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An SBO Company
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Phone: +1-(604)-945-8408 CA
Fax: +1-(604)-464-0103 CA
Covering the areas of feed, food and energy from ligno-cellulose feedstock or biomass, a primary area of interest at Skye Blue, we have recently opened up three (3) new areas worth a look into further: (1) Ionic liquid pre-treatment deconstruction of the ligno-cellulose backbone fibre to the saccharolific fractions and ligninous fraction with further downstream processing with process flow engineering technology and recycle of the ionic fluid, considered v. effective and advanced as a separation pre-treatment technology for developing countries at this time, (2) The saccharification and subsequent fermentation of sugars from switchgrass, wheat straw, sorghum straw and stover, and bagasse, etc. from ionic liquid treatment, using open algal vat solar fermentors in large arrays in ISO-environmental zones producing ISO-butanol and ISO-pentenol monomers and biodiesel polymer fatty acid chains, and (3) Convert-starch production using enzyme technology that 'reverses' the production of beta-glucan polymers chains for alpha-glucan as an economical commodity in countries like the Philippines where farm byproducts are plentiful as fibre and yet starches in flours have a premium in supply and demand from crops such as rice, corn and wheat imports.
Technologies involving bagasse would help diversify the production of cane sugar milling towards energy from iso-monomers (iso-butanol and iso-pentenol) as opposed to direct fermentation of sucrose sugars to potable ethanol as petrol substitute in sugarcane growing areas and where algal cultures can be maintained through ISO-envirofermentative conditions, at this time in locales such as Queensland AU, New Mexico USA and the Central Asian 'Stans in open solar arrayed vats.
SKYEVIEW: Energy policy and global climate change has been discussed recently (2009) leading to suggestions of zero to negative carbon fingeprinting and use of ligno-cellulosic feedstocks. It has been speculated that a policy be targeted set by 2050 for use of fuels exclusively from non-fossil fuel sources. This policy includes the use of C-sequestration strategies and continuing expansion of solar and wind turbine power. Cellulosic-ethanol as a C-sink source is one example (there are other e.g.s. of higher density polycarboxyols or alcohols that may be fermented) and so with biodiesel for transporation fuels. Heating/cooling may be a combination of solar-based technology and carbon sequestration in addition to C-sink sources. There is also the question of safety of nuclear-based technologies based on advances in this are that is continuing and expansion of geothermal plants where they are needed. There will be a need to derive energy from sustainable means based on agronomic/energy inputs vs. weighing on inputs more directly from the sun's energy source.
SKYEVIEW: The activity for food production will compete with urban clustering of life sciences bio-hubbing, e. g. food mfging., processing/packaging, trade, and biopharma. This is discussed here with SkyeBlue's contention under the topic of the "Fraser Hi-Tech Mainland" (FHTM) of B.C. As with Manitoba CANADA's resource base of a vast and diverse land base, feedstock availability, highly-skilled workforce and value-added processors, it is claimed that the province is positioned to take advantage of a broad array of innovation in future. The same would hold true for the FHTM including a diverse mix of agri-food landbase throughout the island and mainland. In addition to feedstock conversion to high-grade animal feedstock for dairy and meat, feedstock can be shipped in from vast renewable reserves in the prairies, for e. g. pulp from beets and residuals from fermentation and the conversion of ligno-cellulosic fibre to convert starch for wafered/pelleted feeds in aquaculture/capture fisheries or poultry broiler or layer production applications. The above arguments using resource bases from other countries will apply where foreign investments can accrue in places like continental Africa as a future for renewed bio-hubbing in this potentially new breadbasket of the world (e.g. Europe, Japan, China, Korea, Middle East-Arabia, Brazil, USA). (See: also stories covering SkyBlue's contention for an 'SHTM' region in the Soviet-Asia Far East on the Asia-Pacific rim.)
SKYEVIEW: Comparative approaches to feeding and feed improvement are beneficial in the following order: (1) pre-treatments (viz. biological, chemical, physical and thermal, of which chemical is the most cost-effective), (2) genetically modified (GM) pre-biotic approaches to feeds which benefit digestive fermentation (e. g. water-soluble carbohydrates (WSC) and non-structural carbohydrates (NSC), surrogate proteins, ionophores and low-lignin), (3) protein, energy concentrate supplementation (e. g. by-product feeds such as rice pomace, cassava pomace, palm kernel, leucaena leaf meal) and (4) theoretical/practical feeding of GM pro-biotics boosted for digestion of structural carbohydrates and lignin and 'storage' proteins. This subject matter is still new and requires considerable input which we invite from our viewing audience.
SKYEVIEW: The whole movement in biotech cropping and plant protection with pest management has been as recent with Bt corn, wheat resistant to rust infestation and now just in soybean and nematode infection. It was noted by one of our researchers that wheat rust resistance is a world stage issue in food security and time is running to find cultivars resistant with the so-called multiple resistance factors both in wild type and inbred strains and defining their mechanisms at the molecular level. Both foundations in the U.K. and the Bill and Melinda Gates Foundation Inc. USA have undertaken grant funding. Nematode research is beginning at Monsanto with interests in soybean research in places like Brazil and finding ways to assay the egg count in infected planted soils with advanced biotech (there is an invitation from IdeaConnection.com to submit leads for research in this area) and finding mechanisms in natural nematocides in soybeans against the eggs which are the likely target of such agents.
