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SKYENEWS: We are further asking for bioengineering collaborators for organically-synthesizing hi-performing 'branched' fibre into supplements to regulate hypercholesterolemia in the diet although new advances in designing sequences of DNA recombinantly (rec-DNA) using primary-secondary structure via cryo-TEM (cryo state transmission electron microscopy) which gives elaborate structural detail and poses the question of whether proteins can be radically dissected and reassembled, i. e. genetically modified (GM) by computer design for new mechanistic reactions and with individual residues for reactive mechanism; a fructan inulin biozyme reactor similar to existing artificial sweetener synthetic devices and design of a processor for lipase-treated oleo unsaturated 'L-fat' from vegetable sources are planned.
SKYENEWS: It was noted recently at Skye Blue that boosting in microbial systems such as the well-known effect of water-soluble carbohydrates (WSCHOs) on microbial cell protein (MCP) synthetic efficiency (kg OM / moles ATP) and 'catalytic' effects of lactose in whey on biomethanation, for e. g., in animal waste would be, if immediate in time-scale by effects on the unit of metabolism, the enzyme(s), not via allosteric effects as enzymes using entropy as a concept can only be synthesized by residue substitution (and as singlets alone as has been and now found in e.g.s.) although they in comparison can be inhibited only, leaving 'agonist' theories, as has already been used with drug applications.
SKYENEWS: For a similar technology as outlined here with Canadian rights acquired for biological decontamination, see: staff, Ideaconnection.com. 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).
SKYENEWS: We take particular interest in animal research on: reproductive biotechnology & physiology, animal health, animal nutrition, genetics & breeding, meat products and waste management and concerns with increasing calf production and mortality, forage quality, milk production, increasing AI efficiency and socio-economics (e. g. livestock for food, as capital, income amongst small farmers).
SKYENEWS: Recently, (cf.: Ma. Visitacion D. Guingab, Isabela State University, the Philippines.) it was proposed in the Philippines that tropical grasslands which predominate small farmer agriculture be replanted into C4 C-sink crops, sugarcane being a king of crops (cf. sweet sorghum, yellow corn.) for this. This is a v. tantalizing idea for farmers and agri-field workers weighing the benefits of paper production, feeds production en mass and for bioenergy fermentation vs. in situ cropping in grasslands and forested undergrowth and in mixed-farm systems (e.g. livestock cum palm oil plantations, rubber tree plantations) although it is predicted that s. farmers will still predominate in the near future unless urbanization increases at an accelerating rate and jobs are created on the otherhand leading to larger commercialized operations and/or co-ops (albeit, less efficient and profitable ones).
SKYENEWS: Our interests oulined here include food, biologicals and biomass cropping (cereals, fibre crops, grazed pasture, oilseeds, root crops, sugarcane, fruit, tobacco) for feed crops and its improvement including interventive approaches (e. g. pretreatment, probiotics) to improve animal utilization, the problems of identifying and manipulating body mass and composition with nutritional inputs or programming with animal genetics, yet to be further investigated. Also, as tabled, there is a need to institute the ammoniation of straw and other fodders for subsistence small farming practices in developing countries.
Crop improvements as has been discussed elsewhere includes: 1) prebiotic, energy cropping and C-sink strategies to boost biomass for food and energy with increase of secondary cell wall structure with cellulose, xylan and galactans to help ensure food security using biochemical 'switching'; boosting xylans have proven recalcitrant to fermentation in feeding for biofermentation but new enzyme technology and their improvements may be key to the problem; it is unknown as to benefits from these fractions and endproducts or complex or non-structural carbohydrates and water soluble carbohydrates (WSC) from a production standpoint; prebiotic cropping with WSC, pre-formed protected amino acids, anti-protozoal compounds in feed, sequestered fibrolysis in feeds, dual-purpose non-protein nitrogen (NPN) cultivars, leaf-protein surrogates in sugarcane leaf tops and low-lignin crops, which offers new perspectives in feeds pre-treatment and probiotic feeding, 2) feeds pre-treatment (e. g. lignases and other related feed enzyme technology, ureolysis, anaerobic ligninolysis, aerobic ligninolysis including lacasses, yeast bagasse with cellulase), and 3) rumen probiotics in feeding (chromosomal conjugative transgene transposon genetics and fungal cultures); the outstanding issues of rumen turnover (liquid and solid phases) and microbial specific growth rates are to be addressed in terms of minimizing energy expenditure from microbial cell maintenance, association with the particulate or solid phases and any limitations in energy substrate rate of transport given the substrate pool in rumen fluid.
Biofermentation of feeds in addition to pre-treatment, use of probiotics in feed ensilage and the rumen and pre-biotics to improve feed utilization has recently been approached as wet and solid-state or 'dry' fermentation processes in the Federal Republic of Germany (FRG) and an approach using wet fermentation where there has been proven proliferation of microbes as with yeast introduced in the rumen (which reduces CH4 emission and conserves energy for volatile fatty acid production) and the proposition of another non-recDNA approach periodically innoculating the rumen with a co-culture cohort (see: Flores 1988, M.Appl.Sc.thesis, UNSW, Australia) of Lactobacillus cellobiosus that is facultatively anaerobic with Neocallimastix spp. using RUSITEC (R) be done to test the disinhibition of feedback inhibition (FBI) of fungal cellulases in the rumen. Buffer may have to be used as with feeding wet, well fermented silages which when fed can cause rumen lactic acidosis.
