Survey and properties of Staphylococcus aureus isolated from Japanese-style desserts

We surveyed the contamination of 315 Japanese- and western-style desserts and 247 human hands by Staphylococcus aureus and other staphylococcal bacteria. The most frequently isolated staphylococcal bacterium was S. warneri, followed by S. aureus. Only 1.9% of western-style desserts were contaminated by S. aureus strains, while 19.4% and 13.0% of Japanese-style desserts and human hands respectively were contaminated. Ninety-four isolates of S. aureus were characterized as to their biological properties and enterotoxigenicity. Although staphylococcal enterotoxins (SEs) were detected by enzyme-linked immunosorbent assay in the cultured broth of all S. aureus isolates, the reversed passive latex agglutination method and the polymerase chain reaction showed only 39 (41.5%) and 40 (42.6%) samples respectively as SE-positive. The predominant type of SE was SEB (67.5%), and eight strains produced SEA. None of the S. aureus strains had penicillin-binding protein 2', showing that methicillin-resistant S. aureus was not present in the samples.     

高脂饮食中添加短链菊粉对小鼠肠道菌群的影响

目的:探究高脂饮食中添加短链菊粉对小鼠肠道菌群的影响。方法:选择8周龄雄性小鼠,5只喂食高脂饲料,5只喂食10%菊粉复合型高脂饲料,喂食8周后收集小鼠粪便,检测小鼠粪便中三种主要的短链脂肪酸。同时,提取小鼠粪便中的细菌基因组,对菌群基因组16S rRNA基因V4高变区进行测序,对数据进行PCoA分析、Alpha多样性分析、LEfSe分析和16S功能预测。结果:菊粉添加后,小鼠粪便中含有的细菌DNA量增多,短链脂肪酸增加。菊粉组和对照组PCoA图可以看到明显聚类。菊粉组物种多样性低于对照组。菊粉组小鼠粪便中S24_7菌科丰度上升;Lachnospiraceae(毛螺菌科),Ruminococcaceae(瘤胃菌科)和Deferribacteraceae(脱铁杆菌科)丰度下降。16S基因功能预测发现22个第二层级的KEGG通路发生变化。结论:高脂饮食情况下短链菊粉的添加会改变小鼠肠道菌群,继而影响肠道菌群的功能。     

Effect of co-solute and gelation temperature on milk protein and gum tragacanth interaction in acidified gels

The aim of this study was to investigate the role of process conditions and system composition on the acid-induced gelation of a mixture of milk protein and gum tragacanth. This was studied by determining the effects of co-solute (lactose) addition (3, 5 and 7%) and gelation temperature (25, 37 and 45°C) on the mixture's rheological properties and microstructure using a combination of techniques including small-deformation rheology and scanning electron microscopy. The presence of lactose played an important role in the microstructure formation of gels but did not change most rheological properties. The microstructure of gels formed in the presence of lactose was coarser and more particulate, but less interconnected; this can be explained by lactose's role in improving protein aggregation. Gels prepared at a lower temperature had a high structure strength, as indicated by their high storage modulus, τ(f) and G(f) values. Low gelation temperature also caused a more branched and homogenous microstructure.     

Physical properties of emulsion-based hydroxypropyl methylcellulose films: Effect of their microstructure

