Twin-Screw Extrusion Processing of Feed Blends Containing Distillers Dried Grains with Solubles (DDGS)

Extrusion trials were conducted with varying levels of distillers dried grains with solubles (DDGS) along with soy flour, corn flour, fish meal, vitamin mix, mineral mix, and net protein content adjusted to 28% using a Wenger TX-52 twin-screw extruder. The properties of extrudates were studied in experiments conducted using a full-factorial design with three levels of DDGS content, two levels of moisture content, and two levels of screw speed. Increasing the DDGS content from 20 to 60% resulted in a 36.7% decrease in the radial expansion, leading to a 159 and 61.4% increase in the unit density and bulk density of the extrudates, respectively. Increasing the DDGS content resulted in a significant increase in the water absorption index (WAI) but a significant decrease in the water solubility index (WSI) of the extrudates. Changing the screw speed and moisture content had no significant effect on the radial expansion ratio but resulted in a significant difference in the bulk density of the extrudates, which may be due to the occurrence of longitudinal expansion. Even though changing the moisture content and screw speed had no significant effect on the WSI of the extrudates, significant differences in the WAI of the extrudates were observed. The ingredient components in the blend and moisture content had an influence on the color changes of the extrudates, as did the biochemical changes occurring inside the barrel during processing. Overall, it was determined that DDGS could be included at a rate of up to 60% using twin-screw extrusion, and that viable pelleted floating feeds can be produced.     

Effect of Extrusion Process Parameters on the Quality of Buckwheat Flour Mixes

A response surface analysis using a second-order central composite design was used to study the effect of extrusion process parameters on the extrudate quality of three blends containing buckwheat flour. The extrudates were prepared as three blends. Blend 1 was a 55:40:5 (w/w) mix of light buckwheat flour, wheat flour, and nonfat dry milk (NFDM). Blend 2 was a 40:55:5 mix of light buckwheat flour, corn meal, and NFDM. Blend 3 was a 30:60:10 mix of light buckwheat flour, corn meal, and NFDM. The blends were processed in a twin-screw extruder with factorial combinations of the parameters including: process temperatures of 95–150°C, dough moisture of 15–22%, and screw speeds of 260–390 rpm. The linear components alone significantly explained most of the variation of expansion index, bulk density, water absorption, and breaking strength. The greatest amount of variability was explained by process temperature for blend 1. Dough moisture accounted for the greatest amount of variation for blends 2 and 3. Maximum predicted expansion index values and high water absorption percentages were obtained at low dough moisture levels. Dough moisture and process temperatures were the most important factors predicting bulk density. Sensory evaluation of texture, color, flavor, and general acceptability scores of selected samples ranked blend 3 > blend 2 > blend 1. The in vitro protein digestibility values ranked blend 1 > blend 2 > blend 3. An increase of up to 9.5% units in the protein digestibility values was observed when compared to the nonextruded raw blends.     

Effects of Calcium Hydroxide and Processing Conditions on Corn Meal Extrudates

The effects of added calcium hydroxide (0.0, 0.15, 0.25, and 0.35%) and processing conditions, feed moisture content (mc) (16, 18, and 20%) and barrel temperature (130 and 150°C) on characteristics of corn meal extrudates were studied. Extruder screw speed was maintained at 130 rpm. Corn meal was extruded with a single-screw extruder (Brabender model GNF 1014/2) with a screw compression ratio of 3:1. The highest values (P < 0.05) for radial expansion and the lowest values for density and breaking force of extrudates were found for the treatment with 0.00% calcium hydroxide extruded at 16% feed mc and 130°C barrel temperature. This treatment was statistically different from the other treatments. Best values for radial expansion of samples extruded with added calcium hydroxide were for the samples with 0.15% calcium hydroxide at 18% feed mc and 130°C barrel temperature, followed by the sample with 0.35% calcium hydroxide at 16% feed mc and 130°C barrel temperature. Water absorption index and water solubility index were affected by calcium hydroxide and extrusion conditions evaluated. Extrudates had large numbers of flattened and sheared granules. Increases in calcium hydroxide increased extrudate yellowness. The combined action of calcium hydroxide and extrusion conditions completely modified the organized structure of the starch and suggest the formation of a starch-calcium complex (crystalline region). The texture of the extruded products was crispy after puffing.     

