Development of an indirect competitive ELISA for flumequine residues in raw milk using chicken egg yolk antibodies

To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA-flumequine) and to cationized ovalbumin (cOVA-flumequine). For the immunization of chickens, cBSA-flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA-flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 microg/kg). No cross-reactivity was noticed with various related quinolones.     

Does nectar reward affect rarity and extinction probabilities of orchid species? An assessment using historical records from Belgium and the Netherlands

The long-term persistence of plant populations may partly depend on pollination processes and seed production. The exact role of pollination in determining plant population viability still remains largely unclear. Orchid species have often been shown to be pollinator limited. Especially, non-rewarding species are characterized by infrequent pollinator visits and low seed set. Therefore, it can be hypothesized that, if overall population fitness of orchid species is influenced by seed production, non-rewarding species should be more prone to local extinction than rewarding species. To study the importance of nectar reward on orchid persistence, we collected historical records about the distribution of 32 orchid species in Flanders (Belgium) and 37 orchid species in the Netherlands. For both regions, present distribution patterns were compared with historical data. Between 1930 (Flanders) or 1950 (the Netherlands) and 2000, 26 (81%) species showed decreases in distribution range in Flanders, eight of which had gone extinct, whereas 29 (78%) species showed declines in distribution area in the Netherlands, five of which went extinct. Contrary to previously reported results, orchid distribution patterns were not related to nectar reward. There was also no significant relationship between nectar reward and extinction for both regions. Orchids typically occurring in wet grasslands and heathlands suffered greater losses than orchids typically confined to forest habitats or calcareous grasslands. We conclude that the production of nectar does not represent a safeguard for local extinction of orchid species. Habitat loss and other deterministic threats associated with habitat fragmentation and deterioration are more important determinants of orchid persistence.     

Sediments in the Catchment-coast Continuum (7 pp)

-  Dedicated to Prof. Dr. Ulrich Förstner on his 65th birthdayBackground   Sediments pose problems at their deposition sites when there is too little sediment (e.g. wetlands) or too much sediment (e.g. navigable waterways) and, additionally, when they are contaminated. These problems often have their origin upstream in the river catchment. Objective   Global aspects of changes concerning sediment quantity and quality, as they affect downstream areas, are reviewed. A case study of estuarine sediments demonstrates how a holistic approach helps in understanding and predicting their present and future quality. Results and Conclusions   Globally, large reservoirs intercept between 25 and 30% of the sediment and consequently supply the coast with impacts on wetlands and coastal morphology. In estuaries, the composition of sediments is determined by the mixing of marine sediments supplied from the coast and the supply by rivers. Natural tracers can be used to predict mixing ratios of marine to fluvial sediments in estuaries, and hence their contamination. Scenarios on implementation of pollution abatement, the implementation of regulations as well as climate change are needed to predict future sediment quality in downstream areas. The results show, even for a ‘green’ scenario, that sediment quality in the Rhine catchment will pose future problems due to the temporal storage of contaminants in soils and sediments.Recommendations and Outlook   The current methods applied make use of existing models linking sediment transport with point and diffuse sources in the river catchments and scenarios on the development of strength of point and diffuse sources. However, more effort is needed to come to a uniform framework which includes land use changes and links with more advanced scenario methodology for long to medium-term management of sediment quality and quantity in river catchments.     

Comparison of measured and simulated changes in base cation amounts using a one-layer and a multi-layer soil acidification model

Biogeochemical processes describing nutrient cycling strongly affect the development of different soil horizons and soil acidification. However, in many soil acidification models these processes are not directly simulated, and spatially distributed soil properties are lumped into average values. This paper describes the extension of a one-layer soil acidification model (SMART) to a multi-layer model (SMARTEX) by including nutrient cycling, and the performance of both models regarding their ability for the assessment of the effects of acid deposition on forest soil. This was done by comparing simulation results of the two models with respect to changes in base cation amounts to historical data from sites in south Sweden. The results showed that the multi-layer model gives better results for the top soil, whereas the one-layer model gives better results for the rootzone and the total profile. Since the top soil is most important, the extended model is more suited to relate acid deposition to forest growth than the more general results of the one-layer model. However, the sensitivity of the multi-layer model to some parameters is large. A change in these parameters had strong effects on the separate layers, whereas the effects were cancelled out over the total profile. This sensitivity requires a careful parameterization of the model which might be troublesome to lack of data.     

