Uppsala Method Research Articles

Freezability of Dog Semen after Collection in Field Conditions and Cooled Transport.

Simple SummaryDog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. To avoid animal movement, it seemed interesting to investigate whether freezing dog semen after 24 or 48 h of cooled transport to an equipped laboratory was possible when semen collection was performed in the field such as in local breeding kennels. The influence of two pre-freezing holding times (i.e., 24 or 48 h) and two holding diluents (solutions used to dilute semen before freezing) was evaluated. Post-thaw morphofunctional sperm features, such as motility, morphological integrity, and ability to bind female gametes, were assessed. No differences between times or diluents were observed, but motility tended to decrease in the samples frozen at 48 h. Since the insemination dose was based on the number of motile spermatozoa, a shorter pre-freezing time is advisable. Yet, considering that the rest of the morphofunctional parameters remained comparable between samples frozen after collection or after 24/48 h of transport, freezing after cooled transport is a good option for avoiding animal stress and for promoting a greater diffusion of semen cryopreservation.Dog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. The aim of this study was to evaluate whether freezing dog semen after 24 or 48 h of cooled transport to an equipped laboratory was possible when semen collection was performed in the field such as in local breeding kennels. Single ejaculates from different dogs (mixed breeds and ages) were collected. In Experiment I, 10 ejaculates were conventionally frozen using the Uppsala method or frozen after 24 or 48 h of storage in a Styrofoam transport box cooled by icepacks. In Experiment II, 10 ejaculates were used to assess the influence of two extenders (Uppsala chilling extender or freezing extender 1) used for semen dilution during the 24 or 48 h storage. Motility, morphology, membrane, and acrosome integrity were analyzed as well as spermatozoa zona-binding ability. No significant differences were observed among the frozen groups, regardless of freezing time (Experiment I) or extender (Experiment II). Motility at thawing, however, decreased in absolute value at 48 h. Freezing of freshly collected semen is the gold standard, but the results obtained in this study prompt the application of freezing after cooled transport for the long-term preservation of dog semen, especially if the transport can be organized in 24 h.

Lignin is the main determinant of total dietary fiber differences between date fruit (Phoenix dactylifera L.) varieties

Date fruits (Phoenix dactylifera) of ten varieties, collected in the United Arab Emirates, were studied to determine their dietary fiber content and composition. Fourier transform infrared (FTIR) spectroscopy indicated that the dietary fiber components in all the date fruit varieties was similar. The major dietary fiber components, including cellulose, hemicellulosic components, lignin, and pectin, were analyzed by the Uppsala method. The total dietary fiber content in the date fruits analyzed (5.2%–8.3%) is comparable to commonly consumed dried fruits and is correlated with the content of lignin. The lignin was the main determinant of the total dietary fiber content in dates and its content was higher in semi-hard and hard fruit varieties.

Side Streams of Broccoli Leaves: A Climate Smart and Healthy Food Ingredient.

Human consumption of fruits and vegetables are generally below recommended levels. Waste from the production, e.g., of un-used parts such as broccoli leaves and stem when producing broccoli florets for food, is a sustainability issue. In this study, broccoli leaves were analyzed for the content of various dietary fibre and phenolics, applying the Uppsala method and HPLC analyses, respectively. The results showed that broccoli leaves had comparable levels of dietary fibre (26%–32% of dry weight (DW)) and phenolic compounds (6.3–15.2 mg/g DW) to many other food and vegetables considered valuable in the human diet from a health perspective. A significant positive correlation was found among soluble dietary fibre and phenolic acids indicating possible bindings between these components. Seasonal variations affected mainly the content of conjugated phenolics, and the content of insoluble dietary fibre. This study verified the importance of the use of broccoli production side streams (leaves) as they may contribute with health promoting components to the human diet and also socio-economic and environmental benefits to the bioeconomic development in the society.

Effects of rapid cooling prior to freezing on the quality of canine cryopreserved spermatozoa.

The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality. In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid, cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM), progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability (spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM, PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated glycerol.