Extrusion of Cross‐Linked Hydroxypropylated Corn Starches II. Morphological and Molecular Characterization

A series of cross‐linked hydroxypropylated corn starches were extruded with a Leistritz micro‐18 co‐rotating extruder. Extrusion process variables including moisture (30, 35, and 40%), barrel temperature (60, 80, and 100°C), and screw design (low, medium, and high shear) were investigated. Scanning electron microscopy (SEM) of extruded starches showed a gel phase with distorted granules and granule fragments after extrusion at 60°C. After extrusion at 100°C only a gel phase was observed with no granular structures remaining. High performance size exclusion chromatography (HPSEC) equipped with multiangle laser light‐scattering (MALLS) and refractive index (RI) detectors showed extruded starches degraded to different extents, depending on extrusion conditions. The average molecular weight of the amylopectin of unextruded native corn starch was 7.7 × 10⁸. Extrusion at 30% moisture, 100°C, and high shear reduced the molecular weight of amylopectin to 1.0 × 10⁸. Hydroxypropylated normal corn starch extruded at identical conditions showed greater decreases in amylopectin molecular weight. With the addition of cross‐linking, the amylopectin fractions of the extruded starches were less degraded than those of their native and hydroxypropylated corn starch counterparts. Similarly, increasing moisture content during extrusion lowered amylopectin degradation in the extruded starches. Increasing temperature during extrusion of cross‐linked hydroxypropylated starches at high moisture content (e.g., 40%) lowered amylopectin molecular weights of the extruded starches, whereas increasing extrusion temperature at low moisture content (30%) resulted in less degraded molecules. This difference was attributed to the higher glass transition temperatures of the cross‐linked starches.     

Nutritional and Economic Implications of Protein Variance and Application of Statistical Process Control in the Chinese Feed Industry

The adoption of Statistical Process Control (SPC) in the Chinese feed industry to reduce finished feed protein and moisture content variation can improve feed quality and profitability. Two studies were conducted to characterize protein variation in the Chinese feed industry. Study I involved four participating feed mills where we collected three samples per bag from four bags per lot of soybean meal, rapeseed meal, fish meal, corn gluten meal, meat & bone meal, and cottonseed meal. We analyzed these ingredients for protein and moisture content. Cottonseed meal and meat & bone meal displayed the highest protein variability between bags and within bags compared with other protein ingredients. We found no significant difference for within‐bag protein variance of the other feed ingredients; however, soybean meal had a significantly lower between‐bag variance (0.04%). Study II assessed the effect of adopting SPC. Five feed mills provided data for one year of finished feed protein and moisture content before and after adopting SPC. Three of the five feed mills experienced a significant (P < 0.10) reduction (0.20, 0.52, and 0.70%) in the absolute finished feed protein variation after adopting SPC. The two feed mills that did not display a significant reduction in absolute protein variation were already manufacturing feed with a low (<0.4% protein) absolute deviation from the target. These data demonstrate that feed mill managers can control finished feed variability through a number of management techniques including SPC.     

Preparation of High‐Quality Protein‐Based Extruded Pellets Expanded by Microwave Oven

The aim of this work was to study the effects of extrusion barrel temperature (75–140°C) and feed moisture (16–30%) on the production of third‐generation snacks expanded by microwave heating. A blend of potato starch (50%), quality protein maize (QPM) (35%), and soybean meal (SM) (15%) was used in the preparation of the snacks. A laboratory single extruder with a 1.5 × 20.0 × 100 mm die‐nozzle and a central composite routable experimental design were used. Expansion index (EI) and bulk density (BD) were measured in expanded pellets, viscosity at 83°C (V₈₃), thermal properties, and relative crystallinity were measured in extruded pellets. EI increased and BD decreased when the barrel temperature was increased, while the feed moisture effect was not significant. V₈₃ increased when feed moisture increased. Extrusion modified the crystalline structures of the pellets and the X‐ray data suggests the formation of new structures, probably due to the development of amylose‐lipid complexes. The maximum expansion of pellets was found at barrel temperatures of 123–140°C, and feed moisture of 24.5–30%. It is possible to obtain a functional third‐generation snack with good expansion characteristics using a microwave oven, and this snack has health benefits due to the addition of QPM and SM.     

