Cross-linking and rheological changes of whey proteins treated with microbial transglutaminase

Modification of the functionality of whey proteins using microbial transglutaminase (TGase) has been the subject of recent studies. However, changes in rheological properties of whey proteins as affected by extensive cross-linking with TGase are not well studied. The factors affecting cross-linking of whey protein isolate (WPI) using both soluble and immobilized TGase were examined, and the rheological properties of the modified proteins were characterized. The enzyme was immobilized on aminopropyl glass beads (CPG-3000) by selective adsorption of the biotinylated enzyme on avidin that had been previously immobilized. WPI (4 and 8% w/w) in deionized water, pH 7.5, containing 10 mM dithiothreitol was cross-linked using enzyme/substrate ratios of 0.12-10 units of activity/g WPI. The reaction was carried out in a jacketed bioreactor for 8 h at 40 degrees C with continuous circulation. The gel point temperature of WPI solutions treated with 0.12 unit of immobilized TGase/g was slightly decreased, but the gel strength was unaffected. However, increasing the enzyme/substrate ratio resulted in extensive cross-linking of WPI that was manifested by increases in apparent viscosity and changes in the gelation properties. For example, using 10 units of soluble TGase/g resulted in extensive cross-linking of alpha-lactalbumin and beta-lactoglobulin in WPI, as evidenced by SDS-PAGE and Western blotting results. Interestingly, the gelling point of WPI solutions increased from 68 to 94 degrees C after a 4-h reaction, and the gel strength was drastically decreased (lower storage modulus, G'). Thus, extensive intra- and interchain cross-linking probably caused formation of polymers that were too large for effective network development. These results suggest that a process could be developed to produce heat-stable whey proteins for various food applications.     

Foliar and soil deposition of pesticide sprays in peanuts and their washoff and runoff under simulated worst-case rainfall conditions

There are few studies that relate the timing and amounts of pesticide washoff from plant foliage during rainfall to runoff losses at the edge of the field. We hypothesized that foliar deposits, if washed onto the soil slowly during rainfall, may then undergo less leaching during the period of infiltration that occurs prior to soil saturation and runoff, thus exhibiting larger runoff losses than pesticides on/in the soil at the beginning of rain. We measured the runoff of ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT dye using simulated rainfall on 450 m2 mesoplots planted in peanut. Ethalfluralin was applied preplant incorporated, and metolachlor was applied preemergence on bare soil. Chlorothalonil and rhodamine WT were applied to the peanut canopy at maturity. Rainfall was simulated 24 h after each chemical application (in May and July, 1998, and May and August, 1999) using raindrop sprinklers, applying 5.5 +/- 0.5 cm over a 2 h period to create reasonable worst-case conditions; between 3 and 9 mm of runoff was generated. Volume-weighted average concentrations of chemicals in runoff were 7, 104, 163, and 179 ug L(-1) for ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT, respectively. The total amounts of chemicals lost in the runoff events were 0.04 +/- 0.01, 0.2 +/- 0.1, 0.6 +/- 0.5, and 0.2 +/- 0.1, as percents of amounts applied, respectively. Rhodamine WT formed a vivid red solution on wetting and provided visual clues to the dynamics of chemical washoff/runoff. The washoff from rain-exposed peanut foliage appeared to be complete within a few minutes of the beginning of rainfall, and disappearance of dye from rain-exposed soil surface occurred within the first 10 min of rainfall. However, dye was present in runoff water at near-constant concentrations throughout the 2 h runoff event, indicating that near-constant amounts of chemical remained in the soil extraction zone. These results confirm earlier studies showing that soil incorporation at application significantly reduces runoff losses and that a majority of foliar residues can be washable if rainfall occurs within a few days after application. Runoff losses of foliar-applied pesticides were small relative to washoff amounts but were sensitive to runoff timing relative to washoff.     

Antioxidant activity of betalains from plants of the amaranthaceae

Antioxidant activity of betalain pigments (seven pure compounds and four combined fractions) from plants of the family Amaranthaceae was evaluated using the modified DPPH(*) (1,1-diphenyl-2-picrylhydrazyl) method. All tested betalains exhibited strong antioxidant activity. Their EC(50) values ranged from 3.4 to 8.4 microM. Gomphrenin type betacyanins (mean = 3.7 microM) and betaxanthins (mean = 4.2 microM) demonstrated the strongest antioxidant activity, 3-4-fold stronger than ascorbic acid (13.9 microM) and also stronger than rutin (6.1 microM) and catechin (7.2 microM). Antioxidant activity of the tested betalains decreased in the following order: simple gomphrenins > acylated gomphrenins > dopamine-betaxanthin > (S)-tryptophan-betaxanthin > 3-methoxytyramine-betaxanthin > betanin/isobetanin > celosianins > iresinins > amaranthine/isoamaranthine. This study also investigated and discussed the relationship between the chemical structure and the activity of the betalains. The free radical scavenging activity of the betalains usually increased with the numbers of hydroxyl/imino groups and, moreover, depended on the position of hydroxyl groups and glycosylation of aglycones in the betalain molecules.     

