Resistant Starch Alters the Microbiota-Gut Brain Axis: Implications for Dietary Modulation of Behavior

The increasing recognition that the gut microbiota plays a central role in behavior and cognition suggests that the manipulation of microbial taxa through diet may provide a means by which behavior may be altered in a reproducible and consistent manner in order to achieve a beneficial outcome for the host. Resistant starch continues to receive attention as a dietary intervention that can benefit the host through mechanisms that include altering the intestinal microbiota. Given the interest in dietary approaches to improve health, the aim of this study was to investigate whether the use of dietary resistant starch in mice to alter the gut microbiota also results in a change in behavior. Forty-eight 6 week-old male Swiss-Webster mice were randomly assigned to 3 treatment groups (n = 16 per group) and fed either a normal corn starch diet (NCS) or diets rich in resistant starches HA7 diet (HA7) or octenyl-succinate HA7 diet (OS-HA7) for 6 week and monitored for weight, behavior and fecal microbiota composition. Animals fed an HA7 diet displayed comparable weight gain over the feeding period to that recorded for NCS-fed animals while OS-HA7 displayed a lower weight gain as compared to either NCS or HA7 animals (ANOVA p = 0.0001; NCS:HA7 p = 0.244; HA7:OS-HA7 p<0.0001; NCS:OS-HA7 p<0.0001). Analysis of fecal microbiota using 16s rRNA gene taxonomic profiling revealed that each diet corresponded with a unique gut microbiota. The distribution of taxonomic classes was dynamic over the 6 week feeding period for each of the diets. At the end of the feeding periods, the distribution of taxa included statistically significant increases in members of the phylum Proteobacteria in OS-HA7 fed mice, while the Verrucomicrobia increased in HA7 fed mice over that of mice fed OS-HA7. At the class level, members of the class Bacilli decreased in the OS-HA7 fed group, and Actinobacteria, which includes the genus Bifidobacteria, was enriched in the HA7 fed group compared to the control diet. Behavioral analysis revealed that animals demonstrated profound anxiety-like behavior as observed by performance on the elevated-plus maze with time spent by the mice in the open arm (ANOVA p = 0.000; NCS:HA7 p = 0.004; NCS:OS-HA7 p = 1.000; HA7:OS-HA7 p = 0.0001) as well as entries in the open arm (ANOVA p = 0.039; NCS:HA7 p = 0.041; HA7:OS-HA7 p = 0.221; NCS:OS-HA7 p = 1.000). Open-field behavior, a measure of general locomotion and exploration, revealed statistically significant differences between groups in locomotion as a measure of transitions across quadrant boundaries. Additionally, the open-field assay revealed decreased exploration as well as decreased rearing in HA7 and OS-HA7 fed mice demonstrating a consistent pattern of increased anxiety-like behavior among these groups. Critically, behavior was not correlated with weight. These results indicate that diets based on resistant starch can be utilized to produce quantifiable changes in the gut microbiota and should be useful to “dial-in” a specific microbiome that is unique to a particular starch composition. However, undesirable effects can also be associated with resistant starch, including lack of weight gain and increased anxiety-like behaviors. These observations warrant careful consideration when developing diets rich in resistant starch in humans and animal models.     

Process optimization for the synthesis of octenyl succinyl derivative of waxy corn and amaranth starches

Modification of starch by dicarboxylic acid anhydrides to starch esters, containing both hydrophilic and hydrophobic groups are known to improve its emulsification properties, and can also be used for encapsulation after hydrolysis. Reports on the effect of process conditions on the extent of modification of starches by using n-octenyl succinic anhydride (n-OSA) are not readily available. In the present study, the process of manufacturing of OSA starches from waxy corn and amaranth starch were studied with respect to the OSA/starch ratio, pH, temperature and time of the reaction. The effects of these parameters were evaluated on the basis of degree of substitution (DS). The concluding conditions for amaranth-OSA starches was a reaction time of 6 h at 3% OSA/starch ratio at 30 °C and pH 8.0 at 25% starch concentration. For waxy corn-OSA starch, all parameters were identical except for the reaction time of 24 h. The maximum DS achieved for both the starches was 0.02. Emulsification capacity and oil absorption capacity of the OSA-modified starches were more or less similar within the parameter chosen and also independent of starch type.     