Isoflavones in soy-based foods consumed in Brazil: levels,distribution, and estimated intake

The isoflavone content and profile in processed soy-based products consumed in Brazil were determined by high-performance liquid chromatography and photodiode array detection of the intact isoflavones (naturally occurring aglycons, malonyl, acetyl, and beta-glycosides derivatives). Total isoflavone content varied significantly among products, from 2 to 100 mg/100 g (wet basis, expressed as aglycons), with the lowest content being found for soy-based enteral/oral diets and the highest found for textured soy proteins. For soy beverages isoflavone content varied from 12 to 83 mg/L. Soy sauce, miso, and tofu had isoflavone contents of 5.7 mg/L, 20 mg/100 g, and 7 mg/100 g, respectively. The beta-glycosides were the predominant form of the isoflavones in the products analyzed, except for miso, shoyu, and "Diet Shake" in which the aglycons were present in the highest proportions. On the basis of these data, the daily intake of isoflavone from soy products was estimated: the highest values were found for infants fed soy-based formulas, from 1.6 to 6.6 mg/kg of body weight.     

Distribution profiles of isoflavone isomers in black bean kojis prepared with various filamentous fungi

This study was conducted to compare the transformation of both isoflavone derivatives (aglycones, beta-glucosides, and acetyl and malonyl gluclucosides) and beta-glucosidase activity in kojis fermented with various generally recognized as safe filamentous fungi including Aspergillus awamori, Aspergillus oryzae, Aspergillus sojae, Rhizopus azygosporus, and Rhizopus sp. no. 2. Solid fermentation was performed to prepare the kojis by inoculating the steamed black beans with starter organism and culturing at 30 degrees C for 3 days. Results revealed that fermentation caused a marked increase in the content of aglycone (daidzein, glycitein, and genistein), the bioactive isoflavone, and a significant reduction in the content of beta-glucoside isoflavone (daidzin, glycitin, and genistin), compared with the unfermented steamed black bean. The extent of increased aglycone and reduced beta-glucoside isoflavone content varied with the starter organism used. Among the various black bean kojis prepared, the Rhizopus sp. no. 2 koji showed the highest level of enhancement in aglycone content. In the Rhizopus sp. no. 2 koji, the percentage of aglycone to total isoflavone increased from an initial approximately 2.9 to approximately 58.9% after fermentation. In comparison, the percentages found in kojis prepared with other starter organisms ranged from 18.9 to 38.9% after fermentation. Further preparations of black bean kojis with A. awamori at different cultivation temperatures (25, 30, and 35 degrees C) and various fermentation periods (1-5 days) revealed that koji prepared at 30 degrees C for 4 days showed the highest content of aglycones, with 7.7-, 5.7-, and 4.8-fold increases in the content of daidzein, genistein, and glycitein, respectively. In addition, the increase of aglycone content and the increase of beta-glucosidase activity during the fermentation of this koji showed a similar trend.     

Effect of Die Dimensions on Extrusion Processing Parameters and Properties of DDGS-Based Aquaculture Feeds

The goal of this study was to investigate the effect of die nozzle dimensions, barrel temperature profile, and moisture content on DDGS-based extrudate properties and extruder processing parameters. An ingredient blend containing 40% distillers dried grains with solubles (DDGS), along with soy flour, corn flour, fish meal, whey, mineral and vitamin mix, with a net protein content adjusted to 28% was extruded in a single-screw laboratory extruder using seven different die nozzles. Increasing moisture content of the ingredient mix from 15 to 25% resulted in a 2.0, 16.0, 16.3, 22.9, 18.5, 32.5, and 63.7% decrease, respectively, in bulk density, water-solubility index, sinking velocity, L*, b*, mass flow rate, and absolute pressure, as well as 11.6, 16.2, and 7% increases, respectively, in pellet durability, water-absorption index, and a*. Increasing the temperature from 100 to 140°C resulted in 17.0, 5.9, 35.4, 50.6, 28.8, 33.9, and 33.9% decreases, respectively, in unit density, pellet durability, sinking velocity, absolute pressure, specific mechanical energy, torque, and apparent viscosity, but a 49.1 and 16.9% increase, respectively, in dough temperature and water-absorption index. Increasing the L/D ratio of the die nozzle resulted in an increase in bulk density, L*, a*, and torque, but a decrease in unit density, pellet durability, water-absorption index, sinking velocity, b*, mass flow rate, dough temperature, and apparent viscosity. As demonstrated in this study, the selection of an appropriate die geometry, in addition to the selection of suitable temperature and moisture content levels, are critical for producing DDGS-based extrudates with optimum properties.     

