Saccharomyces cerevisiae and phytase co-fermented wheat bran (FWB) on the growth performance and health status of broilers.The aim of this study was to investigate the effects of different amounts of wheat bran (WB) inclusion and postbiotics form by Study randomly allocated a total of 300 male broilers to a control and 4 treatment groups  with each pen having 20 broilers and 3 pens per treatment.Lactobacillus spp. number in caecum; and both 5% and 10% FWB can increase ash content in femurs. Compared to control group, all treatments increase mucin 2, and tight junction (TJ), such as occludin, claudin-1, zonula occludens-1, and mRNA expression in ileum by at least 5 folds. In chicken peripheral blood mononuclear cells, nicotinamide adenine dinucleotide phosphate-oxidase-1 mRNA expression decreases from 2 to 5 times, and glutamate-cysteine ligase catalytic subunit mRNA expression also increases in all treatment groups compared to control group. The mRNA expression of pro-inflammatory cytokines, including interleukin-6 (IL-6), nuclear factor-κB, and IL-1β, decreases in 5% and 10% FWB groups compared to control group.The WB does not contain enzymes, but there are 152.8, 549.2, 289.5, and 147.1 U/g dry matter xylanase, protease, cellulase and β-glucanase in FWB, respectively. Furthermore, FWB can decrease nitric oxide release of lipopolysaccharide stimulated chicken peripheral blood mononuclear cells by about two times. Results show that 10% FWB inclusion had significantly the highest weight gain (WG) at 1 to 21 d; 5% FWB had the lowest feed conversion rate at 22 to 35 d; 10% WB and 10% FWB inclusion have the highest villus height and To summarize, both WB and FWB inclusion in broilers diets increase TJ mRNA expression and anti-oxidation and anti-inflammation, but up to 10% FWB groups have better WG in different stages of broiler development. As climate change and global crude prices rise, corn, the main ingredient of polysaccharide source of poultry diet and regarded as one of the materials for biomass energy is becoming more expensive. Furthermore, the outbreak of COVID-19 spreading worldwide has caused a decrease of available labor that affects the production of corn and soybeans leading to feed deficiencies. Therefore, it is important to find alternative sources of carbohydrates while safeguarding the health of animals . BecauseAs a major by-product of wheat production, wheat bran (WB), comprises about 15% crude protein (CP), 28% carbohydrate, and 42% insoluble fiber with many anti-oxidation compounds included, such as ferulic acid, tocopherol, and lutein . TherefoAmong the restrictions of WB utilization, fermentation by probiotics and phytase addition can solve the problem of high phytate acids and fiber content. Cowieson et al  and HamdSaccharomyces cerevisiae (S. cerevisiae) mainly secrets fiber-related enzymes, such as glucanase, mannanase, cellulase and xylanase, etc. Therefore, fermentation by S. cerevisiae can degrade fiber in WB, while it cannot decrease the phytase content in WB. Therefore, co-fermentation of WB by S. cerevisiae and phytase may be better than using S. cerevisiae only. Nevertheless, few studies have carried out co-fermentation of WB with probiotic and a specific enzyme. In theory, the postbiotic form from S. cerevisiae and phytase co-fermentation wheat bran (FWB) should increase the nutrient utilization and decrease the phytate content, and increase the growth performance of broilers. Furthermore, from the performance of S. cerevisiae, phytase and WB shown in previous studies, we predict that the FWB can increase the antioxidation function, enhance the tight junction (TJ) protein presentation, and decrease the inflammatory-related cytokine in broilers. We supposed that the use of FWB and WB to replace part of the feed will not have a negative impact on the growth performance of the broilers, and will improve their intestinal health. Therefore, we measured the broilers’ growth performance, intestinal morphology, microbiota composition, blood biochemical analysis, and mRNA expression in chicken peripheral blood mononuclear cells (chPBMCs).In recent years, the new concept of “postbiotic” has arisen. Unlike probiotic, prebiotic, or synbiotic, postbiotic emphasizes more on the metabolic changes due to probiotic in the fermentation matrix . BecauseSaccharomyces cerevisiae and phytase co-FWB was created by the method as follows: Add 10,000 units phytase-6 and 100 mL 108 colony-forming unit (CFU)/mL S. cerevisiae to 1 kg WB and let ferment at 30°C for 3 days. The FWB was moved from the incubator to a 50°C oven and staved for 1 day. The FWB was collected and stored at a 4°C refrigerator before use.We analyzed the xylanase , proteasExtractable phosphorus measurement was modified from the previous study . BrieflyThe methods of pentose and hexose measurement were modified from the phenol-sulfuric acid method. Briefly, add 1 mL 5% phenol solution and 5 mL sulfuric acid to FWB water extra. After incubation of 15 minutes, detect the absorbance at 480 nm for pentose and 490 nm for hexose. Take the xylose and glucose as the standard curve for pentose and hexose, respectively.Add deionized water to FWB and stir it for 1 hour to measure the xylooligosaccharide. After stirring, centrifuge it at 3,000 rpm for 10 minutes and filter through Advantec No. 1 filter paper and 0.22 μm filter successively. Then, analyzed the sample by high performance liquid chromatography (HPLC)  equipped with a pump (L-2130), a column , a RI-detector (L-2490), an autosampler (L-2200) and a computer system with HPLC D-2000 Elite.7 cells/mL.The methods of chPBMCs isolation were done according to Kaiser et al . The who2 for 2 hours. The 10 μL 1 ng/mL lipopolysaccharides (LPS) and 10 μL bacteria-free sample solution were added in the 107 chPBMCs and co-incubated for 24 hours. After culturing, add 100 μL Griess reagent and detect the absorbance at 540 nm.chPBMCs were cultured in the RPMI-1640 and 10% FBS at 37°C and 5% CO2 for 2 hours. The 10 μL 1 ng/mL LPS and 10 μL bacteria-free sample solution were added in the 107 chPBMCs and co-incubated for 48 hours. After co-incubating, adding 20 μL 0.5% 3--2,5-diphenyltetrazolium bromide (MTT) solution and culture for 4 h. After culturing, add 100 μL dimethyl sulfoxide and detect the absorbance at 570 nm.chPBMCs were cultured in the RPMI-1640 and 10% FBS at 37°C and 5% CORoss 308) were divided into 5 groups: basal diet (control), 5% WB inclusion (5% WB), 5% FWB inclusion (5% FWB), 10% WB inclusion (10% WB), and 10% FWB inclusion (10% FWB). There were 20 broilers in each pen and 3 replicates for each treatment . The average initial body weight (49.0±0.5 grams/birds) was similar in each pen. A temperature-controlled house (33°C±0.5°C for one-day-old chicks and 21°C±1°C for chickens after 4 weeks old) was provided for this experiment during the whole rearing stage. In the whole rearing stage, there was 23-hour lighting and 1 hour dark per day, and free of additional medical treatment. Furthermore, feed and water were ad libitum. In order to standardise metabolic energy and CP in each treatment, the diet of each treatment group was recalculated to meet or exceed the nutrient requirements (NRC) [The experiments were carried out at National Chung Hsing University, Taiwan, and all of the protocols followed those of the Animal Care and Use Committee (IACUC: 107-013) at October 2018. Animal experiment design methods followed by Teng et al [ts (NRC) . The staThirty 35-day-old broilers  were used for the intestinal morphology test. The middle of the jejunum and the middle ileum of fasting one-day broilers were removed and fixed in formalin for 3 days. The samples were embedded in paraffin and stained with haematoxylin and eosin. The stained paraffin was sliced and observed under a light microscope, and a Motic Image Plus 2.0 analysis system  was used to measure the villus height, crypt depth, and tunica muscular.The blood of forty-five 35-day-old broilers  was collected for the blood and serum characteristic analysis. The 5 mL blood samples were stored at 4°C for 4 to 5 hours and centrifuged at 3,000 rpm for 10 minutes at 4°C to separate the blood cells and serum . Blood cThirty femurs  were taken from the 35-day-old broilers, and the ash content in a 600°C oven was measured. The calcium and phosphorus contents were measured by the methods described by AOAC . BrieflyLactobacillus spp. and by tryptose sulfite cycloserine agar  for Clostridium perfringens at 37°C and anaerobic incubator for 48 hours. The CFU on the agars was counted after culturing.Thirty 35-day-old broilers  were used for the intestinal microbial content measurement. The chyme in the ileum and the chyme in the cecum were squeezed out and diluted by sterilized PBS and cultured by the De Man, Rogosa, and Sharpe agar (Difco Lactobacilli MRS Agar) for Fifty micrograms of ileum of 35-day-old broilers  were removed and soaked in 1 mL RNAzol . The ileum tissue was homogenous and stored in a −20°C refrigerator before being used.