The initial characteristics of emulsions and the rearrangement of the oil droplets in the film matrix during film drying, which defines its microstructure, has an important role in the physical properties of the emulsion-based films. The objective of this work was to study the effect of the microstructure (two droplet size distributions) and stability (with or without surfactant) of HPMC oil-in-water emulsions over physical properties of HPMC emulsion-based edible films. HPMC was used to prepare sunflower oil-in-water emulsions containing 0.3 or 1.0% (w/w) of oil with or without SDS, as surfactant, using an ultrasonic homogenizer. Microstructure, rheological properties and stability of emulsions (creaming) were measured. In addition, microstructure, coalescence of oil droplets, surface free energy, optical and mechanical properties and water vapor transfer of HPMC films were evaluated. Image analysis did not show differences among droplet size distributions of emulsions prepared at different oil contents; however, by using SDS the droplet size distributions were shifted to lower values. Volume mean diameters were 3.79 and 3.77μm for emulsions containing 0.3 and 1.0% without surfactant, respectively, and 2.72 and 2.71μm for emulsions with SDS. Emulsions formulated with 1.0% of oil presented higher stability, with almost no change during 5 and 3 days of storage, for emulsions with and without SDS, respectively. Internal and surface microstructure of emulsion-based films was influenced by the degree of coalescence and creaming of the oil droplets. No effect of microstructure over the surface free energy of films was found. The incorporation of oil impaired the optical properties of films due to light scattering of light. Addition of oil and SDS decreased the stress at break of the emulsion-based films. The replace of HPMC by oil and SDS produce a lower "amount" of network structure in the films, leading to a weakening of their structure. The oil content and SDS addition had an effect over the microstructure and physical properties of HPMC-based emulsions which lead to different microstructures during film formation. The way that oil droplets were structured into the film had an enormous influence over the physical properties of HPMC films.     

Rheological and microstructural investigation of oat β-glucan isolates varying in molecular weight

The rheological properties and microstructure of aqueous oat β-glucan solutions varying in molecular weight were investigated. The structural features and molecular weights (MW) were characterized by ¹³C NMR spectroscopy and high performance size-exclusion chromatography (HPSEC), respectively. The microstructure of the β-glucans dispersions was also examined by atomic force microscopy (AFM). The samples with β-glucan content between 78 and 86% on a dry weight basis had MW, intrinsic viscosity ([η]) and critical concentration (c*) in the range of 142-2800 × 10³ g/mol, 1.7-7.2 dl/g and 0.25-1.10 g/dl, respectively. The flow and viscoelastic behaviour was highly dependent on MW and on the concentration of the β-glucans dispersions. Pseudoplastic behaviour was exhibited at high concentrations and Newtonian behaviour was evident at low concentrations. At the same concentration, the viscosity was higher for higher MW samples. The Cox-Merz rule was applicable for the lower molecular weight samples at higher concentrations whereas the high molecular weight sample deviated at concentrations greater than 1.0%, w/v. The mechanical spectra with variation of both MW and concentration were typical of entangled biopolymer solutions. AFM images revealed the formation of clusters or aggregates linked via individual polymer chains scattered heterogeneously throughout the system. The aggregate size increased with the molecular weight of the samples investigated and has been linked to the rheological behaviour of the samples.     

Development of reduced-fat mayonnaise using 4αGTase-modified rice starch and xanthan gum

In this study a disproportionating enzyme, 4-α-glucanotransferase (4αGTase), was used to modify the structural properties of rice starch to produce a suitable fat substitute in reduced-fat (RF) mayonnaise. The mayonnaise fat was partially substituted with the 4αGTase-treated starch paste at levels up to 50% in combination with xanthan gum and the physical and rheological properties of the modified RF mayonnaise samples were investigated. All mayonnaises prepared in this study exhibited shear thinning behavior and yield stress. Viscoelastic properties of mayonnaise were characterized using dynamic oscillatory shear test and it was observed that mayonnaises exhibited weak gel-like properties. The magnitude of elastic and loss moduli was also affected by 4αGTase-treated starch concentration and presence of xanthan gum. In relation to microstructure, RF mayonnaise prepared with 3.8 or 5.6 wt% of 4αGTase-treated starch and xanthan gum showed smaller droplets. The use of 5.6 wt% of 4αGTase-treated starch and 0.1 wt% of xanthan gum produced a RF mayonnaise with similar rheological properties and appearances as FF mayonnaise with gum. This study demonstrated a high feasibility for using 4αGTase-treated rice starch as a viable fat replacer in mayonnaise.     