SKYEVIEW: The renewed use of proteomics should be directed to a holy grail of cereal biology regards the grain and its constitutive parts and protein products that can accelerate productive output comparing known 'models' from earlier research on cancer and development applied to plant crops and their growth as well as species in weeds and other plants such as the daffodil, bamboo or the sunflower. Gene-specific promoters and specificity will play a key role in providing means for search of such mechanisms. It is with great speculation as to the fecundity that can be obtained from results of such research on hyper-accelerated crop yield and production and 'pleiotropy' or its effects on the metabolosome and with its perturbed regulation. The yield increment vs inputs [urea for nitrogen-phosphorous-potassium (N-P-K) and other energy inputs] ratios will measure the sustainabilities obtained with innovation.
SKYEVIEW: The bioengineering of plants for biomass for both food & energy (viz. in marginalized lands) (e. g. root crops, corn, sorghum, sugarcane, miscanthus & switchgrass) is now projected. Crops in temperate climes where photosynthetic activity is boosted for production and for less H2O loss (transpiration) in the more humid, night hours, allowing for diminished rainfall in America's breadbasket by 2080 will be possible with CAM (Crassulacean acid metabolism) photosynthesis, a type of photosynthesis that evolved in plants that grow in arid conditions where transpiration occurs during the night (rather than day time) and around the Rubisco protein, for greater photosynthesis.
SKYEVIEW: Recent finds studying cellulolytics in the rumen and their interaction with pre-formed amino acid nitrogen (PFAA-N) and attempts to tie in treatments to slow protein degradation and make them more available for rumen microbial uptake during digestion show in certain cases stimulation of PFAA-N uptake by cellulolytics and also for water soluble carbohydrates (WSCs) in perennial rye grass varieties where digestibility and later lactation is significantly improved accompanied by increased dry matter digestibility (%DMD), neutral detergent fibre (%NDF) and acid detergent fibre (%ADF) digestibilities.
SKYEVIEW: There is an indication from the published literature that such anaerobic "ligninolytic" enzymes are beginning to be revealed, identified and characterized as to their mechanism and kinetics to indicate how possibly effective they are. The enzymes for facultative anaerobic rumen fungi and strictly anaerobic fungi are esterases and dearomatization "ring" enzymes contributing to separating lignin from crystalline cellulose and leading to ring fissure. There is also a need for additional energy (ATP) for lignin oxidation contributing perhaps to its relative recalcitrance to degradation.
SKYEVIEW: The current approach to chemically-controlled ensilage in grasses and legume fodders independent of weather conditions at harvest has been via the addition of sugar substrates such as molasses and concentrate (e. g. milled grain) although genetically modified biocrops are a possibility using current tools of maternally mitochondrial segregated DNA 'encapsulated' from genetic drift and safely with biolistic transformative techniques that only act when released at harvest, i. e. those enzymes that degrade buffering capacity from citrate, malate and glycerate. Malate dehydrogenase is one example of such an enzyme. Other microbial systems are currently being investigated to find and annotate via bioinformatic genomics the correct genes for the enzyme(s) responsible for buffer degradation. (The other approach to decreasing buffering capacity in ensilage is to down-regulate synthetic genes, not mentioned here.)
SKYEVIEW: At Skye Blue we are proposing a product as a spin-off of the humeral response mechanism is a Hygienic Spray Delivery System (HSDS) against bacterial and odour contamination on solid surfaces, solid wastes and soils. Monoclonal mixtures (idiotypic specific) to strains in humans and the environment will be screened for IgG, IgA, IgD and IgM production that have been lyophilized (stored prior at -5 degrees C) and reconstituted and delivered on a carrier of buffered-saline and preservative. Organic farming and increased use of animal body fluids and wastes for horticultural gardening will ensure against exposure to pathogens in the environment including waste handlers, solid waste treatment operators and decontamination of physical plant areas, equipment and materials. One approach already in the market is an in vivo delivered vaccine against E. coli contamination in meat and dairy products. This fine-grade industry cleaner can be used to decontaminate farm equipment, implements and materials from farm handlers used in nematode infected soils designed to kill their eggs and cysts in roots by washing, a current problem being addressed (see above). For a similar technology as outlined here with Canadian rights acquired for biological decontamination, see: staff, at Ideaconnection.com.
SKYEVIEW: The search for idiotypic determinants on protozoan cell surfaces to control them immunologically in the rumen will be used first with use of fluorescent-tagged anti-bodies (Abs) in an assay for transport, structural and enzymatic surface proteins. Those closest to the surface with greatest surface area and affinity will be chosen and possibly porins or ion channel and other transport system proteins. Monoclonals will be used for this assay. The question arises what correlates between in vivo and in vitro studies of this reaction, i. e. an assay with fluorescent tagged immunoglobulins (Igs) derived from rumen fluid will be used for this correlation.
SKYEVIEW: A new development at Skye Blue with pre-biosis and biocropping programming for ensilage with subsistence traits that are so-called pro-poor or friendly such as down-regulated protease activity and up-regulated water-soluble carbohydrates (WSCs) would lead to better microbial efficiency in terms of improving silage amino acid profile (to be further explored), improved volatile fatty acids (VFAs) profile, decreased protein and amino acids degradation and NH3 production resulting in better silage overall quality and improved nutritive value.