−ΔΔCt method was used to calculate the relative mRNA expression level, and β-actin was used as the housekeeping gene for normalization. Gene-specific primers were utilized according to the genes of Gallus gallus (chicken) and according to Genbank given as The methods of mRNA isolation were done according to the manufacturer’s protocol of SuperScript FirstStrand Synthesis System reagent . 2× SYBR GREEN PCR Master Mix-ROX , cDNA, deionized water, and each primer were mixed at the ratio of 5:1.2:1.8:1. StepOnePlus Real-Time PCR System  was used to detect qRT-PCR performance. The 2The data collected were statistically analyzed using general linear models procedure of SAS software  following a completely randomized design. Data on the dietary treatments were subjected to analysis of variance using Statistical Analysis System Institute Package (SAS) and the mean values were compared using Tukey test with a significant level at p<0.05.The mathematic model was:ij is the measurement on average of birds in pen j, dietary treatment i; μ is the overall mean; Di is the fixed effect of dietary treatment i; ɛij is the residual term that ɛij∩N . The experimental units were different depend on the experiments, including in vitro test (WB and FWB product or well of chPBMCs culturing plate), and in vivo test (per chicken).Where YS. cerevisiae, phytase, and S. cerevisiae with phytase, the extractable inorganic phosphorus increased to 79.2, 198, and 337 μmol/g DM  the NO production of 107 chPBMCs, which were stimulated by 1 ng LPS  production in creased to about 120 μmol on 101 ng LPS . Further1 ng LPS .In the starter stage (1 to 21 d), the 10% FWB group had the best WG (p<0.05), and the 5% FWB and 10% FWB groups had the lowest FCR in data  . There wVillus height increased in the 10% WB and 10% FWB groups . However, there were no significant differences in crypt depth, tunica muscularis, and the villus:crypt ratio in the jejunum. In the ileum, there were no significant differences in villus height, crypt depth, and tunica muscularis; however, the 5% FWB group had the highest villus:crypt ratio (6.35 compared to 5.65 in the control group) . SupplemFrom the data in From the data in Lactobacillus spp. had a significant increase in the 10% WB and 10% FWB groups. Although there were no significant differences in the number of caecum Clostridium perfringens, it was positively correlated with the interferon-gamma mRNA expression in the chPBMCs test  mRNA expression increased in all treatment groups compared to the control group (p<0.05). The ileum mRNA expression of TJ occludin, claudin-1, and zonula occludens-1 (ZO-1) are also shown in In the expression of anti-oxidation-related mRNA on chPBMCs, nuclear factor erythroid 2-related factor 2 (Nrf-2) increased significantly in the 5% FWB group (p<0.05), and the 5% WB and 10% FWB groups increased significantly on the heme oxygenase-1 (HO-1) mRNA expression (p<0.05). Nicotinamide adenine dinucleotide phosphate-oxidase-1 (NOX-1) and glutamate-cysteine ligase catalytic (GCLC) have complementary results in all treatments. The lower NOX-1 and higher GCLC mRNA expressions were in the 5% WB and 10% WB groups compared to the control group (p<0.05) .The data of pro-inflammatory cytokines, such as interleu kin-6 (IL-6), induciblenitric oxide synthases (iNOS), nuclear factor-κB (NF-κB), IL-1β, cyclooxygenase-2 (COX-2), and interferon-gamma (IFN-γ), are shown in in vitro, containing a variety of substrates, probiotics, and microbes secretion by the fermentation [Previous studies pointed out that fiber can improve the intestinal health of animals . Fibers entation . Many enentation ,25. In aentation , includientation .S. cerevisiae could only produce fiber or protein-related enzymes such as cellulase, xylanase, protease, and glucanase but not phytase. Therefore, fermentation WB by S. cerevisiae is less helpful for the release of inorganic phosphorus. However, S. cerevisiae fermentation can increase the phytic acid release and increase the function of phytase. Therefore, co-fermentation with S. cerevisiae and phytase will increase the release of inorganic phosphorus.Because the phytic acid in WB is present in complex fibers , and S. Bacillus amyloliquefaciens, and S. cerevisiae fermented WB inclusion will not increase broilers’ growth performances. Nevertheless, Santos et al [Because of the fermentation, FWB has more simple sugars, oligosaccharide, enzyme, and inorganic phosphorus content than WB, and these can increase the nutrient absorption of broilers . Furtheros et al  indicateos et al –5. The rFrom the above data, it can be inferred that the use of 10% FWB in the early stage and the use of 5% FWB in the finisher stage may be better for the body weight gain of broilers. Furthermore, FWB has a better effect than WB inclusion.Lactobacillus spp. in the cecum of broilers. In a previous study, fiber addition to a diet has a positive effect on Lactobacillus spp. growth [As for the higher fiber treatment, 10% WB and 10% FWB significantly increased the number of . growth . In addi. growth . Althoug. growth .Similar results can also be seen in the blood biochemical analysis. The hemoglobin (Hb) content is related to the oxygen-carrying capacity of the animal and is positively correlated with the efficiency of nutrient utilization. Therefore, the 5% FWB group has better results on improving nutrient utilization at the 35-day-old. In addition, elevated blood sugar levels are associated with fiber content and fermentation. Both the reduction of BUN and UA in the blood are related to the utilization of protein . Fiber iAs an important component supporting the body of the animal, the skeleton is mainly composed of minerals such as calcium and phosphorus and organic substances such as proteins . FWB canThe poultry industry often encounters climatic or environmental challenges. One of the common problems is the response to oxidative stress, inflammatory response, and intestinal epithelial damage of poultry . The mucS. cerevisiae being able to promote the health of intestinal epithelial cells and the performance of TJ protein [From the results of this report, the TJ-related mRNA ex pression exhibits a significant increase, indicating that both WB and FWB can significantly improve the performance of TJ. Furthermore, the effects of FWB treatment on occludin mRNA expression are higher than WB treatment. The results may be due to short-chain fatty acids (SCFAs) produced by  protein .The health of intestinal epithelial cells is also related to the expression of inflammatory factors in animals, and the role of inflammation is not only related to immune regulation, but also the decreased oxidation stress of animals. Therefore, the health of intestinal epithelial cells is closely related to the regulation of antioxidant capacity and the regulation of immunity in the blood of animals ,36. The S. cerevisiae is mainly composed of β-glucan and mannan, both of which have the function of encapsulating toxins and can reduce the damage caused by LPS to cells [In addition to oxidative stress, another protective mechanism in animals is their inflammatory response. An inflammatory response protects the host from pathogens, but it can also cause apoptosis in the host due to excessive inflammatory reactions . The celto cells , and theThe main regulators of inflammation are NF-κB and in terleukin family . NF-κB iS. cerevisiae cell walls, which can also inhibit IL-1β function. In addition, WB and FWB will also affect intestinal microbiota can lead to different inflammatory responses in the host. 10% WB and 10% FWB inclusion will significantly increase the Lactobacillus spp. number in the caecum, and Hegazy and El-Bedewy [10 CFU of Lactobacillus delbruekii and L. fermentum can decrease the IL-6 and NF-κB mRNA expressions.When an animal is traumatized or infected, it causes a large secretion of COX-2, promotes the conversion of arachidonic acid to prostaglandin E2 and prostaglandin F2α, induces macrophage aggregation, and induces an inflammatory response . The IFNl-Bedewy  indicateThe above results show that both FWB and WB can achieve an anti-inflammatory effect by inhibiting the expression of IL-1β, wherein the effect of FWB is better than that of WB, but the amount of substitution has little effect on the degree of inclusion.Based on the above results, a high WB diet does increase the health of animals. However, although both WB and FWB promote the performance of TJ mRNA expression and improve the antioxidant capacity of broilers, FWB is better in decreasing inflammatory response. In addition, data also showed that, 10% FWB inclusion show better growth performance than other treatments in starter stage but finisher stage of broilers. Therefore, it is confirmed that, as a postbiotic, there was higher nutrition value in FWB than WB and worthy to be a feed inclusion strategy.