Currently, order in for a publication featuring anaerobic lignases and their role as feed enzymes and fibroin digestion discussed in areas such as improving lean body mass and composition and improving ensilage composition and utilization. This a novel area involving dietary treatment effects and nutrient intake on genetic variance response which is being further investigated with use of feed enzymes. It is known in the Philippines that Zebu cattle fed on roadside bunds and their green cuttings are lean and tender in lean body mass and so it can be argued with lamb production in countries like Ireland and New Zealand. Silage and limitations in fermentable water-soluble carbohydrate sugars as has been shown (see Flores et al., 1986) with fresh frozen herbage versus the ensiled mass as indicated by rumen pH can lead to a better match between energy availability from fermentable carbohydrate and nitrogen degradation and recapture for microbial cell protein (MCP) synthesis and increased nitrogen (N) utilization making lignases revolutionary with the ensiling process.
We are also further investigating the role of fibroin enzymes such as hemicellulases or xylanases and pectinases in rumen digestive nutrition and microbial metabolism, body composition and growth or output (e. g. body composition in lean body mass (LBM) and milk solids). The traditional practice of cut-and-carry of green, lush grass and other pasture high in hemicellulose in neutral detergent fibre (NDF) and pectin with livestock such as water buffalo, zebu cattle and goats should be further investigated in terms of balancing rations fed for productive outputs such as body composition and weight gain. There are applications to improving the efficiency of feeding pasture and intensive lamb production in the West such as New Zealand with the addition of feed enzymes such as cellulases, hemicellulases and pectinases. There is much to be believed about characteristic nutritional inputs and outputs such as with water buffalo and LBM and milk solids composition and zebu cattle which are characteristically lean in meat quality. In bio-pharma drug discovery anabolic cell signalling and phosphorylation of amino acids for LBM accretion would be an area for further investigation including other already studied mechanisms of musculo-tissue accretion which are future non-steroidal or bovine somatotropin (BST) with possible nutritional biochemicals or anabolic signalling growth factors such as insulin growth factor-1 (IGF-1) and products of Wnt7a gene family which has reversed muscular dystrophy, all approaches to increasing efficiency in body weight gain.
There should be further discussion and research among meat scientists regards grass-feeding versus grain-feeding and nutrient interactions with genetic variance and production of leaner beef or meat and markers to genes responsible for this, cholesterol levels in meats in leaner animals, growth factors in tissue involved with cholesterol levels in tissue inherent as genetic traits in such animals, Vit D and its involvement with cholesterolemia and sterol metabolism in research as a nutritional and higher omega-3-fatty acids resulting and cholesterol metabolism in meat animals and/or consumers of meat products. It is tantalizing to think that leaner beef is related to lower cholesterol and markers between the two traits used to investigate them further as there is lower cholesterol in water buffalo carabeef which is leaner and the same with the more distant background of turkey and whether it is just by coincidence or whether there is a direct link or interaction between the two traits and also whether there is a link found if cholesterol levels and leaness shows that same pattern in grass-fed versus grain-fed beef.
The issue of lean body tissue growth and fattening and roles of specific enzymes (e. g. marbling) and/or partitioning functions yet to be investigated physiologically and metabolically is being explored. Leaner grass-fed beef has improved commercial retail value from an increase in omega-3-fatty acid levels in those consuming leaner beef and should be a strong marketing factor including those preferring red meat in their diets but protecting against certain cancers and cardiovascular disease.
At the Univ. of New South Wales Sydney there is interest in both involving biotechnology with biomolecular sciences or molecular biology. With metabolism in general, and in particular, for example, anabolism and cell proliferation and cell signaling factors (e. g. growth factors and proliferation and differentiation), little is understood together with the understandable complexities of metabolic assay and so with the opportunity to introduce 'new pharma' in meat production. Biomolecular sciences (like genetics & genetic engineering with biotechnology) will have a significant role in furthering developments in future in meat science. Myostatin is a known gene responsible for inhibiting muscle growth and is mutated in a breed Belgian Blue which is more amply muscular in live weight and much leaner.
Probiotics (i. e. molecular agents in metabolism) to nutritionals (e. g. Vit D) have recently been found [See: Biomolecular and Medical J., Skye Blue Publications 2012 (c)] as such that are apparently coincidental, that are in place metabolically that serve to treat deranged states of disease. This could serve to predict what would appear to be a gold mine of drug discovery that could ensue and including small organic molecular drugs and the low molecular weight (LMW) proteome. Nutritionals could also predictably possess such probiotic relationships metabolically as far as growth and development is concerned in muscular tissue (e. g. actin & myosin) such as in specific processes of cell proliferation, cell differentiation and cell signaling.