The non–starch polysaccharides quantity changes in pastry products where Jerusalem artichoke (Helianthus tuberosus L.) added

Non-starch polysaccharides (NSP) constitute the main part of dietary fiber (DF). There are two kinds of NSP - insoluble and soluble. Most plant foods contain both types although proportions vary. A good source of NSP/fiber is Jerusalem artichoke (Helianthus tuberosus L.), vegetable - rich in fiber, vitamins and minerals. Jerusalem artichoke (JA) is used for managing cholesterol, lipids in the blood, can also help with weight control, not through better metabolism of fats but through better management of sugars. The addition of Jerusalem artichoke powder (JAP) increases the fiber content of the pastry products, making it a healthier food. Several methods are available for the determination of total dietary fiber in products. The Uppsala method is the most appropriate when information of the DF components is required. Since different DF components have a range of beneficial effects on human health this approach of analyzing DF should become more widely used when characterizing foods. The aim of the study was to determine quantity changes in individual NSP of DF in raw material JAP and such pastry products as cakes, butter biscuits and honey biscuits where JAP was added. The study identified quantity and types of NSP in JAP and pastry products with JAP in concentration 30% and 50% of total amount of flour, but as control are samples without JAP. Results showed that the major NSP monomers of all pastry kinds are, in decreasing manner, glucose, galactose, mannose, arabinose, xylose, rhamnose, and fucose. Additionally, NSP content increases substantially with increasing concentration of JAP in cakes, honey biscuits and butter biscuits.

Prediction of cell wall polysaccharide and lignin concentrations of alfalfa stems from detergent fiber analysis

Identification of superior biomass feedstocks for conversion to ethanol requires accurate, rapid, and inexpensive measurement of cell wall polysaccharide and lignin concentrations. A study was undertaken to determine if cell wall characteristics measured with the Uppsala dietary fiber method could be accurately predicted from detergent fiber data. Spring and summer growth alfalfa (Medicago sativa L.) stems were harvested from two germplasm sources for 2-years. Samples were analyzed for neutral sugars, uronic acids, and Klason lignin using the Uppsala dietary fiber method, and neutral detergent fiber, acid detergent fiber, and acid detergent lignin by sequential detergent analysis. Cell wall, hemicellulose, cellulose, and lignin concentrations derived from these two methods were positively correlated, but detergent estimates of cell wall and lignin concentration were lower and hemicellulose and cellulose concentrations were higher than for the Uppsala method. These patterns were attributed to solubility of pectin in neutral detergent solution and poor lignin recovery in acid detergent lignin. Regression equations for prediction of cell wall characteristics using detergent results were significant and accounted for 35–92% of the variation in the data. Significant deviations between predicted and observed cell wall results were observed when the equations were tested using an independent sample set, although the deviation was very small for cellulose concentration. It does not appear that most cell wall characteristics of alfalfa stems obtained using the Uppsala dietary fiber method can be accurately predicted using detergent fiber results; however, cellulose concentration might be predicted with acceptable accuracy.

Dietary Fibre Content and Composition of Some Edible Fungi Determined by Two Methods of Analysis

Six common edible fungi from four orders of Basidiomycetes, namely Agaricus bisporus (Agaricales), Auricularia auricula and Auricularia polytricha (Auriculariales), Tremella fuciformis (Tremellates), Ganoderma lucidum and Poria cocos (Aphyllophorales), were analysed for their total dietary fibre (TDF) content using the Association of Official Analytical Chemists (AOAC) method and for dietary fibre content and composition using the Uppsala method. The non-protein nitrogen contributed from fungal chitin was corrected in the fibre residue obtained by the AOAC method. The TDF content measured by the AOAC method was always greater than that of the Uppsala method. The TDF content determined by the AOAC and Uppsala methods ranged from 182 and 132 g kg-1 (dry matter) in A bisporus to 735 and 711 g kg-1 (dry matter) in P cocos, respectively. Neutral and amino sugars were the dominant sugars in all the fungi. Sugar composition of the TDF reflected that the major cell wall polysaccharides in most fungi were hemicelluloses, such as β-glucan and glucuronoxylomannan, pectic substances and chitin. Judging from their high fibre content and unique fibre composition, edible fungi have considerable value as sources of dietary fibre in human nutrition. © 1997 SCI.

