Development of an immunochromatographic strip test for rapid detection of melamine in raw milk, milk products and animal feed

A simple, rapid and sensitive immunogold chromatographic strip test based on a monoclonal antibody was developed for the detection of melamine (MEL) residues in raw milk, milk products and animal feed. The limit of detection was estimated to be 0.05 μg/mL in raw milk, since the detection test line on the strip test completely disappeared at this concentration. The limit of detection was 2 μg/mL (or 2 μg/g) for milk drinks, yogurt, condensed milk, cheese, and animal feed and 1 μg/g for milk powder. Sample pretreatment was simple and rapid, and the results can be obtained within 3-10 min. A parallel analysis of MEL in 52 blind raw milk samples conducted by gas chromatography-mass spectrometry showed comparable results to those obtained from the strip test. The results demonstrate that the developed method is suitable for the onsite determination of MEL residues in a large number of samples.     

Resistance to Marssonina rosae in Rosa L. Seedlings Obtained from Controlled Crosses Including Germplasm L83

Blackspot is a serious disease on cultivated roses and resistance is rare; however, some wild Rosa species exhibit a high level of resistance. Disease incidence in field-grown offspring from three seedling families was evaluated as percentage of infected leaves (regardless of the size and number of spots) on each plant. Dolly, Queen Elizabeth and Friesia were used as pistillate parents and the resistant germplasm L83 (derived from the amphidiploid R     

Can field populations of the enchytraeid,Cognettia sphagnetorum,adapt to increased drought stress?

The ability to evolve increased drought tolerance in response to climate change was investigated in the enchytraeid, Cognettia sphagnetorum. Populations exposed to reduced precipitation or increased night time temperature for more than six years were collected in mixed Calluna/grass heathland at the Mols Laboratory, Denmark. The level of prolonged drought and increased temperature corresponded to a predicted climate change scenario and has been applied since 1999. In autumn 2005, enchytraeids were sampled in 3 cm intervals down to 9 cm depth and total number, biomass, diversity and soil organic matter were determined. The drought treatment resulted in a significant reduction of the density and biomass of enchytraeids, as well as changes in the species composition. In total, five different genera were found at the site in all three treatments (control, temperature and drought). C. sphagnetorum was the dominant species, especially in the upper 0–3 cm, and was clearly affected by the drought treatment. C. sphagnetorum from all plots were cultured in the laboratory to rear second or third generation adults. Results showed that populations of drought treated plots had not developed an increased drought resistance compared to populations of control or warming plots even after several years of a putative severe selection. Lack of adaptive potential in C. sphagnetorum suggests that more frequent periods with drought in the future will have a very strong negative influence on enchytraeid density, biomass and diversity.     

Identification of different BLV provirus isolates by PCR, RFLPA and DNA sequencing

A first attempt for the investigation of molecular epidemiology of BLV was carried out. PCR amplicons of a part of the env gene of BLV isolated from 309 cattle of different geographical origin were compared with known BLV env sequences. Using RFLPA most of the PCR products can be assigned to the Australian, the Japanese or the Belgian subgroup. A phylogenetic tree resulting from the comparison of the sequences of these env fragments demonstrates the relations and differences between and within the subgroups.     

Assessment of urban forestry research and research needs in Nordic and Baltic countries

A review of research and research needs in urban forestry was carried out in Denmark, Estonia, Finland, Latvia, Norway and Sweden during 2005. A questionnaire addressing post-2000 and ongoing research was sent to 146 researchers and generated 76 completed questionnaires. Universities were found to lead urban forestry research, while municipalities headed funding organisations in terms of number of projects funded. Planning, ecological and management aspects were the most common research themes, but socially oriented research also played an important role. The research needs questionnaire was sent to 192 key research actors (assignors, users and researchers), resulting in 63 completed needs assessments. The research themes of ‘urban forest management’, ‘social and cultural values’ and ‘urban forest and green planning’ were prioritised for future research. Comparison of ongoing research and research needs showed discrepancies, as ongoing research does not always cover the same themes identified as primary research needs. Priorities for future research as identified by the research community respective those assigning and using research also differed. Economic assessment of benefits, for example, scored much higher as a need among researchers than other respondents. In terms of present weaknesses in the research ‘infrastructure’, research actors emphasised lack of funding, fragmentation of research and insufficient critical mass. The region's urban forestry research can be enhanced and made more meaningful by strengthening national and international networking within the research community, across disciplines, as well as between researchers and those commissioning and using research.     