Based on FTIR, there were two new peaks at 1729 and 1568 cm−1 of the HMT-OSA starch, which proved that the hydroxyl group of the HMT starch molecule had been substituted with the carbonyl and carboxyl ester groups of OSA.To elucidate the pretreatment of a heat moisture treatment that could increase the DS and hydrophobicity of OSA starch, the effect of the moisture level of the HMT process on the physicochemical properties was investigated. The higher moisture content (MC) in the HMT process led to a decreasing degree of crystallinity and gelatinization enthalpy and also produced surface damage and cracking of the granules. HMT pretreatment with the right moisture content resulted in OSA starch with the maximum DS value and reaction efficiency. Pre-treatment HMT at 25% MC (HMT-25) followed by OSA esterification exhibited the highest DS value (0.0086) and reaction efficiency (35.86%). H25-OSA starch has been shown to have good water resistance (OAC 1.03%, WVP 4.92 × 10 Metroxylon sp.) because of its abundance, low cost, it can easily be gelatinized, has high viscosity, is easy to form, and low syneresis [Starch is a carbohydrate source that is widely used for food and non-food applications. One source of starch that has high potential to be used in various food and non-food industries is sago  but below the gelatinization temperature [Heat moisture treatment (HMT) is a physical modification that involves incubating starch granules at a moisture content of <35% perature . HMT hasperature ,23,24. Tperature . Previous studies have reported that HMT before OSA modification increased DS and reaction efficiency of sweet potato, cassava, and corn OSA-starches by the reduction in starch crystallinity and the formation of channels, voids, and hollows inside the granules, which facilitate the entry and reaction of OSA inside the granules ,18,25. HMetroxylon sp.) was obtained from the local market in Manokwari, West Papua, Indonesia. The starch was washed and filtered three times to remove impurities and then dried in a cabinet dryer at 50 °C for 24 h. Dried sago starch was ground with a mortar and pestle then sieved through a 0.149 mm sieve and then stored in an airtight container until further use. 2-Octen-1-ylsuccinic anhydride (OSA) was purchased from Sigma-Aldrich Corporation , and all the reagents used in this study were of analytical grade.Sago starch  by spraying the calculated amount of distilled water onto the starch and then homogenized for 20 min. The exact moisture content of the mixture was measured with a moisture analyzer . The moist starch was then placed in a 500 mL Duran glass bottle equipped with a screw cap and incubated for 1 h before being placed in an autoclave and heated for 1 h at 120 °C . The glav/v) within a period of 2 h while maintaining the pH and constant stirring. Esterification was conducted for 4 h at 35 °C, and then the pH was adjusted to 6.5 with 3% HCl solution at the end of the reaction. The OSA starch was collected by vacuum filter, washed two times with distilled water, two times with 70% aqueous alcohol, and then dried at 40 °C for 24 h.The preparation of OSA starch followed the method of Jiranuntakul et al. . Native Swelling power, solubility, water absorption capacity, and amylose contents were determined to characterize the physicochemical properties of native, HMT, and HMT-OSA starch. The swelling power and solubility of the starches were determined following the method of Zavareze et al. . To deteThe pasting property of native and HMT starch samples was evaluated using a Rapid Visco Analyzer . The starch sample  was added to 25 mL of distilled water in the RVA canister. The sample was held at 50 °C for 1 min, then the starch suspension was heated from 50 °C to 95 °C at a rate of 5.2 °C/min, held at 95 °C for 5 min, cooled to 50 °C at 5.2 °C/min, and held at 50 °C for 2 min. The paddle speed was 960 rpm for the first 10 s, and 160 rpm for the rest of the experiment. Parameters including pasting temperature, peak viscosity, trough viscosity, final viscosity, breakdown viscosity, and setback viscosity were recorded.w/v) of the starch slurry was heated in a boiling water bath for 30 min with intermittent mixing, then cooled to room temperature. The light transmittance (%T) was measured at 650 nm against distilled water as the blank using a UV–Visible Spectrophotometer . The samples were stored at 4 °C and percent transmittance (%T) was measured at 0 h, 24 h, 72 h, and 168 h.Paste clarity of starch solutions determined following the method of Mehboob et al. . A samplThermal characteristics of the native and modified starches were determined using a differential scanning calorimeter . Starch  was loaded into an aluminum pan and 5 µL of distilled water was added using a micro-syringe. Samples were hermetically sealed and allowed to stand for 1 h at room temperature before heating in the DSC. Sample pans were heated at a rate of 10 °C/min from 30 to 110 °C and an empty pan was used as the reference.Ac is the area of the crystalline peak and At is the total area under the curve.The X-ray patterns were obtained using an X-ray diffractometer  with Cu Kα radiation (λ = 0.154 nm). The scanning region of the diffraction angle (2θ) was from 4°–35° at 25 °C. The total area under the curve and the area under each prominent peak  were determined using Origin Pro software  . The totThe surface morphology of native and modified starch granules was examined using scanning electron microscopy . Native and modified starch samples were mounted individually on the stubs using double-sided adhesive tape and the starch samples were coated with gold. Micrographs were taken at 500× magnification and 5 kV accelerating potential.Vb is the volume of HCl required for blank titration; Vs is the volume of HCl required to titrate the sample; W is the weight (g) of sample; and N is the normality of the HCl solutionThe determination of the degree of substitution (DS) was based on the study by Bhosale and Singhal  by alkalRE) was calculated as follows:Theoretically DS is calculated by assuming that all the added OSA reacted with starch to form the ester derivative.The reaction efficiency  of the native and modified starch was measured according to the ASTM E96-92 method . Native w/w) and scanned in the wavenumber range of 400–4000 cm−1 with a resolution of 8 cm−1.FTIR spectra of native and modified starches were determined using a FTIR spectrophotometer . Before the preparation of the pellet, the starch samples were dried at 105 °C for 12 h to remove any moisture. To make a thin pellet, the dried sample was finely ground with KBr in a ratio of 1:100 , medium amylose content (20–25%), and high amylose content (>25%) . The amyThe pasting properties of native and HMT sago starches are shown in p < 0.05). Reduction in the light transmission in HMT starch might be due to increased flexibility of the starch chain in the amorphous region and amylose leaching during pasting [The paste clarity of native starch and HMT starch during storage was presented in  pasting . During  pasting . The incThermal properties of native and HMT sago starches were analyzed using DSC . ModificXRD pattern and relative crystallinity of native and HMT sago starches are shown in The morphology and granule structure of the native sago starch and HMT starches were observed using SEM and are presented in Based on the crystallinity and enthalpy data of HMT starches , HMT staMoisture level treatment during HMT affected the DS and RE of OSA starch, where the highest DS and RE were obtained from the HMT treatment sample with 25% moisture content treatment (HMT25-OSA). There was a reduction in the DS value and reaction efficiency of H30-OSA, indicating excessive moisture content while the HMT process was not effective in esterification of the OSA reaction. The higher moisture content makes starch granules active due to water absorption throughout treatment, promoting their expansion, providing morphological changes, and also reducing double helices in the crystalline and amorphous regions of starch granules driven by the thermal force, which produced granule agglomeration and crush, consistent with the SEM images, decreasing enthalpy and gelatinization temperature , and incWater contact angle measurement enables the qualitative estimation of changes in the hydrophobicity of modified sago starch. The water contact angle values of native sago starch and HMT-OSA sago starch are presented in The oil absorption capacity of native, HMT 25, OSA, and H25-OSA starches are shown in Dual modification by HMT and OSA affected the water solubility of sago starches. Water vapor permeability of native and modified sago starches is shown in −1, which is related to intra- and intermolecular hydrogen bonds, while the peak at about 2932 cm−1 is associated with –CH2 stretching vibration. The peak at the 800–1200 cm−1 region of the IR spectrum could be used to analyze the crystalline structure and the short-range order of starch. The absorption bands at 1047 cm−1 and 1022 cm−1 were associated with the crystalline and amorphous structures of starch, respectively, and the band at 995 cm−1 was related to the intramolecular hydrogen bonds of the hydroxyl group at C-6 [−1 [1022/995  modification affects the physicochemical properties of sago starch where the changes are influenced by the moisture content during the foraging process. HMT caused a reduction in swelling power, solubility, relative crystallinity, gelatinization enthalpy, and paste clarity, but increased WAC, final viscosity, setback, pasting temperature, To, Tp, and Tc gelatinization. The shape of the granules after HMT did not change, but there was roughness, cracks on the surface of the granules, and it was also hollow in the center of the granules with increasing moisture content treatment. The excess moisture content of the HMT process (30% MC) led to agglomeration of the starch granules. Pre-treatment of HMT before OSA esterification increased DS, reaction efficiency, and hydrophobicity of the OSA starch. The combination of HMT25 (25% moisture content) and OSA (H25-OSA) resulted in the highest DS, reaction efficiency, water contact angle, oil absorption capacity, and water barrier. The results obtained from this study suggest that HMT enhances the entry of OSA into starch granules and increases the reaction between OSA and hydroxyl groups of starch via an increased amorphous area; moreover, channels or hollows in the starch granules generated after HMT could facilitate the entry of OSA into starch molecules in the granules.


