Background: Cellulose, the major component of the plant cell wall, is the most abundant and cheap polymer on earth. It can be used by varieties of cellulolytic enzymes. Cellulases can hydrolyze cellulose to its glucose monomers, which can be fermented to many biotechnological products, such as biochemicals, bioplastics, and biofuels. Actinomycetes are potential sources of cellulases. Objective: The current study sheds light on the cellulolytic activity of Thermobifida cellulosilytica, a previously isolated thermophilic actinomycete, and the analysis of the lignocellulases produced in the secretome as a result of induction by different carbon sources. Methods: The cellulolytic activity was qualitatively confirmed by Congo red method showing a large halo zone around the colonies. The activity was also assayed using the 3,5-dinitrosalicylic acid (DNS) method. The secretome analysis was conducted by liquid chromatography-tandem mass spectroscopy (LC-MS/MS) based proteomic approach. Results: The cellulolytic activity increased by two folds upon the growth of T. cellulosilytica on rice straw (RS) as a complex substrate comparatively to Carboxymethylcellulose (CMC) as a simple one. These results were highly assured by LC-MS/MS. Where more proteins (n=31) were produced in the RS secretome, CMC produced only six proteins, including only one cellulase. Different classes of proteins produced in the RS secretome were cellulases (26%), hemicellulases (16%), proteases (10%), and others (48%). Conclusion: Lignocellulases are inducible enzymes. RS as a complex substrate induced T. cellulosilytica for the expression of more lignocellulolytic enzymes than CMC.

Recent developments in targeted genome editing accelerated genetic research and opened new potentials to improve the crops for better yields and quality. Genome editing techniques like Zinc Finger Nucleases (ZFN) and Transcription Activator-Like Effector Nucleases (TALENs) have been accustomed to target any gene of interest. However, these systems have some drawbacks as they are very expensive and time consuming with labor-intensive protein construction protocol. A new era of genome editing technology has a user-friendly tool which is termed as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated protein9 (Cas9), is an RNA based genome editing system involving a simple and cost-effective design of constructs. CRISPR/Cas9 system has been successfully applied in diverse crops for various genome editing approaches. In this review, we highlight the application of the CRISPR/Cas9 system in cereal crops including rice, wheat, maize, and sorghum to improve these crops for better yield and quality. Since cereal crops supply a major source of food to world populations, their improvement using recent genome editing tools like CRISPR/Cas9 is timely and crucial. The genome editing of cereal crops using the CRISPR/Cas9 system would help to overcome the adverse effects of agriculture and may aid in conserving food security in developing countries.

Objective: The study aimed at increasing the sensitivity of immunochromatographic tests for the control of toxic contaminants (on the examples of aflatoxin B1 and T-2 toxin) in agricultural products. Methods: For reliable immunochromatographic detection of low concentrations of analytes, a replacement of the (specific antibodies – gold nanoparticle) conjugate by a combination of native specific antibodies and anti-species antibodies conjugated with gold nanoparticles was proposed. Different variants of test systems based on the principle of indirect labeling were realized and compared. Results: Immunochromatographic assays with indirect labeling for aflatoxin B1 and T-2 toxin were implemented experimentally. A reduction in the detection limit by one to two orders of magnitude was demonstrated. Conclusion: The presented results confirm that indirect labeling of specific antibodies overcomes the limitations of the competitive immunochromatographic analysis and can be used to detect analytes of different chemical nature.

Modern society has easy access to a vast informational database. The pursuit of sustainable green and healthy lifestyle leads to a series of food choices. Therefore, it is of importance to provide reliable, comprehensive and up-to-date information about food content including both nutritional and antinutritional elements.Nutrients are associated with positive effects on human health. Antinutrients, on the other hand, are far less popular for the contemporary man. They are highly bioactive, capable of deleterious effects as well as some beneficial health effects in man, and vastly available in plant-based foods. These compounds are of natural or synthetic origin, interfere with the absorption of nutrients, and can be responsible for some mischievous effects related to the nutrient absorption. Some of the common symptoms exhibited by a large amount of antinutrients in the body can be nausea, bloating, headaches, rashes, nutritional deficiencies,etc. Phytates, oxalates, and lectins are few of the well-known antinutrients.Science has acknowledged several ways in order to alter the negative influence antinutrients exhibiting on human health. Mechanical, thermal and biochemical approaches act synergistically to provide food with lower antinutritional levels.The purpose of this review was to synthesize the availability of antinutrients, clear their effect on the human body, and commemorate possible paths to disable them. This review provides links to the available literature as well as enables a systematic view of the recently published research on the topic of plant-based antinutrients.