Effect of Free Fatty Acids Addition on Corn Grits Extrusion Cooking

The influence of added fatty acids on extrusion cooking of corn grits and extrudate properties was studied. Samples with three average corn grits particle sizes were processed in a twin-screw extruder; fatty acids content (0.2–0.8%, wb) varied, and experimental conditions were kept constant (moisture content 18.4% wb, barrel temperature 150°C, screw speed 165 rpm). The effect of adding fatty acids was studied by analyzing water solubility and absorption indices, expansion indices, and mechanical (puncture test) and structural (computerized image analysis) extrudate properties. When fatty acids were added, water solubility and absorption indices decreased, radial expansion index decreased, longitudinal expansion index increased, number of cells increased, and mechanical resistance of extrudates decreased. The influence of added fatty acids was attributed to the formation of fatty acids-amylose complexes.     

Functional Characteristics of Extruded Blends of Whey Protein Concentrate and Corn Starch

The aim of this work was to study the effects of extrusion barrel temperature (70–180°C), feed moisture (18–30%), pH (3–8), different proportions of corn starch (75–95%), and whey protein concentrate (WPC, 80% protein concentration) (25–5%) on the preparation of functional blends. Expansion index (EI), bulk density (BD), compression force (CF), color, water absorption index (WAI), water solubility index (WSI), gel strength (GS), syneresis of the gel, and in vitro digestibility were evaluated. Barrel temperature and the proportion of WPC had significant effects on BD; at higher temperatures, BD was lower. Feed moisture and pH had significant effects on EI; with lower moisture and higher pH, the EI increased. An interaction of barrel temperature and feed moisture had an effect on WAI; at lower moisture content, the temperature effect was nonexistent, whereas at higher temperatures and feed moisture content, the WAI increased. The pH level had a significant effect on WSI, showing high WSI when lower pH levels were used. Color analysis showed that higher protein content and pH generated higher δE values; low feed moisture and low pH resulted in gel syneresis. Higher in vitro digestibility was obtained when a higher WPC proportion and pH were used. Extruded WPC-CS blends under alkaline and acidic conditions were affected by the preparation of diverse formulations that potentially can be used in foods to improve some functional and protein content.     

Extrusion of Cross‐Linked Hydroxypropylated Corn Starches I. Pasting Properties

A series of cross‐linked (0, 0.014, 0.018, 0.024, and 0.028% POCl₃, dry starch basis) hydroxypropylated (8%) corn starches were extruded using a Leistritz micro‐18 co‐rotating extruder. Process variables included moisture, barrel temperature, and screw design. Differential scanning calorimetry and X‐ray diffraction studies showed the level of starch crystallinity decreased with increasing severity of extrusion conditions. Pasting properties of the extruded starches were examined using a Rapid Visco Analyser. Pasting profiles of starches extruded at different conditions displayed different hot paste viscosity and final viscosity. Increasing starch moisture content during extrusion and level of cross‐linking increased starch viscosity (P < 0.0001), whereas increasing extrusion temperature and shear decreased starch viscosity (P < 0.0001). Interactions were found between level of cross‐linking and screw design and between extrusion temperature and starch moisture content (P < 0.0001).     

Red clover Trifolium pratense L. phytoestrogens: UV-B radiation increases isoflavone yield, and postharvest drying methods change the glucoside conjugate profiles

Isoflavone extracts of red clover Trifolium pratense L. (cv. Pawera) with dissimilar glucoside conjugate profiles were obtained by employing different postharvest drying methods. The most prominent isoflavones found were formononetin and biochanin A and their corresponding glucosides and malonyl glucoside esters. Postharvest freeze drying inhibited the conversion of the glycosides to the aglycones, while vacuum drying allowed for maximum conversion of the glycosides to their corresponding aglycones. Air drying produced a low level of the aglycones formononetin and biochanin A, and oven drying promoted decarboxylation of the malonyl glucosides to the acetyl glucosides. Exposure to enhanced UV-B radiation resulted in an increase in total formononetin and biochanin A isoflavone levels, indicating that harvest during a period of high ambient UV-B radiation may be appropriate for maximum yield. The levels of caffeic acid and flavonols also increased by about 40 and 250%, respectively, on exposure to enhanced UV-B radiation.     