Horisenbada, prepared by the soaking, steaming, and baking of millets, is a traditional Mongolian food and is characterized by its long shelf life, convenience, and nutrition. In this study, the effect of processing on the starch structure, textural, and digestive property of millets was investigated. Compared to the soaking treatment, steaming and baking significantly reduced the molecular size and crystallinity of the millet starch, while baking increased the proportion of long amylose chains, partially destroyed starch granules, and formed a closely packed granular structure. Soaking and steaming significantly reduced the hardness of the millets, while the hardness of baked millets is comparable to that of raw millet grains. By fitting digestive curves with a first-order model and logarithm of the slope (LOS) plot, it showed that the baking treatment significantly reduced the digestibility of millets, the steaming treatment increased the digestibility of millets, while the soaked millets displayed a similar digestive property with raw millets, in terms of both digestion rate and digestion degree. This study could improve the understanding of the effects of processing on the palatability and health benefits of Horisenbada. Horisenbada, prepared by the soaking, short-time steaming, and high temperature baking of millets, is a traditional Mongolian food. Horisenbada is usually consumed as porridge with tea or yogurt, having good palatability, strong satiety, and a unique flavor. It is an important daily staple-food for Mongolian steppe herdsmen due to its long shelf life, convenience, and nutrition. Millets are one of the major cereal grains, mainly distributed in arid and semi-arid areas of Africa and Asia (China and India) . In genew/v), or sodium bicarbonate , did not affect the in vitro starch digestion  as functions of (ti+1-ti-1)/2 for all points except the first and last point [The value of st point .All the data was analyzed with analysis of variance (ANOVA) with Tukey’s pairwise comparisons for a statistical significance and all values were expressed as mean ± standard deviation. SPSS 22.0 software  was used for the above data analysis.h, denoted by h ≤ 100 nm) and amylopectin  [h > 100 nm for steamed and baked millet starch was decreased. In Typical SEC weight molecular distributions of wholly branched millet starch are presented in  100 nm) . As showbranches .w(logX), obtained from the DRI signal were plotted against DP X. As described elsewhere [X ≤ 100 and 100 < X ≤ 10,000 were defined as AP chains and AM chains, respectively. For the amylopectin CLDs, it showed the usual features of two large amylopectin peaks, while for the amylose CLDs, the millet starch also displayed two peaks with one at X~200–300 and the other at X~700 [X ≤ 1000, 1000 < X ≤ 5000, and 5000 < X ≤ 20,000, which were defined as short, medium, and long amylose chains, respectively [X~100–1000 of baked and steamed starch was significantly increased, especially for that of baked starch, which almost doubled.lsewhere , the popat X~700 . Clearlyectively . InteresX~100–1000 and X~1000–5000. This might be caused by the degradation of long amylose chains and/or the repolymerization of short starch chains [X~100–1000 and X ≤ 100. On the other hand, with the heating temperature continuously increasing, the short amylopectin chains were also repolymerized by forming new glycosidic bonds and presented a nonlinear structure with a higher hydrodynamic radius [X~100–1000.Compared with raw and other processed millet, the chain length distribution of baked millet starch had a higher proportion of chains with h chains . It was c radius , contribThe XRD patterns and relative crystallinity of starches from raw and different processed millets are illustrated in The morphologies of raw and different processed millets were analyzed using SEM. As shown in As shown in In vitro digestion curves of raw and different processed millets are shown in k and C∞, were calculated from LOS plot. k is the starch digestion rate coefficient, and C∞ is the estimated percentage of starch digested at the end-point of the reaction. As shown in C∞ values. In addition, the k and C∞ values of raw and soaked millet had no significant differences, keeping consistent with the observation in Further parameterizing the digestion between millet samples, LOS plots and model fitting curves are shown in k and C∞ values. The increased millet digestion may be caused by the gelatinization of millet starch by the steaming process. The soaking process did not significantly change the molecular, crystalline, and granular structure of the millets, which explains the similar digestion property between the raw and soaked millet. Interestingly, the baked millet displayed the most desirable digestive property in terms of the lowest k and C∞ values, whereas the molecular size and the degree of gelatinization of the baked millet starch were the lowest, indicating that the degree of gelatinization is not the only factor affecting cereal digestion. On one hand, it is reported that the baking process made starch granules packed tightly and blocked the enzyme approaching and hydrolyzing pathways [The soaking, steaming, and baking processes significantly changed the digestive property of the millet grains, while the variation trends remained consistent between all three cultivars. For details, the steamed millet showed the rapid digestive property, which was consistent with the highest pathways , which wpathways .This study investigated the effect of various processes on starch structure, texture, and digestibility of millet during the preparation of “Horisenbada”. Between these different processing methods, the baking treatment affected the molecular structure, crystalline structure, and granular structure most significantly, which explained the obviously increased hardness and retarded digestibility. The possible links between starch structure, textural, and digestive properties of millets as affected by different treatments were also proposed, which provided fundamental evidence for Horisenbada as a nutritional food with low digestibility. The study could be beneficial to understanding the structural reasons for the palatability and heathy benefits of Horisenbada, the traditional Mongolian food.


Objectives: The objective of this study was to investigate the mechanical properties of high-impact (HI) heat-cured acrylic resin (PMMA) reinforced with silane-treated zirconia nanoparticles. Methods: Forty-five PMMA specimens reinforced with zirconia were fabricated and divided into three groups: Pure HI PMMA (control group), PMMA reinforced with 3 wt.% of non-silanised zirconia nanoparticles and PMMA reinforced with 3 wt.% of silanised zirconia nanoparticles. Silanised and non-silanised zirconia nanoparticles were analysed with Fourier Transform Infrared (FTIR) Spectroscopy. For measuring the flexural modulus and strength, a Zwick universal tester was used, and for surface hardness, a Vickers hardness tester were used. Furthermore, raw materials and fractured surfaces were analysed using Scanning Electron Microscopy (SEM). A one-way ANOVA test followed by a post-hoc Bonferroni test was employed to analyse the data. Results: The results showed that the mean values for flexural strength (83.5 ± 6.2 MPa) and surface hardness (20.1 ± 2.3 kg/mm2) of the group containing 3 wt.% treated zirconia increased significantly (p < 0.05) in comparison to the specimens in the group containing non-treated zirconia  and the control group . However, the group with silanised zirconia showed an increase in flexural modulus (2313 ± 161 MPa) but was not significantly different (p > 0.05) from the non-silanised group (2207 ± 252 MPa) and the control group (1971 ± 235 MPa). Conclusion: Silane-treated zirconia nano-filler improves the surface hardness and flexural strength of HI PMMA-zirconia nanocomposites, giving a potentially longer service life of the denture base. Acrylic resins remain the preferential choice for complete dentures as they possess satisfactory properties  and provThe incorporation of inorganic nanoparticle fillers into HI PMMA acrylic resin has been suggested as a potential solution to this problem. The incorporation of nanoparticles into polymers has created a novel set of materials that perform better than polymers filled with micro-particles . The eff2) is a metal oxide that has attracted interest as an inorganic filler material for developing polymer nanocomposites [1/2). It is also resistant to corrosion and is biocompatible, making it an excellent option for polymer reinforcement in denture base applications [Attempts have been made to enhance the mechanical properties of acrylic resins by incorporating different metal oxide nanoparticles including titanium oxide ,11, alummposites . The incmposites ,16,17,18ications .As nanoparticles exhibit high surface energy, high polarity and hydrophilic surfaces, uniform distribution throughout the polymer matrix can be challenging, causing some phase separation and aggregation ,21,22. TAccording to previous research, which evaluated conventional PMMA acrylic resin reinforced with metal oxides, silanisation improved the properties ,18. TherThe hypothesis was that there would be no significant difference between the flexural strength, flexural modulus and surface hardness of silanised and non-silanised PMMA-zirconia nanocomposites.2) nanoparticles  were used as the filler material for manufacturing the experimental specimens.High-impact, heat-cured, acrylic resin polymethyl methacrylate (PMMA) powder and methyl methacrylate liquid monomer   were used for the denture base material. Yttria-stabilised zirconia  as explained in the Author’s previous publication .According to previous studies, a small percentage addition (1% to 7.5 wt.%) of different metal oxide nanoparticles can enhance the mechanical properties of PMMA denture base resin ,14,15,18An electronic balance  was used to weigh the silane-treated and non-silane zirconia and acrylic resin powders according to In order to understand their behaviours, a comparison between silanised and non-silanised zirconia nanoparticles was assessed in methyl methacrylate monomer (MMA) and in water. In order to ensure an equal number of particles for this test in both liquids, an equal weight of nanoparticles was silanised. In both liquids (MMA and water), silanised particles were homogeneously distributed and remained suspended, and this was in contrast to the non-silanised particles, which appeared to settle at the bottom of the containers . A simil−1 at a resolution of 4 cm−1 at room temperature [An Energy Dispersive X-ray Spectroscopy (EDX) was used to characterise the elemental compositions of PMMA and zirconia particles. The particles were randomly selected and then loaded into the SEM/EDX  for imaging using the secondary electron detector at an acceleration voltage of 20.0 kV. Fourier Transform Infrared spectroscopic (FTIR)  were used to find the functional groups and bonds in the silanised zirconia nanoparticles powders and non-silanised zirconia. Spectra were obtained with the wavenumber range from 4000 to 400 cmperature .The flexural strength of the specimens was determined using a 3-point bending test in a universal testing machine (Zwick/Roell Z020 Leominster) in accordance with the British International Standard for Denture Base Polymers (2487:1989) . Ten spe0.05) of the specimens was evaluated using a micro-hardness testing machine . Fifteen specimens were divided into five specimens for each group and the dimensions of the specimens were 65 ± 1.0 mm length × 10 ± 0.1 mm width × 2.50 ± 0.1 mm thickness, and the test load was fixed at 50 g for 30 s. During measurement, three indents were taken at different points on one side of each specimen. The distances between the indentations were determined by multiplying the average indentation diagonal length by four (4×D) to ensure adequate distance was maintained between the indentations. The Vickers hardness was calculated by measuring the diagonals  of the pyramid-shaped indentation impressed on the specimen under the applied load. The hardness values were averaged from five specimens for a particular group. Further details of the test procedures are described in [The Vickers hardness . The surfaces of fractured of the nanocomposites from the 3-point bending test were also studied using SEM. The particles from the raw materials and the fractured nanocomposite specimens were mounted onto aluminium stubs and sputter-coated with a thin layer of gold. The specimens were then loaded into the SEM for imaging using the secondary electron detector at an acceleration voltage of 2.0 kV.p < 0.05).The mechanical tests results were recorded and analysed using a SPSS . Descriptive statistics were conducted for each of the tests to determine if the data were normally distributed. According to tests on normality with Kolmogorov–Smirnov and Shapiro–Wilk, the data were normally distributed for all tests. The data from the flexural strength, flexural modulus and hardness tests were analysed by using a one-way analysis of variance (ANOVA) with post-hoc Tukey significant difference test  on the surface of the zirconia nanoparticles, [rticles, ,17, with−1 exhibited the asymmetric and symmetric vibrations of CH3 and CH2 groups. In contrast, the curve relating to silanised zirconia demonstrated absorption peaks around 2966 and 2844 cm−1. The absorption peak in the γ-MPS curve at approximately 1717 cm−1 was attached to the C=O group (carbonyl) vibrations of the (γ-MPS) molecules [−1 indicated the formation of C=O group vibrations. The absorption peak at 1638 cm−1 was attributed to the C=C group in the γ-MPS on the silanised zirconia surface. The characteristic peaks between 1160 and 1078 cm−1 in the γ-MPS spectrum represented vibration absorption bands of Si-O-C and Si-O-Si, respectively. A strong peak at 813 cm−1 was denoted by the vibration of Si-C [−1 matched Zr-O, while the peaks at 1330 and 1054 cm−1 indicated Zr-O-Si. Silane hydrolysis produced silanol groups that condensed with surface hydroxyl groups on the inorganic particles (ZrO2) to form covalent bonds [In the spectral curve of γ-MPS, the peaks between 2944 and 2841cmThe mechanism by which the 3-trimethoxysilyl propyl methacrylate silane coupling agent reacts with the zirconia nanoparticles’ surface and the bonding with PMMA is presented in p < 0.001) with the Tukey test showing a significant difference (p < 0.05) between test groups of silanised and non-silanised nanocomposites and the control group. However, flexural modulus values showed no significant difference (p > 0.05) between silanised and non-silanised specimens. The mean and standard deviation values of flexural strength and modulus are given in 2 representing a 5% increase in the modulus compared to the non-silanised group (2207.0 ± 252.7 MPa) but was not significantly different (p < 0.05). When compared to the control, this represented an increase in the flexural modulus of 17%. The highest mean value of flexural strength was (83.5 ± 6.2 MPa) for the silanised group compared to the non-silanised group (59.9 ± 7.1 MPa). The reinforcement (3 wt.