SKYEVIEW: Flores, et al. (1986), McGill U. in animal experiments with sheep probing methods to measure intestinal flows of amino acids using various dietary treatments such as grasses fresh and those that had undergone degradation and processing through the rumen stomach demonstrated that digesta solids collected and analyzed for total amino acids showed significant differences in amino acids levels in diets that had degraded reflecting dietary influences and despite microbial resynthesis to protein from microbial cell protein (MCP). It was concluded that continuing studies were needed to support evidence that nitrogen (N) status of animals affect their intake in kind of the corresponding diet. Here we reveal a point of controversy regarding Skye Blue's interests in studying the digestive tract as a endocrine organ that has a larger yet to be discovered role in mind-body axis function (see: MEDBIOS). The duodenal digest samples collected were thrown out shortly after subsampling without collection of the liquid supernatant, viz.: (1) definition of what is the liquid phase of duodenal and other parts of the digestive tract, (2) what mixing and propulsion means in terms of bulk flow and use of so-called solid-phase and liquid-phase markers to correct for random effects and (3) use of concentrations of duodenal amino acids in the 'liquid' phase that might impact physiologically the digestive tract, viz. intake response and/or feedback loops to the CNS. Two theories are to be tested here: what blood vessels (viz. endocrine) or neural circuitry similar to liver that respond to amino acids/peptide levels in the digestive tract.
SKYEVIEW: The method of re-entrant cannulation and sample collection and infusion with the s. intestines would be at points along the duodenum, jejunum, and ileum and involve measuring samples at each of these points for whatever parameters are required at these points [viz. free amino acids (FAAs)] at the duodenum, jejunum and ileum impacting that section of the digestive tract, or controlled studies measuring and confirming their impact by infusement of FAA mixtures at these points. It is thus possible to corroborate findings from duodenal FAA flow measurements (viz. concentrations) and their possible impact on intake at the level of the digestive tract or s. intestines with controlled infusion data. The findings, for example, would be interpreted as ff., if the intake response were significantly higher at infusion points further down it can be assumed that there are 'receptors' there; at points, if lower, then 'receptors' further upstream from those points. The practical benefits of studying possible linkages between FAA levels and dietary treatments is to find the ideal protein solubility point, for example, in grass forages or grains and other introduced hi-essential amino acid proteins recombinant in these feeds or as processed bypass sources.
We are a company with an interest in Biomedical/Biotech research. Enzyme bioengineering for food/feed applications and use of probiotics in the rumen is on the drawing board. So is Bio-crop (Gen) (generic) development defined as a class of commercial plant crops with practical scope for both crops in feeds, food and bioenergy production. We also have an interest in biomedical instrumentation/device and manufacture for electrophoresis, functional food and applications and are being offered at this time for further collaboration. Bioenergy for economic and eco-friendly, sustainable development is a goal.
Livestock byproduct feeding and processing are under development. These include probiotics for feeds (e. g. tropical and temperate ensilage) including ground-breaking research on lignase enzymes currently undertaken at Skye Blue and ruminal probiotics including conjugal transposon technology and with fungal culture (reported from Australia).
Bioengineered cellulases are currently under development with research on their catalytic domains and site-specific mutagenesis and structure/mechanism, cooperativity of amongst these class of carbohydrases and their 'scaffolding' with carbohydrate dockerin molecules to be applied to feed biomass processing of ligno-cellulose together with lignase enzymes to produce glucose converted to fructose/glucose feedstock with molasses (from beet or sugarcane) for bio-butanol production and for animal feeding converted to glycogen or starch, a hypothetical process at this time, using known enzymes, Candida utilis (var. Major) and Trichoderma viride cellulases applied with yeast bagasse and/or lignase-cellulases in combination as top dressed enzymes or in microbial Lactobacillus plantarum culture for temperate ensilage.
The issue of probiotics in the rumen vis-a-vis other means of supplementation on low-quality feed byproducts such as pre-treatment and energy/protein byproduct concentrate feeding presents the following comparisons: 1) the use of boosting fibrolysis (e. g. with cellulases/hemicellulases and lignases) is of use especially with low planes of nutrition (e. g. with v. fibrous feeds or diets and water buffaloes who characteristically consume them), 2) the use of SCP in yeast fermentation and essential amino acid nitrogen (E.A.A.-N) and with probiotics with boosted branched-chain amino acids (br-A.A.s) can boost microbial cell protein (MCP) synthesis and 3) total nitrogen (TN) which includes non-protein-N (NPN, A.A.-N and other N) in comparison presents no added benefit at certain levels with ammoniated straw and straw supplemented with protein concentrate, except, as has been hypothesized when the protein A.A.-N is protected which apparently effects more efficient uptake with greater efficiency of MCP synthesis and stimulation of cellulolytic activity and fibre digestion in the rumen.
The means of increasing digestibility, and especially, fibre digestibility would be first to remove the first barrier of enzymatic attack on fibre, lignin, either by: decreasing lignin content genetically as has been done with certain varieties of forage, physically dissociating bonds between lignin and fibre (specifically hemicellulosic moieties) by steam explosion (elevated moist temperature and pressure), ammoniation pre-treatment of fibrous residues, bio-bleaching with aerobic type-III lignase or lacasse, as they are called, and finally use of anaerobic lignases (there are outstanding issues of the level of enzyme production and the efficiency or Kcat of the enzyme) from rumen stomachs which participate in digestion of fibrous ingested feeds and from human waste municipal collection sites. After the first action here there is then the issue of the addition of the cellulase gene as with genetically modified (GM) processed feeds as with: temperate silage, yeast bagasse fermentation and compounding and ammoniation with symbiotic species. (Solid-substrate fermentation (SSF) using Basidiomycetes might require genetically improved aerobic lignases that act quicker to depolymerize lignin so much as to spare organic matter loss due to fungal spp.). Freeing fibre from lignin by physical treatment or decreasing the lignin content by about half has led to a 15-20% increase in digestibility points and it is only estimable at this point what cellulase (and other fibrolytic enzymes) would do to further improve performance.