Dietary Fibre Content and Composition of Some Edible Fungi Determined by Two Methods of Analysis

Six common edible fungi from four orders of Basidiomycetes, namely Agaricus bisporus (Agaricales), Auricularia auricula and Auricularia polytricha (Auriculariales), Tremella fuciformis (Tremellates), Ganoderma lucidum and Poria cocos (Aphyllophorales), were analysed for their total dietary fibre (TDF) content using the Association of Official Analytical Chemists (AOAC) method and for dietary fibre content and composition using the Uppsala method. The non-protein nitrogen contributed from fungal chitin was corrected in the fibre residue obtained by the AOAC method. The TDF content measured by the AOAC method was always greater than that of the Uppsala method. The TDF content determined by the AOAC and Uppsala methods ranged from 182 and 132 g kg-1 (dry matter) in A bisporus to 735 and 711 g kg-1 (dry matter) in P cocos, respectively. Neutral and amino sugars were the dominant sugars in all the fungi. Sugar composition of the TDF reflected that the major cell wall polysaccharides in most fungi were hemicelluloses, such as β-glucan and glucuronoxylomannan, pectic substances and chitin. Judging from their high fibre content and unique fibre composition, edible fungi have considerable value as sources of dietary fibre in human nutrition. © 1997 SCI.

Dietary Fiber Content and Composition of Some Cultivated Edible Mushroom Fruiting Bodies and Mycelia

The mycelia and the caps and stalks of fruiting bodies of four edible mushrooms (Lentinus edodes, Lycophyllum shimeji, Pleurotus sajor-caju, and Volvariella volvacea) were analyzed for their total dietary fiber (TDF) content according to the Association of Official Analytical Chemists (AOAC) method and for TDF content and composition according to the Uppsala method. The nonprotein nitrogen attributable to chitin was corrected in the fiber residue obtained with the AOAC method. The TDF contents of all of the mushrooms as measured by the AOAC method were considerably greater than those determined by using the Uppsala method. Mushroom mycelia had higher TDF values than did the fruiting bodies as measured by both methods. The TDF composition of the mycelia and the TDF composition of the caps and stalks of the mushroom fruiting bodies were similar. Sugar composition reflected that β-glucans were the major fiber polysaccharide with chitin, hemicelluloses, and polyuronides as minor ones. Notably, one-third of th...

Total Dietary Fiber Determined as Neutral Sugar Residues, Uronic Acid Residues, and Klason Lignin (The Uppsala Method): Collaborative Study

A joint AOAC/American Association of Cereal Chemists (AACC) collaborative study was conducted to determine by the Uppsala method the dietary fiber content and its composition in various foods. The method includes preparation of a residue by treatment with thermostable alpha-amylase and amyloglucosidase and then ethanol precipitation of solubilized dietary fiber components while leaving low-molecular weight carbohydrates in solution. After acid hydrolysis of residue, neutral polysaccharide residues are determined as alditol acetates by gas-liquid chromatography, uronic acid residues are determined by colorimetry, and ash-free acid-insoluble residue (Klason lignin) is determined gravimetrically. Total dietary fiber, including enzyme-resistant starch, is calculated as the sum of nonstarch polysaccharide residues and Klason lignin. Nine laboratories completed the study, analyzing in duplicate 8 unknown dried products that included 4 cereal products, green peas, potato fiber, carrots, and apples. Total dietary fiber contents of products tested ranged from 4.6 to 84.3%, with an average RSDR value of 8.4% (range, 4.8-11.1%). Total neutral polysaccharide residues ranged from 3.8 to 64.1%, with an average RSDR value of 7.5% (range, 5.4-10.5%). Individual neutral sugars (rhamnose, arabinose, xylose, mannose, galactose, and glucose) and uronic acid residues present at more than 1% generally had good RSDR values (3.3-22.8%), whereas, as expected for Klason lignin, only the wheat bran sample with a high content (16%) had an excellent RSDR value (5.0%). The gas chromatographic-colorimetric-gravimetric method (Uppsala method) for determination of total dietary fiber (as neutral sugar residues, uronic acid residues, and Klason lignin) has been adopted first action by AOAC INTERNATIONAL.

