Animal Feed Supplement Research Articles

Enhancing Fatty Acid Production of Saccharomyces cerevisiae as an Animal Feed Supplement.

Saccharomyces cerevisiae is used for edible purposes, such as human food or as an animal feed supplement. Fatty acids are also beneficial as feed supplements, but S. cerevisiae produces small amounts of fatty acids. In this study, we enhanced fatty acid production of S. cerevisiae by overexpressing acetyl-CoA carboxylase, thioesterase, and malic enzyme associated with fatty acid metabolism. The enhanced strain pAMT showed 2.4-fold higher fatty acids than the wild-type strain. To further increase the fatty acids, various nitrogen sources were analyzed and calcium nitrate was selected as an optimal nitrogen source for fatty acid production. By concentration optimization, 672 mg/L of fatty acids was produced, which was 4.7-fold higher than wild-type strain. These results complement the low level fatty acid production and make it possible to obtain the benefits of fatty acids as an animal feed supplement while, simultaneously, maintaining the advantages of S. cerevisiae.

Synthesis of hydrophilic and hydrophobic carbon quantum dots from waste of wine fermentation.

Wine lees are one of the main residues formed in vast quantities during the fermentation of wine. While toxic when applied to plants and wetlands, it is a biodegradable material, and several alternatives have been proposed for its valorization as: dietary supplement in animal feed, source for various yeast extracts and bioconversion feedstock. The implementation of stricter environment protection regulations resulted in increasing costs for wineries as their treatment process constitutes an unavoidable and expensive step in wine production. We propose here an alternative method to reduce waste and add value to wine production by exploiting this rich carbon source and use it as a raw material for producing carbon quantum dots (CQDs). A complete synthetic pathway is discussed, comprising the carbonization of the starting material, the screening of the most suitable solvent for the extraction of CQDs from the carbonized mass and their hydrophobic or hydrophilic functionalization. CQDs synthesized with the reported procedure show a bright blue emission (λmax = 433 ± 13 nm) when irradiated at 366 nm, which is strongly shifted when the wavelength is increased (e.g. emission at around 515 nm when excited at 460 nm). Yields and luminescent properties of CQDs, obtained with two different methods, namely microwave and ultrasound-based extraction, are discussed and compared. This study shows how easy a residue can be converted into an added-value material, thus not only reducing waste and saving costs for the wine-manufacturing industry but also providing a reliable, affordable and sustainable source for valuable materials.

Complete Genome Sequence of Lactobacillus acidophilus Strain ATCC 53544.

ABSTRACTHere we present the complete genome sequence of Lactobacillus acidophilus ATCC 53544. The assembly contains 1,991,906 bp and is 99.7% similar to L. acidophilus NCFM. This strain was isolated from a rectal swab specimen of an infant and has previously been used as a feed supplement for animals.

Concept for Recycling Waste Biomass from the Sugar Industry for Chemical and Biotechnological Purposes.

The objective of this study was to develop a method for the thermally-assisted acidic hydrolysis of waste biomass from the sugar industry (sugar beet pulp and leaves) for chemical and biotechnological purposes. The distillates, containing furfural, can be catalytically reduced directly into furfurayl alcohol or tetrahydrofurfuryl alcohol. The sugars present in the hydrolysates can be converted by lactic bacteria into lactic acid, which, by catalytic reduction, leads to propylene glycol. The sugars may also be utilized by microorganisms in the process of cell proliferation, and the biomass obtained used as a protein supplement in animal feed. Our study also considered the effects of the mode and length of preservation (fresh, ensilage, and drying) on the yields of furfural and monosaccharides. The yield of furfural in the distillates was measured using gas chromatography with flame ionization detector (GC-FID). The content of monosaccharides in the hydrolysates was measured spectrophotometrically using enzymatic kits. Biomass preserved under all tested conditions produced high yields of furfural, comparable to those for fresh material. Long-term storage of ensiled waste biomass did not result in loss of furfural productivity. However, there were significant reductions in the amounts of monosaccharides in the hydrolysates.

Composition and Nutrient Value Proposition of Brewers Spent Grain.