Bio-crops (Gen) varieties still have not been produced for markets and has at this time been regulated for use only with pesticide control and herbicide tolerance although it also has been proposed that widespread research activity on boosting sugars beneficial to microbial digestion and other prebiotic effects (e. g. anti-protozoal forage, low-protease forages wilted when harvested and ionophoric peptides coded feeds) with animals be produced with forage spp. using both marker identification and mapping to important gene determinants and used in molecular assisted selection (MAS) or breeding and genetic modification (GM).
SKYENEWS: Low-lignin forage varieties their regulatory approval and use in livestock productions will be used in future in the upcoming 2nd edition of the book: "A Compilation of Ligno-cellulose Feedstock and Related Research for Feed, Food and Energy." There are two ways of genetic modification to low-lignin crops or cultivars. (1) There are the long-standing and naturally bred brown mid rib (bmr) mutants which are signficantly more productive with livestock in milk productions or output in dairy cows with corn and sorghum varieties. (2) And there are the genetically modified (GM) alfalfa varieties of which have been approved by the U. S. Dept. of Agriculture (USDA) which use alfalfa-derived CCOMT genes, that metabolically disrupt lignin's biosynthetic pathways effectively and thus total lignin biosynthesis, rather than other varieties that use bacterial-derived CCOMT gene copies. Further, low-lignin manipulations for optimum use with various lignocellulose feed bioprocesses are outlined: (1) sulfur dioxide (SO2)/steam explosion (SE), (2)urea-ammoniation with anaerobic lignase (type-II) (yet to be further studied, characterized and defined), (3) solid substrate fermentation (SSF) and newly discovered fungal lacasse spp., (4) "Yeast Bagasse" Process and (5) pulping with bio-bleaching. Lastly, we will address manipulating lignin biosynthesis and structure and composition of monolignols for production ends. These include (1) silage fermentation, (2) rumen fermentation, (3) pulping of paper and animal feeding and (4) saccharification for food and energy.
An area that has now reached considerable attention with hopes that a robust crop that is safe, pest resistant, environmentally adaptable, possesses good seed nutrient composition and has good heritability can be developed. The seed is already a reality and shows no adverse pleiotropic effects (due to possible imbalance of metabolic fluxes) on seed development. The high free threonine (Thr) soybean developing seeds was developed by introducing bacterial XbAK [aspartate kinase in the amino acid branched pathways with Thr, Lysine (Lys), Methionine (Met) and Isoleucine (Ile)] gene variants that was site-directed mutated with the replacement of Thr-359 for Ile and Glutamic acid-257 (Glu-257) for Lys showing less allosteric feedback inhibition but no changes in other kinetic parameters except a lower KmATP. The alleles called XbAK-T359I and XbAK-E257K with the use of high speed genomics and tagging and gene markers where introduced into seeds using seed-specific promoters 7Sa' or USP997 and a chloroplast plastid targetting sequence CTP1. The bacterial gene XbAK was recovered by HPLC.
There is an outstanding issue that is to be addressed in future likewise regarding essential amino acid with food consumption in food/feed sources that supply commensurate EAA content in addition to increasing caloric intake in food production which, in actual cases at this time, would be: Lys and Trytophane (Trp) in maize, Met and Thr in soybeans, Met, Cysteine (Cys) and Ile in potatoes and Lys and Thr in rice.
Genetically modified (GM) and non-GM feed processing and cropping for small farms in tropical developing countries is being called of at this time with the spectre of overpopulation and pressure to produce more of what is already stretched food resources such as with India.
The use of alkali was earlier proposed to treat ligno-cellulose in fibre feed residues but lost acceptance due to its lower cost-effectiveness compared to feeding supplements (e. g. protein and energy concentrates from farming, co-op and industrial feeds and food processing). High-tech or GM technologies with crops (lowering lignin content and boosting water-soluble carbohydrate (WSC) and non-structural carbohydrate (NSC) (that are shorter oligomers and ferment faster resulting in more rapid, efficient energy yield) has proven to be comparable in nutritive value compared to supplement feeding.
It has been proposed that GM feed processing together with GM crops might present new approaches to improved feeding practices including the ammoniation with GM symbionts of GM low-ligin forages and their residue byproducts which would be more cost-effective to another alternative of feeding energy supplements and in addition supply the requirement for crude protein-N for microbial fermentation to be used with GM-feeds with bypass or escape protein. Also, introduction of WSC and NSC in the GM biocrop would improve microbial efficiency and yield on the energy in biomass fermented further, as was mentioned.
It is expected that it would be possible to approach the problem of boosting chorophyll a & b content and the prospect of boosting production of biomass in crops for feed and biofuel feedstock such as has been proposed with high efficiency sugarcane and sorghum species. The multiple trait-linked expression of chlorophyll production and its overproduction will prove a daunting challenge to scientists including possible pleiotropic effects. The photosynthetic action of chlorophyll is believed like energy metabolic pathways to be boosted in turnover when genetically expressed. Further to this are the phototropic pigments in photosynthetic organisms like algae which have a greater efficiency in photo conversion which can be architecturally (i. e. structurally) stacked to complement plant photosynthetic activity.
