Two stomacher accessories.

Two useful and easily constructed accessories for the Colworth Stomacher blender are described. The first automatically opens and holds Stomacher bags above a balance of weighing in the sample. The second stores bags on the bench between manipulations and opens and holds them as required for pipetting, etc.     

Usefulness of the stomacher in a microbiological regulatory laboratory.

The relative efficiency of the Waring blender, the Stomacher 400, and the Stomacher 3500 for preparing food samples for microbiological analysis was studied. Comparative aerobic plate count (APC) values were determined on 671 samples, representing 30 categories of foods. Of the 26 categories of nonfatty foods, the blender gave significantly higher geometric mean APC values than those given by the Stomacher 400 and the Stomacher 3500 in 65 and 69 percent of the categories, respectively. In a comparison of the two Stomacher models, the Stomacher 400 gave significantly higher geometric mean APC values than these given by the Stomacher 3500 in 73 percent of the food categories. Addition of Tween 80 to four categories of fatty foods at concentrations of 0.5, 1.0, and 2.0 percent did not raise the APC values given by either model of stomacher to the levels given by the Waring blender. Overall, the efficiency of both models of Stomacher, relative to the blender and to each other, was specific and depended upon the particular food being analyzed.     

Usefulness of the stomacher in a microbiological regulatory laboratory

The relative efficiency of the Waring blender, the Stomacher 400, and the Stomacher 3500 for preparing food samples for microbiological analysis was studied. Comparative aerobic plate count (APC) values were determined on 671 samples, representing 30 categories of foods. Of the 26 categories of nonfatty foods, the blender gave significantly higher geometric mean APC values than those given by the Stomacher 400 and the Stomacher 3500 in 65 and 69 percent of the categories, respectively. In a comparison of the two Stomacher models, the Stomacher 400 gave significantly higher geometric mean APC values than these given by the Stomacher 3500 in 73 percent of the food categories. Addition of Tween 80 to four categories of fatty foods at concentrations of 0.5, 1.0, and 2.0 percent did not raise the APC values given by either model of stomacher to the levels given by the Waring blender. Overall, the efficiency of both models of Stomacher, relative to the blender and to each other, was specific and depended upon the particular food being analyzed.     

Comparison of bacterial counts obtained from naturally contaminated foods by means of Stomacher and blender

Four Regional Health Protection Branch laboratories each compared aerobic colony counts obtained after "stomaching" and blending, for a minimum of 10 samples in each of the seven food groups: dry pastas; chocolate and cocoa powders; frozen entrees (macaroni and cheese, chow mein, chop suey, fried rice, seafood casseroles, and Salisbury steak); nonfat dry milk; shrimp and crabmeats; spices; and breakfast sausages. Overall, counts obtained after using the Stomacher were equivalent to or higher than counts obtained after using the blender in 73% of the comparisons (alpha = 0.05). Where differences existed, counts obtained after using the Stomacher tended to be higher than counts obtained after using the blender from milk powder and lower from sausage. Aerobic colony counts from these foods are not unacceptably biased when obtained by Stomacher.     

Development and Characterization of Multifunctional Gelatin-Lysozyme Films Via the Oligomeric Proanthocyanidins (OPCs) Crosslinking Approach

Herein, we report firstly the development of sustained antimicrobial and antioxidant gelatin-lysozyme films crosslinked by the oligomeric proanthocyanidins (OPCs), a duel-functional agent. Lysozyme release kinetic studies were performed at neutral and acidic pH, and they could be described as a biphasic process. OPCs crosslinking retarded lysozyme release at pH 7.0, in a dose dependent manner, and the inhibition zone tests confirmed that the sustained release of lysozyme was realized upon weak crosslinking with OPCs. OPCs crosslinking enhanced thermal stability of the gelatin films, and gave them the ability to barrier ultraviolet light. OPCs loadings endowed the films excellent antioxidant activities, the DPPH radical scavenging activity of the films increased linearly to 93.97% upon increasing OPCs loadings from 0 to 2.0%. Concomitantly, the reducing powder of the films increased linearly from 6.08 ± 0.09 to 45.53 ± 2.74 μmol Asc Acid/g film. Additionally, the antioxidant properties of gelatin films against lipid oxidation in edible oils were evaluated. Lipid hydroperoxides of algal oils in the gelatin bags were approximately a quarter of that in low-density polyethylene (LDPE)-based bags, and the malondialdehyde (MDA) values of algal oils were lower than that in LDPE-based bags by 1–2 orders of magnitude. Regrettably, the incorporation of OPCs did not enhance the antioxidant capability of gelatin films against lipid oxidation in wrapped edible oils, possibly due to the limited release toward algal oils in term of its oil-soluble attribute. This study opens a promising pathway for producing sustained antimicrobial and antioxidant gelatin films using a bi-functional agent.     