Brewer's spent grain (BSG), a major brewing industry byproduct, is generated in large quantities annually. This review summarizes research into the composition and preservation of BSG, different extraction techniques for BSG proteins and phenolic acids, and the bioactivities of these phenolic components. Moreover, this article also highlights BSG integration into foodstuff for human consumption and animal feed supplements. BSG is considered a rich source of fiber, protein, and phenolic compounds. The phenolic acids present in BSG are hydroxycinnamic acids (ferulic, p-coumaric, and caffeic acids), which have many biofunctions, such as antioxidant, anticarcinogenic, antiatherogenic, and antiinflammatory activities. Previously, attempts have been made to integrate BSG into human food, such as ready-to-eat snacks, cookies and bread, to increase fiber and protein contents. The addition of BSG to animal feed leads to increased milk yields, higher fat contents in milk, and is a good source of essential amino acids. Therefore, many studies have concluded that integrating the biofunctional compounds in BSG into human food and animal feed has various health benefits.

A Bacillus subtilis strain as probiotic in poultry: selection based on in vitro functional properties and enzymatic potentialities.

We have proposed and validate an in vitro probiotic selection, based on enzymatic potentialities associated to well-established probiotic functional properties. A new Bacillus subtilis HB2 isolate, selected based on its high extracellular enzyme production, was chosen as a probiotic candidate for application as animal feed supplement. The HB2 strain showed an excellent acid and bile salts tolerance, a strong adhesion to chick enterocytes and produced antimicrobials against pathogens. An in vivo trial in poultry farming was conducted to evaluate the HB2 probiotic performance. After 35days, HB2 achieved the higher growth performance than the control groups. The mortality and the feed conversion ratio were significantly decreased. Finally, the HB2 treated group showed wet litter and less severe ammonia odor in the atmosphere. Our study provides new insights into the importance of enzymatic potentialities, associated with the common functional properties, as a novel approach for probiotic selection.

Fish Scales as Potential Substrate for Production of Alkaline Protease and Amino Acid Rich Aqua Hydrolyzate by Bacillus altitudinis GVC11.

Fish processing industries generate large quantities of fish scales as processing waste, if not treated leading to environmental pollution. Fish scales are hard to degrade, hence cause difficulty in waste management. In this context present study was made to utilize fish scales as substrate for the production of alkaline protease by Bacillus altitudinis GVC11 and subsequently amino acid rich aqua hydrolyzate. B. altitudinis GVC11 efficiently utilized five types of fish scales as substrates and produced maximum alkaline protease using Labeo rohita (28,150 U/mL) followed by Catla catla (23,320 U/mL) at 48h and Cyprinus carpio (17,146 U/mL) Mugil cephalus (18,917 U/mL), Cirrhinus mrigala (12,430 U/mL) at 72h. The HPLC analysis of protein hydrolyzate obtained after fermentation was enriched in essential amino acids, leucine, isoleucine, phenylalanine, lysine and non-essential amino acids, tyrosine, arginine and cysteine which can be used as animal feed supplement and organic fertilizer.

Effect of feeding garlic leaves on rumen fermentation, methane emission, plasma glucose kinetics, and nitrogen utilization in sheep

BackgroundGarlic and its constituents are reported to have been effective in reducing methane emission and also influence glucose metabolism in body; however, studies in ruminants using garlic leaves are scarce. Garlic leaves contain similar compounds as garlic bulbs, but are discarded in field after garlic bulb harvest. We speculate that feeding garlic leaves might show similar effect as garlic constituents in sheep and could be potential animal feed supplement. Thus, we examined the effect of freeze dried garlic leaves (FDGL) on rumen fermentation, methane emission, plasma glucose kinetics and nitrogen utilization in sheep.MethodsSix sheep were fed Control diet (mixed hay and concentrate (60:40)) or FDGL diet (Control diet supplemented with FDGL at 2.5 g/kg BW0.75 of sheep) using a crossover design. Methane gas emission was measured using open-circuit respiratory chamber. Plasma glucose turnover rate was measured using isotope dilution technique of [U-13C]glucose. Rumen fluid, feces and urine were collected to measure rumen fermentation characteristics and nitrogen utilization.ResultNo significant difference in rumen fermentation parameters was noticed except for rumen ammonia tended to be higher (0.05 < P < 0.1) in FDGL diet. Methane emission per kg dry matter ingested and methane emission per kg dry matter digested were lower (P < 0.05) in FDGL diet. Plasma glucose concentration was similar between diets and plasma glucose turnover rate tended to be higher in FDGL diet (0.05 < P < 0.1). Nitrogen retention was higher (P < 0.05) and microbial nitrogen supply tended to be higher (0.05 < P < 0.1) in FDGL diet.ConclusionFDGL diet did not impair rumen fermentation, improved nitrogen retention; while absence of significant results in reduction of methane emission, glucose turnover rate and microbial nitrogen supply, further studies at higher dose would be necessary to conclude the merit of FDGL as supplement in ruminant feedstuff.