Lipase-coupling esterification of starch with octenyl succinic anhydride

Enzymatic modification of starch was conducted by lipase-coupling esterification with octenyl succinic anhydride (OSA). Parameters affecting the esterification were systematically studied. Products were characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), X-ray diffraction, differential scanning calorimetry (DSC) and viscosity analysis (VA). Optimum condition for lipase-coupling OSA starch preparation was as follows: starch pretreatment at 65 °C for 15 min, starch concentration 35%, amount of lipase and OSA, 0.6% and 3%, reaction pH, temperature and time, 8.0, 40 °C and 30 min respectively, which resulted in 0.0195 of the degree of substitution and 84.05 ± 2.07% of the reaction efficiency. FT-IR spectroscopy confirmed the formation of OSA starch. SEM and X-ray diffraction showed apparent surface change, but no crystalline change. DSC and VA results indicated the synthesized OSA starch gelatinized rapidly with high viscosity. Attractively, reaction time drastically reduced to 30 min, showing vast potential for scale production of OSA starch.     

The role of cotyledon cell structure during in vitro digestion of starch in navy beans

Studies on the physico-chemical, microstructural characteristics and in vitro (under simulated gastric and small intestine conditions) starch digestibility of navy beans were carried out. The microstructure of raw and cooked beans observed through scanning electron microscopy (SEM) showed the presence of hexagonal or angular shaped cotyledon cells (50-100 μm size) containing starch granules with a size ranging between 10 and 50 μm. The extent of starch hydrolysis (%) after 120 min of in vitro gastro-intestinal digestion differed between whole navy beans (∼60%) and milled bean flour and bean starch (85-90%) after they were cooked under similar conditions. Starch hydrolysis (%) increased significantly when the cotyledon cells in the cooked whole navy beans were disrupted using high pressure treatment (French press). The storage of freshly cooked whole beans resulted in a lower (40-45%) starch hydrolysis whereas re-heating increased the same to 70-80% during in vitro small intestinal digestion. The SEM pictures of cooked navy bean digesta after different intervals of in vitro gastric and small intestinal digestion showed that the cotyledon cell structure is maintained well throughout the digestion period. However cotyledon cells appear shrunken and developed wrinkles during in vitro digestion. Particle size analysis of cooked bean paste taken before and after the in vitro gastro-intestinal digestion showed similar particle size distributions.     

Measurement of resistant starch content in cooked rice and analysis of gelatinization and retrogradation characteristics

Digestion-resistant starch (RS) has many physiologic functions. The RS content is measured by enzymatically degrading flour samples according to the method of the Association of Official Analytical Chemists. Experiments have been performed with wheat, corn, and other grains, but there are no data for cooked rice grains in the form ingested by humans. Thus, we investigated a method to measure RS that is suitable for cooked rice grains using rice cultivars that are reported to differentially increase postprandial blood glucose in humans. Using a method for cooking individual rice grains and optimized enzyme reaction conditions, we established an RS measurement method. We also found that the amylopectin crystal condition affects the RS content measured using our method.     

Physicochemical properties and in vitro digestibility of flour and starch from pea (Pisum sativum L.) cultivars

Flours and isolated starches from three different cultivars (1544-8, 1658-11 and 1760-8) of pea grown under identical environmental conditions were evaluated for their physicochemical properties and in vitro digestibility. The protein content, total starch content and apparent amylose content of pea flour ranged from 24.4 to 26.3%, 48.8 to 50.2%, and 13.9 to 16.7%, respectively. In pea starches, the 1760-8 showed higher apparent amylose content and total starch content than the other cultivars. Pea starch granules were irregularly shaped, ranging from oval to round with a smooth surface. All pea starches showed C-type X-ray diffraction pattern with relative crystallinity ranging between 23.7 and 24.7%. Pea starch had only a single endothermic transition (12.1-14.2 J/g) in the DSC thermogram, whereas pea flour showed two separate endothermic transitions corresponding to starch gelatinization (4.54-4.71 J/g) and disruption of the amylose-lipid complex (0.36-0.78 J/g). In pea cultivars, the 1760-8 had significantly higher setback and final viscosity than the other cultivars in both pea flour (672 and 1170 cP, respectively) and isolated starch (2901 and 4811 cP). The average branch chain length of pea starches ranged from 20.1 to 20.3. The 1760-8 displayed a larger proportion of short branch chains, DP (degree of polymerization) 6-12 (21.1%), and a smaller proportion of long branch chains, DP ≥ 37 (8.4%). The RDS, SDS and RS contents of pea flour ranged from 23.7 to 24.1%, 11.3 to 12.8%, and 13.2 to 14.8%, respectively. In pea starches, the 1760-8 showed a lower RDS content but higher SDS and RS contents. The expected glycemic index (eGI), based on the hydrolysis index, ranged from 36.9 to 37.7 and 69.8 to 70.7 for pea flour and isolated pea starch, respectively.     