Overview of SHOT hardware capabilities and range of cell biology experiment designs

Cell science hardware is currently available for lease or production by SHOT(R). The SHOT Avian Development Facility (ADF) is a single middeck locker experiment containing two carousels capable of rotating at variable g. Each carousel accommodates 18 40-ml vessels that can be adapted for the cultivation of avian or reptilian eggs, small invertebrates, zebra fish, small plants, seeds, spores, cells or tissues. Cellcult Cassettes contain a small (20-50 ml) rotating cylindrical culture vessel with feed media reservoir, pumps, waste bag and sample collection bags. Cultures can be rotated, perfused, fed, aerated and sampled automatically or on remote command. Fluid Processing Cassettes contain a flexible combination of feed, culture and fixative bags with valves and pumps programmable for various feeding and fixing protocols. Dynacult Cassettes contain a cylindrical bioreactor with differentially rotating walls, pumps and valves for feeding and sampling, and pH and oxygen measurement. All cassettes are doubly contained and are accommodated by a computerized, thermally controlled processing facility, the Advanced Space Experiment Processor, which processes three cassettes, or the Biotechnology Thermal Environment Carrier, which holds up to six cassettes.     

Vacuum packing: a model system for laboratory-scale silage fermentations

AIMS: To determine the utility of vacuum-packed polythene bags as a convenient, flexible and cost-effective alternative to fixed volume glass vessels for lab-scale silage studies. METHODS AND RESULTS: Using perennial ryegrass or red clover forage, similar fermentations (as assessed by pH measurement) occurred in glass tube and vacuum-packed silos over a 35-day period. As vacuum-packing devices allow modification of initial packing density, the effect of four different settings (initial packing densities of 0.397, 0.435, 0.492 and 0.534 g cm(-3)) on the silage fermentation over 16 days was examined. Significant differences in pH decline and lactate accumulation were observed at different vacuum settings. Gas accumulation was apparent within all bags and changes in bag volume with time was observed to vary according to initial packing density. CONCLUSIONS: Vacuum-packed silos do provide a realistic model system for lab-scale silage fermentations. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of vacuum-packed silos holds potential for lab-scale evaluations of silage fermentations, allowing higher throughput of samples, more consistent packing as well as the possibility of investigating the effects of different initial packing densities and use of different wrapping materials.     

Controversy fuels trafficking of GPI-anchored proteins

The model that glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) take a direct transport route to the apical membrane of epithelial cells has recently been challenged. In this issue, Paladino et al. (p. 1023) and Hua et al. (p. 1035) show that the original view nevertheless holds. This closes a chapter in the winding story of GPI-AP trafficking but opens another phase, as the controversy has stimulated the development of new methodology.     

Long-term culture of normal human colonic epithelial cells in vitro.

Studies of normal cellular function as well as the understanding of cellular mechanisms of carcinogenesis and other diseases of the large intestine have been limited, particularly due to the lack of long-term culture of normal human large intestinal epithelial cells (NHLIEC). Using the epithelia from surgically resected human colon, we have dissociated a sufficient number of viable NHLIEC and maintained them in in vitro culture for up to 5 months. Normal-appearing human large intestinal mucosal fragments (1 mm2) were treated with 0.01 mg/ml trypsin, 0.2 mg/ml collagenase + 0.1 mM EGTA or 0.1 mg/ml trypsin + 0.1 mM EGTA in a Stomacher laboratory blender to isolate the cells. Compared with other methods, the use of the Stomacher blender combined with low concentrations of proteolytic enzymes yielded greater numbers of cells per gram of tissue, with up to 84% viable cells. Primary and serially passaged NHLIEC were cultured in CMRL-1066, MEM with 5% serum, and serum-free KGM. These media were all supplemented with insulin, hydrocortisone, epithelial growth factor, and bovine pituitary extract. CMRL-1066 was found to be the best medium for NHLIEC. Contaminating fibroblasts were selectively removed by briefly allowing the cells to adhere to the culture vessel and adding 25 U/ml collagenase to the culture media at the first subculture treatment. The epithelial nature and secretory function of the established cells were confirmed by morphological criteria (light microscopy, phase contrast microscopy and electron microscopy), immunoreactivity to cytokeratin, and positive mucin cytochemistry. We propose that using this methodology for the culture and maintenance of NHLIEC for an extended period of time would serve as a valuable model for a variety of investigations.     