Study on the Parameters of Extrusion Pretreatment of Corn Germ with Semi‐Wet Milling in Solvent Oil Extraction

Efficiency of oil extraction from corn germ was improved by the extrusion pretreatment, and residual oil was taken as the index of this oil extraction process. An orthogonal rotation combination test design of five levels and four factors, consisting of the moisture of material, screw speed, barrel temperature, and die nozzle diameter, was employed to optimize the model and reaction condition. The optimum parameters of extrusion were as follows: moisture of material, W = 12%; extrusion temperature, T = 105°C; screw speed, n = 185 rpm; die nozzle diameter, Φ = 9 mm × 3. Under these optimum conditions, residual oil of this process was 0.61–0.66%, which was lower than residual oil by pressing alone (residual oil of 5–6%) or by a combination of prepressing and hexane extraction (residual oil of nearly 2%). The corn oil obtained by the optimum extrusion parameters was analyzed, and iodine value, acid value, peroxide value, and saponification value were 1,250 g/kg, 400 mg/kg, 5.1 mmol/kg, and 189%, respectively.     

Determination of 15 isoflavone isomers in soy foods and supplements by high-performance liquid chromatography

Soy isoflavone is the generic name for the isoflavones found in soy. We determined the concentrations of 15 soy isoflavone species, including 3 succinyl glucosides, in 22 soy foods and isoflavone supplements by high-performance liquid chromatography (HPLC). The total isoflavone contents in 14 soy foods and 8 supplements ranged from 45 to 735 μg/g and from 1,304 to 90,224 μg/g, respectively. Higher amounts of succinyl glucosides were detected in natto, a typical fermented soy product in Japan; these ranged from 30 to 80 μg/g and comprised 4.1-10.9% of the total isoflavone content. In soy powder, 59 μg/g of succinyl glucosides were detected, equivalent to 4.6% of the total isoflavone content. These data suggest that the total isoflavone contents may be underestimated in the previous studies that have not included succinyl glucosides, especially for Bacillus subtilis -fermented soy food products.     

Addition of Sodium Stearoyl Lactylate to Corn and Sorghum Starch Extrudates Enhances the Performance of Pregelatinized Beer Adjuncts

Double mashing for wort production is a time‐consuming process that can be reduced if pregelatinized adjuncts are used. Optimal extruding conditions were determined to obtain brewing adjuncts from corn and sorghum starch. For corn starch extrusion, a Box–Behnken design was devised in which moisture, screw speed, temperature of the barrel, and concentrations of sodium stearoyl lactylate (SSL) were varied, and sorghum starch was extruded according to a 2³ model in which the modified variables were moisture, SSL concentration, and temperature. The aim was to maximize starch damage and minimize resistant starch and final viscosity as determined with a Rapid Visco Analyzer. The treatments that satisfied these requirements were mashed, and wort extract yield was determined. Glucose, maltose, and maltotriose concentrations in the resulting worts were determined by HPLC with a refractive index detector. Feedstock tempering and SSL content were the most important factors affecting the response; for corn starch, treatments with lower moisture (20%) and middle levels of SSL (0.5%) or with high levels of both moisture (40%) and SSL (1%) produced the most desirable samples for mashing, whereas for sorghum starch the best treatment was tempering to 20% moisture and containing middle levels of SSL (0.5%). No statistical differences were found between these experimental treatments and the control.     