Pasting viscosity and in vitro digestibility of retrograded waxy and normal corn starch powders

Pasting viscosity and in vitro digestibility of oven-dried powders of waxy and normal corn starch gels (40% solids) retrograded under an isothermal (4 °C) or temperature cycled (4/30 °C) storage were investigated. Temperature cycling induced higher onset temperature for melting of amylopectin crystals than isothermal storage under a differential scanning calorimeter whereas little difference in crystalline type was observed under X-ray diffraction analysis. Temperature cycling caused higher pasting temperature and viscosity for the retrograded starches than isothermal storage. The retrograded waxy corn starch powders exhibited pasting behaviors similar to that of native waxy corn starch. However, the retrograded normal corn starch powders showed very much different pasting patterns with lower pasting viscosity but higher pasting temperature than native starch counterpart. The retrogradation increased slowly digestible starch content without changing resistant starch content, more effectively by the temperature cycling than the isothermal storage.     

Feed intake patterns nor growth rates of pigs are affected by dietary resistant starch,despite marked differences in digestion

Current feed evaluation systems often assume that fermented starch (i.e. resistant starch (RS)) yields less energy than digested starch. However, growth rates of pigs fed low and high RS diets are often the same when feed is available ad libitum. This may be explained by its effect on digestive processes changing feeding behavior, and consequently energy utilization. This study aims to investigate the effect of RS on nutrient digestion and digesta passage rate in pigs, in combination with its effect on feeding behavior and growth performance under ad libitum conditions. In experiment 1, 20 male pigs (40 ± 2.82 kg) were fed diets containing either 50% waxy maize starch (low in RS (LRS)) or high-amylose maize starch (high in RS (HRS)), and soluble and insoluble indigestible markers. After 14 days of adaptation to the diets, pigs were fed hourly to reach steady state (6 h), dissected, and digesta were collected from eight segments. From the collected samples, nutrient digestion and passage rate of the solid and liquid digesta fraction were determined. In experiment 2, 288 pigs (80 ± 0.48 kg; sex ratio per pen 1 : 1; boar : gilt) were housed in groups of 6. Pigs were ad libitum-fed one of the experimental diets, and slaughtered at approximately 115 kg. Feed intake, growth and carcass parameters were measured. Ileal starch digestibility was greater for LRS-fed than for HRS-fed pigs (98.0% v. 74.0%; P < 0.001), where the additional undigested starch in HRS-fed pigs was fermented in the large intestine. No effects of RS on digesta passage rate of the solid or liquid digesta fraction and on feeding behavior were observed. Growth rate and feed intake did not differ between diets, whereas feed efficiency of HRS-fed pigs was 1%-unit higher than that of LRS-fed pigs (P = 0.041). The efficiency of feed used for carcass gain did not differ between diets indicating that the difference in feed efficiency was determined by the non-carcass fraction. Despite a 30% greater RS intake (of total starch) with HRS than with LRS, carcass gain and feed efficiency used for carcass gain were unaffected. RS did not affect digesta passage rate nor feeding behavior suggesting that the difference in energy intake between fermented and digested starch is compensated for post-absorptively. Our results indicate that the net energy value of fermented starch currently used in pig feed evaluation systems is underestimated and should be reconsidered.     

Comparative hormonal profile of two newly developed obese congenic rat strains

1. The effect of feeding diets containing either 54% sucrose or cooked corn starch for 12 weeks on levels of fasting plasma insulin, corticosterone, growth hormone and glucagon were compared in two newly developed genetically obese rat strains--the normoglycemic LA/N-cp and the diabetic SHR/N-cp. 2. In corpulent rats of either strain, levels of plasma insulin and corticosterone were greater when compared to the lean littermates. Corpulent LA/N-cp rats had lower levels of plasma glucagon and higher levels of plasma growth hormone than did lean LA/N-cp rats. 3. SHR/N-cp rats fed sucrose had greater levels of corticosterone and glucagon than did SHR/N-cp rats fed starch.     