Leaching of plasticizers from and surface characterization of PVC blood platelet bags.

The leaching of phthalate plasticizers from four types of blood platelet bags was investigated. The anticoagulant solutions used in the blood collection bags had pH values of 5.64 +/- .04 and contained no detectable amounts of phthalates. Platelet bag materials from each bag were soaked in normal salines for up to 5 days. The salines were tested for the leached phthalates from the bags but none could be found. However, di-(2-ethylhexyl) phthalate (DEHP) leached out of the PL-146 and Terumo bags into bovine calf serum used for soaking the bag materials. There was an increase in the amount of DEHP leached from about 1.1 mg at the end of one day to about 3.3 mg per gm of bag material at the end of a five day extraction with the serum. In PL-732 sets, a platelet bag made of a specialty polyolefin, the amount of DEHP leached out was less than 0.02 mg per g of bag material. CLX bags, which contained tri-(2-ethylhexyl) trimelliate (TETM) as a plasticizer, showed a negligible amount of it leaching into the calf serum. Infra-red spectra showed that PL-146 bags had been coated with a layer of a fatty acid amide while the Terumo bags contained a layer of a silicone fluid on their inner surfaces. CLX bags showed a coating of stearates, which were probably soaps of calcium or zinc. Scanning electron microscopy (SEM) showed that the inner surfaces of each brand of the bag were distinctly different morphologically. The two PVC bags were very similar whereas the surfaces morphology of PL-732 was rougher. Terumo bag had a different surface morphology than those of the other bags whereas the CLX bags had a very regular surface pattern. The exact significance of the surface morphology is not certain but excessively rough surfaces may not be desirable for the bags.     

Application of a flow-type antibody sensor to the detection of Escherichia coli in various foods

A flow-type biosensor system which uses a broad-spectrum anti-Escherichia coli antibody and quartz crystal microbalance as biological component and transducer was developed. Biosensor responses were initiated by injecting viable E. coli suspensions through a flow cell and the sensor system was optimized for response time according to flow rate and injection time, followed by the measurement of responses for various E. coli strains. As expected, the sensor system showed a characteristic broad binding feature against E. coli strains. A linear sensor response in double-logarithmic scale was observed for the microbial suspensions ranging from 1.7 x 10(5) to 8.7 x 10(7) CFU/ml. Sample measurements could be done within 20-30 min after Stomacher treatment followed by spiking or enrichment.     

Storage of apheresis platelets in ethylene-vinyl acetate copolymer bags: relationship between the bag size and the number of platelets maintaining aerobic metabolism

The present study was designed to determine whether a bag made from ethylene-vinyl acetate copolymer (EV) with superior flexibility at subzero temperature is suitable for a storage container of single-donor apheresis platelets. Apheresis platelets were stored with 100 ml plasma in 1-liter bags made of EV or standard polyvinyl chloride (PVC) plastic at 22 degrees C with constant agitation. The oxygen permeability of the 1-liter EV bag averaged 1447 nmol/min/atm, which was about 1.5 times higher than that of PVC bags. The partial oxygen tension (PO2) of platelet concentrates (PC) has linearly decreased to 16 mm Hg with increasing platelet counts. The level of the partial carbon dioxide was always higher in EV bags than in PVC bags. Oxygen consumption rates of platelets stored in EV and PVC bags with a sufficient oxygen supply averaged 1.25 and 1.20 nmol/min/10(9) platelets, respectively. The rates of glucose consumption and lactate production were not changed in two bags. Ninety percent of the total ATP production of about 8 nmol/min/10(9) platelets were generated through the aerobic metabolism. The platelet counts in the 1-liter EV and PVC bags, at which PO2 is 16 mm Hg, were 2.2 and 1.5 x 10(11) platelets, respectively. The study indicates that apheresis platelets stored in EV bags at 22 degrees C have no different metabolic changes when compared with those of PVC bags. In addition, the number of platelets maintaining the aerobic metabolism is 1.5 times higher than that of PVC bags.     

A novel method for the transport and long-term storage of cultures and samples in an anaerobic atmosphere

Nylon-aluminium-polyethylene laminated, sealable bags provided a more economical and flexible method of transportation, incubation and storage than anaerobic jars. The bags were used with a special heat sealer and 'Anaerocult-A' sachets to remove oxygen. The environment inside the bags remained oxygen-free for at least 3 years.     