Currently, at Skye Blue we are intent on furthering advances in ammoniation of straw (and stover and legume haulms) as fodders with isolation, identification and manipulation of plasmid DNA in autochthonous bacteria for increasing expression of urease (hydrolysis of urea to ammonia and carbon dioxide) and carbonic anhydrase (solubilization of CO2 to bicarbonate) enzymes to boost presence of permeants ammonium carbonate, bicarbonate and hydroxide in the lignocellulose/hemicellulose matrix and bring about ligninolysis. We will also add bacterial species that are viable anaerobically which can metabolize the end-products of ligninolysis to minimize any deleterious effects on ruminal digestion post-treatment. Cellulase from Trichoderma reesei will also be combined and the optimal expression dose response found between ureolysis and cellulolysis over the time course of sealed bag en silo, with the right temperature, moisture and partial pressure of ammonia. The extent to which ammoniation can be boosted en silo and still be viable for microbial growth is open to question. Ammoniation of straw with implementation, collection of field residues and technological enablement perhaps with economic incentives for use in India via Australian aegis with personnel and technology from their institutions as recently recommended is expected to be mandated soon including the hastening by pressure for fodder use for bioethanol with India being the sixth largest consumer of oil in the world. Ammoniation of straw with non-protein-nitrogen (NPN) has led to increased average daily gains, nitrogen retention, in part due to greater amino acid flow from the rumen and, it is found, that NPN can replace N in grain for utilization; research shows that at certain levels of supplementation that NPN is just as nutritive as amino acid-N and supplementation with concentrate, at high levels of inclusion, tends to minimize the effect derived from ammonia-N with straw.
The long-standing problem of balancing nitrogen utilization and energy availability in temperate forages for ensilage such as grasses and legumes can be approached by field-wilting or drying to protect protein with irradiation resulting in better assimilation into microbial protein rather than being lost to microbial protein degradation together with what is bypassed and at a higher plane of protein nutrition with use of protein bypass supplements which result in more optimal assimilation into microbial protein synthesis and bypass. Grazed ration nitrogen (N) can be diluted with more mature haylage to balance N degradation and energy (E) availability. Energy availability on forages would be improved, although it is up to speculation what results are with varying conditions, where total nitrogen (TN) is in excess, by increasing fibre digestibility using lignase enzyme addition to diets and other fibroin enzymes (e. g. cellulases, hemicellulases and pectinases). To increase E availability in grazed high N forages concentrates with readily available carbohydrates can be fed with grazed pasture.
We would like your valued feedback on a research area we are furthering, viz. enzyme technology and bioengineering and feed technology. We would like to put it forth that there is growing interest in enzyme bioengineering and feeds processing (e. g. ensilo, pre-treatment and feedstock conversion) and their role as future commodities in the marketplace together with grain and forages that are sold inter-alia between smallholder farmers.
June 22, 2013 / rev. February 21, 2014
Dear: Friends, Respondents and Guests to Skye Blue!
We recently got a letter from the Editor, J. of Biotech. (FRG) regards our mini-review proposal for a paper. We share here with you an outline of the paper manuscript that the editor may consider for publication as long as it demonstrates scope and depth as expected by editorial review by the editor. The title: "GMO Prebiotic, Proteinogenic Approaches in Rumen Digestion of Forages: Fructan and Amino Acid Metabolism".
In addition to this submission in this separate format we are also considering a Monograph which is accepted and being considered by Skye Blue to one of Cambridge Journals' Advances in Animal Biosciences on both basic and strategic science in whole animals and farming systems as it impacts productivity, product quality, food security, the environment, climate change or humans.
The journal is supported by the British Society of Animal Science, INRA and EAAP.
I. Introduction to Forages - Microbial cell protein (MCP) is a major source of protein with livestock. - Increasing productivity determined by increase in dietary intake, in turn determined by metabolizable energy (ME) and microbial protein flows (duodenal amino acids, DAAs); protein/energy (P/E) ratios in diets in the tropics are important in improving intake of animals undergoing heat stress. - Targeted productive ends are for growth, lactation and epidermal tissue and hair. - The focus of the paper is on protein as the most limiting nutrient towards animal productivity and also the fact of protein degradation and digestive utilization in the rumen of PFAAs and WSCs can be now prebiotically manipulated through biocrops as major approaches to directly boost the differential in g MCP/mol ATP, as part of the nexgen in GM crops, after the first generation in Bt and herbicide tolerant wheat and corn crops, as examples. Other approaches used for manipulating microbial protein intake (MPI) currently available is use of shrub or tree fodders with defaunating qualities and non-GMO probiotic yeast and fungi fed continuously to sheep. These approaches have not met widespread use.
II. Genetically Modified (GM) Techniques. - Water-soluble carbohydrates (WSCs), pre-formed amino acids (PFAAs) and their metabolic manipulation in forages. - Conservation of grasses and legumes and their ruminal digestion with respect to MCP synthesis vs that of grazed forages.
III. Fructan Metabolism. - Levans in GM plants and dilution rate determine MCP efficiency, especially in C4 plants.
IV. Down-regulation of Proteases. - Limit proteolysis in grazed forages and conserved silages - grasses and legumes. - But not to be used in C4 plants of low degradable nitrogen (N).
V. Improving Fermentation Characteristics of Conserved Forages. - Improving nitrogen utilization, increasing protein flow to the intestines of duodenal amino acids (DAAs). - Decreasing buffering capacity with sequestered enzymes for breaking down malate, citrate and glycerate. - Decreasing lignin content and increasing cellulose content in plants. - And use of sequestered enzyme lignases, cellulases, hemicellulases and pectinases as prebiotic aides to rumen digestion. VI. Conclusion. It is important to communicate to the Editor, at this time, that the efficiency differential offered by WSCs and PFAAs is primary first of all and in support of the more secondary effects on energy ‘pool’ requirements: 1) their increased transport across the cell membrane by genetic modification (GM) in probiotics (we assume that membrane transport is the most limiting step here) and 2) the fact that ATP from sugars from cellulolytics from feeding protein supplements of limiting solubility increasing amino acid 'pools' in the rumen stimulating cellulolytics and their activities providing more sugar for the ATP 'pool', should suggest these manipulations.