Resistant starch reduces colonic and urinary p-cresol in rats fed a tyrosine-supplemented diet,whereas konjac mannan does not

The effect of resistant starch (RS) and konjac mannan (KM) to maintain and improve the large intestinal environment was compared. Wistar SPF rats were fed the following diets for 4 weeks: negative control diet (C diet), tyrosine-supplemented positive control diet (T diet), and luminacoid supplemented diets containing either high-molecular konjac mannan A (KMAT diet), low-molecular konjac mannan B (KMBT diet), high-amylose cornstarch (HAST diet), or heat-moisture-treated starch (HMTST diet). The luminacoid-fed group had an increased content of short-chain fatty acids in the cecum. HAS caused a significant decrease in p-cresol content in the cecum, whereas KM did not. Urinary p-cresol was reduced in the HAST group compared with the T group, but not the KM fed groups. Deterioration in the large intestinal environment was only improved completely in the HAST and HMTST groups, suggesting that RS is considerably more effective than KM in maintaining the large intestinal environment.     

芙蓉鲤鲫幼鱼饲料适宜淀粉含量

实验以鱼粉和酪蛋白为主要蛋白源、以鱼油和豆油为主要脂肪源(鱼油/豆油, 1/1), 设计了7组不同玉米淀粉水平(6%、12%、18%、24%、30%、36%和42%)的等氮(35%)等脂(8%)饲料来饲喂芙蓉鲤鲫(Carassiusauratus var Furong carp ♀Cyprinus carpio red crucian carp ♂)幼鱼(8.940.59) g以确定其最适饲料淀粉水平。实验周期为61d。结果表明, 随饲料玉米淀粉水平的增加实验鱼增重率、特定生长率和饲料效率先上升(P0.05)而后趋于稳定, 18%42%饲料玉米淀粉组之间差异不显著(P0.05)。鱼体蛋白沉积率随饲料玉米淀粉水平的增加而增加, 30%和42%处理组显著高于6%和12%处理组(P0.05)。血浆葡萄糖没有受饲料玉米淀粉水平的影响(P0.05), 血浆总胆固醇含量在30%处理组最高(P0.05), 血浆甘油三酯随饲料玉米淀粉水平的增加而上升(P0.05)。肝糖原随饲料玉米淀粉水平的增加而上升, 36%处理组显著高于6%和12%处理组(P0.05)。肌糖原和肠道淀粉酶活性随饲料玉米淀粉水平增加而显著上升(P0.05)。通过特定生长率和饲料淀粉水平的折线法分析表明, 芙蓉鲤鲫幼鱼达到最大生长时饲料淀粉水平为27.47%。     

亚东鲑幼鱼饲料中适宜淀粉种类与水平的研究

为考察淀粉种类与水平对亚东鲑(Salmo trutta)幼鱼生长性能、饲料利用、消化酶活性、肝脏生化指标和组织学的影响, 实验采用2×3双因素设计, 选取玉米淀粉和木薯淀粉, 分别以5%、10%和15%水平添加, 共配制6种等氮等脂饲料, 饲喂初始体重为(0.50±0.03) g的亚东鲑幼鱼84d。结果表明, 随着饲料中玉米淀粉和木薯淀粉水平的提高, 增重率呈现先上升后下降的趋势, 饲料系数则先下降后上升(P<0.05), 其中10%木薯淀粉组增重率最高(518.8%), 饲料系数最低(1.32)。各组成活率、脏体比、肥满度和全鱼水分、粗蛋白质、粗脂肪和粗灰分含量均没有显著差异(P>0.05), 粗蛋白沉积率随淀粉水平的提高呈下降趋势, 其中15%水平组显著低于其他水平组(P<0.05), 而脂肪沉积率则随着淀粉水平的升高先上升后下降, 且10%木薯淀粉组显著高于玉米淀粉组(P<0.05); 淀粉种类和水平对胃蛋白酶和胃淀粉酶无显著影响(P>0.05), 15%淀粉水平组肠淀粉酶和肠蛋白酶活性显著高于其他水平组(P<0.05)。饲料中淀粉种类和水平对肝脏谷丙转氨酶、谷草转氨酶、总胆固醇和甘油三酯均无显著影响(P>0.05), 15%淀粉组的肝糖原含量显著高于其他水平组(P<0.05)。在肝脏组织学方面, 15%水平组较其余两个水平组表现出明显的细胞核移位和细胞空泡化现象。上述结果表明, 在实验条件下, 亚东鲑幼鱼饲料中淀粉的适宜添加水平为10%, 木薯淀粉的效果优于玉米淀粉。     

Lipoprotein status in Sprague-Dawley and LA/N-corpulent rats as affected by dietary carbohydrates.