Rehabilitating Seagrass by Facilitating Recruitment: Improving Chances for Success

Attempts to arrest seagrass loss through numerous rehabilitation methods have traditionally produced inconsistent results. On Australia's southern coast, hessian bags made from biodegradable jute fibers show promise for rehabilitating Amphibolis antarctica by facilitating recruitment of seedlings in situ. Testing ways to improve the performance of bags (i.e. increasing seagrass recruitment and establishment) showed that bags with a coarse outer weave of hessian facilitated greater seedling densities (approximately 1700 individuals/m²) than bags with a fine outer weave, but the content of bags (sand vs. sand and rubble mixture) had little effect. Isolated bags facilitated greater longer term densities than bags grouped together, while similar densities were sampled up to 80 m away from a natural meadow. Lastly, bags that had spent less time in situ initially facilitated more recruits than older bags, but longer term (21–32 months) retention was similar among bag ages. Collectively, the results suggest hessian bags can be a relatively simple, cost‐effective, and environmentally friendly method for rehabilitating Amphibolis seagrass, with few considerations in their use other than their physical architecture and arrangement (e.g. isolated coarse‐weave bags).     

Does the ‘Old Bag’ Make a Good ‘Wind Bag’?: Comparison of Four Fabrics Commonly Used as Exclusion Bags in Studies of Pollination and Reproductive Biology

BACKGROUND AND AIMS: Fabrics used in pollination bags may exclude pollen carried by biotic vectors, but have varying degrees of permeability to wind-borne pollen. The permeability of bags to wind-borne pollen may have important consequences in studies of pollination and reproductive biology. The permeability of four fabrics commonly used in the construction of pollination bags was examined. METHODS: Deposition of wind-borne pollen on horizontally and vertically oriented microscope slides was assessed on slides enclosed in pollination bags, as well as on control slides. KEY RESULTS: It was found that the permeability of fabrics to wind-borne pollen, as measured by deposition on both horizontally and vertically oriented slides, decreased with pore size. However, deposition on horizontal slides was always greater than on vertical slides for a given fabric; this could manifest itself as differential success of pollination of flowers in bags-dependent on flower orientation. CONCLUSIONS: Obviously, bags with mesh size smaller than most pollen grains are impermeable to pollen. However, material for such bags is very expensive. In addition, it was also observed that bags with even moderately small pore size, such as pores (approx. 200 microm) in twisted fibre cotton muslin, offered highly significant barriers to passage of wind-borne pollen. Such bags are sufficiently effective in most large-sample-size reproductive biology studies.     

Influence of Packaging and Processing Conditions on the Decontamination of Laboratory Biomedical Waste by Steam Sterilization

The conditions for optimal steam decontamination of polypropylene bags half loaded with laboratory biomedical waste were studied (276 bags were processed). Controls were single-closed bags without water added or incisions made in the top, standing freely in an autoclave set at 121°C. The average time required to reach 121°C at the load center was 46 min for controls. A significant increase in this time occurred following addition of water to bags without incisions (60 min), with double bagging (60 min), or when using vertical containers (82 min). A significant decrease occurred when bags were slashed (37 min) or processed at 123°C (32 min) or 132°C (19 min). Horizontal containers or addition of water to slashed bags had no significant effect.     

套袋苹果不同类型果袋内温、湿度变化特征及其对果实外观品质的影响

研究了套袋苹果果实发育过程中不同类型果袋内温、湿度的变化以及果袋对果实外观品质的影响.结果表明:果袋内的温度和湿度变化呈负相关,一天中袋内温度从9:00开始迅速升高,最高温度在春秋季一般出现在13:00-14:00,盛夏一般出现在13:00-17:00,最低温度出现在5:00前后,最高湿度出现在6:00前后.不同类型的果袋温、湿度变化不同,特别在高温和阴雨天差异明显,最高温度表现为双层袋》单层袋,而最高湿度表现为单层袋》双层袋.高温往往伴随极度干燥,当袋内温度超过35 ℃时,微域湿度一般低于40%.套袋对苹果果实品质有重要影响,果实外观品质以双层袋较好、单层袋较差;而双层袋中又以XL袋效果最好.     

