Nutritional Value And Efficacy Of Biotransfulation Fulvic Acid

Humic acid (HA) is a class of natural polymer macromolecular organic compoundsproduced by biodegradation, decomposition and conversion and by geophysical andchemical reaction and accumulation of mineralized animal and plant residues.Biotransfulation fulvic acid (BFA) is a mixture produced by one or more friendly bacteria toferment and convert straw or wood chips, etc., combined with soybean meal, wheat branand other fermented raw materials at the right temperature and humidity. BFA is a complexvariety of substances, and naturally, is a synbiotic microecological agent.BFA used in this study is made through an innovative process route with fermentationengineering and enzyme engineering as core technology, combined with superior probioticstrains, distillers grains as the main raw material, through probiotics-prebiotic enzymescoupled biological transformation as main while physical and chemical treatment assupplemented methods. BFA, a microecological agent comprising of probiotics fermentedfulvic acid (FA) and participating probiotics, organically combines the functions of humicacid and probiotics. Through probiotics playing the role of competitive exclusion to harmfulbacteria, BFA thereby regulates the balance of intestinal flora, assists the host animals toestablish a favorable gastrointestinal microflora, enhances immunity system, preventsdiarrhea and promotes digestion and absorption of nutrients, and promotes body growth,and in such way, finally improves feed efficiency. Meanwhile, the nucleic acids, aminoacids, inositol, vitamins, polysaccharides and other substances in the BFA can be directlyinvolved in the body’s metabolism as good nutrients in one side, and effectively promoteslivestock growth in the other side. BFA can efficiently convert a variety of macromolecularnutrients in the feed into small molecular nutrients, thereby promotes the absorption ofnutrients and improves production performance. Quinone groups, contained in the BFAactive ingredient humic acid, are involved in the oxidation-reduction reaction of the body,and promote metabolism and accelerate cell division and proliferation. Currently, BFA has been widely used as feed additives, drugs synergist, soil conditioner, organic fertilizerstarter, plant growth regulator and other products, throughout all areas of agriculture, andshows a bright market prospect in the application of efficient ecological agriculture.In this study, we planed to study the standardization of BFA source in the first section,namely to test product quality stability. In the second section, we would proportionally addBFA to the conventional feeds which met the proposed standards and had been standardlyanalyzed, and take mammals (SD rats) as the research object in order to determine the effectof BFA on the general living conditions, growth performance, physiological andbiochemical status, intestinal microflora, immune system and other indicators. So, wewould preliminarily study the safety and ergonomic value of BFA used as the nutritionadditive for functional feed.Methods1. Standardization of BFA source and its nutritional valueBFA source standardization was conducted on unfermented BFA (distills grains) and4other biological humic acid bacteria agents, including lactic acid fermentation BFA, bacillusfermentation BFA, yeast fermentation BFA, and fermented BFA by above3mixed bacteriain order to establish a testing system for product quality stability. The measurementindicators included: sensory analysis, total humic acid (HA), fulvic acid (FA), the number ofviable probiotics, crude protein (CP), crude fiber (CF), ether extract (EE), crude ash (ASH),nitrogen-free extract (NFE), phosphorus (P), calcium (Ca), dry matter and amino acids, etc..Among them, the humic acid content was determined by volumetric method, viableprobiotic count by plate count method, feed nutrients detection based on national standardmethods to number and name, that is, crude protein by GB/T5009.6-2003, crude fiber byGB/T5009.10-2003, crude fat by GB5009.3-2010, ash by GB5009.4-2010; NFE by GB/T5009.5-2010, P by GB/T6436-2002; Ca by GB/T5009.87-2003, and amino acids byHitachi L-8800automatic amino acid analyzer. The results of above mentioned integrateswere summarized.2. Efficacy of BFA as a feed additiveFour biological humic acid bacteria agents (all standardized, qualified and scaled-classagent, including lactic acid fermentation BFA, bacillus fermentation BFA, yeastfermentation BFA, and fermented BFA by above3mixed bacteria) were respectively added by a proportion of1.5%to the conventional feed. Then the SD rats were fed by normal feed(without adding BFA, as control group), and above feeds with4different BFA (asexperimental groups) for4weeks.After4weeks’ feeding, the general living conditions, growth performance,physiological and biochemical status, intestinal microflora, immune system and otherindicator were observed to study the impact of BFA added in the feed.Results1. Results of standardization of BFA source and its nutritional valueThis study proposed the contents of basic indicators of BFA: fulvic acid (FA) content≥16%; probiotic counts≥2×108cfu/g;1.1. Content analysis of total HA and FA: the content of BFA in the distillers grainswas (41.63±0.21)%, and that