1. To compare the impact of type of carbohydrate, genotype and phenotype on the synthesis and levels of plasma lipoprotein protein. Sprague-Dawley rats and carbohydrate-sensitive LA/N-corpulent (cp) rats were fasted (2 days) and then fed diets containing 54% carbohydrate as either sucrose, fructose or cooked cornstarch for 2 days. 2. The amount of 3H-protein present in the VLDL + chylomicron fraction of Sprague-Dawley rats 2 hr after injection of 3H-leucine was affected by type of dietary carbohydrate: sucrose greater than fructose greater than starch. 3. Obese and lean LA/N-cp rats fed diets containing sucrose or fructose had lower concentrations of HDL protein and higher levels of 3H-protein in VLDL + chylomicron fraction than those fed starch. 4. Obese LA/N-cp rats had more HDL protein and higher levels of 3H-protein in VLDL + chylomicron fraction than their lean littermates.     

汶川大地震灾后不同滑坡体上柏木体内非结构性碳水化合物的特性

"5.12"汶川大地震不仅直接摧毁森林17.3万hm2,还通过对土壤和林内生境的剧烈改变对现存森林造成潜在威胁。以地震重灾区北川的主要分布树种柏木为研究对象,在2009年6月初(夏初)和10月初(秋季)两个时期,对不同类型柏木林的叶、枝、根可溶性糖和淀粉含量进行测定分析,旨在认识震后初期柏木对逆境胁迫的生理响应,为研究大地震对植物体内碳水化合物的影响和对逆境的适应性提供科学依据。结果表明:地震对柏木体内碳水化合物的含量有较大影响,总体表现为严重滑坡类型具有较高的可溶性糖含量,较低的根系淀粉含量。夏初严重滑坡类型柏木叶、枝、根可溶性糖含量分别为(11.44±1.08)%、(4.64±0.42)%、(5.48±0.51)%。就不同器官而言,可溶性糖含量叶>根>枝,淀粉含量叶>枝>根;夏初碳水化合物含量均高于秋季,且仅叶片可溶性糖和淀粉含量在不同季节存在显著性差异(P<0.05),说明柏木叶片对地震反应最为敏感。地震带来的灾害如山体滑坡等对植物的生理活动产生巨大影响,不利于林木生长。     

Effects of chronic modification of dietary fat and carbohydrate on the insulin, corticosterone and metabolic responses of rats fed acutely with glucose, fructose or ethanol

1. Male rats were fed for 14 days on powdered diets containing (by weight) 53% of starch, or on diets in which 20g of starch per 100g of diet was replaced by lard or corn oil. They were then fed acutely by stomach tube with a single dose of glucose, fructose or ethanol of equivalent energy contents, or with 0.15m-NaCl. The serum concentrations of corticosterone, insulin, glucose, glycerol, triacylglycerol and cholesterol were measured up to 6h after this treatment. 2. Feeding saline (0.9% NaCl) acutely to the rats maintained on the three powdered diets produced a small transient increase in circulating corticosterone that was similar to that in rats maintained on the normal 41B pelleted diet. 3. Feeding glucose acutely to the rats on the powdered diets produced peak concentrations of corticosterone that were 2–3-fold higher than those seen in rats maintained on the 41B diet. The duration of this response increased in the order starch diet<lard diet<corn-oil diet. This abnormal corticosterone response to glucose feeding appeared to be responsible for an increased activity in phosphatidate phosphohydrolase in the livers of rats fed the starch and lard diets of 2.9- and 4.9-fold respectively. The latter increase was similar to that produced by ethanol, whereas glucose did not increase the phosphohydrolase activity in the liver of rats maintained on the 41B diet. 4. Feeding fructose acutely produced even more marked increases than glucose in the concentrations of circulating corticosterone in rats given the powdered diets, but unlike glucose did not increase circulating insulin. The duration of the corticosterone response again increased in the order starch diet<lard diet<corn-oil diet. The concentrations of circulating glucose were increased by fructose feeding in rats maintained on these diets, but they were not altered in the rats maintained on the 41B pellets. A prolonged increase in serum corticosterone concentrations was also observed when fructose was fed to rats maintained on pelleted diets enriched with corn oil or beef tallow rather than with starch or sucrose. However, these effects were less marked than those seen with rats fed on the powdered diets. 5. These results are discussed in relation to the mechanism whereby high dietary fat exaggerates the effects of ethanol, fructose and sorbitol in stimulating triacylglycerol synthesis in the liver.     

