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Supplemental Defatted Microalgae Nannochloropsis oceanica Affected Apparent Retention and Ileal Digestibility of Nutrients in a Corn-Soybean Meal Based Diet for Broiler Chickens

Objective: This study was to assess impacts of including 10% defatted microalgae (Nannochloropsis oceanica, 45% crude protein and 3.8% ether extract) into a corn-soybean meal basal diet for broiler chickens on apparent retention and ileal digestibility of nutrients. Procedures: Day-old hatchling Cornish Giant cockerels were divided into two groups (6 cages/group, 6 chicks/cage) and fed either the basal (control) or the microalgae-supplemented (microalgae) diet for 6 weeks. Starting week 3, the two experimental diets were mixed with 0.2% chromium oxide as an indigestible marker. Total excreta of individual cages were collected daily for 3 consecutive days during week 6. At the end of week 6, chicks were euthanized to collect ileal digesta from 1 chick/cage. Energy and concentrations of dry matter, crude protein, ether extract, minerals, and amino acids in digesta, excreta, and diets were assayed. Apparent retentions and ileal digestibilities were calculated using the direct collection and indirect indicator methods. Data were analyzed by the Student’s t-test and the significant level of difference was at p < 0.05. Results: The microalgae diet had elevated (4.2% and 8.1% by the two methods, p < 0.05) apparent retention of ether extract, but similar apparent retention and ileal digestibility of dry matter and apparent metabolizable energy, compared with the control diet. Supplemental defatted microalgae decreased (p < 0.05) the apparent retention and ileal digestibility of dietary crude protein by 13% and 13%. The supplementation also decreased (p < 0.05) ileal digestibilities and apparent retentions of multiple amino acids ranging from 0.6% to 26%. Apparent retentions of sodium and total phosphorus were decreased (p < 0.05) by 13% and 71%, respectively, by the microalgae inclusion compared with the control. Conclusion: The inclusion of 10% defatted N. oceanica in the corn-soybean meal basal diet for broiler chickens led moderate increases in apparent retention and ileal digestibility of ether extract, but decreased those of crude protein, amino acids, sodium, and phosphorus.

60 APPLIED LIFE SCIENCES↗

Supplemental defatted microalgae affects egg and tissue fatty acid composition differently in laying hens fed diets containing corn and flaxseed oil

Effects of defatted microalgae (DFA) on fatty acid composition of eggs and tissues of hens have not been well studied. This experiment was to determine how low concentrations of supplemental DFA in the presence or absence of flaxseed oil (FO) altered fatty acid profiles of egg yolk, liver, adipose tissue, and plasma of layers. A total of 60 Shaver Leghorn layer hens (individually caged, 20-wk old) were allotted into 6 groups (n = 10) and fed a corn/soybean meal basal diet containing 0, 3, or 5% DFA and 0 (1.5% corn oil, CO) or 1.5% FO for 6 wk. BW, ADFI, egg production rate, and egg component weights were not affected by any of the dietary treatments at various time points, but egg yolk color was changed (P < 0.05) with increasing concentrations of DFA. The concentration of n-3 fatty acids in egg yolk and plasma were elevated by FO up to 9-fold (P < 0.05) starting at wk 1 or 2, and 39 to 83% (P < 0.05) by DFA. Supplemental DFA improved (P < 0.05) n-6/n-3 fatty acid ratios in egg yolk and plasma from 13 to 23 and 7 to 13, respectively, only in hens fed diets without FO. Fatty acid profiles of liver and adipose tissue (at wk 6) displayed responses to supplemental DFA and FO similar to those of egg yolk or plasma. In conclusion, supplemental 3 or 5% DFA caused moderate enrichments of n-3 fatty acids and decreased the n6/n3 fatty acid ratios of egg yolk and tissues in hens fed diets without FO.