Chapt. Outlines, PART II.: " Ligno-cellulose Feedstock Biotechnology for Feed, Food and Energy." for 2018-21.
E.g. Chap. Anaerobic Lignases: Protocols and Possible Progress. In the current use of technologies to crack the lignin barrier with 'anaerobic tools', microbiology as it applies to anaerobic bioremediative processes (e. g. municipal activated waste sludge and marine ecohabitats) and the rumen pose scenarios where fibrolytics can be further explored. Polymeric fragmentation with LC/MS (liquid chromatography/mass spectrophotometry) and dimerized fluorometric assays to isolate, identify and further characterize enzymes for fibrolysis of ligno-cellulose and genomics to construct probes for hybridization studies to enumerate and identify microbial isolates with superior lignase activity and further genomics to resolve further "homologues" amongst enzyme activities. Polymeric fragmentation will require timed sampling and "distributional fingerprinting" of digested peaks on a homogeneous substrate of lignin (e. g. poplar).TFs (transcription factors) assays will further identify regulatory elements for lignase production. There is an indication from the published literature that such anaerobic "ligninolytic" enzymes are beginning to be revealed, 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 de-aromatization "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. The application of lignozymes (or lignases as they are formally or systematically known, from their substrate lignin) that belong to the lignase type-II group (anaerobic) (type-I, are aerobic from fungi in solid substrate fermentation, SSF, type-III, aerobic Cu3+-lacasses) are being investigated as to their rate or half-life of substrate breakdown for microbiotic spp. with lignases to be isolated, identified and characterized from soils or composts, municipal waste activated sludge (this shows most promise in terms of rate of depolymerization and mineralization of lignin backbones and ring fissures), from rumen microbiota of livestock including the carabao, Nili-Ravi or Murrah, and marine eco-habitats; the uses of type-II lignases (e. g. the depolymerases such as lyases, etherases, esterases and phenyl oxidases, and those enzymes that remove by dehydroxylation and demethoxylation of group substitutions on the aromatic rings of the phenyl propanoid subunits that constitute the building blocks and the 'larger networked chain of lignan', reduction of the benzene ring to the cyclohexane (2-hydroxycyclohexanecarboxyl coenzyme A dehydrogenase) and then ring fissure leading eventually to mineralization to CO2 + H20, thus far having been investigated in lignan's breakdown) will have far-reaching implications in both aerobic (they are stable, it is presumed to atmospheric O2) and anaerobic applications competing on all fronts with both the fungi in SSF of feeds biomass as well as with the practice of: 1) top dressing, 2) bio-cropping by sequestration until release when the cell dies upon harvesting or chewing by the grazing animal, 3) conventional ensilage to follow harvesting and 4) post-harvest technology such as release of sequestered enzymes with urea-ammoniation, yeast bagasse or with other feedstock or with steam explosion (SE)/sulfur dioxide (SO2) pre-treatment. There is much to be learned about the gains or improvements in production parameters with these new treatments and in combination with other treatments used in feeds post-harvest technology.
E.g. Chap.: Bio-cropping Self-subsistence Traits. Genetic modification of crops for improvement of utilization traits in bio-crops. The rural, small farmer needs better tools to increase productive ends with greater efficiency to keep up food production and growing population demands using cost-effective ones including in-built recombinant crops. 1) High-Sugar Grasses and Legumes. Clover and alfalfa, MAS breeding, recombinant approaches (maternal segregation) and biosafety, rumen simulation techniques to study efficiency and feeding trials with dairy cattle. 2) Low-Protease Crops. This facilitates greater utilization of nitrogen (N) in forage digested in the rumen in particular with the ensiled herbage and but with the grazed herbage the exception. 3) Dual-purpose Crops. Non-protein-nitrogen (NPN) and the value for feeding fibrous crop residues (FCR) for food and animal feeds. 4) Recombinant approaches to using surrogate proteins (e. g. albumins from legume seeds) in corn and rice, as example. Feeding trials comparing dual-purpose NPN and urea-supplemented diets.
E.g. Chap. Pre-biotic Manipulations in Crops. - Lignin. Lowering lignin, physiological implications to manipulation, recent developments in their manipulation. - C sink. Boosting C4-sink, from C4 grasses, increasing cellulose deposition in the cell wall, boosting water soluble carbohydrates (WSCs) and complex structural carbohydrates to improve output of plant crops and increase plant C uptake. Implications with livestock production and food security. - Genetically engineering proteases in plant crops and implications of their activity with ensilage or harvested and grazed forages. Fructan as a predictive case study for improving nutritional value of crops; health and environmentally sound genetic engineering (GE) using biolistic transformation with plasmid DNA via homologous recombination in calli using somatic embryogenesis with maternal side segregated chloroplastid DNA succession against field dispersal of foreign DNA using genetic boosting and/or TFs involved in increasing fructan content in crops. - Corn as a crop usually is characterized by having a lower crude protein (CP) content than is optimal for microbial cell protein (MCP) synthesis in the rumen and deficient in particular essential amino acids (EAAs). It would be desirable to predict the use of 'designer' crops that have been enabled with optimal EAA profiles or patterns (see: below, for discussion on Flores' hypothesis') which could be a revolution in animal feeding with concentrate as corn grain in the United States.