Metabolic Design of Corynebacterium glutamicum for Production of l-Cysteine with Consideration of Sulfur-Supplemented Animal Feed.

l-Cysteine is a valuable sulfur-containing amino acid widely used as a nutrition supplement in industrial food production, agriculture, and animal feed. However, this amino acid is mostly produced by acid hydrolysis and extraction from human or animal hairs. In this study, we constructed recombinant Corynebacterium glutamicum strains that overexpress combinatorial genes for l-cysteine production. The aims of this work were to investigate the effect of the combined overexpression of serine acetyltransferase (CysE), O-acetylserine sulfhydrylase (CysK), and the transcriptional regulator CysR on l-cysteine production. The CysR-overexpressing strain accumulated approximately 2.7-fold more intracellular sulfide than the control strain (empty pMT-tac vector). Moreover, in the resulting CysEKR recombinant strain, combinatorial overexpression of genes involved in l-cysteine production successfully enhanced its production by approximately 3.0-fold relative to that in the control strain. This study demonstrates a biotechnological model for the production of animal feed supplements such as l-cysteine using metabolically engineered C.glutamicum.

Production of feather protein hydrolyzed by B. subtilis AMR and its application in a blend with cornmeal by extrusion

The poultry industry produces approximately 6 million tons of feathers as a by-product annually. The major constituent of feather is protein (80–90% of which is keratin). In order to use feather protein in animal feedstuffs, it must be hydrolyzed by chemical, physical or enzymatic processes. The aim of this study was to improve the production of chicken feather hydrolysates using keratinolytic Bacillus subtilis AMR and evaluate the effectiveness of a blend of this hydrolysate with cornmeal produced by extrusion. The addition of sucrose (0.5 g/L) to the feather medium increased the keratinolytic activity of B. subtilis AMR by 1.3 fold. The highest enzymatic activity and production of soluble protein were achieved at pH 8.0. The fermented feather medium presented large amounts of protein with low molecular weight that provided greater nutrient bioavailability. The physicochemical properties of the extrudates showed that the addition of hydrolyzed feather improved the ash and total N content. All essential amino acids were detected in the corn–feather hydrolysate extrudate. This work represents the first study that describes the use of microbial feather hydrolysate in a blend with cornmeal to make an extruded product. These results demonstrated the potential use of this hydrolysate as a supplement in animal feed.

Valorizing guava (Psidium guajava L.) seeds through germination-induced carbohydrate changes.

Guava seeds are produced as a waste product by the guava processing industry. Their high carbohydrate contents may suit the carbohydrate needs of the feed sector but their high dietary fiber content limits their feed value. The feed values of fruit seeds can be improved through germination, which involves the mobilization of nutrients through seed enzymes and alters the seed carbohydrate composition. The changes of selected carbohydrates in guava (Psidium guajava L.) seeds brought by germination to those in red bean (Vigna angularis) and winter wheat (Triticum aestivum L.) were compared. The contents of soluble carbohydrates, digestible starch, resistant starch and cellulose in the seeds were determined. The radial diffusion method was used to detect carbohydrate-degrading enzymes in the seed extracts. Guava seeds were rich in cellulose (402.2mg/g), which decreased progressively during germination, probably through the action of cellulase. Winter wheat contained the highest starch content (412.2mg/g) and also distinct quantities of α-amylase and cellulase. The starch contents of all the seeds decreased, but the soluble carbohydrate contents in red beans and guava seeds increased significantly by the end of germination, suggesting the transient oversupply of reserve metabolites. The content of hydrolyzed polysaccharides increased in the germinated seeds with detectable amounts of cellulose-degrading enzymes present, indicating improved value as feed. Further research is warranted to explore the potential of guava seeds as a source of low-cost animal feed supplements.