Effects of soluble corn bran arabinoxylans on cecal digestion, lipid metabolism, and mineral balance (Ca, Mg) in rats

The effects of soluble corn bran arabinoxylans on cecal digestion, lipid metabolism, and mineral utilization [calcium (Ca) and magnesium (Mg)] were investigated in rats adapted to semipurified diets. The diets provided either 710 g/kg wheat starch alone (control) or 610 g/kg wheat starch plus 100 g/kg corn soluble fiber (arabinoxylans) and either 0 or 2 g/kg cholesterol (control + cholesterol and arabinoxylans + cholesterol, respectively). Compared with rats fed the control diets, rats fed the arabinoxylan diets had significant cecal hypertrophy (+50% after 3 days of the fiber adaptation) and an accumulation of short-chain fatty acids, especially propionic acid (up to 45% in molar percentage). Arabinoxylans enhanced the cecal absorption of Ca and Mg (from 0.07 to 0.19 micromol/min for Ca and from 0.05 to 0.23 micromol/min for Mg). Mg balance was enhanced by arabinoxylans (+25%). The arabinoxylan diet markedly reduced the cholesterol absorption from 50% of ingested cholesterol in controls up to approximately 15% in rats adapted to the arabinoxylans diet. Arabinoxylans were effective in lowering plasma cholesterol (approximately -20%). There was practically no effect of the diets on cholesterol in d > 1.040 lipoproteins (high density lipoproteins) whereas arabinoxylans were very effective in depressing cholesterol in d < 1.040 lipoproteins (especially in triglyceride-rich lipoproteins). Corn fermentable fiber decreased the accumulation of cholesterol in the liver. In parallel, the arabinoxylan diet counteracted the downregulation of 3-hydroxy-3-methylglutaryl-CoA by cholesterol. These data suggest that arabinoxylans may have a great impact on intestinal fermentation, mineral utilization, and cholesterol metabolism.     

Influence of incubation temperature and time on resistant starch type III formation from autoclaved and acid-hydrolysed cassava starch

Raw cassava starch, having 74.94 and 0.44 g/100 g resistant starch type II and III (RS II and RS III), respectively, was autoclaved at 121 °C in water, 1, 10 or 100 mmol/L lactic acid. The formation of RS III was evaluated in relation to variable incubation temperature (−20 to 100 °C), incubation time (6–48 h) and autoclaving time (15–90 min). Negligible to low quantities of RS III (0.59–2.42 g/100 g) were formed from autoclaved starch suspended in 100 mmol/L lactic acid, whereas intermediate to high quantities (2.68–9.97 g/100 g) were formed from autoclaved starch suspended in water, 1 or 10 mmol/L lactic acid, except for treatments with water or 10 mmol/L lactic acid incubated at 100 °C for 6 h (1.74 g/100 g). Autoclaving times corresponding to maximum RS III contents were 15 and 45 min for water and 10 mmol/L lactic acid, respectively. Whereas, the RS III fractions from cassava starch suspended in water had melt transitions between 158 and 175 °C with low endothermic enthalpies (0.2–1.6 J/g), the thermal transitions of the acid treated samples were indistinct.     