E.g. Chap. Processing/Storage. - Lignases and top dressing/rumen digestion of feeds: new and proposed developments in enzyme technology. - Steam explosion and SO2 pre-treatment of ligno-cellulose feeds. - Silage-making in the tropics/subtropics and temperate climes and their issues. - Ammoniation of silage: new developments to improve nutritive value. -The feeds pre-treatment and supplementation with single cell protein (SCP) of agro-industrial residue feedstocks. Yeast and other microbiotic fermentative processing to breakdown and saccharify fibre to sugars and supplement feedstock with protein an e. g. is that of making "Yeast Bagasse"proposed for the Philippines by a Japanese corporation. -Temperate vs. tropical climes and feedstock process management is characteristic of practices in their climes, i. e. seasonal with harvest, processing/storage, and feeding management year round vs. tropical seasonal year round operations of cropping and harvest, processing/storage including use of perennials, annuals and/or seasonals.
E.g. Chap. Probiotic Manipulations. - The current state of technological know-how with probiotics for the rumen. Use of insertional mutation or 'knock-out' of genes with rumen gram negative plasmid vectors with transposable (Tn) elements that mobilize and do not maintain in the recipient outside their genera (e. g. enteric bacteria) (i. e. termed suicide vectors) with protein elements that act to block expression on the operon when end-products are at a limit causing a run-a-way expression of enzymes (assuming no expenditure of energy above maintenance) or insertional mutation with genetically engineered enzymes (cf. Novozyme's CTec3 cellulase). - Fungal probiotics appear at this time to be in the front running gate amongst feasible approaches that are to be used and those apropos where there is a given need, that is, with the utilization of low quality sustainable feeding systems (viz. tropical, grasses, farm wastes, agro-industrial byproducts) and direct probiotic feeding (note: feeding as additives and special supplementation are alternative approaches) and their effect on ligno-cellulose digestibility, intake and productivity. Preliminary findings indicate possible feasibility with studies that compare geographic structure vs host in transfer studies and persistence although it was indicated that persistence in hosts was for a 6 week period while another showed an effect over the growth or observation period of the experiment. Natural transfer/persistence is another approach that can be used and what it will be determined what is to be found of its signficance or extent of persistence. - Yeast probiotics and their effects on protein output in the rumen intake. - 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 the vector or donor plasmid), 3) evolution of improved GM transport porins with increased capacity to transport pre-formed amino acids (i.e. amino acids and peptides) to improve efficiency of microbial cell protein (MCP) synthesis, 4) selection of donor hosts with recombinant enzymes that bind effectively with and bind the particulate phase in rumen digesta and have increased sustained half-life of retention, 5) genetic modification (GM) by single residue replacements with X-ray crystallographic studies of structure-function mechanisms in the components of the cellulosome and for improved cellulases amongst the family of cellulases and transcription factor (TF) structure-function analysis of the cellulosome and their modification to increase efficiency (component adaptation or multi-functionality), rate of processing, avidity and half-life, and 6) there is an issue of the role of amino-peptides (APP) (ie. pre-formed amino acids=free amino acids+peptides) and the role of profiling dietary amino acids with those appearing in total whole intestinal flows especially certain chosen essential amino acids (EAAs) such as threonine, lysine and methionine due to their preponderance in dietary profile, their subsequent "enrichment" in microbial cell protein (MCP) synthesis in the rumen, a hypothesis for Ph.D. research by our P. I. D. A. Flores at SkyeBlue, and their similar preponderant profiling in duodenal amino acids (DAAs) (see: Flores et al., 1986a: a 1.7x factor in dietary fresh feed versus one that had undergone degradation in those EAAs to start, finishing in a 2.4x factor comparatively in the duodenal flow with enrichment of these 3 EAAs). Applications in future may entail the use of "designer" proteins with a specific structure-functional relationship towards this end and designer drugs as feed additives to boost, the so-called, "expansion factor", posited with Flores' hypothesis.
E.g. Chap. Fibre-based Technologies for Food and Energy.-Sugarcane (beet & cane), a cash crop and major agro-industrial commodity in developing economies (cane sugar); -note: grains will be the major focus in food supplies to balance out food security as a priority and including subsistence needs of the developing world; crop by-products (fibre-based) may now be converted to starch amylose using two industry-grade technologies: ionic liquid decontructive fractionation (ILDF) pre-treatment and convert starch amylose conversion for feeds such as for sustainable livestock protein production (poultry and fish, aquaculture/capture) and agro-industrial wastes [e. g. sugar beet pulp waste and MCP (microbial cell protein) from residuals of bioenergy fermentation production to produce alcohols] can be used to produced compounded flaked wafers and pelleted feeds; root crops where indigenously grown are now serving as emerging sources of starch flours in the African continent and may compete as a more sustainable source against convert amylose starch where available and may allow to spare for further diversification towards livestock production feeding.
E.g. Chap. Tropical ensilage additives. –Silage types covered are 1) maize, 2) grasses, 3)legumes, 4) whole plant silage (e.g. high moisture ear corn, HMEC) and 5) sorghum in this discussion. -Lignases type II (anaerobic lignases from said eco-habitats) may prove to be the ‘breakthrough’ in providing an ‘even playing’ field rather than relative fractionation in terms of lignification and inaccurate estimates that results and inconsistency in results in digestibility, utilization and productivity found in studies. - There may be a need to investigate further and address the matter of what metabolizes to completion lignin polymers to remove toxicity towards microbiota and to animal hosts from its end-products. –In regards to management of ensilage there is the same parallel that exists between the discussion of prebiotic effects and underlying and the mixing of readily fermentable carbohydrate/sugars such as molasses and grains and sources of protein supplementation (i. e. sources of crude protein, CP) as feed supplements or additives. There is the principle of adding enough CP to fulfill needs for nitrogen (N) for cellulolytics, i. e. effective fibrolysis in digestion in addition to minimal N requirements for microbial cell protein synthesis for protein from rumen digestion when undergoing supplementation of basal forage rations for tropical ensilage with CP from non-protein-nitrogen (NPN).