59 BASIC BIOLOGICAL SCIENCES↗

A new type of defatted green microalgae exerts dose-dependent nutritional, metabolic, and environmental impacts in broiler chicks

The potential of defatted microalgae from biofuel production in animal feeding remains unclear. The objective of this experiment was to determine the metabolic and environmental impacts of a defatted green microalgal biomass (DGA, Nannochloropsis oceanica, Cellana, Kailua-Kona, HI) in broiler chick diets. Day-old chicks (total = 180) were fed the control diet containing zero, 2, 4, 8, or 16% of DGA for 6 wk to determine the optimal level of DGA inclusion. Compared with chicks fed the control diet, only those fed the 16% DGA diet had lower (P < 0.05) ADG and feed use efficiency. The DGA feeding produced dose-dependent (linear) increases in starter period water intake (P < 0.0001, R 2 = 0.76) and in wk 6 relative weights of heart (P < 0.0001, R 2 = 0.45), liver (P = 0.09, R 2 = 0.19), and intestinal tract (P = 0.02, R 2 = 0.19), retentions of soluble inorganic phosphorus (P = 0.001, R 2 = 0.39) and DNA retention (P = 0.001, R 2 = 0.46), and ileal DNA content (P < 0.0001, R 2 = 0.50). Meanwhile, the DGA feeding led to weak linear decreases (P < 0.05, R 2 = 0.14 to 0.27) in wk 6 tibia weight and length and excretion of soluble inorganic phosphorus. In contrast, the DGA feeding did not affect relative weights of breast, proventriculus, or gizzard, tibial bone strength, or plasma DNA concentrations. In conclusion, this new type of DGA biomass may be supplemented into diets for broilers at 8% without adverse effects if the salt content is decreased.

59 BASIC BIOLOGICAL SCIENCES↗

Supplemental Microalgal Iron Helps Replete Blood Hemoglobin in Moderately Anemic Mice Fed a Rice-Based Diet

Iron deficiency anemia affects 1.2 billion people globally. Our objectives were to determine if (1) supplemental iron extracted from defatted microalgae (Nannochloropsis oceanica, DGM) and (2) a combination of minute amount of plant phytase and inulin could help replete hemoglobin in anemic mice. Mice (7 weeks old) were fed a control diet (6 mg Fe/kg). After 10 weeks, the mice were assigned to three treatments: control, control + DGM iron (Fe-DGM, 39 mg Fe/kg), or control + 1% inulin + 250 units of phytase/kg (INU–PHY, 6 mg Fe/kg). The mice had free access to diets and water for 6 weeks. The Fe-DGM group had elevated blood hemoglobin (p < 0.01) and a two-fold greater (p < 0.0001) liver non-heme iron over the control. Strikingly, the INU-PHY group had 34% greater non-heme iron than the control, despite the same concentrations of iron in their diets. Fe-DGM group had altered (p < 0.05) mRNA levels of hepcidin, divalent metal transporter 1, transferrin and transferrin receptor 1. Iron extracted from defatted microalgae seemed to be effective in alleviating moderate anemia, and INU-PHY enhanced utilization of intrinsic iron present in the rice diet. Our findings may lead to a novel formulation of these ingredients to develop safer and bioavailable iron supplements for iron-deficient populations.

60 APPLIED LIFE SCIENCES↗

Marine Algae Industrialization Consortium (MAGIC): Combining biofuel and high-value bioproducts to meet the RFS