Starch grain morphology of the seagrasses Halodule wrightii, Ruppia maritima, Syringodium filiforme, and Thalassia testudinum

Starch grains are a ubiquitous component of plants that have been used in tandem with phytoliths, pollen, and macrofossils to reconstruct past floral diversity. This tool has yet to be fully explored for aquatic plants, specifically seagrasses, which lack phytoliths and are rarely preserved as macrofossils or pollen. If starch grains in seagrasses are morphologically distinct, this method has the potential to improve seagrass identification in the fossil record in such cases where its starch is preserved (e.g. scratches and occlusal surfaces of tooth enamel from seagrass consumers). The goals of this study were twofold: (1) to determine if starch is present in seagrass material and (2) to assess how starch grain morphology differs between different seagrasses.This study focused on four abundant and ecologically distinct seagrasses from the Caribbean: Halodule wrightii, Ruppia maritima, Syringodium filiforme, and Thalassia testudinum. Starch grains were observed in all species except S. filiforme. Grains from H. wrightii are typically observed in side-on orientation, are sub-round to angular, and are fairly small (3-19 μm, end-on). Grains of R. maritima are small spherical grains (4-8 μm) that have a centric hilum and a straight extinction cross with a median angle between the arms of 90°. Grains from T. testudinum are large (9-31 μm, end-on), conical in side-on and round/sub-round in end-on orientation, have a slightly eccentric hilum with an obvious particle, and prominent lamellae.Visual assessment and comparative statistics demonstrate that the morphology of starch grains from T. testudinum, R. maritima, and H. wrightii are significantly different. With more extensive research, there is potential for the positive identification of starch grains from an unknown seagrass. The ability to identify seagrass from starch grains could facilitate the identification of seagrasses in the fossil record and supply information on seagrass evolution and distribution, climate effects on seagrass distribution, and the diets of seagrass consumers.     

Dietary starch sources affect net portal appearance of amino acids and glucose in growing pigs

Four male pigs (Duroc × Landrace × Yorkshire; average initial (mean ± SEM) BW = 22.5 ± 1.1 kg), fitted with permanent catheters in the portal vein, ileal vein and carotid artery, were used in a 4 × 4 Latin square experimental design to measure the effect of dietary starch sources on the net portal appearance of glucose and amino acids. Dietary starch sources were resistant starch (RS), maize, sticky rice and brown rice. Diets were provided at 0730, 1530 and 2330 h during a 6-day adjustment period and 1-day collection period. On day 7 of each period, blood samples were collected from the portal vein and carotid artery at 0730 h (prior to feeding) and hourly up to 8 h after meal. Blood samples were used to determine glucose, amino acid, packed cell volume and partial pressure of oxygen (pO2). When calculated per 100 g feed intake, cumulative portal glucose appearance was lower (P < 0.05) for resistant starch than for maize, sticky rice or brown rice up to 8 h after the meal. Cumulative portal glucose appearance was higher (P < 0.05) for sticky rice and brown rice than for other diets until 4 h after the meal, but maize had higher cumulative glucose appearance after 4 h. Net cumulative portal concentrations of most amino acids for resistant starch were also reduced (P < 0.05) than for the other starch sources. Cumulative portal appearance of amino acid represented 48.39%, 63.76%, 61.80% and 59.18% of dietary intake for resistant starch, maize, sticky rice and brown rice, respectively. Collectively, our results indicate that dietary starch sources substantially affect the appearance of amino acids and glucose in the portal circulation.     

Phenoloxidase and trypsin in germ-free larvae of Artemia fed with cooked unicellular diets: Examining the alimentary and protective effects of putative beneficial bacterium, yeast and microalgae against vibriosis

Three putative beneficial unicellular organisms, the marine bacterium Roseobacter sp, the yeast Saccharomyces cerevisiae mnn9 strain (SC-mnn9) and the microalga Tetraselmis suecica were cooked and offered separately as diets to developing germ-free (GF) Artemia larvae, in order to analyze their alimentary and protective effects. GF Artemia larvae were able to grow with cooked Tetraselmis and SC-mnn9 but failed with cooked Roseobacter. In spite of its high dietary quality, Tetraselmis failed to provide protection against Vibrio proteolyticus infection, while Roseobacter failed as food as well to provide protection. Cooked SC-mnn9 appeared to possess both values, dietary for growth and protective against Vibrio infection. GF Artemia larvae were apparently rapid adapted to dietary swaps; from yeast to algal and from algal to yeast. While the diets swap from algal or yeast, to bacterial diet appeared to be detrimental. Phenoloxidase-L (PO-L) and trypsin-L were used as biochemical indicators of defense and digestive functions, respectively. Developmental trypsin-L patterns were similar when fed on yeast and microalgae diets, suggesting a good digestive adaptation to plant or fungal substrates at early stages. On the contrary, diets swap or Vibrio infection affected PO-L and trypsin-L suggesting a sort of ‘alteration’ of digestive and defense functions.