of FA was (14.36±0.19)%, with the latter below the proposedstandards. So in the subsequent nutrient analyses and efficacy study, the distillers grainswere not studied any more. All4kinds of BFA complied with proposed standards of totalHA and FA. And the contents of total HA and FA were significantly higher in the mixedfermented BFA than3other fermentation BFA (lactic acid bacteria, yeast, and bacillus)(P<0.01);1.2. Viable probiotic count:4kinds of fermentation BFA (lactobacillus BFA, yeastBFA, bacillus BFA, and mixed fermentation BFA) are in line with proposed standards;1.3. Sensory analysis: the color, mixing uniformity, no mildew or agglomeration in the4kinds of fermentation BFA were in line with national standards;1.4. Feed nutrients analysis: In the detection of crude protein, crude fiber, crude fat,crude ash, Ca, P, dry matter and nitrogen-free extract, the mixed fermentation-BFA hadsignificantly higher crude protein and crude fat than the other3kinds of fermentation BFA(lactic acid BFA, yeast BFA, and bacillus BFA)(P<0.01), and also had significantly highercontents of P than lactic acid BFA group and bacillus BFA group (P<0.01), but there was nosignificant difference in the content with yeast BFA group;1.5. Amino acid analysis:4kinds of fermentation BFA all contained17kinds of aminoacids, with great variety and covering the growth and development of animals required.2. Efficacy studies of BFA as a feed additive2.1. General condition of the animals: the SD rats from the experimental groups (4 kinds BFA fermented group) and control group had generally good life, eating and drinkingwell, with normal mental status, good activity, no sickness, death or others.2.2. Blood routine: The counts of neutrophils, white blood cells, and monocytes wassignificant lower in the lactobacillus BFA group than the control group (P<0.05); bacillusBFA group had reduced neutrophil than the control group with significant difference(P<0.05).2.3. Blood biochemical indicators: Lactobacillus BFA group had significantlyincreased total protein, albumin, and albumin/globulin ratio (P<0.05), and obviouslydecreased total bilirubin (P<0.05) compared with the control group. The total bilirubin wasvery significantly lower in the Bacillus BFA group than the control group (P<0.01).2.4. Weight gain and feed/gain ratio (F/G): the mixed fermentation BFA group hadsignificantly increased total weight gain and daily gain than the control group (P<0.05); Forthe feed conversion rates in each group, the mixed fermentation BFA group had the highestfeed efficiency, increased by10.04%than the control group, followed by the yeast-BFAgroup, with an increase by4.14%.2.5. Intestinal flora analysis: the mixed fermentation BFA group and lactobacillusfermentation BFA group had significant reduced number of E. coli than the control group(P<0.05), while the number of lactobacilli were significantly increased than the controlgroup (P<0.05).2.6. Analysis of immune system parameters: thymus index and spleen index: thethymus index was significantly higher in the mixed fermentation BFA group andLactobacillus BFA group than the control group (P<0.05); while significant difference wasseen in the spleen index between the mixed fermentation BFA group and the control group(P<0.01). Immu-serological indicators: the mixed fermentation group had significantincreased serum levels of IgA, IgG, IgM and IL-2compared with the control group(P<0.05); the Lactobacillus BFA group had obviously increased levels of IgA, IgG and IL-2than the control group (P<0.05).Conclusion1. Standard testing for BFA: the lactobacillus BFA, yeast BFA, bacillus BFA, andmixed fermentation-BFA all meet our proposed standards in the FA contents and probioticcounts. However, the FA content in the unfermented BFA (distillers grains) is lower than the proposed standards, and no probiotics is found. So in the subsequent nutrient analyses andefficacy study, the distillers grains are not studied any more.2. Nutritional studies of BFA as a feed additive: our obtained BFA is in line withnational standards in sensory analysis; nutrient analysis shows that the mixed fermentationBFA achieved the highest contents in crude protein and crude fat; amino acid analysisindicates that all our4kinds of BFA contained various amino acids, covering the growthand development of animals required.3. Feeding efficiency of BFA as a feed additive: after BFA feeding, SD rats are in goodcondition of general life, suggesting that it has a high safety; BFA can promote the growthof each group of rats, and the mixed fermentation BFA get the highest in the total weightgain, average daily gain, feed efficiency and other indicators, presenting the features of2fast-growing period.4. Physiological effect of BFA: BFA can improve intestinal flora, suppress harmfulbacteria breeding, and promote the growth of probiotics in SD rats; It also improveshumoral and cellular immune function, and enhances the body’s overall resistance todisease. Among the4kinds of BFA, the mixed fermentation BFA and Lactobacillus BFA aresuperior to the control group in increasing the thymus index, spleen index andimmunoglobulins (IgA, IgG and IgM); Our results indicate that BFA can enhance proteinsynthesis in the rat liver.5. BFA does not have adverse effects on the liver and kidney functions in rats.