The Marine Algae Industrialization Consortium (MAGIC) was formed to address pressing challenges in the commercialization of microalgae as a source of biofuel. The “Marine Algae Industrialization Consortium (MAGIC): Combining biofuel and high-value bioproducts to meet the RFS” project formally addressed two US Department of Energy Bioenergy Technologies Office (BETO) goals: (1) Model the sustainable supply of 1 million metric tonnes ash free dry weight (AFDW) cultivated algal biomass and (2) Demonstrate valuable co-products produced along with biofuel intermediates to increase value of algal biomass by 30%. To achieve these goals, the project demonstrated and validated high-value co-products to drive down the cost of biofuel by increasing the value of algae “co-products” towards increasing the selling price of total algae biomass as one of the key drivers of economics and adoption. This was accomplished through five core, interdependent tasks including: (1) strain selection to identify and deliver strains for mass culture, (2) mass culture using a hybrid cultivation system and following key operating parameters for downstream applications to provide algae feedstock, (3) recovery and conversion to evaluate two alternative methods to separate dry algae biomass into oil and residuals for downstream testing, (4) product assessment to determine biofuel, aquafeed or poultry feed product efficacy using algae biomass fractions as well as to provide critical performance data for valuation and (5) commercialization to use technoeconomic and life cycle assessments (TEA/LCA) as iterative design and assessment tools including consideration of target markets, competitors, and distribution channels to guide product assessment, development and valuation. A total of 46 peer-review publications, many open-access, provide detail of much of the work carried out and the results of the tasks. Additional reports and presentations provide other technical and public engagement material. At a high level, using a variety of approaches, more than 1000 marine microalgae strains were evaluated to ultimately identify the seven winners that were down-selected to be grown in mass culture. Strain selection demonstrated that there were no ‘super strains’ and that each candidate had strengths and limitations for specific products, growth conditions or operational considerations. Mass culture growth of these seven strains at >5000 L / 29 m 2 scale found that four them were suitable for product assessment. More than 250 kg of biomass was produced across hundreds of pond runs along with thousands of cultivation entries on the growth and biomass characteristics as well as environmental parameters. In the process, dozens of standard operating procedures were generated as was custom software to process and analyze cultivation data. Recovery and conversion of algae biomass demonstrated that a hexane solvent based extraction protocol was most effective at recovering oil (biocrude) from algae and four strains were processed to produce oil and lipid extracted algae (residuals) for downstream testing. Membrane-based oil separation was less successful, but may still be applicable to other commercial applications in the future. Product testing demonstrated that algae biocrude is of high quality and hydrotreating generated numerous fractions of high quality composition for fuel and lubricate based applications. Aquafeed studies performed at a variety of scales showed that both whole and defatted (lipid extracted algae) microalgae were suitable as a feed ingredient, but that the specifics of the fed animal and biochemical composition of the algae are critical factors when determining formulation. Similarly, poultry studies on whole and defatted microalgae generally showed positive outcomes on animal growth and health, with some microalgae providing enhanced nutritional composition of the animal product. Economic and life cycle assessments covered a wide range of possible commercialization and sustainability scenarios. Replacement value, improved product value added, consumer values marketing added valuation and improved animal health were considered as alternatives for microalgae valuation. Using the open pond system, algae productivity was identified as the key driver of commercialization economics, but combination of co-products (e.g. animal feed) with biofuel production substantially increased the total selling price of algae. Modeled microalgae selling price exceeded $\$$1500/tonne and could generate competitive biofuel selling prices below $\$$5 gallon gas equivalents using realistic algal productivities. Short (process scale) and longer (decadal trends) sustainability assessments show that marine microalgae can enhance the sustainability of energy production and lead to other realized benefits in water, fertilizer and land use for other sectors (e.g. agriculture). This project successfully demonstrated all of the components of an end-to-end process from mass microalgae cultivation and dewatering, to recovery and conversion of algae biomass components, to final product demonstration and process valuation; the combined results provide a framework for future commercialization of algae based biofuels.

09 BIOMASS FUELS↗

Supplemental dietary full-fatted and defatted Desmodesmus sp. exerted similar effects on growth performance, gut health, and excreta hydrothermal liquefaction of broiler chicks

This study determined effects of full-fatted (C046, 41% crude protein) and lipid extracted (LEA, 39% crude protein) microalgae Desmodesmus sp. on growth performance, gut health, and excreta hydrothermal liquefaction of broiler chickens. Two experiments were conducted by feeding day-old Cornish male chicks: 1) a corn-soybean meal basal diet (control) 2) control +5% C046, or 3) control +5% LEA for 2 weeks. Growth performance was measured weekly in both experiments. Blood, liver, and duodenum samples of chickens were collected for biochemical analyses, and excreta samples were collected for hydrothermal liquefaction (300 °C, 60 min) at the end of Experiment 2. Compared with the control, supplemental dietary C046 and LEA enhanced average daily gain (13% to 40%, p < 0.05) in both experiments, and improved feed efficiency (15% and 11%, p < 0.01) in Experiment 2. The C046 and LEA diets enhanced the duodenum (19% to –2 fold, p < 0.05), but decreased the liver (17 to 78%, p < 0.05) mRNA abundances of interleukins-1, 6 and 8 and (or) tumor necrosis factor-α, respectively, compared with the controls. Meanwhile, the two types of biomass enhanced the protein amounts of claudin-1 in the duodenum (5–34%, p < 0.05) than the controls. Hydrothermal liquefaction of the excreta derived from the three diets showed no major differences in total bio-crude oil (average of 23%), hydrochar yield (average of 12%), or elemental compositions. However, the heating values of the resultant two bio-crude oils from the microalgae-fed chicken excreta (average of 34 MJ/kg) were 16% greater than those from the control chickens (29 MJ/kg). In conclusion, supplementing the full- and de-fatted Desmodesmus sp. microalgal biomass in the starter diets for broiler chickens produced largely similar improvements of growth performance, gut health, and excreta-derived oil quality.

09 BIOMASS FUELS↗