Purification and characterization of a protease-resistant phytase of Aspergillus oryzae SBS50 whose properties make it exceptionally useful as a feed supplement

An extracellular phytase of Aspergillus oryzae SBS50 was purified to homogeneity using ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. Purified phytase has a monomeric molecular mass of ∼80kDa exhibiting its optimal activity at pH 5.0 and 50°C with a T 1/2 of 300min at 50°C. Phytase of A. oryzae displayed broad substrate specificity with Vmax and Km values of 58.82μmol/ml/min and 1.14mM, respectively, for calcium phytate. Purity and homogeneity of the phytase was confirmed by high performance liquid chromatography and MALDI-TOF analysis revealed the identification of a peptide showing homology with acid phosphatase of Aspergillus oryzae RIB40. Among the inhibitors, 2,3-butanedione and sodium molybdate significantly inhibited the enzyme activity. Phytase of A. oryzae showed protease-resistance and was more stable during storage at 4°C and −20°C as compared to room temperature. Among all the feed samples, mustard oil cake was dephytinized more efficiently than other feed samples. These unique properties suggested that the phytase has the potential to be useful as an animal feed supplement.

Vitamin Fortification of Fluid Milk.

Vitamin concentrates with vitamins A and D are used for fortification of fluid milk. Although many of the degradation components of vitamins A and D have an important role in flavor/fragrance applications, they may also be source(s) of off-flavor(s) in vitamin fortified milk due to their heat, oxygen, and the light sensitivity. It is very important for the dairy industry to understand how vitamin concentrates can impact flavor and flavor stability of fluid milk. Currently, little research on vitamin degradation products can be found with respect to flavor contributions. In this review, the history, regulations, processing, and storage stability of vitamins in fluid milk are addressed along with some hypotheses for the role of vitamin A and D fortification on flavor and stability of fluid milk.

Potential probiotic properties of lactic acid bacteria isolated from the intestinal mucosa of healthy piglets

In the present study, the probiotic properties of 52 lactic acid bacteria strains, isolated from the intestinal mucosa of 60-day-old healthy piglets, were evaluated in vitro in order to acquire probiotics of potential application. Based on acidic and bile salt resistance, 11 lactic acid bacteria strains were selected, among which 1 was identified as Pediococcus acidilactici, 3 as Enterococcus faecium, 3 as Lactobacillus rhamnosus, 2 as Lactobacillus brevis, and 2 as Lactobacillus plantarum by 16S rRNA gene sequencing. All selected strains were further investigated for transit tolerance in simulated upper gastrointestinal tract, for adhesion capacity to swine intestinal epithelial cells J2 (IPEC-J2), for cell surface characteristics including hydrophobicity, co-aggregation and auto-aggregation, and for antimicrobial activities. Moreover, hemolytic, bile salt hydrolase and biogenic amine-producing abilities were investigated for safety assessment. Two E. faecium (WEI-9 and WEI-10) and one L. plantarum (WEI-51) exhibited good simulated upper gastrointestinal tract tolerance, and showed high auto-aggregation and co-aggregation with Escherichia coli 1570. The strains WEI-9 and WEI-10 demonstrated the highest adherence capacity. The 11 selected strains mentioned above exhibited strong antimicrobial activity against E. coli CVCC1570, Staphylococcus aureus CVCC1882 and Salmonella pullorum AS1.1859. None of the 11 selected strains, except WEI-9 and WEI-33, exhibited bile salt hydrolase, hemolytic or biogenic amine-producing abilities. This work showed that the E. faecium WEI-10 and L. plantarum WEI-51were found to have the probiotic properties required for use as potential probiotics in animal feed supplements.

Utilization of Phytase to Overcome Phytic Acid in Broiler Diet

&lt;p class="awabstrak2"&gt;Phytic acid has been considered as an antinutrient in broiler diet due to its strong chelator of divalent minerals. Phytic acid has ability for binding positively charged proteins, amino acids, and/or multivalent cations or minerals. The resulting complexes are insoluble, difficult to be hydrolyzed during digestion in poultry, and thus, nutritionally less available for absorption. The reduction of phytate activity can be carried out by phytase supplementation. The application of phytase with respect to animal feed supplement is reviewed in this paper. Application of phytase in broiler diet may liberate cations and other nutrients bound by phytate-P complexes resulting in improved production parameters in broilers. This is because phytase supplement increased nutrient (protein, minerals, amino acid, energy, and carbohydrate) digestibilities and availabilities. Overall, phytase could increase nutrient utilization in broiler, hence, increase the economic efficiency of broiler production and reduce the phosphor pollution to the environment.&lt;/p&gt;&lt;p class="awabstrak3"&gt; &lt;/p&gt;

In vitro evaluation of candidate Bacillus spp. for animal feed.