E.g. Chap. Maize Fibre Components. -Can be fractionally manipulated in the plant producing different silage types including the grain as such with varying management requirements and styles. -Genomics and the identification and manipulation of specific genes to control lignin and other fibre components is still at its infancy or perhaps at an inception we may presume. -Prediction of the nutritive value of various feed types produced from silage and grain composition and their digestion, utilization and production will be discussed.
E.g. Chap. Farming & Renewable Resources Management. The farming for food amongst small-farmers in land reserves will be managed in the same way as rural and urban stakeholders using the rudiments of mixed-farming practices, e. g. pig operations with chickens and layers or goats, sheep and milking cattle with cash and vegetables crops and medium-large sized co-op or commercial farming operations, e.g. practices with monoculture or organic and other sustainable practices and intensive animal rearing practices, viz. grain-fed/silage or hay-fed with for e. g. beef cattle with corn grain-fed, soy beans with chickens, alfalfa or timothy with dairy, concentrate and forage with goats or sheep, either be it in Mozambique, Africa or the Fraser Valley, British Columbia CANADA. The activity for food production will compete with urban clustering of agro-food life sciences bio-hubbing, e. g. food mfging, processing/packaging, trade, and bio-pharma life sciences bio-hubbing. This will be discussed further here at SkyeBlue under a topic of contention, the "Fraser Hi-Tech Mainland" (FHTM) of B. C. As with Manitoba, CANADA's resource base of 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 innovative activities in future. The same would hold true for FHTM in B.C. CANADA including a diverse mix of agri-food landbase throughout the island and provincial mainland. In addition to feedstock conversion to hi-grade animal feedstock for dairy and meat, feedstock can be shipped in from vast renewable reserves in the Prairie provinces, for e. g. pulp from beets and residuals from ethanol fermentation and the conversion of residual fibre to convert starch for wafered and pelleted feeds for aquaculture/capture fisheries and poultry meat production. The above arguments using resource bases for other countries will apply where foreign investments accrues in places like continental Africa as a future of renewed bio-hubbing in potentialy such a new breadbasket, it is predicted.
Farms of the future for beef vs. dairy will involve assets of feedstock vs. livestock and hardwired (computers, electronics and communications, mechanized equipment) vs. free-standing infrastructure (glass, steel & concrete). Feedstocks include: 1) pasture: low-lignin, low-protease, high-fructan; 2) silage: low-lignin, top-dressed solid substrate fermentation (SSF) lacasses (type III lignases); 3) haylage: low-lignin, top-dressed SSF lacasse; 4) ligno-cellulose (L-C) feedstock: SSF, "Yeast Bagasse Process", ureolysis (providing also non-protein nitrogen, NPN); 5) L-C feedstock: bio-bleaching (L-C feedstock is used also for bioenergy fermentation to biogas, EtOH and ButOH). Livestock is classed as either: 1) beef: feed type would be for e. g. low-protease, high-fructan and 2) dairy: feed type would be for e. g. low-lignin (see: paper in literature).These are envisioned for the 'FHTM' Bio-Hub in British Columbia to be fed by the Western Prairie provinces for their resources in various feedstocks to be diversified (in this way) for manufacturing and farming for food production out West. Abundant rail will provide for transport of commodities with shipment from port. Use of SSF and its recent developments with agri-wastes (e.g. rice and wheat straw in India) and its research possibilities for bioenergy production will spur its adaptation in feeds production in the Western world including North America. This is one form of feed processing that may prove feasible in the near future depending on competing grains & forages (pasture, haylage, silage) and other demographic and environmental factors bearing on the land-food systems.
Knowledge-based, innovation-driven economies are related as a direct function it is hypothesized to driving investment and empowering the "economic machinery" of agricultural and food development and production; this as claimed has been understood and will be applied in life sciences bio-hubbing including agri-food and rural development in Western Canada; the hi-tech nature of the knowledge-based, innovation-driven economies include the R&D hatcheries that will be derived from biogenomics or computational biology research and will lead to advances in countries like Canada in crop technology & feeding, animal health and reproduction, irrigation, mechanization and energy advances, for e. g. in wind and solar gathering available at the farmer's "pump".
E.g. Chap. Feed Resources and Mixed Livestock Farming Systems: Africa a future bread basket of the world. Food deficit areas can be balanced out with food surplus area eventually resulting in less need and cost to import such as rice commodities. A new paradigm in organized agrarian reform by public and private (including multinational, albeit, guarded against neo-colonialistic or imperialistic overtones) interests with med.-larger commercial farmings operations such as was organized by Pro Savanna in Zambezi Village, Mozambique as a development planning management project (med. size 3-10 ha. farms) with knowledge, technology and equipment like a larger Brazilian cousin or model and thus have made for farmers in developing country settings feasible developments in farming middle-size farms large enough to make money and not loose their land.
E.g. Chap. non-Genetically Modified Organismal (non-GMO) Feedstock Pre-treatment Technologies: State-of-knowledge. Steam explosion (SE)/Sulfur dioxide (SO2); Ionic Liquid Deconstructive Fractionation (ILDF); Convert Starch Production from Fibre Biomass for Feed and Food.