%) of HI PMMA with silanised zirconia nanoparticles increased flexural strength by 40% and 15% compared to the non-silanised group and control group, respectively. The highest mean value of flexural modulus was found (2313.0 ± 161.3 MPa) for the group with 3 wt.% of silanised ZrOp < 0.05) between the silanised and non-silanised nanocomposites and control group. The highest value of Vickers hardness was obtained (20.1 ± 2.3 kg/mm2) for the group with 3 wt.% silanised zirconia, and was significantly different (p< 0.05) in comparison to the non-silanised (15.0 ± 0.2 kg/mm2) and control (17.1 ± 0.9 kg/mm2) groups, representing a 33% and 17% increase in the surface hardness, respectively.The mean and standard deviation surface hardness values for each experimental nanocomposite are listed in SEM micrographs in Silanisation of zirconia nanoparticles can enhance the mechanical properties of HI PMMA-zirconia nanocomposites. The hypothesis was therefore rejected, as significant differences in the surface hardness and flexural strength of the nanocomposites with silanised zirconia were found compared to both the nanocomposite with non-silanised zirconia and the control group. However, the hypothesis was partially accepted as there was no significant difference in flexural modulus between silanised and non-silanised groups.According to the British International Standard for Denture Base Polymers (2487: 1989) , flexuraA similar study by Zhang et al. investigated reinforcement of PMMA denture bases with silanised zirconia nanoparticles and aluminium borate at different concentrations and their effects on flexural strength . They fo3 functional groups in silane molecules were hydrolysed to silanol groups by moisture containing solvent. These silanol groups formed covalent bonds upon condensing with surface hydroxyl groups on the silica [The increase in flexural strength noted in the present study might be explained by the fact that the reinforcement of the HI PMMA acrylic resin matrix with silanised zirconia nanoparticles caused better adhesion of the particles and the matrix facilitated by favourable chemical bonding due to silanisation. Silane (γ-MPS) has a bifunctional molecule that can react by its alkoxysilane groups with the filler and itself, and with the resin due to its methacrylate functional group . Accordie silica . During e silica . In addie silica .Lung et al. concluded that the rate of silane hydrolysis is dependent on multiple factors such as temperature and type of solvent used for hydrolysis activation . In the 2O3/YSZ particles at varying concentrations  into a PMMA denture base. The findings demonstrated that an increase in fillers improved the flexural modulus [The flexural modulus values varying between 2207 MPa to 2313 MPa for both silanised and non-silanised nanocomposites was acceptable according to the British Standard. However, the flexural modulus value was 1971 MPa for the control group and was not acceptable. The modulus value is slightly lower than the recommended values and the reason for the lower value could be explained by the fact that PMMA reinforced with rubber particles increases the impact strength but reduces the flexural modulus . Alhareb modulus . In addi modulus ,35,36. THigh hardness values commonly correspond to higher resistance to abrasion . It is i2 nanoparticles with different concentrations (wt.%). The finding showed that the 3–5 wt.% of ZrO2 nanoparticles increased flexural strength and surface hardness, which was in agreement with the current study [2 nanoparticles to the PMMA matrix to achieve higher strength [Alhotan et al. evaluated the flexural strength and surface hardness of heat-cured PMMA modified by the addition of ZrOnt study . In relastrength .One of the limitations of the present report could be that the ageing effect on the stability of the chemical bonding between the zirconia nanoparticles and acrylic resin after immersion in water for a long time was not evaluated by measuring the flexural strength or hardness. Some variables could alter the results of the present work. In fact, wear  or long-Silanisation of zirconia nanoparticles is important in the clinical practice for denture base application to enhance the mechanical properties of HI PMMA-zirconia nanocomposites through strong filler bonding with the resin matrix. This study suggested that HI PMMA incorporating 3 wt.% of silanised zirconia nanoparticles could additionally improve the flexural strength and hardness possibly due to the uniform dispersion of the nanoparticles and good bonding with the PMMA matrix in clinical use when compared with the non-silanised zirconia, leading to a longer operating life of the denture.2) nanoparticles were treated with a silane (γ-MPS) coupling agent, and PMMA-based nanocomposites were prepared with 3wt.% treated and non-treated nanoparticles. Silanised and non-silanised zirconia nanoparticles were analysed with Fourier Transform Infrared (FTIR) Spectroscopy and mechanical properties of the nanocomposites were evaluated. Within the limitations of this study, the following conclusions can be drawn.Zirconia  were improved significantly (p < 0.05) compared to the nanocomposite reinforced with non-treated zirconia and pure PMMA (control group) possibly due to uniform distribution of the nanoparticles and good bonding between the nanoparticles and PMMA.Surface hardness and flexural strength of the denture base nanocomposite based on PMMA and silane-treated 3 wt.% zirconia (ZrOThe flexural modulus of nanocomposites with treated zirconia was significantly higher than the control. However, no significant difference was found when compared to the nanocomposite with non-treated zirconia.Therefore, in order to improve the clinical service life of complete denture, PMMA resin should be reinforced with silane-treated zirconia nanoparticles.


The aim of the study was to determine how the molecular structure of porcine fat-in-water type emulsions stabilised with potato starch affected their rheomechanical properties. Dynamic mechanical analysis (DMA) and instrumental analysis of the texture were the method used in experiments. Starch gels with concentrations corresponding to the water starch concentration of the examined emulsions were used as control systems. The analysis of the starch and starch–fat systems showed that the values characterising their rheomechanical and textural properties reflected the spatial reaction of the amylose matrix to dynamic mechanical interactions. Changes in their values resulted from conformational changes in the structure of segments and nodes of the lattice, conditioned by the concentration of starch and the presence of fat. As a result of these changes, starch–fat emulsions are distinguished by greater densities of network segments and nearly two times greater functionalities of nodes than starch gels. The instrumental analysis of the texture showed that the values of the texture parameters in the starch gels were greater than in the starch–fat emulsions. The high values of the correlation coefficients (R~0.9) between the texture determinants and the rheological parameters proved that there was a strong correlation between the textural properties of the tested systems and their rheomechanical properties. The fat phase constitutes an important component of restructured meat products designed following traditional recipes . An extrThis refers both to potato starch as well as to starch obtained from cereals. Starch in the presence of water, and at elevated temperature, forms permanent spatial structures. For this sake, if used as an additive, it can modify the structure and texture of food products ,10. StarStarch—in particular, its amylose fraction—is capable of forming complexes with various organic and inorganic compounds  in whose presence amylose assumes the form of a spiral and then it forms a crystalline structure . InclusiAmong articles connected with the impact of fats on starch system properties, the majority concern fats of plant origin. Few researchers investigated interrelationships between starch systems and animal-derived fats, especially as the structure of fatty acids derived from animals is very complex in comparison with plant fatty acids. This refers not only to the content of saturated fatty acids, which clearly prevail in animal fats, but also to their molecular structure. If such investigations are undertaken, then the majority of them are focused on issues associated with structure stabilisation of the continuous phase in low-fat food articles confining themselves to investigations of final product properties.The rheological properties of materials, such as elasticity, strength, and plasticity, are manifested under short-term mechanical stresses ,16. TheyTherefore, parallel analysis of rheological and textural properties has been the subject of many studies, including those investigating how the substitution of starch for fat affects the rheological and textural properties of meat products. Barbut  identifiSo far there have been few articles on the determinants of molecular mechanisms responsible for the rheomechanical and thus the textural properties of emulsified low-fat meat products. There have been even fewer studies on the interrelations between changes in the molecular structure of starch-, fat-, and protein-containing systems and their rheomechanical properties. Available studies only show that the values characterising the rheomechanical and textural properties of emulsified sausages reflect the reaction of the spatial protein matrix to the mechanical interaction and the physical state of the meat emulsion being the continuous phase of the system ,23,24. TIn view of the circumstances discussed above, extensive research was undertaken to investigate the structure-forming and water-binding effects resulting from the introduction of native as well as physically and chemically modified potato starch into the animal (porcine) fat emulsion system in water, modelling the continuous phase in low-fat comminuted meat products. The results presented in this study are part of this research. The aim of this study was to determine the influence of the molecular structure of starch–water–animal (porcine) fat systems on their rheomechanical and textural properties. The model of the continuous phase used in the low-fat comminuted meat products was not complete because it did not contain proteins. The results concerning this model will be the subject of another publication.G′), loss modulus (G′′), and loss tangent (tgδ) are particularly important rheomechanical parameters. The storage modulus is related with the part of the potential deformation energy which is retained during periodic deformations. The loss modulus is related with the part of energy dissipated as heat. The loss tangent is a measure of internal friction; it determines the relative amount of energy dissipated in the material during one deformation cycle. These parameters are often used to correlate the rheomechanical and textural sensory properties of various products to which starch is added to improve their textural properties.In the case of poorly cross-linked starch and starch–fat systems, the delicate network of intermolecular links guarantees the shape stability of gels in conditions of weak non-destructive mechanical interaction. Oscillation rheometry expanded to the technique of dynamic-mechanical analysis (DMA) is one of the most universal and objective methods which can be applied in investigations of these types of systems, since it affords possibilities of determining values of elasticity coefficients, coefficients of internal friction (viscosity) as well as other rheomechanical parameters at any frequency of non-destructive interactions