial culture and reagents required for soil treatment. Therefore, the purpose of the present study was to investigate soil improvement using the biocementation activity of poly-extremophile, calcium carbonate crystal- producing bacterial strain, Bhargavaea cecembensis N1, in whey as an inexpensive medium. This strain was isolated and molecularly identified from sandy soils in our previous research, and its 16S rRNA gene sequences was deposited in the NCBI Gene Bank with an accession number MK420385. This strain exhibited a high level of urease activity (8.16 U/ml) and produced a large amount of calcium carbonate (4.1 mg/ ml). It was able to improve the soil by increasing the compressive strength up to 205 kPa and reducing permeability by 36%, with 20% of the improvement attributable of calcium carbonate production. This was achieved using this strain in a whey culture medium. This strain can be an eco-friendly and economical alternative to conventional methods in soil stabilization, and other MICP related applications.

Keywords: biocementation, Bhargavaea cecembensis, soil improvement, whey culture medium

Procedia PDF Downloads 32

Authors: Rishi Richa, N. C. Shahi, J. P. Pandey, S. S. Kautkar

Abstract:

Citrus fruits rank third in area and production after banana and mango in India. Sweet oranges are the second largest citrus fruits cultivated in the country. Andhra Pradesh, Maharashtra, Karnataka, Punjab, Haryana, Rajasthan, and Uttarakhand are the main sweet orange-growing states. Citrus fruits occupy a leading position in the fruit trade of Uttarakhand, is casing about 14.38% of the total area under fruits and contributing nearly 17.75 % to the total fruit production. Malta is grown in most of the hill districts of the Uttarakhand. Malta common is having high acceptability due to its attractive colour, distinctive flavour, and taste. The excellent quality fruits are generally available for only one or two months. However due to its less shelf-life, Malta can not be stored for longer time under ambient conditions and cannot be transported to distant places. Continuous loss of water adversely affects the quality of Malta during storage and transportation. Method of picking, packaging, and cold storage has detrimental effects on moisture loss. The climatic condition such as ambient temperature, relative humidity, wind condition (aeration) and microbial attack greatly influences the rate of moisture loss and quality. Therefore, different agro-climatic zone will have different moisture loss pattern. The rate of moisture loss can be taken as one of the quality parameters in combination of one or more parameter such as RH, and aeration. The moisture contents of the fruits and vegetables determine their freshness. Hence, it is important to maintain initial moisture status of fruits and vegetable for prolonged period after the harvest. Keeping all points in views, effort was made to store Malta at ambient condition. In this study, the response surface method and experimental design were applied for optimization of independent variables to enhance the shelf life of four months stored malta. Box-Benkhen design, with, 12 factorial points and 5 replicates at the centre point were used to build a model for predicting and optimizing storage process parameters. The independent parameters, viz., scavenger (3, 4 and 5g), polythene thickness (75, 100 and 125 gauge) and fungicide concentration (100, 150 and 200ppm) were selected and analyzed. 5g scavenger, 125 gauge and 200ppm solution of fungicide are the optimized value for storage which may enhance life up to 4months.

Keywords: Malta fruit, scavenger, packaging, shelf life

Procedia PDF Downloads 266

Authors: M. El-haj, Z. Olama, H. Holail

Abstract:

Single cell oils (SCOs) accumulated by oleaginous fungi have emerged as a potential alternative feedstock for biodiesel production. Five fungal strains were isolated from the Lebanese environment namely Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger that have been selected among 39 oleaginous strains for their potential ability to accumulate lipids (lipid content was more than 40% on dry weight basis). Wide variations were recorded in the environmental factors that lead to maximum lipid production by fungi under test and were cultivated under submerged fermentation on medium containing glucose as a carbon source. The maximum lipid production was attained within 6-8 days, at pH range 6-7, 24 to 48 hours age of seed culture, 4 to 6.107 spores/ml inoculum level and 100 ml culture volume. Eleven culture conditions were examined for their significance on lipid production using Plackett-Burman factorial design. Reducing sugars and nitrogen source were the most significant factors affecting lipid production process. Maximum lipid yield was noticed with 15.62, 14.48, 12.75, 13.68 and 20.41g/l for Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger respectively. A verification experiment was carried out to examine model validation and revealed more than 94% validity. The profile of extracted lipids from each fungal isolate was studied using thin layer chromatography (TLC) indicating the presence of monoacylglycerols, diaacylglycerols, free fatty acids, triacylglycerols and sterol esters. The fatty acids profiles were also determined by gas-chromatography coupled with flame ionization detector (GC-FID). Data revealed the presence of significant amount of oleic acid (29-36%), palmitic acid (18-24%), linoleic acid (26.8-35%), and low amount of other fatty acids in the extracted fungal oils which indicate that the fatty acid profiles were quite similar to that of conventional vegetable oil. The cost of lipid production could be further reduced with acid-pretreated lignocellulotic corncob waste, whey and date molasses to be utilized as the raw material for the oleaginous fungi. The results showed that the microbial lipid from the studied fungi was a potential alternative resource for biodiesel production.

Keywords: agro-industrial waste products, biodiesel, fatty acid, single cell oil, Lebanese environment, oleaginous fungi

Procedia PDF Downloads 386

Authors: Blessing C. Nwachukwu, Michael O. Taiwo, Wasiu A. Abibu, Isaac O. Ayodeji

Abstract:

Milk is an important source of micro and macronutrients for humans. Soft Cheese (Wara) is an example of a by-product of milk. In addition, water is considered as one of the most vital resources in cattle farms. Due to the high consumption rate of milk and soft cheese and the traditional techniques involved in their production in Nigeria, there was a need for a microbiological assessment which will be of utmost public health importance. The study thus investigated microbial risk assessments associated with consumption of milk and soft cheese (Wara). It also investigated common pathogens present in dairy water in farms and antibiotic sensitivity profiling for implicated pathogens were conducted. Samples were collected from three different Fulani dairy herds in Ido local government, Ibadan, Oyo State, Nigeria and subjected to microbiological evaluation and antimicrobial susceptibility testing. Aspergillus flavus was the only isolated fungal isolate from Wara while Staphylococcus aureus, Vibro cholera, Hafnia alvei, Proteus mirabilis, Escherishia coli, Psuedomonas aeuroginosa, Citrobacter freundii, and Klebsiella pneumonia were the bacteria genera isolated from Wara, dairy milk and dairy drinking water. Bacterial counts from Wara from the three selected farms A, B and C were 3.5×105 CFU/ml, 4.0×105 CFU/ml and 5.3×105 CFU/ml respectively while the fungal count was 3CFU/100µl. The total bacteria count from dairy milk from the three selected farms A, B and C were Farms 2.0 ×105 CFU/ml, 3.5 × 105 CFU/ml and 6.5 × 105 CFU/ml respectively. 1.4×105 CFU/ml, 1.9×105 CFU/ml and 4.9×105 CFU/ml were the recorded bacterial counts from dairy water from farms A, B and C respectively. The highest antimicrobial resistance of 100% was recorded in Wara with Enrofloxacin, Gentamycin, Cefatriaxone and Colistin. The highest antimicrobial susceptibility of 100% was recorded in Raw milk with Enrofloxacin and Gentamicin. Highest antimicrobial intermediate response of 100% was recorded in Raw milk with Streptomycin. The study revealed that most of the cheeses sold at Ido local Government are contaminated with pathogens. Further research is needed on standardizing the production method to prevent pathogens from gaining access. The presence of bacteria in raw milk indicated contamination due to poor handling and unhygienic practices. Thus, drinking unpasteurized milk is hazardous as it increases the risk of zoonoses. Also, the Provision of quality drinking water is crucial for optimum productivity of dairy. Health education programs aiming at increasing awareness of the importance of clean water for animal health will be helpful.

Keywords: dairy, raw milk, soft cheese, Wara

Procedia PDF Downloads 152