Determination of ractopamine in cattle and sheep urine samples using an optical biosensor analysis: comparative study with HPLC and ELISA

A biosensor method, using the surface plasmon resonance (SPR) principle, was developed for the determination of ractopamine in cattle and sheep urine. A monoclonal antibody was used to compete with ractopamine in the sample and ractopamine immobilized on the sensor chip. Addition of bovine serum albumin (BSA, 1 mg/mL) as an antibody stabilizer to the incubation buffer was required to achieve a stable biosensor response throughout each sample set. The calibration curve gave a mean IC(50) of 4.7 +/- 0.21 ng/mL (n = 7). Over sample concentrations from 2.5 to 10 ng/mL recoveries were typically approximately 100-110%, whereas inter- and intra-assay reproducibilities (% CV) were usually less than 10 and 6%, respectively. Comparison of biosensor results with results obtained from high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assays (ELISA) using enzyme-hydrolyzed urine (to convert ractopamine conjugates to free ractopamine) gave correlation coefficients of 0.94 for sheep and 0.86 for cattle. Slopes of the lines, with zero intercepts, equaled 0.80 for sheep and 0.74 for cattle. For untreated (nonhydrolyzed) urine samples, the correlations between biosensor and HPLC results were 0.95 for sheep and 0.72 for cattle with slopes of 1.18 (sheep) and 1.69 (cattle). The slopes greater than unity indicate that the biosensor responded to ractopamine metabolites in addition to free ractopamine. The biosensor assay is an excellent analytical tool to screen ractopamine residues in sheep or cattle urine, and the results should be extendible to other species with suitable validation.     

Direct analysis of pesticide residues in olive oil by on-line reversed phase liquid chromatography-gas chromatography using an automated through oven transfer adsorption desorption (TOTAD) interface

A fully automated on-line reversed phase liquid chromatography-gas chromatography system is described. The system uses a prototype of the automated through oven transfer adsorption desorption interface. The system is demonstrated by presenting a new rapid method for the determination of pesticide residue in olive oil, which is injected directly with no sample pretreatment step other than filtration. Methanol:water is used as the eluent in the LC preseparation step, while the LC fraction containing the pesticide is automatically transferred to the gas chromatograph. Detection limits of pesticides varied from 0.18 to 0.44 mg/L when a flame ionization detector was used. As an example, relative standard deviation and linear calibration are presented for terbutryne.     

Evidence from tag recapture experiments that fish learn to avoid fishing gear

We found evidence that tagging induced trap shyness in snapper (Pagrus auratus), i.e., tagged fish had a reduced probability of recapture by the method by which they had originally been caught. Tagging experiments in 1985 and 1994 involving single release over a short period and single recapture were conducted on a closed population (SNA 1: East Northland-Hauraki Gulf-Bay of Plenty). Initial capture was by trawling and by line fishing, while recapture over an extended period included other methods. A test for trap shyness that removed the possible effects of spatial and fish size heterogeneity gave a significant result for both years. The data suggested that the trap shyness effect might have been smaller for trawl-released fish than for line-released fish. However, we estimated a single trap shyness factor (0.71). There was also some evidence for attenuation of trap shyness over time.     

Comparison of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States

A number of different beverage products claim to have antioxidant potency due to their perceived high content of polyphenols. Basic and applied research indicates that pomegranate juice (PJ), produced from the Wonderful variety of Punica granatum fruits, has strong antioxidant activity and related health benefits. Although consumers are familiar with the concept of free radicals and antioxidants, they are often misled by claims of superior antioxidant activity of different beverages, which are usually based only on testing of a limited spectrum of antioxidant activities. There is no available direct comparison of PJ's antioxidant activity to those of other widely available polyphenol-rich beverage products using a comprehensive variety of antioxidant tests. The present study applied (1) four tests of antioxidant potency [Trolox equivalent antioxidant capacity (TEAC), total oxygen radical absorbance capacity (ORAC), free radical scavenging capacity by 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP)]; (2) a test of antioxidant functionality, that is, inhibition of low-density lipoprotein (LDL) oxidation by peroxides and malondialdehyde methods; and (3) evaluation of the total polyphenol content [by gallic acid equivalents (GAEs)] of polyphenol-rich beverages in the marketplace. The beverages included several different brands as follows: apple juice (3), a?aí juice (3), black cherry juice (3), blueberry juice (3), cranberry juice (3), Concord grape juice (3), orange juice (3), red wines (3), iced tea beverages (10) [black tea (3), green tea (4), white tea (3)], and a major PJ available in the U.S. market. An overall antioxidant potency composite index was calculated by assigning each test equal weight. PJ had the greatest antioxidant potency composite index among the beverages tested and was at least 20% greater than any of the other beverages tested. Antioxidant potency, ability to inhibit LDL oxidation, and total polyphenol content were consistent in classifying the antioxidant capacity of the polyphenol-rich beverages in the following order: PJ>red wine>Concord grape juice>blueberry juice>black cherry juice, a?aí juice, cranberry juice>orange juice, iced tea beverages, apple juice. Although in vitro antioxidant potency does not prove in vivo biological activity, there is also consistent clinical evidence of antioxidant potency for the most potent beverages including both PJ and red wine.     