The aim of this study was to select aerobic spore-formers for animal feed based on their in vitro probiotic potential, including their enzyme-producing ability and safety assessment. Seven isolates out of 187 spore-forming bacteria were selected for their ability to produce cellulase (89.21-1668.32 U/ml), xylanase (1399.68-4351.10 U/ml), and phytase (2.72-28.70 U/ml). Among seven isolates, five had activities towards a broad range of p-nitrophenyl esters with acyl chain lengths from C2 to C12. The probiotic properties of all selected isolates varied with respect to their acid and bile salt tolerance under simulated gastrointestinal tract (GIT) conditions, and their adherence ability to human intestinal cell lines (Caco-2 and HT-29). The safety assessment revealed that the isolate CM40 was not cytotoxic to Caco-2 and HT-29, did not exhibit hemolytic activity, carried no enterotoxin or emetic toxin genes, and was susceptible to ten antibiotics, including six key antibiotics (chloramphenicol, erythromycin, gentamicin, tetracycline, streptomycin, and kanamycin) as recommended by the European Food Safety Authority (EFSA). Co-incubation of isolate CM40 with enteric bacteria (Salmonella Typhi, Salmonella Enteritidis 1781, and Escherichia coli) demonstrated that CM40 significantly decreased the number of pathogens (about 30-48%) adhering to Caco-2 and HT-29 (P < 0.05). Analysis of gene encoding 16S rRNA, gyrase A (gyrA) and the cheA histidine kinase revealed that CM40 belongs to Bacillus subtilis. On the basis of probiotic properties and basic safety aspects, the B. subtilis strain CM40 was found to possess desirable in vitro probiotic properties, and may be a potential candidate for supplementation of animal feed.

Efficacy of L-Cysteine as an Anti-Oxidant in Papain Catalyzed Synthesis of Oligopeptides in Organic Solvent System

Enzymatic peptide synthesis has drawn considerable attention for synthesis of high by-pass oligopeptide feed supplements in animal nutrition. A hardy protease, (papain) with a cysteine moiety at the active site requires the presence of an anti-oxidant in the reaction medium to ensure that the thiol group remains intact. Free cysteine has been the antioxidant of choice for papain-catalyzed synthesis of oligopeptides in aqueous systems. However, due to limited solubility of cysteine in organic solvents, it is generally not a suitable antioxidant for the synthesis of oligopeptides in biphasic solvent systems; instead, mercaptoethanol is often used. Lysine and Methionine is couple of well-known limiting amino acids that find application in cattle feed and poultry. Synthesis of co-oligopeptides of Lysine and Methionine has generally been attempted in bi-phasic solvent systems with Mercaptoethanol as an anti-oxidant. The inherent toxicity of mercaptoethanol when present even in trace amounts make its use undesirable during synthesis of oligopeptides to be used as feed supplements. Use of non-toxic antioxidants such as Cysteine would make the end product more amenable as a nutritional supplement for animal feed. Therefore efficacy of L-cysteine as an anti-oxidant was investigated during papain catalyzed oligomerization of Lys, Arg, Glu and Asp in two organic systems: a three phase micro-aqueous media consisting of n-octane, DFP and water, as well as homogeneous ACN/water mixtures. Reactions were also carried out under an argon atmosphere in the presence and absence of anti-oxidants. The results of the experiments showed that L-cysteine facilitated oligomer synthesis in both the three phase system and the ACN/water mixture. The overall oligomer yields were found to be better than 75% in the presence of L-Cysteine. Oligopeptide yields obtained through reactions carried out under the argon atmosphere were less than 20%.