用于浸油的大豆挤压膨化研究

研究了大豆挤压膨化系统诸参数（模孔孔径、挤压温度、物料含水率、螺杆转速）对大豆粗脂肪、豆粕残油率、蛋白质含量、及其氮可溶解指数、脲酶活性的影响规律和挤压膨化系统最佳参数。结果表明,只要系统参数选择合适,可使膨化后大豆粗脂肪基本不减少,粕残油小于１％,使原设备浸出能力增加１５０％。且膨化前后大豆粗蛋白含量基本不变,氨基酸没有损失,膨化后氮可溶解指数和脲酶活性均略有下降。同时省去目前大豆膨化浸油和传统浸油过程的破碎、软化、轧胚、蒸炒等工序。需增加粉碎、膨化工序。     

Effect of extrusion on isoflavone content and antiproliferative bioactivity of soy/corn mixtures

The present studies were conducted to determine changes in the quantities of select isoflavones and in the bioactivity (ability to inhibit proliferation of human breast cancer cell lines) of extracts from blends of soy protein and cornmeal during extrusion processing. The extrusion of samples resulted in an average 24% decrease in the concentration of total isoflavones for all samples. Although the amounts of specific genistein-derived and daidzein-derived forms changed following extrusion, the content of the aglycones genistein and daidzein per g sample generally did not change. The extrusion of samples generally resulted in decreased antiproliferative action toward breast cancer cells, although antiproliferative activity was not eliminated. Therefore, extrusion of soy protein/cornmeal-containing foods are likely to retain a considerable portion of their isoflavone content and some of the health benefits associated with soy.     

Evaluation of isoflavone aglycon and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector

An ultrafast HPLC/UV-vis DAD method working at 254 nm was applied for the determination of isoflavone aglycons and glycosides (genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin, and biochanin A) in roots, stems, leaves, and soy pods of soy plants and in soybeans of five varieties (Korada, Quito, Rita, OAC Erin, and OAC Vison). An Atlantis dC18 ultrafast RP chromatographic column (20 mm x 2.1 mm, 3 microm particle size) was applied for separation of the isoflavone aglycons and glycosides. A flow rate of the mobile phase (0.1% (v/v) acetic acid, pH 3.75-solvent A and methanol-solvent B) was 0.35 mL min(-1), and the column temperature was 36 degrees C. A linear gradient profile from 13 up to 22% B (v/v) from zero to 2.5 min, up to 30% B to 3.21 min, up to 35% B to 4 min, up to 40% B to 4.5 min, up to 50% B to 5.14 min, and followed by negative gradient up to 13% B to 7.71 min was used. The absolute limits of detection per sample injection (5 microL) were the highest for biochanin A (166.2 fmol) and the lowest for genistin (17.0 fmol), respectively. An accelerated solvent extraction (ASE) in combination with sonication was applied for isolation of biologically active compounds. A solid-phase extraction procedure was used to purify the extracts in the case of analysis of soy plants parts. The recoveries of 96-106% were obtained for the different concentrations of the isoflavone aglycons and glycosides and the different matrixes (overall RSDs 2-9%). The highest isoflavone concentrations were found in roots (12.5 microg g(-1) dry weight), while the amounts were about 3-1100 microg g(-1) fresh weight in different varieties of soybeans.     

Variations in isoflavone levels in soy foods and soy protein isolates and issues related to isoflavone databases and food labeling

The reliability of databases on the isoflavone composition of foods designed to estimate dietary intakes is contingent on the assumption that soy foods are consistent in their isoflavone content. To validate this, total and individual isoflavone compositions were determined by HPLC for two different soy protein isolates used in the commercial manufacture of soy foods over a 3-year period (n = 30/isolate) and 85 samples of 40 different brands of soy milks. Total isoflavone concentrations differed markedly between the soy protein isolates, varying by 200-300% over 3 years, whereas the protein content varied by only 3%. Total isoflavone content varied by up to 5-fold among different commercial soy milks and was not consistent between repeat purchases. Whole soybean milks had significantly higher isoflavone levels than those made from soy protein isolates (mean +/- SD, 63.6 +/- 21.9 mg/L, n = 43, vs 30.2 +/- 5.8 mg/L, n = 38, respectively, p < 0.0001), although some isolated soy protein-based milks were similar in content to "whole bean" varieties. The ratio of genistein to daidzein isoflavone forms was higher in isolated soy protein-based versus "whole bean" soy milks (2.72 +/- 0.24 vs 1.62 +/- 0.47, respectively, p < 0.0001), and the greatest variability in isoflavone content was observed among brands of whole bean soy milks. These studies illustrate large variability in the isoflavone content of isolated soy proteins used in food manufacture and in commercial soy milks and reinforce the need to accurately determine the isoflavone content of foods used in dietary intervention studies while exposing the limitations of food databases for estimating daily isoflavone intakes.