Dietary Fiber Content of Cereal and Grain Products Determined by Enzymatic–Chemical and Enzymatic–Gravimetric Methods

The two objectives of this research were to compare total dietary fiber values obtained by the AOAC enzymatic–gravimetric method and the Uppsala enzymatic–chemical method for 50 grain and cereal products and to compare the fiber content and composition of foods that are likely to be grouped during nutrient intake studies. Mean fiber contents of 44 refined grain products determined by the two methods were significantly different ( P < 0.01), 2.5 ± 1.2 g/100 g (fresh wt) by the AOAC method and 2.0 ± 0.7 g/100 g by the Uppsala method. Mean fiber contents of 6 higher fiber foods also were different ( P < 0.01), 9.1 ± 1.6 and 7.9 ± 1.2 g/100 g, respectively. Data from the two methods of analysis were strongly correlated, r 2 = 0.93 ( P < 0.001). The soluble fiber content of the refined products was an average of 25% of total fiber, of the higher fiber products, 20%. Ingredients influenced the fiber content and composition of the samples; some such as fat and sugar diluted the fiber concentration and others such as fruit, nuts, coconut, and chocolate increased it. We conclude that complete descriptions should be obtained if food intake records are to reflect fiber intake; grouping some foods prior to calculating fiber intake is likely to lead to errors and fiber intakes calculated from AOAC fiber data will be higher than those determined by the Uppsala fiber method. Because a more specific set of analyses would be likely to provide more accurate results, we recommend that data obtained using an enzymatic–chemical method be used to calculate fiber intake in nutritional studies.

Dietary fiber content and composition of different forms of fruits

The dietary fiber contents of 19 different forms of seven fruits were analyzed by two methods and compared. Fiber content measured by the AOAC method was always greater than that obtained by the Uppsala method; the two sets of fiber data were significantly different ( P < 0.002). Fiber contents of different forms of oranges, peaches and plums were similar. Fiber contents of four berries (blackberries, cranberries, red respberries, strawberries) ranged from 1.0 to 7.0% fresh weight. Peeling or canning changed the uronic acid and neutral sugar contents of the soluble and insoluble fiber fractions of several fruits. Measurement of more fiber in fruits by the AOAC versus the Uppsala method is similar to the results obtained when grain products and legumes, but not vegetables, were analyzed.

Dietary fiber content and composition of vegetables determined by two methods of analysis

Forty-one samples representing different forms of 12 types of vegetables were analyzed for dietary fiber content using the Association of Official Analytical Chemists method and for fiber content and composition using the Uppsala method. Mean fiber content measured by the former method was 2.1 +/- 0.9 g/100 g and by the latter method, 1.8 +/- 0.8 g/100 g. The mean soluble fiber concentration determined by the Uppsala method was 15% of total vegetable fiber. Insoluble fiber contained more neutral sugars than uronic acid; soluble vegetable fiber was a mixture of uronic acid and polysaccharides containing galactose and arabinose. Cooking either had no effect or increased the proportion of total fiber extracted as soluble fiber. Some vegetables that are frequently treated as similar foods in nutritional studies (e.g., potatoes and cooked greens) contained different amounts of fiber. The results indicate that for some vegetables the source, preparation method, variety, and analytical method should be considered when daily intakes of total dietary fiber or fiber components are calculated.

Comparison of Two Methods of Fiber Analysis of 58 Foods

Nine fruits, 8 grains, 9 legumes, 19 vegetables, 6 nuts and seeds, and 7 miscellaneous foods were analyzed for dietary fiber content using two methods of analysis. Fiber values determined using the AOAC method were either similar to or greater than those measured using the Uppsala method. The total fiber contents of the samples were generally similar to what have been reported for other foods in the same food group. Soluble fiber was an average of ∼15–20% of the total fiber in fruits, vegetables and grains, and <10% of total fiber in legumes, nuts, and seeds. Pectin and hemicelluloses were major soluble fiber components of fruits and vegetables. Grains contained little pectin. Glucose, representing cellulose, was usually the major (≥50%) neutral sugar in the insoluble fiber fractions of fruits, legumes, and vegetables. Neutral sugars in soluble fiber fractions suggested the presence of arabinogalactan hemicelluloses in fruits and vegetables. Arabinose and xylose were usually the major hemicellulosic sugars in grains. Within a food group the dietary fiber content and composition of the 58 foods were generally similar to those previously analyzed in the same food groups.