REVIEW ON PHORBOL ESTER DEGRADATION OF JATROPHA SEED CAKE FOR ITS USE AS ANIMAL FEED

&lt;em&gt;Jatropha curcas&lt;/em&gt; is an oil-seed plant with good adaptability to grow in unfavourable conditions like infertile soil with scanty rainfall. It had been exploited for the extraction of oil for bio-diesel. The compressed seed cake, after the oil extraction, is a rich source of protein with certain toxic and anti-nutritional factors. The major toxins in the seed cake are phorbol esters and trypsin inhibitors that lead to various health problems if ingested. Even though the application of the various extracts carries a lot of beneficial advantages, yet the toxicity in oil and the compressed cake does not allow the by-products and the oil to be used elsewhere. Various physicochemical and biological methods have been described for the detoxification of &lt;em&gt;Jatropha&lt;/em&gt; seed cake and oil of which the chemical extraction with methanol and ethanol have shown promising results in reducing the toxin contents by 97-100% while UV-irradiation reduced the phorbol esters completely. Submerged fermentation by&lt;em&gt; Bacillus&lt;/em&gt; sp. achieved complete detoxification of phorbol esters within a week. A new strain was found to degrade the phorbol esters to phorbol, myristic acid and acetic acid within 12 h of incubation in submerged fermentation process. The detoxified products, in future, can be used as animal feed and food supplement to help utilize the by-products as a healthy diet.

Investigation of the mineralogical petrographic, geochemical and physical properties of carbonate rocks from Ileia Prefecture and assesment for industrial applications

Research on carbonate rocks has been increasingly important during the last few decades, due to their numerous applications. In this paper, we examined the properties of Cretaceous carbonate rocks from the Olonos-Pindos Zone from Ileia Prefecture. The relationships between various physical properties and between petrographic features and physical properties were defined. This may lead to preliminary evaluation of carbonate rocks using microscopic investigation, which unlike the elaboration of a series of physical and mechanical properties, is a fast and inexpensive method. In addition, these properties, along with their geochemical characteristics, were used to assess the suitability of these carbonate rocks as fillers in various applications. Detailed petrographic study of the collected samples revealed the occurrence of biomicrite, intramicrite (mudstone και wackestone), biosparite (packstones and grainstones), intrasparite and rudstone. From the lower to the upper members, a decrease of the grain size of calcite is observed. X-ray diffraction study of the insoluble residue revealed the presence of quartz, chlorite, illite, smectite, and mixed layer illite-chlorite. The colour index of the investigated powdered samples is negatively influenced by the increase of organic and insoluble residue contents. The water absorption is inversely correlated with CaO content while moisture is positively correlated with insoluble residue. The grainstone and rudstone are characterized as ultra-high pure calcium limestones, except for Mg-rich crystalline dolomitic limestones. The packstones are calcium limestones of high to medium purity. Mudstones and wackestones are characterized as low calcium limestones. The results suggest that all the examined lithotypes can be used as soil conditioners and animal feed supplements; however only grainstones and rudstones are suitable for desulphurization of exhaust gasses and cement production.

Yield and nutrient content of hydroponics cowpea sprouts at various stages of growth

Hydroponics cowpea (Vigna unguiculata) (NB-4) sprouts were produced daily for 15 days in a hydroponics chamber (greenhouse) equipped with automatic sprayer irrigation of tap water. Cowpea (NB-4) with seed rate of 2.54 kg/m2 with replicates of 15 was soaked in tap water individually for 4 h.The seeds were strained and spread in the greenhouse trays. Inside the greenhouse, each replicate was allowed to grow for 9 d, after which it was harvested. The fresh yield (kg/kg seed) of the hydroponically sprouted cowpea (NB-4) increased with the advancement of growing period and remained similar and highest from day 6 (6.34) to 9 (6.63) growing period. Simultaneously, with the growth of the hydroponics cowpea sprouts, the DM content (%) decreased and remained similar and lowest from day 8 (6.91) to day 9 (6.49) growing period. The CP(%) of the hydroponics cowpea fodder was lowest on the day 1 (22.30), then increased and remained similar during seventh (30.10) to ninth (31.23) day sprouting period. The EE content increased and remained similar during day 5 (2.31%) to 8 (2.47%) of sprouting, however, it was highest on day 9 (3.22) of growth. During the growth period of the hydroponics cowpea sprouts, the CF(%) and NFE(%) had an increasing and decreasing trend and was highest and lowest, respectively, on day 9 (21.59 and 38.59). There was an increasing trend in the TA and AIA content of the hydroponics cowpea sprouts with the advancement of the growth period, which was lowest on day 1 (3.99) and highest (5.38) on day 9. It can be concluded that based on the yield and different nutrient contents, cowpea (NB-4) can be sprouted hydroponically for 6–9 days to use as feed supplement for dairy animals.