Soil microbial community characteristics along an elevation gradient in the Laguna Mountains of Southern California

We sampled soil at four sites in the Laguna Mountains in the western Sonoran Desert to test the effects of site and sample location (between or beneath plants) on fatty acid methyl ester (FAME) and carbon substrate ulilization (Biolog) profiles. The four sites differed in elevation, soil type, plant community composition, and plant percent cover. Soil pH decreased and plant density increased with elevation. Fertile islands, defined as areas beneath plants with greater soil resources than bare areas, are present at all sites, but are most pronounced at lower elevations. Consistent with this pattern, fertile islands had the greatest influence on FAME and Biolog profiles at lower elevations. Based on the use of FAME biomarker and principal components analyses, we found that soil microbial communities between plants at the lowest elevation had proportionally more Gram-negative bacteria than all other soils. At the higher elevation sites there were few differences in FAME profiles of soils sampled between vs. beneath plants. Differences in FAME profiles under plants among the four sites were small, suggesting that the plant influence per se is more important than plant type in controlling FAME profiles. Since microbial biomass carbon was correlated with FAME number (r=0.85,P<0.0001) and with FAME named (r=0.88,P<0.0001) and total areas (r=0.84,P<0.0001), we standardized the FAME data to ensure that differences in FAME profiles among samples were not the result of differences in microbial biomass. Differences in microbial substrate utilization profiles among sampling locations were greatest between samples taken under vs. between plants at the two lower elevation sites. Microbial substrate utilization profiles, therefore, also seem to be influenced more by the presence of plants than by specific plant type.     

Effects of net height of crab entangling nets on the capture of targeted economically important portunid species and non-target species

The aim of this study was to determine the effects of net height on the capture performance of crab entangling nets. Fishing trials were conducted using nets at varying net heights (1) 12 meshes down (MD), (2) 24 MD and (3) 50 MD. A total of 1290 individuals comprising 87 species belonging to 53 families were caught. One-way analysis of variance showed that net height significantly affected the various catch parameters, including catch per unit effort (CPUE) of the total and target catch, amount of non-target catch, size of catch and species richness. The use of appropriate net height is a potential technical measure for a selective but still efficient crab entangling net fishery. Lower net height significantly reduced non-target catch by up to 70%. Lower net height also decreased the CPUE of target catch such as Portunus pelagicus and Charybdis feriatus by up to 65% at 12 MD, but catch at 24 MD was not significantly different than that at 50 MD. The use of a net height of 24 MD also resulted in the capture of larger-sized P. pelagicus. The richness of the catch species decreased by up to 58% in lower nets. These results are useful to fishery managers and government institutions when developing and/or improving existing regulations towards a sustainable crab fishery, particularly blue swimming crabs.     

Seeding and propagation of untransformed mouse mammary cells in the lung

The acquisition of metastatic ability by tumor cells is considered a late event in the evolution of malignant tumors. We report that untransformed mouse mammary cells that have been engineered to express the inducible oncogenic transgenes MYC and Kras(D12), or polyoma middle T, and introduced into the systemic circulation of a mouse can bypass transformation at the primary site and develop into metastatic pulmonary lesions upon immediate or delayed oncogene induction. Therefore, previously untransformed mammary cells may establish residence in the lung once they have entered the bloodstream and may assume malignant growth upon oncogene activation. Mammary cells lacking oncogenic transgenes displayed a similar capacity for long-term residence in the lungs but did not form ectopic tumors.     

Detection of Campylobacter jejuni strains in the water lines of a commercial broiler house and their relationship to the strains that colonized the chickens

Campylobacter jejuni is frequently present in the intestinal tract of commercial broiler chickens, and their drinking water has been proposed to be an initial source of bacteria for newly hatched chicks. We studied three sequential commercial broiler flocks raised in a house from which we had cultured C. jejuni from the nipple waters prior to placement of the first flock. Campylobacter cells were detected by immunofluorescence in the biofilm of the drinking nipples during the weeks when the flock was colonized with C. jejuni but not during weeks when the birds were negative. Campylobacter jejuni was isolated from the drinking water during the growth of the first flock and was present in the birds from all three flocks. Randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing with primer OPA11 indicated that seven distinct strains were present within the broiler house. One strain found in drinking water was similar to a strain found in birds in the second flock; however, RAPD-PCR with primer HLW85 showed that the strains were not identical. These results suggest that although the watering system is a potential source of C. jejuni in broiler flocks, the waterborne strain in this study was not detected in the birds.     

Antisporulant activity of 8-hydroxyquinoline and its 5- and 6-n-alkyl derivatives against Gloeosporium fructigenum berk

The antisporulant activities of 8-hydroxyquinoline (oxine) and members of the homologous 5- and 6-n-alkyloxine series against Gloeosporium fructigenum at varying ambient pH values and in the presence or absence of added Cu²⁺, Zn²⁺ and Fe²⁺ are described. Whilst changes in the pH value only influenced the antisporulant activity of oxine at low concentrations, lowering the ambient pH increased considerably the activities of the 5- and 6-alkyl isomers at all concentrations. Below pH 7.0 the addition of these ions to the oxines resulted in decreased antisporulant activity (Fe²⁺ > Zn²⁺ > Cu²⁺) whereas, at pH 8.0, the addition of Cu²⁺ increased the activity of some oxines. It is suggested that at low pH (3.2) the oxine cation is an important toxicant.