Microorganisms are the great sources of Natural Products (NPs); these are imperative to their survival apart from conferring competitiveness amongst each other within their environmental niches. Primary and secondary metabolites are the two major classes of NPs that help in cell development, where antimicrobial activity is closely linked with secondary metabolites. To capitalize on the effects of secondary metabolites, co-culture methods have been often used to develop an artificial microbial community that promotes the action of these metabolites. Different analytical techniques will subsequently be employed based on the metabolite specificity and sensitivity to further enhance the metabolite induction. Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography (GC)-MS are commonly used for metabolite separation while Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS) have been used as tools to elucidate the structure of compounds. This review intends to discuss current systems in use for co-culture in addition to its advantages, with discourse into the investigation of specific techniques in use for the detailed study of secondary metabolites. Further advancements and focus on co-culture technologies are required to fully realize the massive potential in synthetic biological systems.

Background:This paper would be a starting point addressed to a methodology to minimize the effects on livings of man made Electromagnetic Fields (EMFs) pollution.Methods:Given that previous literature highlighted that the most relevant EMFs effects on biological systems can be due to resonance phenomena between electromagnetic field and organic matter, it was proposed here an algorithm to obtain values of frequencies of an applied electromagnetic field far from resonant frequencies, depending on the natural frequencies and viscous damper of a biological system. These frequencies have been named non-resonant frequencies.Results:The displacement of the α-helices in cellular membrane channels due to EMFs has been proposed as a relevant parameter for quantifying the result of the interaction between an applied EMF and organic matter, in order to find both the natural frequencies of a biological system and the resonant frequencies at which α-helices displacement should be maximum.Conclusion:The non-resonant frequencies can be obtained using the algorithm proposed here.

Background:Mercury is a toxic metal that is present in small amounts in the environment, but its level is rising steadily, due to different human activities, such as industrialization. It can reach humans through the food chain, amalgam fillings, and other sources, causing different neurological disorders, memory loss, vision impairment, and may even lead to death; making its detoxification an urgent task.Methods:Various physical and chemical mercury remediation techniques are available, which generally aim at: (i) reducing its mobility or solubility; (ii) causing its vaporization or condensation; (iii) its separation from contaminated soils. Biological remediation techniques, commonly known as bioremediation, are also another possible alternative, which is considered as cheaper than the conventional means and can be accomplished using either (i) organisms harboring themeroperon genes (merB,merA,merR,merP,merT,merD,merF,merC,merE,merHandmerG), or (ii) plants expressing metal-binding proteins. Recently, differentmerdeterminants have been genetically engineered into several organisms, including bacteria and plants, to aid in detoxification of both ionic and organic forms of mercury.Results:Bacteria that are resistant to mercury compounds have at least a mercuric reductase enzyme (MerA) that reduces Hg+2to volatile Hg0, a membrane-bound protein (MerT) for Hg+2uptake and an additional enzyme, MerB, that degrades organomercurials by protonolysis. Presence of bothmerA andmerB genes confer broad-spectrum mercury resistance. However,merA alone confers narrow spectrum inorganic mercury resistance.Conclusion:To conclude, this review discusses the importance of mercury-resistance genes in mercury bioremediation. Functional analysis ofmeroperon genes and the recent advances in genetic engineering techniques could provide the most environmental friendly, safe, effective and fantastic solution to overcome mercuric toxicity.