E.g. Chap. Genetically Manipulated (GM) and Genetically Modified Organismal (GMO) Feeds Technologies: the Future Challenge. GMO Feeds / MAS Bred Feeds. 1) Low-lignin – hi-energy feeds for dairy; 2) heat-treated / low-protease – hi-prutein (protein from rumen) feeds for beef; 3) hi-fructan – hi-prutein (protein from rumen) feeds for beef production; 4) Sile-pro system using low-lignin and microbial cellulase innoculant piggy-backed SSF-lacasse enzyme technology; 5) hi-sugar pasture (e. g. ryegrass). Pre-treatment of L-C (ligno-cellulose) Feeds: 1) “Yeast Bagasse” Process cellulase piggy-backed with type II lignase, 2) Urea-ammoniation with piggy-backed type II lignase and 3) Solid Substrate Fermentation (SSF) (see: paper on breakthrough with lacasse-bearing host).
E.g. Chap. Energy Policy and Global Climate Change: a strategy of mixing energy sources will be used in future including: solar, wind - which both can be significantly expanded on, nuclear, C-sequestration (a newer technology ready for scale-up), shale gas, biodiesel and cellulosic ethanol (of interest here); - sustainable energy development will hopefully lead to depoliciticizing the discussions and agreements on oil and gas fossil fuel reserves; - C-capture strategies to decarbonize the atmosphere with zero to negative carbon fingerprinting and use ligno-cellulosic feedstocks are being proposed; 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 transportation fuels; heating/cooling may be a combination of solar-based technology and C-sequestration in addition to C-sink sources; there is also the question of safety of nuclear-based technologies based on advances in this area that is continuing; expansion of geothermal plants where they are feasible and needed; energy needs to be derived from sustainable means based on agronomic/energy inputs vs. weighing on inputs more directly from the sun's energy source; diversification with energy demand and the development for power infrastructure and hi-technology (one e. g. in diversification here is with agri-food & pharma with biotech) will apply to "mega bio-hubs" (e. g. Western Canada- B. C. and Priairie provinces, India-Far East Asia, Globally-African Continent, Europe-C. Asia) around the world; -there has been the suggestion that countries in Africa like Nigeria and Mozambique, to follow, where energy is becoming more available will help fuel diversification as led by S. Africa's example not only with minerals but with agro-processing and pharmaceuticals (this time, it is now suggested) with food and energy using for example an external link of Indian transfer of technology as a trading clause equally as useful in a different proposed scenario between India and the Philippine republic claiming soverign rights and continuing its activities of reclaiming what will be sizeable oil and gas in the Western sea off its coast line. The Philippines should launch on a similar strategy of involving itself with domesticated oil reserves use for industry development such as a national past time of food processing/manufacturing and going "green" on biorenewable energy.
December 25-27, 2017.
Dear: Friends, Respondents and Guests to Skye Blue! We have decided at Skye Blue to enact two new editions for the older one of: "A Compilation of Ligno-cellulose Feedstock and Related Research for Feed, Food and Energy," as an: 1) Australian Edition 2019. D. A. Flores (c). The Preface will include practices that have evolved with the use of low-quality feeds used in grazing and feedlot raising in and outside the Australian Outback region. Preliminarily, the Dedication will include: The book is also dedicated to all residents who reside with me in the Tri-Cities (Port Moody, Coquitlam, Port Coquitlam) / New Westminster / Burnaby and to their senior communities. May there be peace, and may they have an inspired time among the greenscapes of their respective cities. Chap. 1. Includes anaerobic lignases applied to ensilage of grasses, legumes, grains and high-moisture ear corn (HMEC). Chap. 3. Includes the tropical and new temperate (colder acclimated) hi-sugar grasses for grazers or ensilage and probiotic use gastroenterically, in future, which would be ideal in synergy. Also, the tropical and new temperate (colder climed) hi-sugar grass applications hydroponically grown in piled aquaria-based blocking. Chap. 12. Fibroin and New Textiles Technologies: (a) Hi-Tech & Bamboo-based Fibroin (e. g. sheeting tarp, tents and linens, etc.), (b) Wool Fibroin & Bamboo Technology (Anti-bacterial "Protek" Textile Technology.). Chap. 13. For the production of wool the traditional most-limiting amino acid ae the sulfur-containing amino acids to be supplemented by yeast over-producers of MET and CYS as escape SCP as a nutraceutical to supplement protein nutrition. Also, it would be interesting to observe whether HIS supplementation with ensilage diets where HIS is likely limiting productivity including wool growth could stimulate further protein accretion vis growth of wool hair in sheep. 2) USA / Int'l Edition 2018. D. A. Flores (c). Chap. 13. Hydroponics & Marine Grass for Pharma. (a) High-degree esters and alcohols and their chemistry for use in pharma as a precipitant to enable technology development. (b) Chap. 13. Pasture Grass, Legume and Grain Forages for Pharma and Fodders.