Growth areas in plant-based products include energy biofuels [e. g. bioethanol, n-butanol (ABE), biogas (CH4) and biodiesel and phyto-olefins]. Also, there are biofine resources from biomass sources including paints, solvents, adhesives, resins and finishes, construction and packaging material including those from polymeric resins, plastics and polysynthetics for clothing, outerwear for fashion, recreation or sportswear and a vast area of other environmental applications.
In lastest developments with algal biomass production for cosmetics, pharma, feed, food and bioenergy (oil) applications, photobioreactors that maintain temperature with mixing/heating with power plants (e. g. CH4), a branched design which offers maximum light exposure with use of specialized plastics preventing biofilm formation and cost-effective use of CO2 and nutrients including waste water and on-going selection of fast-growing spp, will allow practical delivery of systems several hundred hectares in area. Areas that have been used for their ideal weather conditions are New Mexico U.S.A. and Queensland Australia.
In studies in the Philippines there could be cultural, socio-economic and technology transfer advantages for the production of increased dairy lactation from the increased practice of feeding fibrous residues, byproduct generation of whey from cheese dairy production and its use further in biogas production by the rapid process natural gas (RPNG) and biodiesel production (methyl esters converted from triglyceride or lipid) from photosynthetic algae (and soon also to be developed from seaweed in seawater field tanks) in tandem with dual sugar fermentation to n-butanol (acetone, butanol, ethanol)(ABE).
The use of wet fermentation versus solid-state fermentation as a point of reference has identified rumen or silage fermentation in addition to biopharma and biofuel fermentation in the wet category and feed treatment with fungal culture (e, g. Basidiomycetes) in the 'dry' category. There is a breakthrough with the latter with the prospect of using lignases to a limited extent to 'top dress' fungal fermentative lignocellulolysis of fibre making energy more available and making available their natural detoxifying biofermentative action. Feed residue feeding practices in the Philippines with water buffalo with high producers of caramilk would make this an ideal area to make it more practicable and a score a breakthrough in caramilk production in the country.
A study particular to Skye Blue and under publication (see: Flores et al., 1986) is the role of simple sugars on rumen fermentation and volatile fatty acid (VFA) production and degradation of proteins and their rumen lability and contribution to proteinogenesis microbially. There is data available that relates these two factors to proteinogenic efficiency as possible candidates for further research.
An area we are involved with research at the moment with ensilage (tropical and temperate) and rumen digestion and its manipulation is biotechnology applied to ensilage and probiotics in rumen digestion. In the tropics haymaking would be the method of choice. But also in the tropics ensilage, where there is a marked wet season (e. g. China and Australia). Also limited forage supplies during the dry season is making ensilage a practice to consider of choice for throughout the year. The meteorological seasons for planting, byproduct production and use, C4 output or yield and potential to treat fibre and improved ensilage practices would make world-wide production of crops for ensilage of increasing choice. The use of fibrolytics to increase the sugar availability should result in decreased NH3-N (nitrogen) and increased VFAs (volatile fatty acids) production. Preliminary data shows that osmotic pressure restriction and protection of nitrogen (N) would be called for with tropical ensilage. Transport processes of amino acids (AAs) may be limiting the efficiency of MCP (microbial) synthesis. Decreased NH3-N and increased VFAs would be consistent with increased N utilization for MCP synthesis and increased output of VFAs from increased availability of sugars and carbohydrates from fibrolytic enzymes. Future work with ligninolytics will be a major breakthrough with fibrolysis in feeds and rumen digestion. The annexation of lignases with ammoniation of straw, is an application with feeds pre-treatment and use of this biofermentation process. Yeast and fungal probiotics are the only probiotics that sucessfully persist in the rumen and have demonstrated benefits to rumen digestion and are commercially viable propositions. RNAi small molecular techniques with low energy of maintenance conjugative transposons with operon elements for controlling enzymes (viz. cellulases, hemicellulases, pectinases and lignases) are possible applications. There are no known studies of operon control mechanisms/modeling with rumen microbial species. It will be assumed, for now, that a lac operon-like model where cellobiose together with other low molecular weight (LMW) mRNA product(s) acts as an indicator for fibre digestion and acts on the operator to prevent expression of cellulase allowing for RNAi to be introduced in microbial DNA and blocking LMW products from expression acting to cause 'run-a-way' production of fibrolytic enzyme(s).
SKYEVIEW: Probiotics with rumen digestion to supplement poor quality residue-based (PQR-based) feeding have been further advanced with rec-DNA bio/technology.The use of conjugative transposal technology with bioengineered cellulases, solids-phase persistent microbial species and further exploration of growth-limiting membrane transport processes have been mentioned in the literature as further technicalities. Approaches that have advanced feeding: use of lignases in tropical/ temperate ensilage to optimizing nitrogen and energy availability for proteinogenesis from the rumen, the use of fungal solid substrate fermentation (SSF) with Basidiomycetes with improved aerobic lignases and their efficiency in tropical/subtropical climes, the use of lignocellulosic heat co-generation and SO2/steam pre-treatment of the bagasse substrate to optimize biofermentation with fungal spp. Candida utilis and Trichoderma viride and applications of lacasse type-III lignase bio- and/or chemical bleaching to substitute fibroin for grain in animal feeding, and even further the 'reverse synthesis' of starch from fibre, are options being investigated.