of the developed system structure. The storage modulus  presented in With the passage of time counted from the moment of attaining the temperature of surroundings at different starch concentrations, the research results obtained with the assistance of the DMA technique of the experimental starch gel  and starch–fat emulsion  systems are illustrated by model runs of dependences of the storage modulus o1G indicate that already during the cooling process a spatial network developed in the examined systems with the concentration of os(cs)n segments dependent on the concentration of starch (sc). The approximate correlation between the storage modulus of highly elastic polymer networks and the concentration of segments in these networks is determined by equation in the following [The initial values different from zero of the dynamic rigidity modulus ollowing   in the examined systems. In emulsions: m ≅ 1 for 0.17 g/g < sc < 0.2 g/g and m ≅ 2 for sc < 0.17 g/g, whereas in starch gels of starch concentrations corresponding to the concentration of the starch in water of the examined emulsions: m ≅ 1 at the interval 0.14 g/g > sc < 0.25 g/g and m ≅ 3 for sc < 0.14 g/g. The nature of changes of the k constant . This indicates diversification of the dispersive structure of starch gels and starch–fat emulsion developing as a result of starch retrogradation.The values are expressed as means ± SD (w) and the functionality of these nodes (f):Bearing in mind the fact that segment concentration in the gel network is determined by the product of the node concentration (Equation (3) can be presented in the following form:sc: for starch gels and emulsions of ≅ 7.9 and 8, respectively. This means that the initial spatial structure of the two examined systems is determined by nodes formed by at least two tetrafunctional structures which are probably bispiral associations of chains. This provides evidence that already during the cooling process of the examined systems a spatial network is formed of structure parameters dependent on the polymer concentration. Together with aging of the gels and emulsions, this network undergoes gradual reconstruction involving, mainly, the increase of the functionality of initiating nodes in the result of adsorption of successive segments of macromolecules. This is confirmed by high values of mean final functionalities kf ≅ 120 in starch gels and kf ≅ 220 in emulsion systems. These high values of mean final functionalities kf make it possible to assume that the structure of spatial network of the examined systems is determined by nodes which are hexagonal associations of bispiral structures. Earlier, on the basis of measurements of dynamic storage modulus G’  and merged into bihelical associates. This form is characteristic of native and retrograded starch.CI (complex index) on the basis of the reduction in the iodine binding capacity of amylose [CI value increases together with the decrease of starch content in systems   .Porcine fat is mostly composed of long-chain fatty acids . It is clear from chromatographic investigations  that satCI value was affected by the structure of fatty acids. On the one hand, the fatty acids containing a long carbon chain formed complexes with amylose less effectively because of their amphiphilic nature, dispersion states, and steric hindrances [CI values also tended to increase as the number of double bonds in the fatty acid increased [Generally speaking, the ndrances . On the ncreased ,37.sc = 0.17 g/g) did not result in significant differences in the retrogradation process of the two examined systems (emulsions and gels). This was confirmed by the time power exponent m which was earlier determined from the fitting of Equation (2) for the data obtained with the assistance of the DMA method. The value of this exponent (m ≅ 1) indicates that the re-crystallisation of the examined systems occurred at the initial nucleation phase of crystalline form nuclei.Bearing in mind the complex structure of the applied fat, it is not possible to determine unequivocally which of the fatty acids exerted the strongest impact on the process of formation of amylose–fat complexes. The presence of starch–fat complexes in the range of high starch concentrations .On the other hand, in the case of starch–fat emulsions, the additional increase in the degree of crosslinking is the outcome of development of bonds between amylose–fat complexes. This was reflected in greater values of both equilibrium concentrations of network segments  as well sc < 0.17 g/g, that the interactions polymer–fat began to predominate and the molecular structure of starch–fat complexes was found to determine, in a significant manner, the spatial structure of starch gels in emulsions.At high starch content, polymer–polymer interactions dominated in the examined systems with polymer–fat ones observed much less frequently. This explains the determined similarity in the spatial structure of starch emulsions and gels. It was only when the starch content sc < 0.17 g/g, the exponent of time was m = 2. This suggests that the starch chain associates are mostly well-ordered. They are partially organised in lamellar structures. This organisation system may be largely forced by the molecular structure of starch–fat complexes and by uncomplexed ligands of fatty acids [In recent years, investigations on the molecular structure of starch–fat complexes have been carried out quite intensively by many researchers ,39. The ty acids ,43.Such starch aggregate conformation in emulsions is additionally favoured by the fact that porcine fat crystallizes below starch gelation temperature. Hence, its solidification takes place within the developing gel network which consolidates its structure even more and confines further starch crystallisation.sc < 0.14 g/g, exhibits the value of m ≅ 3 indicating that three-dimensional crystalline structures were developing in them. Starch aggregates assume conformations of a statistical bundle. Due to steric reasons, such conformation of starch associates was impossible at higher starch contents in gels. On the other hand, in starch–fat emulsions, the existing lamellar structure and crosslink bonds between amylose–fat complexes associated with it, made the helix-bundle transfer impossible.The time power exponent, in starch gels for starch concentration of G’ . As a result, the setting’s ability to energy dissipation rises  after 24 h and after 7 days of storage  indicated a strong correlation between the textural and rheomechanical properties of the tested systems. The results of the analysis of the starch and starch–fat systems let us assume that the main factors affecting the texture and rheomechanical properties of starch gels are: the density of the continuous phase spatial lattice formed by amylose diffusing from starch granules during pasting, swollen, pasted granules consisting of amylopectin as well as their interactions [The high values of the correlation coefficient , where ms was starch mass, wm was water mass and mf was fat mass. Starch gels with the following concentrations corresponding to the concentration of starch in water were used as control systems in the emulsions: 0.25, 0.21, 0.17, 0.14, 0.12, and 0.06 g/g.The research was conducted on fat-in-water emulsions prepared at a 1:3 fat:water ratio (g). They were supplemented with unmodified potato starch produced by Trzemeszno, Poland added at amounts of 1, 0.8, 0.6, 0.5, 0.4, and 0.2 of the fat mass. This corresponded to starch concentrations of 0.2, 0.17, 0.13, 0.11, 0.09, and 0.05 g/g, respectively. Starch concentration in the emulsion was defined as The chemical composition of starch was as follows: moisture, 13.2%; protein, 0.2%; fat, 0.1%; ash, 0.35%; amylose, 25%. Amylose content was determined by the iodine-binding procedure  through Liquid porcine fat  with a temperature of 40 °C was applied into the experimental system. Emulsion samples weighing 100 g were subjected to a 1 h thermal treatment at a constant temperature of 90 °C in an LWC2 M water bath . They were stirred continuously with an IKA Eurostar Power Control-Visc 6000 .G′ (storage modulus) and loss tangent (tgδ) were determined. G’ is related with part of potential deformation energy which is maintained in the course of periodical deformations. The loss tangent (tgδ) is a measure of internal friction; it and determines the relative amount of energy dissipated in the material during one deformation cycle. The frequency of vibrations in the system was 1.2 Hz. The starch structuralisation kinetics was measure at a set temperature of 25 °C (with an accuracy up to 0.1 °C). Additionally, the starch gels and emulsions were measured after 24 h and 7 days of storage at 25 °C. The samples were prepared for tests as described in The rheomechanical properties were measured with a DMWT dynamic mechanical thermal analyser . In vibration rheometers the viscoelastic properties and the examined systems are calculated by analysing and characterising the curve of the pendulum—free, damped vibrations with and without the sample . A Hewlett-Packard 5890 SII apparatus equipped with a Supelcowax10 capillary column (30 m × 0.25 mm × 0.25 μm) and a flame ionisation detector (FID) was used for chromatographic separation. The analysis was conducted at a programmed furnace temperature ranging from 60 °C to 220 °C, with an increment of 12 °C per minute. The final temperature was maintained for 25 min [CI) as described in other studies [g, and 0.4 mL of the supernatant was mixed with 8.6 mL of deionized water and 1 mL of iodine solution (2.0% (w/w KI and 1.3% (w/w) I2 in deionized water) in a 15 mL tube. The absorbance (ABS) values of the sample and a reference (non-additive sample) were measured with a Shimadzu 2001 spectrophotomete at 690 nm. The CI was calculated from the following equation: CI, % = 100 × (ABS of reference-ABS of sample)/ABS of reference. All tests were performed in triplicate and the results were averaged.The extent of the complex formation and the reduction in the iodine starch binding capacity of starch were investigated on the basis of the concept of complex index  and A/BE type attachments. A sample weighing about 80 g was placed inside a cylinder with an internal diameter of 50 mm and compressed by a disc with a diameter of 40 mm to a depth of 30 mm. The use of a disc with a smaller diameter caused part of the emulsion sample to squeeze between the walls of the cylinder and the disc. During the compression and the return movement the disc moved at a speed of 1 mm/s. The instrumental measurement helped to calculate five mechanical determinants: surface destructive force (N), hardness (N), consistency (N∙s), cohesion (N), and the viscosity coefficient) (N∙s).-head speed before compression: 1 mm/s,-head speed during compression: 1 mm/s,-head speed after compression: 1 mm/s,-disc penetration depth: 30 mm,-trigger force: 5 mm,-PPS (points per second): 250.The following texture analyser settings were used:The texture was tested on starch–water and starch–water–fat systems containing native starch (Superior Standard) and pork fat. Double-distilled water was added to weighed starch according to the starch-to-water weight ratios given in k(c), f, n parameters were obtained by the method of computer fitting . One-way ANOVA was also used to determine the correlation between textural and rheological parameters The mean and standard deviations were calculated from the rheological measurements on three newly prepared samples and analysed using a spreadsheet statistical package (Microsoft Excel 2011). The values of changes of To determine the molecular rheomechanical and textural conditions resulting from the introduction of starch into the emulsion system of the animal (porcine) fat type in water, the time-related conditions of the course of the phenomena accompanying the process of formation of the spatial structure of starch and starch–fat systems were analysed.G′ and higher values of the average functionalities of the terminal kf nodes. Despite this, the energy used for binding their structural elements to the network nodes is lower. In consequence, starch–fat systems have greater ability to dissipate mechanical energy (tgδ) than starch gels.The study showed that the rheomechanical and textural properties of the starch–fat gels resulted from conformational changes occurring over time within the structural elements forming their spatial network, which were conditioned by the concentration of starch and the presence of fat. Structural elements are segments formed by complex associates of amylose chains and by amylose–fat complexes. They form interpenetrating networks. Thanks to this, starch–fat gels are characterised by greater equilibrium concentration of the network segments than analogous starch gels. As a result, they have higher values of the storage modulus R~0.9) between the texture determinants and rheological parameters, i.e., between hardness and storage modulus and between the index of viscosity and loss modulus, proved that the textural properties of the tested systems were strongly correlated with their rheomechanical properties. This suggests that both types of properties were conditioned by the same factors.The simultaneous instrumental investigations of the texture of analogous starch and starch–fat systems showed that the starch–fat systems had higher values of texture determinants than the starch systems. The high values of the correlation coefficients .