饲料中添加菊粉对暗纹东方鲀幼鱼生长、消化及非特异性免疫能力的影响

为探讨菊粉对暗纹东方鲀(Takifuguobscures)幼鱼生长、消化及免疫能力的影响,利用单因素实验设计,选取640尾暗纹东方鲀,体重(6.97±1.32)g,随机分为4组,每组设4个重复,在日粮中添加不同浓度的菊粉(0%、0.25%、0.5%、1%),连续投喂8周后测定鱼体生长、肌肉营养成分、肠道消化酶和肝非特异性免疫酶活性。结果显示,各菊粉添加组的生长指标无显著性差异(P>0.05);0.25%组肌肉的粗脂肪含量(0.87%)显著高于0.5%组(0.82%)和1%组(0.76%);0.25%组肌肉的粗蛋白含量(18.75%)显著高于1%组(18.50%),与对照组(18.62%)和0.5%组(18.60%)无显著差异。暗纹东方鲀幼鱼肠道组织的胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶活性均随着菊粉添加量的增加而呈逐渐上升的趋势。0.5%菊粉添加组肝组织的谷胱甘肽过氧化物酶和总超氧化物歧化酶活性极显著(P <0.01)高于其余处理组;0.5%组的过氧化氢酶活性也极显著(P <0.01)高于0.25%和1%组;丙二醛含量随着菊粉添加量的增加而呈递减的趋势。因此,菊粉添加量...     

The Effectiveness of Cryoprotectants in Inhibiting Multiple Freeze-Thaw-Induced Functional and Rheological Changes in the Myofibrillar Proteins of Common Carp (Cyprinus carpio) Surimi

The purpose of the present study was to investigate the effects of cryoprotectants (4 % sucrose?+?4 % sorbitol) on functional and rheological changes, including thermal stability, dynamic rheological and emulsifying properties, gel texture profile, gel whiteness, gel water-binding capacity, and gel microstructure, in the myofibrillar proteins (MP) of common carp (Cyprinus carpio) surimi subjected to multiple freeze-thaw cycles (FT, 0, 1, 3 and 5 times). Increased numbers of FT cycles were accompanied by reduced thermal transition temperature (T _max), reduced enthalpy of denaturation (ΔH) (P?<?0.05), and enhanced susceptibility to thermal aggregation. The observation of the gel microstructure indicated that the FT processes produced empty spaces and changed the aggregate gel structure into a coarser-stranded network. The addition of cryoprotectants significantly prevented the reduction of MP thermal stability (P?<?0.05). The protective effect on protein functionality was demonstrated by the cryoprotectant’s efficacy in maintaining the three-dimensional gel network by small-strain oscillatory rheological testing and increasing the emulsifying capacity and emulsion stability of the MP. The FT process caused a decrease (P?<?0.05) in whiteness, water-binding capacity, and texture (hardness, springiness, and gumminess) of the MP gels, which were reduced by the addition of cryoprotectants. Overall, the FT-induced loss of the abovementioned functional and rheological properties was significantly reduced by the presence of cryoprotectants.     

Lung tissue engineering and preservation of alveolar microstructure using a novel casting method

Abstract

We used a rat model to decellularize and seed alveolar cells on a three-dimensional lung scaffold to preserve alveolar microarchitecture. We verified the preservation of terminal respiratory structure by casting and by scanning electron microscopy (SEM) of the casts after decellularization. Whole lungs were obtained from 12 healthy Sprague-Dawley rats, cannulated through the trachea under sterile conditions, and decellularized using a detergent-based method. Casting of both natural and decellularized lungs was performed to verify preservation of the inner microstructure of scaffolds for further cell seeding. Alveolar cell seeding was performed using green fluorescent protein (GFP) lung cells and non-GFP lung cells, and a peristaltic pump. We assessed cell seeding using histological and immunohistochemical staining, and enzymatic evaluation. All cellular components were removed completely from the scaffolds, and histological staining and SEM of casts were used to verify the preservation of tissue structure. Tensile tests verified conservation of biomechanical properties. The hydroxyproline content of decellularized lungs was similar to native lung. Histological and immunohistochemical evaluations showed effective cell seeding on decellularized matrices. Enzymatic measurement of trypsin and alpha 1 antitrypsin suggested the potential functional properties of the regenerated lungs. Casts produced by our method have satisfactory geometrical properties for further cell seeding of lung scaffolds. Preservation of micro-architecture and terminal alveoli that was confirmed by SEM of lung casts increases the probability of an effective cell seeding process.     