SKYEVIEW: The issue of agricultural byproducts both from farm waste which was burned in fields and/or used as mulch and agro-industrial byproducts (AIBPs) processing (e. g. rubber seed meal, rice bran, oil meals, tofu curds, spilled wheat flour etc.) have been proposed and yet to be implemented in practice and has recently been transferred in concept as a practice to industrialized countries as was recently reported from Japan where feeding studies providing comparative total digestible nutrients (TDN) [a comparative weighting system totalling the nutrient components making up the feed ration digestively available nutrient fractions (%)], weight, carcass composition and radius proves to be comparable to controls. (See also: LIGNO-CEL MILL CENTRAL; AGROTECHNICA)
SKYEVIEW: Industrial pre-treatment (IPT) of poor quality residues (PQRs) with IPT technologies (e. g. including steam explosion (SE) which disrupts the ligno-cellulose and makes it amenable to exposure and permeation by type II lignases and bio-bleaching wood pulp with lacasse-type III lignases, yeast fermentation to increase availability of cellulose and balance nutrients in bagasse, straw, and legume leaf haulms and for saccharification and fermentation to bioethanol; leaf meal pelleting with grain/molasses-based binder and heat extrusion) will increase with adoption or importation of technologies where physical plants are established (e. g. wood pulp, feed and sugarcane mills and cereal granaries, amongst others) that can also supply co-generated energy in electricity from bagasse and straw. Order a copy of Biotechnique Communique or Monthly Newsletter, with e. g. s.: "Yeast Bagasse Process", "Ammoniation of Rice Straw", "Lignase as a Feed Enzyme", "Carbohydrase Starch Synthases for Feed", "Anaerobic Lignases in GMO Autochthonous Ammoniation of Rice Straws" & "Sugarcane as Feed".
SKYEVIEW: Find this book just out in AU: http://www.angusrobertson.com.au/book/a-compilation-of-ligno-cellulose-feedstock-and-related-research-for-feed-food-and-energy/39555482/
Featuring also: http://www.blackbondbooks.com/location2010.html
SKYEVIEW: The feature publication is available on Amazon.ca, hardback, paperback and e-book editions; also in Kindle eBooks with Barnes and Noble and for tablets: iPads, Tivax 7s, Vizios, and Nooks. Sales are also through Xlibris.com stores online and direct call-in.
SKYEVIEW: Sugarcane is the queen of C-sink crops in the future and should be developed for flooding, drought-resistance, nitrogen fertilization, rate of photosynthesis and weeding management, as important agronomic traits to cultivate in places the British should be contracted not in neo-colonialitic style or modern imperialistic overtones but entrepreneurially with inclusive developmental strategy in mind as they have the geo-political spirit and drive and perhaps arguably know-how to develop the "Stans". It is arguable that the British could participate in ventures as these as with soya ventures (Africa, China, and Brazil), coffee trade (N. America vis-a-vis S. America) and now the sugarcane trade for feed, food and energy as in tropical areas like S. China and SE Asia, Indian sub-continent and African continent in the area of the globe to augment the current production of beets in Russia and former Soviet socialist republics for sugar and in N. America, as well.
SKYEVIEW: A recent find reported in the literature were studies in the journals BioResources and Chem. Eng. Res. & Design involving solid substrate fermentation (SSF) in particular wheat straw (WS) and pineapple leaf fibre (PALF), amongst other agri-wastes (e. g. rice and wheat straws, corn stove, banana stalk and sugarcane bagasse), reporting a breakthrough earlier discussed by Flores (2013) on the SSF Process with enzymes and fermentation processing of bagasse (Chap. 11) where as much as 47.2% less Klasson lignin was achieved with no change in cellulose or hemicellulose content with basidiomycetes spp. Ganoderma lucidum. It still remains to be seen if uneconomicabilities in the process can be overcome although in more industrialized settings and where ambient environmental conditions allow this may be feasible. Also, in industrialized settings such as in developed countries lignase enzyme production will of course be on the rise with these type-I (LiP, MnP) and type-III (Lac) enzymes with new sources showing greater potency and higher yields which are substrate dependent (e. g. rice straw shows an advantage amongst agri-wastes mentioned).
SKYEVIEW: In temperate-subtropical climes, the seasonality of post-harvest treatment with SSF (see: above) is suggested by the fact that ambient temperatures of 20-30 deg C is required for fungal growth and thus baling of farm byproduct wastage other than what is mulched be left to the Winter in the field and then processed in the later Spring and following Summer. Because early Fall is the harvest date for many farmers it is not possible to process agri-wastes from harvests that are fermentation-driven as we are discussing here at SkyeBlue using non-GMO and future GMO-improved approaches. Another case in point is urea-ammoniation.
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Future Topics to the Application of Agro-Biotechnology. The future of agrobiotechnology research in various field disciplines.
Chapt 1. Animal Feeds Science and Technology.
- Biotechnology has a role to play in further improving the quality and capability in processing feeds: a) post-harvest technology, b) field-drying.
- Storage of feeds: a) ensilage or feed conservation over seasonal feeding practices; ensilage here is an important topic both for the tropics and temperate countries where infrastructure and mechanization and various inputs are factors where feeding is all year round, b) biofer- mentation, c) feed additive enzyme technologies; feeds are better-utilized forages, more digestible and nutritious.