SKYENEWS: Earlier (see: Lux Esto e-Zine of Kzoo.edu in the USA) it was mentioned that the author and researcher at SkyeBlue had understaken research on GM and non-GM feedstocks, processing and probiotics use as approaches to meet food and energy security in future. This research is ongoing and is occurring as it is revealed through the published literature as we speak including the following approaches: 1) GM fungi in silage resulting in increased lactic acid, 2) GM fungi in the rumen resulting in increased saccharification of fibre fractions, 3) GM yeast with bagasse with cloned endoglucanase, 4) GM yeast in the rumen as a well-practiced probiotic with a cloned endoglucanase, 5) oligosaccharide in silage in plant material prebiotic to silage fermentation increasing lactic acid production and 6) oligosaccharide in the rumen from plant prebiotics resulting in increased microbial cell protein synthesis. (See also: Flores, 1991.Table 2).The use of supplementation with "by-pass rumen" proteins as with grass forages like sugarcane tops (SCT) (see: Spangenberg, 1991; Flores, 2013) using Lysine, Methionine and Cysteine to supplement or balance profiles on these diets and use of "Yeast Bagasse" for processing to add SCP and amalgamate rolled GM corn grain supplemented with Threonine as a basal ration in addition to green forage that has been sizeably supplemented with good quality protein and field-dried for heat protection (see: Charmley and Veira, 1990; Flores, 2013) are approaches we are working on at SkyeBlue with our Principal.
SKYENEWS: The use of grazing and rangelands and those under forests and in plantations with tree cropping and browse trees and shrubs may serve at first glance to be: 1) more ecologically sound in terms of conserving soil structure (carbon sequestration and N-fixation), 2) sound in terms of providing eco-habitats for wildlife and 3) providing biomass feedstocks yielding greater net energy gains than monoculture energy crops when converted to biofuels. As greater demand for energy and food rises better land use will prove this approach to be the choice for biomass feedstocking due to the fact that feedstock does not conflict with feedstock for food vs energy and protection against environmental degradation or any such negative impacts.
SKYENEWS: The recent discovery of a further class (type-IV) of ligninolytic enzymes, viz. ureases in soil and plant microbes that occur naturally and when selected out under the right conditions of feed storage/processing as with urea-ammoniation in tropical ensilage can effectively breakdown the lignin cement in the polymer of ligno-cellulose feedstock; further to this, there is the distinct possibility that the FAO, Rome Italy will mandate, as it did with GM cellulase temperate silage with L. plantarum cultures, the boosting of urease using rec-DNA technology in bacteria where they can be studied now at this time including model hosts such as Klebsiella spp. where two (2) nickel atoms (Ni+) act as catalysts in the hydrolysis of urea to alkali end-product permeants as the chemical active ingredients that act on the lignin component of fibre. It is of course assumed here that ureases with urea as substrate are inadmissable as EFEs for action on feedstock consumed in the rumen due to their need for action at a pH range outside the pH balance of the rumen stomach.
SKYENEWS: Africa as we speak is being earmarked as the new breadbasket of the world come 2050 and soon-to-be a new pharming "niche" cropping and silviculture centres is already envisioned for B.C. and subsidiarized for Africa. Phytobotanics include terraforms and marine life. But crops and forest life produce: leaves, flowers,sap, fruit/juice, bark, seed and roots with extracts as common derived products. There is much to "mine" in the biodiversity and what is hoped the sustainability of wildlife in many eco-habitats around the globe including rainforests and alpine forest areas.
SKYENEWS: The recent news out of SkyeBlue regards any breakthroughs with anaero-lignases (type-II lignases) yet to be characterized, pioneered by workers like Colberg and Young (1982), earlier of Stanford University, and continuing work by other workers which we recently recognized with an award of recognition, the Skye Blue Cabre Award, in a review paper, is that hindrances in rates of recalcitrant enzymes (esterases, etherases, lyases (these haven't been well described in the literature), and ring-related enzymes including those that lead to eventual mineralization) will be compared with enzymes in the mesophyllic rumen strain spp. which are known to be more efficient given the transit time of feed digesta and their breakdown of lignocellulose in the rumen stomachs compared to other 'habitats' as with activated waste sludge or marine eco-habitats that can take months at length. There will be a need to study the structure-functional mechanisms including characteristics of the active sites and what makes these enzyme(s) more rate-limiting and thus efficient including more recent techniques of cryo-electron microscopy imaging of subunit structure and bonding and X-ray crystallography and eventually engineering the enzymes to be more efficiently stable at the optimum pH and T (deg C) for temperate ensilage (20-30 deg C, ensilo), ureolysis (ambient in the tropics), yeast bagasse (ambient in the tropics), solid-substrate fermented feedstock (ambient in the tropics).
SKYENEWS: It is speculated at this point what enzymes are rate-limiting in the delignification process. The dearomatization ring enzymes are less likely compared to the de-branching enzymes considering the bond energies of the aromatic ring structure; the side groups versus the ester, ether and C-C bonds. However, among the latter three whether it matters which amongst the three bond type energies may not be critical in that dissolution of lignin in lignocellulose is significant, ie. fragmentation of large branched structures, it is presumed, accounting for %15 of the disappeared lignin. Flores (1989) highlighted a strain '7-1' first identified by D.E. Akin and colleagues in the rumen and should precipitate other candidates to be studied by atomic powered microscopy to have digested at the molecular level lignous tissues in the ligno-cellulose fractions of plant materials in leaves and stems leaving for example exposed tissues including the holocellulose (cellulose+hemicellulose).
SKYENEWS: Lowering protease activity in soybeans (and other grains) would boost escape protein while decreasing solubility which tends to be too high and needs heat treatment with extrusion. Whether this can be made amenable to optimization is still open to question based on the current evidence from the literature.
SBO for Animal and Crop Research
Last update of this entry: December 27, 2017