Simultaneous saccharification of inulin and ethanol fermentation by recombinantSaccharomyces cerevisiae secreting inulinase

VariousSaccharomyces cerevisiae strains were transformed with a 2 μ-based multicopy expression plasmid, pYIGP, carryingKluyveromyces marxianus inulinase gene under the control ofGAPDH promoter. Among them two strains, SEY2102 and 2805, showed high levels of cell growth and inulinase expression, and were selected to study their fermentation properties on inulin. Jerusalem artichoke inulin was more effective for cell growth (10∼11 g-dry wt./L at 48 hr) and inulinase expression (1.0 units/mL with SEY2102/pYIGP and 2.5 units/mL with 2805/pYIGP) than other inulin sources such as dahlia and chicory. It was also found that maximal ethanol production of 9 g/L was obtained from Jerusalem artichoke inulin at the early stationary phase (around 30 hr), indicating that recombinantS. cerevisiae cells secreting exoinulinase could be used for the simultaneous saccharification of inulin and ethanol fermentation.     

Applications of inulin and oligofructose in health and nutrition

Inulin and oligofructose belong to a class of carbohydrates known as fructans. The main sources of inulin and oligofructose that are used in the food industry are chicory and Jerusalem artichoke. Inulin and oligofructose are considered as functional food ingredients since they affect physiological and biochemical processes in rats and human beings, resulting in better health and reduction in the risk of many diseases. Experimental studies have shown their use as bifidogenic agents, stimulating the immune system of the body, decreasing the levels of pathogenic bacteria in the intestine, relieving constipation, decreasing the risk of osteoporosis by increasing mineral absorption, especially of calcium, reducing the risk of atherosclerosis by lowering the synthesis of triglycerides and fatty acids in the liver and decreasing their level in serum. These fructans modulate the hormonal level of insulin and glucagon, thereby regulating carbohydrate and lipid metabolism by lowering the blood glucose levels; they are also effective in lowering the blood urea and uric acid levels, thereby maintaining the nitrogen balance. Inulin and oligofructose also reduce the incidence of colon cancer. The biochemical basis of these beneficial effects of inulin and oligofructose have been discussed. Oligofructoses are non-cariogenic as they are not used byStreptococcus mutans to form acids and insoluble glucans that are the main culprits in dental caries. Because of the large number of health promoting functions of inulin and oligofructose, these have wide applications in various types of foods like confectionery, fruit preparations, milk desserts, yogurt and fresh cheese, baked goods, chocolate, ice cream and sauces. Inulin can also be used for the preparation of fructose syrups.     

Physiological effects of lactulose and inulin in the caecum of rats

A model experiment was performed on rats to evaluate the effect of partial or total substitution of saccharose (S) and cellulose (C) by preparations of lactulose and inulin on the development and metabolism of the caecum. In the experimental diets given to rats for 4 weeks, the examined preparations were administered either with an equivalent amount of cellulose (each at 4% of the diet) or as sole source of dietary fibre at 8% of the diet. Compared to the saccharose group cellulose had no effect, and low doses of lactulose and inulin in the diet increased to a medium extent the weight of the caecum wall and caecal digesta. The addition of lactulose and inulin at 8% increased significantly the content of caecal digesta (4.62 and 4.11?g/100g BW, respectively) and the weight of the caecal wall (1.10 and 0.86?g/100g BW, respectively), compared to the groups with saccharose and cellulose (0.73, 0.90 and 0.24, 0.28?g/100g BW, respectively). Cellulose and cellulose partially-substituted with lactulose and inulin caused an increase in the dry matter content of caecal digesta (26.5–27.5%), compared to other groups (21.8–22.8%). The administration of lactulose and inulin preparations was accompanied by a significant drop in pH (5.47–5.81), compared to the groups with cellulose or saccharose (6.83–6.91), and a decrease in the ammonia concentration in the caecal digesta, compared to the cellulose control (0.27–0.40 and 0.62?mg/g, respectively). The group with 8% lactulose was characterized by the highest activities of microbiological α- and β-galactosidase and β-glucosidase in the caecal digesta. Cellulose and both preparations significantly decreased the activity of β-glucuronidase, compared to the saccharose group (0.39–0.89 and 1.52?U/g, respectively). The highest concentration of VFA in the caecal digesta was observed in the saccharose group (89.2?μmol/g), and the lowest concentration in the group where cellulose was totally substituted by lactulose and inulin (55.1 and 57.5?μmol/g, respectively). The total production of VFA in the caecum was fourfold higher with 8 % lactulose and inulin (254.7 and 236.4?μmol/100g BW, respectively) than in both controls groups (65.1 and 67.8?μmol/100g BW, respectively). The high dose of inulin and lactulose increased the share of propionic acid in the VFA profile (C₂:C₃:C₄) compared to both control groups. When 4% inulin was added to the diet a significant increase of butyrate concentration in the caecum was observed.     

Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays

A major problem for the effective diagnosis and management of prion diseases is the lack of rapid high-throughput assays to measure low levels of prions. Such measurements have typically required prolonged bioassays in animals. Highly sensitive, but generally non-quantitative, prion detection methods have been developed based on prions' ability to seed the conversion of normally soluble protease-sensitive forms of prion protein to protease-resistant and/or amyloid fibrillar forms. Here we describe an approach for estimating the relative amount of prions using a new prion seeding assay called real-time quaking induced conversion assay (RT-QuIC). The underlying reaction blends aspects of the previously described quaking-induced conversion (QuIC) and amyloid seeding assay (ASA) methods and involves prion-seeded conversion of the alpha helix-rich form of bacterially expressed recombinant PrP(C) to a beta sheet-rich amyloid fibrillar form. The RT-QuIC is as sensitive as the animal bioassay, but can be accomplished in 2 days or less. Analogous to end-point dilution animal bioassays, this approach involves testing of serial dilutions of samples and statistically estimating the seeding dose (SD) giving positive responses in 50% of replicate reactions (SD(50)). Brain tissue from 263K scrapie-affected hamsters gave SD(50) values of 10(11)-10(12)/g, making the RT-QuIC similar in sensitivity to end-point dilution bioassays. Analysis of bioassay-positive nasal lavages from hamsters affected with transmissible mink encephalopathy gave SD(50) values of 10(3.5)-10(5.7)/ml, showing that nasal cavities release substantial prion infectivity that can be rapidly detected. Cerebral spinal fluid from 263K scrapie-affected hamsters contained prion SD(50) values of 10(2.0)-10(2.9)/ml. RT-QuIC assay also discriminated deer chronic wasting disease and sheep scrapie brain samples from normal control samples. In principle, end-point dilution quantitation can be applied to many types of prion and amyloid seeding assays. End point dilution RT-QuIC provides a sensitive, rapid, quantitative, and high throughput assay of prion seeding activity.     

菊苣提取物和菊粉降脂活性研究

以300日龄商品代尼克T粉壳蛋鸡为试验动物,研究了菊苣提取物和菊粉对血清脂质、蛋黄总脂和胆固醇的影响.结果显示,0.1%菊苣提取物组、2.0%菊苣提取物组和0.1%菊粉组血清胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白(LDL-C)均显著低于对照组(P<0.05),其中0.1%菊苣提取物组TC降低36.47%,TG降低40.71%,LDL-C降低36.09%,0.1%菊粉组LDL-C降低16.23%,差异极显著(P<0.01);2.0%菊苣提取物组高密度脂蛋白(HDL-C)比对照组高23.19%,差异极显著(P<0.01);2.0%菊苣提取物组蛋黄总脂和蛋黄胆固醇显著降低(P<0.05).表明菊苣提取物和菊粉均具有降血脂活性,但菊苣提取物比菊粉活性更强;菊苣提取物还具有降低蛋黄总脂和胆固醇的作用.     

The State of the Art in the Production of Fructose from Inulin Enzymatic Hydrolysis

ABSTRACT

The present work reviews the main advancements achieved in the last decades in the study of the fructose production process by inulin enzymatic hydrolysis. With the aim of collecting and clarifying the majority of the knowledge in this area, the research on this subject has been divided in three main parts: a) the characteristics of inulin (the process reactant); b) the properties of the enzyme inulinase and its hydrolytic action; c) the advances in the study of the applications of inulinases in bioreactors for fructose production.

Many vegetable sources of inulin are reported, including information about their yields in terms of inulin. The properties of inulin that appear relevant for the process are also summarized, with reference to their vegetable origin.