Role of transported particulate organic matter in the macroinvertebrate colonization of litter bags in streams

1. The exposure of mesh litter bags has been widely used to investigate the role of benthic macroinvertebrates in leaf processing in freshwaters. In this sense, several studies have related litter bag breakdown rates to the presence of colonizing invertebrates. A possible confounding factor in such experiments is that the litter bags trap suspended particulate organic matter that can itself attract invertebrate colonists, irrespective of the intended experimental treatment.
2. We attempted to quantify the accumulation of particulate organic matter (POM) within litter bags and to investigate its possible impact on macroinvertebrate density and richness. In seven headwater forested streams we exposed mesh bags filled either with beech leaves ( Fagus sylvatica ) or with plastic strips of an equal surface area.
3. Principal component analysis (PCA) showed that bag type and stream were the main explanatory variables for invertebrate colonization and POM accumulation within the bags. In contrast, there was little variation among sampling dates (6.4% of the total inertia).
4. The accumulated POM within the bags was substantial (up to 8.83 g ash-free dry mass (AFDM)) but highly variable among sites (mean from 0.32 to 4.58 g AFDM). At each of the seven sites, both richness and abundance of invertebrates were positively correlated with the mass of accumulated POM in bags. Macroinvertebrate colonization (notably taxon richness) was directly linked with the quantity of POM accumulated.
5. Our findings provide evidence of a potential pitfall in linking invertebrates to litter processing in mesh bags, particularly when large amounts of POM, entrained in stream flow, accumulate within the bags. An evaluation of the POM mass trapped in litter bags could account for the erratic patterns sometimes observed in their colonization by invertebrates.     

不同颜色果袋对‘巨峰’葡萄果实中挥发性成分的影响

以‘巨峰’葡萄为试材,研究白袋、绿袋、红袋、蓝袋4种不同颜色果袋对葡萄成熟期果实中挥发性成分的影响,为葡萄专用果袋的研发提供理论依据.结果表明:不同颜色果袋可为葡萄果实的发育提供特定的光环境,不同套袋处理葡萄成熟期果实中的挥发性成分差异显著.‘巨峰’葡萄成熟期果实中检测到酯、醛、醇、酮类、萜烯类和芳香族化合物,对照、白袋、绿袋、红袋和蓝袋处理成熟期果实检测到的挥发性组分分别为33、37、38、32和34种.与对照相比,白袋处理乙酸乙酯、己酸乙酯等酯类物质含量降低,己醛、反式-2-己烯醛、癸醛含量增加;绿袋处理除3-己烯酸乙酯、反式-2-己烯酸乙酯、3-羟基丁酸乙酯、反式-2-己烯醛、(反,反)-2,4-己二烯醛、癸醛、苯乙醇外,其他共有组分的含量均出现降低;红袋处理乙酸乙酯、己酸乙酯等酯类物质含量降低,己醛、反式-2-己烯醛等的含量出现降低;蓝袋处理乙酸乙酯、己酸乙酯等主要酯类物质的含量未发生较大变化,己醛、反式-2-己烯醛等的含量出现升高.在非共有组分中,套袋处理中醇类组分的种类减少,萜烯类、芳香族化合物的种类增加.总体上,蓝袋处理果实中主要的‘果香型’酯类组分含量最高,白袋处理果实中主要的酯类组分及‘青草香型’的醛类组分含量较高,绿袋、红袋处理果实主要香气组分含量较低.     

Ectomycorrhizal hyphae structure components of the soil bacterial community for decreased phosphatase production

Ectomycorrhizal fungi (EMF) provide nutrients to their hosts by means of hyphae that extend beyond nutrient-depleted rhizosphere soil. Soil bacteria may compete with EMF for nutrients or may act synergistically to enhance nutrient supply to hosts. To assess the interactions between hyphae and bacteria, two types of small, sand-filled mesh bags were incubated in a Pseudotsuga menziesii/Betula papyrifera forest. The bags allowed ingrowth by EMF (35-μm mesh) or excluded hyphae (0.5-μm mesh), while allowing migration of soil bacteria. After incubation, bacteria were isolated from bags using a method to enrich for Gram-positive bacteria. Isolates were assayed for phosphatase and N-acetyl glucosaminidase (NAGase) activities to assess the potential to access organic phosphorus and nitrogen. The average phosphatase activities were higher in exclusion than ingrowth bags, while NAGase activities did not differ. Streptomyces isolates, which are expected to be strong competitors and antagonists of EMF, were more prevalent in ingrowth bags and yet had lower phosphatase activities. Furthermore, there were no indications of antagonism between fungi and Streptomyces, as there were no increases in NAGase activities in ingrowth bags. We conclude that fungal hyphae can structure components of the soil bacterial community for decreased extracellular enzyme production.