- Biocropping now includes an arsenal of: a) adding protein surrogates that are high-quality protected proteins sources used for intensive feeding, b) herbaceous anti-protozoals in feed, c) low-lignin grasses, legumes and corn cob, making for low-lignin high moisture ear corn (HMEC), 4) feeding ionophores as additive supplements, 5) feeding byproduct fibrous agricultural residues (FAR) and agro-industrial by-products (AIBPs) as protein and energy concentrates; they all improve proteinogenesis in rumen digestion and boosting propionate production highlighting this need in producing meat and milk where protein is the first limiting nutrient factor for production.
Chapt 2. Cereal Biology.
Molecular breeding can be used to improve agronomic traits including yield is also being addressed to improve growth and better performance of cereal crops.
- As a protein feed source, food staple and potentially valuable feedstock for energy, cereals are now beginning to be exploited to their fullest potential using plant molecular breeding techniques to manipulate: a) amino acids and protein; a unique example of dramatically increasing protein output per hectare with corn is to cause twinning of the kernel through hormonal manipulation; although the starch content is the same as in a single kernel, the protein content is doubled, b) starch and c) fibre.
- There have been pleiotropic effects found in grain when studies manipulated cereal nutritive composition and it still remains elusive when the ‘holy grail’ of cereal biology is solved as with grain that grow at a higher density per land area with greater grain size or weight and protein and starch content.
Chapt 3. Forage Science.
Forages are a staple in the ruminant’s diet. Agronomic, qualitative and animal utilization traits are beginning to be exploited for better use of this class of feeds.
- The agronomic traits include greater hardiness to environmental conditions (including salt, drought, pestilence, weeds) and inputs including water, energy and land.
- Nutritive quality traits including protein, water-soluble carbohydrates, vitamin and mineral content.
- Animal utilization traits that pertain to rumen factors with feed that act to increase proteinogenesis by rumen microbes and propionic acid; these are examples: herbaceous non-protozoal compounds, protected or relatively insoluble proteins in plant leaves and complex, including water-soluble, carbohydrates. More digestible, nutritious feeds better utilized by the animal have resulted.
- Biomass production in forage crops will not only produce feed but also diversify use of byproducts with development for the production of alternative chemicals, textiles and bioenergy.
Chapt 4. Plant Molecular Breeding.
- Genetic-bearing transfer systems and protocols using plant cells and regenerative methods and breeding have made the accurate, precise and safe transfer of specific genetic qualities between species in plant hosts a reality.
- Marker assisted selection (MAS) has allowed natural breeding at a more efficient rate and done more precisely.
- Mitochondrial sequestered cloning of rec-DNA has allowed for the biological containment of genetically modified organisms (GMOs).
- Biocropping will cause a revolution that will confer more productive, better performing traits to species including faster growth and flexed harvest seasonality, environmental hardiness and friendliness.
Chapt 5. Rumen Microbiology.
- Biotechnology’s role in manipulating the rumen ecosystem effectively through better fibrolysis and microbial protein synthesis at higher efficiencies is a goal that has yet to be attained using the required genetic methods that confers competitive advantage to these foreign, transformed rumen microbial species. It has been considered a major milestone in the attainment of biotechnology and its applications to animal nutrition.
- Technicalities are hurdles that remain in the realization of this technological first. These include: a) low-energy of maintenance vectors such as transposons that insert short sequences of DNA and then abscond as a suicide vector in the chromosomal DNA, b) cellulolytics that subsist in the solid phase and are less likely to washout, improving through nanotechnology & design uptake mechanisms of peptides and amino acids and water-soluble carbohydrates, c) gene-silencing to instigate via transcriptions factors (TFs) run-a-way enzyme production and d) yeast and fungi fed directly in the rumen has resulted in improved protein flow from the rumen and intake; e) the in situ application of host immunocontrol of protozoa has also been investigated as another approach with biotechnology to eliminate the inefficient activity of protozoa in the rumen. The current controversy over the use of probiotics in the rumen and their persistence, albeit, except known inoculants of certain fungi and yeast cultures, is interesting to note in the case of K. Gregg et al. (1989) where they tracked fluorometrically in the rumen species of hosts transformed for enzymes that breakdown fluoroacetate, a toxic substance in plants ingested by sheep, which suggests that microbes bound to particulate feed is a requisite for recombinant microbes compromised by their transformation to persist in a considerably competitively and turning over environment as the rumen’s. A solution to this technical hurdle is to earmark microbial hosts in future that are bound and so with their secreted enzymes either on the microbial surface or feed particles. A recent astounding find from a technical standpoint has been the use of the ‘functional cellulosome’ or units composed of cellulases (e.g. various endocellulases, exocellulases, and cellobiases), dockerins and adhesins, amongst others, that can be recombined recombinantly, producing units that are more physiologically accurate and functional.
Chapt 6. Animal Biotechnology.
- Genetic basis for muscular development and fatness will allow animal molecular breeding in production animals consistent with desired production parameters. Single nuclear polymorphisms (SNPs) with marker assisted selection for specific or accurate reproductive breeding.
- Disease resistance are being studied which can affect morbidity rates, mortality and animal performance. The molecular breeding in this important area for animal production is still in its infancy for disease-resistant livestock.
- There are many other genetic traits in animals such as musculo-skeletal development that affect growth including their hormonal mechanisms and lactation for both the output and the composition of milk that can be used in molecular breeding.
- The techniques of molecular breeding include reproductive technology such as gene replacement or transgene therapy (pre-partum), cloning, multiple ovulation embryo transfer (MOET) and artificial insemination (A.I.).
- Bioinformatics will be used to locate genetic loci and their relative positions such as those gene markers that correlate with marble scoring in meat and the diameter of the longissimus to breed animal of superior meat carcass quality.