Ceiba aesculifolia subsp. parvifolia (CAP) tuber and determinate rheological, thermal, physicochemical and morphological properties of the starch extracted. The CAP tuber weight was 3.66 kg; the edible yield was 82.20%. The tuber presented a high hardness value (249 N). The content of carbohydrates (68.27%), crude fiber (15.61%) and ash (9.27%) from the isolated starch, reported in dry weight, were high. Phenolic compounds and flavonoid content of CAP tuber peel were almost 3-fold higher concerning the pulp. CAP tuber starch exhibited a pseudoplastic behavior and low viscosity at concentrations of 5–15%. Purity percentage and color parameters describe the isolated starch as high purity. Thermal characteristics indicated a higher degree of intermolecular association within the granule. Pasting properties describes starch with greater resistance to heat and shear. CAP tuber starch has X-ray diffraction patterns type A. The starch granules were observed as oval and diameters ranging from 5 to 30 µm. CAP tuber could be a good source of fiber and minerals, while its peel could be used for extracting bioactive compounds. Additionally, the starch separated from this tuber could be employed as a thickening agent in food systems requiring a low viscosity and subjected to high temperatures.This work aimed to evaluate the physical, chemical and antioxidant properties of  Today, 85% of the world’s food is based on only about 20 domesticated species; nevertheless, the nutritional potential of wild species is enormous since many of them are nutritious, easy to obtain and have a pleasant taste . FurtherCeiba aesculifolia subsp. parvifolia is a tree belonging to the Bombacaceae family that has commercial and nutritional importance in some parts of central Mexico; the seeds, roots and flowers are commercialized as food; the thorns and bark are used to make crafts [e crafts ,4. Medice crafts . The tube crafts ,7. To thStudies aimed at assessing the nutritional composition and content of bioactive compounds in wild edible products allow identifying resources with nutritional potential and evaluate the possibility of nutritional adequacy of these foods in the diet ,9.Besides being a food resource, tubers are known to contain significant amounts of starch . Starch This work aimed to evaluate the physical, chemical and functional properties of the CAP tuber. Furthermore, the starch isolated from this tuber was analyzed by rheological studies, differential scanning calorimetry (DSC), rapid viscoanalysis (RVA), X-ray diffractometry (XRD), Fourier-transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) and, thereby, provide information for the use of this resource.Sechium edule [Colocasia esculenta [Sechium edule) tuber, 86% [Canna edulis) rhizome, 68.54% [um edule  and Colosculenta . The ediber, 86% , and it , 68.54% . The wei, 68.54% .Pachyrhizus ahipa tuber (135.4 N) [The parameters derived from TPA curves for CAP tuber are presented in 135.4 N) . The hig135.4 N) ,23; addi135.4 N) .Pachyrhizus erosus, a root very similar to CAP tuber, that presented an ash content of 4.60% [Roscoea purpurea, a wild tuber for which Misra et al. [The results of the proximate composition determined for CAP tuber pulp are shown in of 4.60%  and Rosca et al.  reporteda et al. .Phenolic compounds are the second most abundant organic molecules in plants, and their consumption is important by virtue of their antioxidant capacity and their ability to regulate cellular processes, such as the expression of some genes and protein phosphorylation , whereasResults, expressed in mg of gallic acid equivalents (GAE) and mg of catechine (CE) per gram of ethanolic extract (EE), showing total soluble phenols (TSP), total soluble flavonoids (TSF) and antioxidant activity (AA), are presented in Peel exhibited a higher TSP and TSF content  than that of pulp . Traditional use of CAP bark has been reported for the treatment of tumors, inflammation and infections . Further50, which corresponds to a higher antioxidant capacity, was found in the peel  than in pulp . The EC50 value in pulp was higher than those reported by Park et al. [Smallanthus sonchifolius) tuber extract, with an EC50 value of 675.28 µg/mL by DPPH method. TSP and TSF content of CAP tuber peel was almost 3-fold higher concerning the pulp. This ties in with the strongest antioxidant potential. The texture of this structure is very hard and cannot be eaten; however, it could potentially be used for therapeutic purposes.Owing to the complex reactivity of bioactive compounds, to assess the antioxidant capacities of food or extracts, at least two different assays must be employed . In the k et al.  for yacoColor is an important attribute to designate starch quality . Table 5Sorghum bicolor) with values of 92.91, −0.26 and 3.58 for L*, a* and b*, respectively; similarly, the starch of Canna edulis rhizome, exhibited and L* value of 94.38 and a* and b* values of −0.06 and 1.47, respectively [Similar results were reported by Boundries et al.  for whitThe extraction yield of CAP tuber starch was 23.86%; this is a relatively low value when compared with other tubers, such as Solanum tuberosum (32.1%)  and ColoEnsete ventricosum (14%), Solanum tuberosum (13.5%) and Zea mays (13%) starches by Gebre-Mariam et al. [Dioscorea spp.) [The proximate analysis of CAP tuber starch was performed, and the results are showed in m et al. . Lipid cm et al.  and starm et al. , lipid cm et al. . The prom et al. . Proteinm et al. , Oxalis m et al.  and ñameea spp.) , which pea spp.)  and Bambea spp.) . The tecea spp.) . Ash wasea spp.) .G′, which refers to the material elastic character, elastic energy can be stored and subsequently recovered; G″, associated with the viscous character, viscous energy is dissipated and lost as heat; and tan(δ)=(G″/G′), which relates the previous two parameters and is calculated as the ratio of the strain energy that is stored and lost [G′ values were higher than G″ values; these describe an elastic behavior; on the other hand, tan δ values were bigger than 0.1 ranged from 0.13 to 0.16, this describes the gel formed by this starch as a weak gel since they are not able to store the applied energy and is lost as heat. By comparison, corn starch gel showed strong gel behavior (tan(δ) = 0.03). η* is a measure of the overall resistance to flow and shows the evolution of the viscoelastic behavior of materials subjected to oscillatory shear [The rheological characterization of starches provides information about their structure and behavior under processing conditions, being useful for guiding their applications . When rhand lost . The visry shear ,52. This2 = 0.99) used to evaluate the flow behavior of CAP tuber starch gel. Shear-thinning behavior shows an n < 1, whereas an n > 1 describes a shear-thickening behavior [w/v), CAP tuber starch exhibit a shear-thinning behavior , as well as corn starch at 10% (n = 0.16). K, a parameter that corresponds to the gel viscosity, had a much higher value (122.771 Pa·s) for corn starch than for CAP tuber starch (33.799 Pa·s).To simplify the comparison of flow curves obtained in rheological tests, mathematical models that adjust the numerous measurement points are used, resulting in a small number of parameters that are easily comparable  The powebehavior . At a coThe viscosity patterns of CAP tuber and corn starches are shown in Solanum tuberosum , Zea mays  and Ensete ventricosum  [−1) than reported for Solanum tuberosum (19.8 J·g−1) and Ensete ventricosum (21.6 J·g−1), in the above-mentioned study, and was within the range of reported values for Colocassia esculenta starch (15.8 an 18.2 J·g−1) by Lu et al. [DSC is the most used technique to study the starch gelatinization process. During this procedure, heat flow, which is generated as a result of endothermal and exothermic events, is monitored in the sample . Figure ctively) . The gelctively) . Thermalctively) . Furtheru et al. ,62. The u et al. .temp) represents the temperature at which viscosity starts increasing, the value obtained for the starch of CAP tuber was higher (79.15 °C) than those determined for three potato cultivars starches, ranged from 70.0 to 73.5 °C [Sechium edule, 67.75 °C and Zea mays, 78.55 °C [Sechium edule  and Zea mays  in the above-mentioned works. Higher Ptemp and lower Pv of CAP tuber starch may be related to the presence of minor components, such as lipids, sugars and proteins [The pasting properties of CAP tuber starch are presented in  73.5 °C , Sechium78.55 °C . On the proteins ,65. Highproteins ; hence, proteins . From thX-ray diffractometry allows characterizing the crystalline structure of starch granules; the diffraction pattern can be considered as the fingerprint of each starch ,70. The −1 may be attributed to O−H stretching regions of water molecules hydrogen-bonded [−1 is assigned to –OH bending vibration of free water molecules due to its hygroscopic nature [−1) [−1 is interpreted as the skeletal mode of α 1→4 glycoside bonds [−1 have been assigned