Development of a monoclonal antibody-based cELISA for the analysis of sulfadimethoxine. 1. Development and characterization of monoclonal antibodies and molecular modeling studies of antibody recognition

Sulfonamide antibiotics are used to treat a variety of bacterial and protozoan infections in cattle, swine, and poultry. Current residue methods for the analysis of sulfonamides in animal-based food products include bioassays, chromatographic methods (HPLC, GLC), and immunoassays. Most immunoassays have employed highly specific polyclonal antibodies. In this paper, we describe the isolation of monoclonal antibodies against sulfadimethoxine (SDM) that vary in their sensitivities and cross-reactivities against a large number of sulfonamides. The most sensitive monoclonal antibody, designated SDM-18, exhibits an IC(50) value for SDM of 1.53 ppb. Another monoclonal antibody, designated SDM-44, exhibits IC(50) values for six sulfonamides well below the established threshold level of 100 ppb for animal tissues. Molecular modeling studies of the cross-reactive drugs suggest that, depending on the monoclonal antibody, both steric and electronic features govern antibody binding. Due to the diversity of these monoclonal antibodies, it should be possible to design both compound- and class-specific monoclonal antibody-based immunoassays.     

Effect of sampling density on regional soil organic carbon estimation for cultivated soils

An extensive knowledge of how sampling density affects soil organic C (SOC) estimation at regional scale is imperative to reduce uncertainty to a meaningful confidence level and aid in the development of sampling schemes that are both rational and economical. Using kriging prediction, this paper examined the effect of sampling density on regional SOC‐concentration estimations in cultivated topsoils at six scales in a 990 km² area of Yucheng County, a typical region in the N China Plain. Except the original data set (n = 394), five other sampling densities were recalculated using grids of 8 km × 8 km (n = 28), 8 km × 4 km (n = 44), 4 km × 4 km (n = 82), 4 km × 2 km (n = 142), and 2 km × 2 km (n = 257), respectively. Experimental SOC semivariances and kriging interpolations at six sampling density scales were calculated and modeled to estimate regional SOC variability. Accuracy of the effects of the five sampling densities on regional SOC estimations was assessed using the indices of mean error (ME) and root mean square error (RMSE) with 100 independent validation samples. By comparison with the kriged grid map derived from the 394 samples data set, the relative error (RE,%) was spatially calculated to highlight the spatial variability of prediction errors at five sampling‐density scales due to the intrinsic limitations of ME and RMSE in accuracy assessment. The results indicated that sampling density significantly affected the estimation of regional SOC concentration. Particularly when the sampling density was < 4 km × 4 km, the large spatial variation of SOC was concealed. Semivariance analysis indicated that different sampling density had significant effect on reasonable detection of the dominant factors which influenced SOC spatial variation. Greater sampling density could more exactly reveal regional SOC variation caused by human management. The prediction accuracy for regional SOC estimation increased with the increasing of sampling density. The critical areas with larger RE values should be intensified in the future sampling scheme, and the areas of lower RE values should be decreased relatively. A specific sampling scheme should be considered in accordance with the demand to the estimation accuracy of regional SOC stock at a certain confidence level. Our results will facilitate a better understanding of the effect of sampling density on regional SOC estimation for future sampling schemes by providing meaningful confidence levels.     

Production and characterization of monoclonal antibodies against the antibiotic tilmicosin

Monoclonal antibodies (Mabs) were developed that specifically bind tilmicosin. Keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) conjugates were used for the immunogen and plate coating antigen, respectively. The conjugates were synthesized by different methods, resulting in different linkages. Six hybridoma cell lines were isolated that produced Mabs that competed with tilmicosin, and have IgG1 isotype. The Til-1 and Til-5 Mabs had IC50 values for tilmicosin of 9.6 and 6.4 ng/well (48 and 32 ng/mL), respectively, and limits of detection at IC20 of 1.84 and 0.89 ng/well (9.2 and 4.45 ng/mL), respectively. The Mabs demonstrated high cross-reactivity to the macrolides containing 3,5-dimethylpiperidine at C20 and the amino sugar at C5. No cross-reactivity was observed for tylosin and other macrolides that did not contain 3,5-dimethylpiperidine. A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the antibiotic tilmicosin by use of the developed Mabs. These Mabs may be excellent candidates for the determination and immunolocalization of tilmicosin.