Chapt 7. Meat Science.
The use of molecular animal breeding with biotechnology in order to enhance or improve meat quality is a very powerful approach for animal producers desiring to increase the quality and value of meat products. The variables of the nature of this problem in animal production will be further investigated.
- Growth rate, lean body mass (LBM), marbling, microfibrillar structure and taste and texture qualities are all going to be correlated using bioinformatics tools to identify the gene markers that determine muscle quality traits.
- Cross-breeding will then be carried out to bring about the desired genetic background. Various meat products or cuts will vary in their improved quality with the highest value meat cuts of greatest value added.
Chapt 8. Soil, Water, Air Quality and their Management.
- Soil amendments can be used with a) organic-derived fertilizers as a dual-purpose organic amendment and on the otherhand as a crop from: a) seaweed, b) algae and c) seagrass; they will be cultivated intensively, in low lying meadows, man-made rafts and reefs, for use in bio-oil energy and olefin. These species will be cloned with biolistic methods of transformation with target DNA and maternally segregated in mitrochondrial DNA making it environmentally contained.
- Marginal land use and use of crop rotation or fallow land and to design hardy biocrops that fix nitrogen (N2), water reclamation from watersheds through conservation, desalinization in semi-arid or arid zones with large-scale irrigation systems and the building of wells and bottling/distribution water channels will answer to the growing demand for crop planting and human consumption.
- Continued global warming, population growth, energy expenditure use with coal-fired plants, vehicular transport, offset by measures such as alternative fuels including solar-based systems, wind power, hydro and, with uncertainty, the aging of modern atomic energy generators and the need to rebuild and maintenance, will in future paint uncertainty with the weather and its effect on droughts affecting the food supply and water, flooding and its effect on crop harvests and dry weather, winds and wildfires affecting vast tracts of wildlife and forest habitats, resettlements and air quality.
2006-2007/revised 2013 © Skye Blue Publications by D. A. Flores
ADVERT/NEW ORDER!: In Press. 2016. Friendly, Pro-Poor Rec-DNA Approaches in Small Farmer Crop and Animal Production: High-Sugar Grasses, Dual-Purpose Cropping, GMO Probiotic Cellulolytics and Phytobotanical Dual Cropping - White Paper. I. Introduction. Bio-cropping (i. e. the genetic modification of crops for improvement of utilization traits in biotechnological crops) and new insights and approaches to cellulolytics, one approach in modifying rumen digestion will be considered in this discussion. The rural, small farmer needs better tools to increase productive ends with greater efficiency in keeping up with upward food production and growing population demands using cost-effective ones including in-built recombinant crops with improved utilization (e. g. digestive) traits. II. High-Sugar Grasses & Down-regulated Proteases with Heat Protection/Field-Wilting. Clover and alfalfa, MAS breeding, recombinant approaches (maternal segregation) and biosafety, rumen simulation techniques to study efficiency and feeding trials with dairy cattle. III.Dual-Purpose Crops. Non-protein nitrogen (NPN) and the value for feeding fibrous crop residues (FCR) for food and animal feeds. Recombinant approaches to using surrogate proteins (e. g. albumins from legume seeds) in corn and rice, as examples. Feeding trials comparing dual-purpose NPN and urea-supplemented diets. IV. GMOs in Rumen Digestion. New approaches to cloning in the rumen: 1) the cellulosome (i. e. the complex of binding cohesins and dockerins and the cellulases) with high efficiency (GM)/low energy of maintenance, 2) use of suicide vectors (single-site insertion of short sequences with suicide of vector donor plasmids), 3) evolution of improved GM transport porins with increased capacity to transport pre-formed amino acids (i. e. peptides) to improve efficiency of microbial cell protein (MCP) synthesis, 4) and selection of donor hosts with recombinant enzymes that bind effectively with and that bind the particulate phase in rumen digesta and have better sustained half-life of retention in rumen contents. [Cf. TR Whitehead or see also here: ICAR(New Delhi, India), Hum-molgen.de. March 2014 - ongoing (present). On the latest updates on research in rumen microbiology.] V. The need for increased expertise in the use of GMOs that are environmentally safe on the small farm scale level is required in the future to widen the repertoire of the farmers outputs for the production of phytobotanical drugs or pharma from plants at a much reduced cost compared to bioreactors.
ADVERT/NEW ORDER!: In Press. 2016. GMO Approaches to Feeding High Protein in Lactating Cows - White Paper. There will be growing demand for not only improving the output of milk of which milk proteins is a limiting factor but also milk constituents touted with functional health benefits for the future world market (viz. anti-inflammatory qualities to be found in milk protecting against a number of disease related to chronic inflammation including cancers as has already been speculated for anti-oxidants such as beta-carotene in "golden rice"). Current approaches with hypothetical GMO cows using promising unregulated technology in genome edited genetic engineering are: 1) feeding of energy fibrous byproducts based feeds along with non-protein-nitrogen (NPN) and protein supplements along with proposed GMO probiotic technology that will both, with genome editing, increase the limiting amino acid supply in the metabolism of rumen microbes (see: Flores, 1989) and which can supply precursor to coded pre-formed supplies of "storage" proteins either native or introduced, 2) GMO cropping approaches using low-protease pasture as forage that have been field-dried as post-harvest treatment to increase its protein content before feeding and after the digestive process and 3) as a non-GMO approach the conventional feeding of byproduct protein concentrates.
Last update of this entry: April 06, 2021