to stretching vibration of C–O–H and C–O–C from glycosidic bonds [−1 is related to amorphous and crystalline regions of the starch, respectively; the more pronounced the peak at 1047 cm−1 indicates a reduction in the amorphous area and an increase in the ordered region [−1 was sharper than that observed at 1022 cm−1, indicating a predominance of crystalline areas over the amorphous component. The band’s intensity at 1242 cm−1 is due to CH2OH and C–O–H deformation mode [−1 corresponded to lipids and proteins bounded to the carboxyl and carbonyl groups of starch [−1 could be attributed to protein attached to starch [FT-IR spectra provide information on a short-range order, close to the granule surface, about double-helical order and complement the data obtained through X-ray diffraction, which provides information about long-range order related to the packing of double helices ,75. Figun-bonded , whereasc nature . In the ure [−1) , peaks dure [−1) . The bande bonds ,80. Simiic bonds . Absorptd region . In CAP f starch . In the o starch ,83.The individual morphological characteristics for each type of starch depend on plant physiology and the amount and types of enzymes involved in the biosynthetic process within the amyloplast, which in turn influences the swelling power and water-binding capacity of the granules . The micPlant material of CAP was collected in San Juan Totolcintla, municipality of Mártir de Cuilapan, Guerrero, México . The plant samples were deposited in the herbarium of the Universidad Autónoma de Guerrero and they were taxonomically identified. Later, CAP tubers were collected.Size, weight, color and texture were determined in CAP tubers. The size was evaluated measuring the longitudinal (LD) and transverse diameters (TD); the color of the peel and pulp were registered by employing a portable colorimeter Ci62  as L* (brightness), C* (chromaticity) and °h (Hue angle) at different periods from the determined CIELCH. The illuminant employed was D65, and the standard observer position was 10°. The edible yield of CAP tubers was measured by dividing edible portions from non-food tissues and using the following formula:Texture measurement was made through a texture profile analysis (TPA) with a texture analyzer . The tubers were washed, and the epicarp peeled off; the pulp was then cut into 1 cm cubes. Each sample was compressed with a 30 mm diameter cylindrical probe. The cross-head speed was 2 mm/s, and the maximum extent of deformation was 50% of the original length. The quality attributes measured were hardness (H), fracturability (F), adhesiveness (A), springiness (S), cohesiveness (C) and resilience (R). Results of size, weight, color and edible yield are presented as the average of four repetitions, while the results of TPA represent the average of ten repetitions.The nutrient content was calculated in CAP tuber pulp. Moisture, ash and crude fat content were determined according to AOAC international standard procedures 20.013, 923.03 and 920.39 , respect•+ methods, both proposed by Thaipong et al. [•+ assay, the modifications were from a starting volume of 100 μL of the pulp and peel extracts, and six serial dilutions were made in 50 μL of methanol in a 96-well plate. The extracts reacted with 150 μL of the ABTS•+ solution for 30 min in the dark. The inhibition of the DPPH and ABTS•+ radicals was calculated using Equation (1):The content of TSP was determined following the methodology of Singleton et al. . TSF cong et al.  with som50) was calculated from a graph of inhibition against the extracts’ concentration.The concentration of the extract that provided 50% free radical scavenging activity (ECw/v) citric acid solution for 10 min to prevent oxidation. Next, the pieces were homogenized in distilled water  in a blender. The resulting slurry was passed through fine muslin cloth and then filtered using a 250 μm-mesh sieve. The liquid fraction (starch) could settle for 24 h at 4 °C, and the residue (fiber) was subjected to the extraction process twice again. Later, starch-containing fractions were mixed and centrifuged at 3000 rpm for 10 min. The starch was dried in an air-circulating oven  at 40 °C for 24 h.CAP tuber starch was isolated following methods described by Jiménez-Hernández et al.  with somColor of starch was determined using a portable colorimeter Ci62 , illuminant D65 and 10° standard observer position. L* to represent lightness , a*  and b*  parameters were measured.The moisture, ash and crude fat of isolated starch were determined according to AOAC  international standard procedures  . Crude pThe starch yield was obtained as described by Pascoal et al. .w/v) were prepared as follows: weighed amounts of starch were mixed with distilled water and immersed in a boiling water bath using a magnetic stirrer equipped with a hot plate. Once a homogeneous dispersion was obtained, the temperature was raised until gelatinization point, which was about 85 °C, and then held at this temperature for 15 min to ensure full viscosity development. Rheological characteristics of starch gels were measured using oscillatory shear test following Tarrega et al. [Starch gels at 5, 10 and 15% , loss or viscous modulus (G″) the loss tangent angle (tan(δ)) and the complex viscosity (η*), as a function of frequency, were determined using the Trios v4.5.0.42498 Discovery HR TA instruments software.Viscoelastic properties were measured using oscillatory shear tests. To determine the linear viscoelastic region, stress sweeps were run at 1 Hz. After this, frequency sweeps of starch gels at 10 and 15% were performed using a frequency range of 0.1 to 100 rad/s, with the strain set at 0.1%, whereas in the starch gel at 5%, the frequency range was 0.01 to 100 rad/s and the strain set at 0.01%. Storage or elastic modulus ) model:n) represents the consistency index, γ (s−1) is the value of shear rate, and n (dimensionless) is the flow behavior index.Flow curves of the prepared starch gels were obtained at the range of shear rate between 0.01 and 1000 s−1. To, Tp, Tc and ΔH were determined by the Mettler-Toledo software.The thermal properties of starch were determined with a differential scanning calorimeter . The equipment was calibrated with indium, and an empty pan was used as a reference. Deionized water (10.0 mg) was added to 5.0 mg of starch in an aluminum pan that was sealed. After equilibration at room temperature for 1 h, samples were heated from 30 to 100 °C at 10 °C mintemp, Pv, Tv, Fv, BDv and SBv and were expressed in centipoise (cP).Experiments were performed according to AACC 61-02 method , using aThe starch’s crystalline structure was obtained on an X-ray diffractometer , operated at 30 kV and 16 mA. The X-ray source was Cu Kα radiation with a wavelength of λ = 1.5405 Å. The sample was placed on a glass surface and scanned from 2θ = 5 to 50° (θ standing for the angle of diffraction).−1 was used for scanning at a resolution of 4 cm−1 for 24 scans.The FT-IR spectrum of starch was obtained using an infrared spectrometer , equipped with a diffuse reflectance system. A region from 400 to 4000 cmThe starch’s morphological features were examined with an environmental scanning electron microscope . The starch was spread on a stainless-steel plate and coated with graphite by a sputter-coater . The starch samples were placed in the SEM chamber and examined with a beam of 10–20 kV and using a gaseous secondary electron detector. The morphology of the granules was examined at 150×, 500×, 1000× and 1500×, 1 torr with a spot size of 4.6.p < 0.05) using the statistical program SPSS 20.0.Data reported were an average of triplicate observations and were expressed as mean ± standard deviation. Results of tests for compounds and antioxidant activity were analyzed by Student’s t-test  and lower a* and b* values.The gelatinized CAP tuber starch showed a non-Newtonian and shear-thinning flow behavior; moreover, it behaves as a weak gel with a low viscosity. Physicochemical characterization methods, such as paste properties and FT-IR, reveal the presence of minor components (proteins and lipids) in CAP tuber starch, a feature that can influence the development of low viscosity. XRD patterns showed an A-type crystal structure; for this reason, they exhibited higher resistance and heat stability. This, in turn, relates to its lower melting enthalpy and higher gelatinization temperatures determined by DSC. These results corroborate that CAP tuber starch could be used to control consistency in food systems where low viscosity is required and in foodstuffs subjected to high temperatures, like dairy products, canned goods, and baby food. In addition, pasting properties results revealed the lower retrogradation tendency and greater resistance to heat and shear of this starch. While extraction yield is barely acceptable, the great quantity of tubers produced per plant increases their potential for use. These all properties make CAP tuber starch a good option as a new functional ingredient for the food industry.