The characteristics of the inulinase enzyme that catalyzes inulin hydrolysis, together with the most relevant information for a correct process design and implementation, are described in the paper. An extended collection of data on microorganisms capable of producing inulinase is reported. The following characteristics and properties of inulinase are highlighted: molecular weight, mode of action, activity and stability with respect to changes in temperature and pH, kinetic behavior and effect of inhibitors. The paper describes in detail the main aspects of the enzyme hydrolysis reaction; in particular, how enzyme and reactant properties can affect process performance. The properties of inulinase immobilized on various supports are shown and compared to those of the enzyme in its native state.

Finally, a number of applications of free and immobilized inulinases and whole cells in bioreactors are reported, showing the different operating procedures and reactor types adopted for fructose production from inulin on a laboratory scale.     

The state of the art in the production of fructose from inulin enzymatic hydrolysis

The present work reviews the main advancements achieved in the last decades in the study of the fructose production process by inulin enzymatic hydrolysis. With the aim of collecting and clarifying the majority of the knowledge in this area, the research on this subject has been divided in three main parts: a) the characteristics of inulin (the process reactant); b) the properties of the enzyme inulinase and its hydrolytic action; c) the advances in the study of the applications of inulinases in bioreactors for fructose production. Many vegetable sources of inulin are reported, including information about their yields in terms of inulin. The properties of inulin that appear relevant for the process are also summarized, with reference to their vegetable origin. The characteristics of the inulinase enzyme that catalyzes inulin hydrolysis, together with the most relevant information for a correct process design and implementation, are described in the paper. An extended collection of data on microorganisms capable of producing inulinase is reported. The following characteristics and properties of inulinase are highlighted: molecular weight, mode of action, activity and stability with respect to changes in temperature and pH, kinetic behavior and effect of inhibitors. The paper describes in detail the main aspects of the enzyme hydrolysis reaction; in particular, how enzyme and reactant properties can affect process performance. The properties of inulinase immobilized on various supports are shown and compared to those of the enzyme in its native state. Finally, a number of applications of free and immobilized inulinases and whole cells in bioreactors are reported, showing the different operating procedures and reactor types adopted for fructose production from inulin on a laboratory scale.     

Effect of sucrose,stevia and xylitol on rheological properties of gels from blends of chestnut and rice flours

The development of high quality gluten-free products requires the understanding of the phenomena that dictate the ingredient interactions commonly used in foodstuff. In this work, the main objective was to develop alternative gluten-free gelled desserts from blends of chestnut flour (Cf) and whole (Rw), Agulha (Ra) or Carolino (Rc) rice flours. The impact of sucrose, stevia and xylitol on textural, rheological and structural properties of selected gels was investigated. Texture results indicated that studied gels in the presence of sucrose and xylitol decreased significantly the firmness. Rheological outcomes showed that the temperature ramps on heating of Cf/Rw gels were similar to those obtained for Cf/Ra, whereas Cf/Rc gels presented a particular pattern. The presence of sucrose resulted in a significant decrease in the values of storage and loss moduli. Confocal microscopic images showed that the sugar addition leads to a less aggregated structure with fracture lines well marked.     

The Effect of Manipulating Fat Globule Size on the Stability and Rheological Properties of Dairy Creams

Recombined cream (RC, 23 % fat w/w) and standardised commercial cream (CC, 28 % fat w/w) were studied to understand the effects of manipulating fat globule size at the micron-/nano-scale on the stability and rheological properties of cream. All samples were adjusted to a fat: protein ratio of 5:1 and a fat: emulsifier (Tween 80) ratio of 30:1 to stabilize emulsion. For both CC and RC, different emulsions with droplet sizes covering micron- (3.9 μm), sub-micron (0.5 – 0.6 μm) and nano-metric scales (0.13 – 0.29 μm) were obtained using either the homogeniser (7/3 MPa) or the microfluidiser (85 MPa and 42 MPa). Fat globules from both RC and CC had high zeta potential values (-28 to -43 mV) and maintained their reduced size after 1 month of storage at 4 °C, providing evidence of emulsion stability. Droplet size had a significant effect on rheological characteristics of all creams produced. Nano-sized RC tended to have a rigid structure (solid/gel-like form) as compared to micron-sized RC (liquid-like form) as reflected by higher phase angle. Surprisingly, the rheological properties of CC exhibited an opposite tendency to that of RC. This implies that the observed rheological properties of CC and RC could not be fully explained by the discrepancy in droplet size. Differences in interfacial properties between RC and CC might also play a role in the rheological behaviour of the creams. Results indicated the stable high milk fat emulsions could be successfully created by reducing the globule size. These findings would be useful in understanding how micron-/nano-sized emulsions can be utilised in further application or processing of creams.     