Competition for shifting resources in the southern Benguela upwelling: Seabirds versus purse-seine fisheries

In the southern Benguela upwelling ecosystem off the west coast of South Africa, seabird populations are decreasing dramatically because of reduced availability of pelagic fish. We tested the hypothesis that the west coast fishing industry is competing for the remaining stocks of anchovy and sardine with the largest colony of Vulnerable Cape Gannets (Morus capensis) along the Atlantic coast. Using GPS-tracking of the birds, echo-sounding of pelagic fish, and vessel log books, we located overlap areas between bird foraging ranges, pelagic fish distribution, and fishing activities. We then compared fish catches by gannets and vessels within their joint foraging zones. In October 2007, purse-seine fishing grounds and gannet foraging areas overlapped by only 13%. However, for a 1-month period, the amount of fish removed from this area by purse-seine boats amounted to 41% of the food requirements of the 72,000 gannets breeding on Malgas Island (25% of the world population). The fishery’s catch in this area is significant in terms of its potential impact on gannets, but comprises only 3.6% of total fishery catch. Based on this finding, the rapidly decreasing size of the gannet colony and the stated objectives of South Africa’s Marine Living Resources Act of 1998, the case for considering and experimenting with at-sea ‘no-take’ areas for the purse-seine fishery is strong. Efforts to establish whether ‘no-take’ fishing zones increase food availability for top predators is an important next step in conservation of the southern Benguela Ecosystem.     

Can placement of seed away from relic stubble limit Rhizoctonia root rot in direct-seeded wheat?

Rhizoctonia root rot of wheat can be a problem in no-till systems, especially during the transition from conventional tillage. There are no effective chemical controls or resistant varieties, leaving only cultural methods to manage this disease. In a no-till system, residue and inoculum of soilborne pathogens are not moved by cultivation, therefore the inoculum may be concentrated in the seeding row of the previous year. Using GPS tracking systems with sub-meter accuracy, the seeding row could be placed away from the row of the previous year. We tested the hypothesis that seeding away from the relic row may reduce Rhizoctonia root rot. In two field experiments, plants were sampled at three distances from the seed row, as well as from fumigated plots. Intact soil cores were also removed from the field, planted with seeds at various distances from the previous row, and grown in the greenhouse under controlled conditions. Pasteurized cores served as controls. Disease levels were higher in the field in the second year, but there was no consistent effect of seed row placement on disease or plant parameters. However, soil fumigation and pasteurization had significant effects, indicating that soilborne pathogens were active. Inoculum of Rhizoctonia is not produced in the crowns and lower stems of the plant, but the pathogen survives in living and dead roots of the previous year crop, volunteers, and grassy weeds. Thus, high inoculum densities may be present in between the relic rows, as well as within the rows. If this is the situation with Rhizoctonia, precision placement of seed rows would not be efficacious.     

Hemoxygenase-2 is an oxygen sensor for a calcium-sensitive potassium channel

Modulation of calcium-sensitive potassium (BK) channels by oxygen is important in several mammalian tissues, and in the carotid body it is crucial to respiratory control. However, the identity of the oxygen sensor remains unknown. We demonstrate that hemoxygenase-2 (HO-2) is part of the BK channel complex and enhances channel activity in normoxia. Knockdown of HO-2 expression reduced channel activity, and carbon monoxide, a product of HO-2 activity, rescued this loss of function. Inhibition of BK channels by hypoxia was dependent on HO-2 expression and was augmented by HO-2 stimulation. Furthermore, carotid body cells demonstrated HO-2-dependent hypoxic BK channel inhibition, which indicates that HO-2 is an oxygen sensor that controls channel activity during oxygen deprivation.     

Effects of N, NP, and NPKS fertilizers applied to four-year old hybrid poplar plantations

Mineral fertilizers were applied to adjacent plantations of 2, 4-year old, hybrid poplars: clone 27 (Northwest, P. deltoides × P. balsamifera) and 794 (Brooks, P. deltoides × P. × petrowskyana), in north eastern Alberta. Fertilization was done in May 2003 to see whether growth rates could be increased and rotations shortened. Three fertilizers (N, NP and NPKS + Cu + Zn) were applied at each of three rates (supplying N at 0, 100 and 200 kg ha⁻¹) in a factorial randomized block design to the two separate plantations. Fertilization with 200 kg ha⁻¹ N and 100 kg ha⁻¹ P increased volume growth of clone 27 by 1 m³ ha⁻¹ year⁻¹ over 2 years. Clone 794 showed no volume response to fertilization, but produced 8.7 m³ ha⁻¹ more than clone 27. Leaf area, dry mass and nutrient concentrations of both clones increased in the first year after fertilization, showing that fertilizer uptake occurred. Decrease in leaf size between 2003 and 2004 was affected by fertilizer level in clone 794. There were differences between some nutrient concentrations in the soils occupied by the two clones, and clone 794 had higher leaf concentrations of N, K, Ca, S, Mn, Zn, B and Mo than clone 27. Fertilization of 4-year old plantations of either clone was unwarranted, and planting clone 794 would be likely to provide greater yield than planting and fertilizing clone 27.