Bacteriocins are biologically active compounds produced by a large number of bacteria, including lactic acid bacteria (LAB), which exhibit antimicrobial activity against various saprophytic and pathogenic microorganisms. In recent decades, bacteriocins are increasingly becoming more important in different branches of the industry due to their broad antibacterial and antifungal spectrum - in the food industry for natural food preservation and expiry date extension; in the health sector for preparation of probiotic foods and beverages; in the clinical practice as alternatives of conventional antibiotics; in the agriculture as biocontrol agents of plant pathogens and alternatives of chemical pesticides for plant protection. The broad antimicrobial spectrum of bacteriocins has stimulated the research attention on their application mainly in the food industry as natural preservatives. Most scientific achievements concerning the application food biopreservation are related to bacteriocins produced by LAB. The lactic acid bacteria bacteriocins can be produced in the food substrate during its natural fermentation or can be added in the food products after obtaining byin vitrofermentations under optimal physical and chemical conditions. Moreover, the immobilization of LAB bacteriocins on different matrices of organic and inorganic origin has been proposed as an advanced approach in the natural food preservation for their specific antimicrobial activity, anti-biofilm properties and potential use as tools for pathogen detection.

Background: The present study was designed to evaluate the phytochemicals of Terminalia glaucescens stem extracts and test their antimicrobial potency against water microbial contaminants reported to be multidrug resistant. Method: Dry stem powder was extracted with ethanol, ethyl acetate and chloroform. These fractions were then examined for antimicrobial activity by using disc diffusion assay against typical clinical bacteria and fungal isolates which have been reported as water contaminants. The microbial strains were exposed to five different concentrations of extracts: 500 mg/ml, 250 mg/ml, 100 mg/ml, 50 mg/ml and 25 mg/ml. Result: It was observed in this study that increase in the concentration of extracts correlated with microbial growth inhibition. In-vitro phytochemical screening of plant extracts revealed the presence of alkaloid, flavonoid, saponin, terpenoid, steroid and anthraquinones. Ethanolic extract performs better than ethyl acetate and chloroform extracts, as it recorded the highest zone of inhibition of 20.5 mm against Streptococcus pneumoniae while ethyl acetate and chloroform recorded 17.50 mm each against Streptococcus pneumoniae and Bacillus cereus, respectively. Ethanolic extract also showed the highest antifungal activity against Trichoderma sp. and Aspergillus niger. The antibacterial and antifungal activities of active extracts were observed in the increasing order Ethanol>Chloroform≥ethyl acetate with respect to the maximum zone of inhibition. Activity of crude extract from ethanol, when further compared with commercial antibiotics (Gentamicin, Streptomycin and Nystatin), was significantly higher. Conclusion: This plant crude extracts could therefore serve as potential source of new biocides with application in water research and other biotechnological fields.

Background:The life style and biology of dung beetles offer a significant opportunity for innovation in biofuel production. The larvae of the African dung beetle,Euoniticellus intermedius, feed solely on cow dung, eating and digesting the fibre while adults live on juices found in fresh dung. The larval gut system consists of a small, almost unrecognizable foregut and two distinct chambers; the midgut and hindgut. It is clear that these two chambers are the centres in which the dung material whose composition includes cellulose is processed. The goal of this study was to assess the cellulolytic activities of cultured gut micro-flora derived fromE. intermedius, (Coleoptera: Scarabaeida).Method:Late second to third instar stageE. intermediuslarvae were dissected and the isolated gut micro-flora consortia aerobically cultured in media containing cellulose (filter paper) as the sole carbon source. Genomic DNA isolation was done on the gut consortia cultures after 10 days of culturing, using the ZR Fungal/Bacterial DNA MiniPrep kit (Zymo Research, USA). A complete and unbiased primary cosmid library was then constructed from the isolated genomic DNA using a cloning ready, pWEB-TNC™ Cosmid Cloning kit (EPICENTRE Biotechnologies, USA). The primary cosmid library clones were screened for endo-glucanase and cellobiohydrolase activities using Carboxymethyl Cellulose (CMC) and 4-Methylumbelliferyl-β-D-Cellobioside (MUC) plate assays respectively.Results:Results indicate that a total of 7 colonies out of 160 screened colonies showed positive CMC and MUC activities.Conclusion:This proves thatE. intermediusis a potential source of cellulolytic micro-organisms and enzymes that can be used for cellulose derived biofuel production.