Cholesterol-lowering effects of dietary pomegranate extract and inulin in mice fed an obesogenic diet

BackgroundIt has been demonstrated in animal studies that both polyphenol-rich pomegranate extract (PomX) and the polysaccharide inulin, ameliorate metabolic changes induced by a high-fat diet, but little is known about the specific mechanisms.ObjectiveThis study evaluated the effect of PomX (0.25%) and inulin (9%) alone or in combination on cholesterol and lipid metabolism in mice.MethodsMale C57BL/6 J mice were fed high-fat/high-sucrose [HF/HS (32% energy from fat, 25% energy from sucrose)] diets supplemented with PomX (0.25%) and inulin (9%) alone or in combination for 4 weeks. At the end of intervention, serum and hepatic cholesterol, triglyceride levels, hepatic gene expression of key regulators of cholesterol and lipid metabolism as well as fecal cholesterol and bile acid excretion were determined.ResultsDietary supplementation of the HF/HS diet with PomX and inulin decreased hepatic and serum total cholesterol. Supplementation with PomX and inulin together resulted in lower hepatic and serum total cholesterol compared to individual treatments. Compared to HF/HS control, PomX increased gene expression of Cyp7a1 and Cyp7b1, key regulators of bile acid synthesis pathways. Inulin decreased gene expression of key regulators of cholesterol de novo synthesis Srebf2 and Hmgcr and significantly increased fecal elimination of total bile acids and neutral sterols. Only PomX in combination with inulin reduced liver and lipid weight significantly compared to the HF/HS control group. PomX showed a trend to decrease liver triglyceride (TG) levels, while inulin or PomX-inulin combination had no effect on either serum or liver TG levels.ConclusionDietary PomX and inulin supplementation decreased hepatic and serum total cholesterol by different mechanisms and the combination leading to a significant enhancement of the cholesterol-lowering effect.     

Effects of seeding procedures and water quality on recovery of Cryptosporidium oocysts from stream water by using U.S. Environmental Protection Agency Method 1623

U.S. Environmental Protection Agency method 1623 is widely used to monitor source waters and drinking water supplies for Cryptosporidium oocysts. Matrix spikes, used to determine the effect of the environmental matrix on the method's recovery efficiency for the target organism, require the collection and analysis of two environmental samples, one for analysis of endemic oocysts and the other for analysis of recovery efficiency. A new product, ColorSeed, enables the analyst to determine recovery efficiency by using modified seeded oocysts that can be differentiated from endemic organisms in a single sample. Twenty-nine stream water samples and one untreated effluent sample from a cattle feedlot were collected in triplicate to compare modified seeding procedures to conventional seeding procedures that use viable, unmodified oocysts. Significant negative correlations were found between the average oocyst recovery and turbidity or suspended sediment; this was especially apparent in samples with turbidities greater than 100 nephelometric turbidity units and suspended sediment concentrations greater than 100 mg/liter. Cryptosporidium oocysts were found in 16.7% of the unseeded environmental samples, and concentrations, adjusted for recoveries, ranged from 4 to 80 oocysts per 10 liters. Determining recovery efficiency also provided data to calculate detection limits; these ranged from <2 to <215 oocysts per 10 liters. Recoveries of oocysts ranged from 2.0 to 61% for viable oocysts and from 3.0 to 59% for modified oocysts. The recoveries between the two seeding procedures were highly correlated (r = 0.802) and were not significantly different. Recoveries by using modified oocysts, therefore, were comparable to recoveries by using conventional seeding procedures.