Superficial keratectomy and cryosurgery as therapy for limbal neoplasms in 13 horses

Objective To determine the usefulness and clinical outcome of a combined procedure of superficial keratectomy and cryosurgery as a treatment for limbal neoplasms in horses. Study design Retrospective study. Animals Thirteen horses with 14 limbal tumors. Methods Medical records of all patients with limbal tumors, referred to the Department of Equine Sciences of Utrecht University between 1995 and 2002, were retrieved. Patient data were analyzed with respect to signalment, tumor surface area and histologic diagnosis. Surgery, performed under general anesthesia, included surgical debulking of the tumor followed by cryosurgery. Long‐term follow‐up information was obtained by re‐evaluation in the clinic, by the referring veterinarian, and/or by telephone enquiry. Results Therapy was successful in 9 out of 14 eyes after the first attempt (64%); one eye needed retreatment (7%), and four eyes were eventually enucleated (29%). The mean follow‐up period was 4.8 years. The initial surface area of the tumor significantly influenced outcome (P < 0.01). Squamous cell carcinoma was the most predominant tumor type (79%). Haflinger horses accounted for 69% of the cases whereas their occurrence in the overall hospital population is approximately 5%. Conclusions and clinical relevance The described technique of superficial keratectomy and cryosurgery is a simple procedure for the treatment of limbal tumors in equine patients that does not require sophisticated equipment. Nor is it tampered by legal restriction, and appears to be effective in tumors with a small surface area (< 2 cm²). Haflinger horses seem to be predisposed for the development of ocular squamous cell carcinoma.     

Comparative genetic characterization of Porcine Circovirus type 2 samples from German wild boar populations

PCV-2 is involved in "postweaning multisystemic wasting syndrome" (PMWS), "porcine dermatitis and nephropathy syndrome" (PDNS), respiratory and reproductive disorders, and thereby plays a crucial role in today's swine production world-wide. The virus is apparently ubiquitous in domestic pigs and has also been demonstrated in wild pigs. Up to now, a characterization of PCV-2 samples from wild pigs, which might help to estimate the possible role of wild pigs as sources of domestic pig infection, has not been carried out. Spleen samples from 16 PCV-2-positive wild pigs from hunting grounds of four regions in Germany were used for the analysis of the viral genome. In one sample, the complete sequence of the genome was determined. In the other, a 742 nucleotide fragment from the highly variable capsid sequence of the ORF2 was sequenced. Analysis of the sequences led to the identification of three PCV-2 strains. One strain, representing 14 of the 16 samples, was closely related with Chinese, but not with German strains. The genome of this strain was shortened by one nucleotide by a deletion close to the end of ORF2. The deletion led to a shift of the stop-codon and to the insertion of a further codon. Two further strains differed in up to 4.7% of nucleotides and up to 10.5% of amino acids (aa). These strains were aligned with clusters of PCV-2 samples from mainly French and German domestic origin.     

Screening of common bean (Phaseolus vulgaris) for resistance against temperate root-knot nematodes (Meloidogyne spp.)

BACKGROUND: An important part of the production area of common bean (Phaseolus vulgaris L.) in Belgium is located on the sandy soils of the provinces of Antwerp and Limburg where Meloidogyne chitwoodi (Golden), M. fallax (Karssen) and M. hapla (Chitwood) are present. The host plant status of ten bean cultivars for root‐knot nematodes was determined by evaluating penetration, development and egg mass formation after inoculation with second‐stage juveniles. RESULTS: The tested cultivars were poor to good hosts for M. chitwoodi, non‐hosts or bad hosts for M. fallax and excellent hosts for M. hapla. Significantly fewer M. fallax were found in the roots, and their development was delayed. Penetration of M. hapla took place over a longer period than that of M. chitwoodi and M. fallax. The number of mature females of M. chitwoodi in cv. Polder 6 weeks after inoculation was no different from that in other cultivars, although fewer egg masses were found on this cultivar in the screening test. There was no influence of M. chitwoodi on vegetative growth of cv. Polder. CONCLUSION: The differences found in host plant status of bean cultivars stress the importance of a correct diagnosis of the Meloidogyne species in agricultural fields. Cultivar Polder showed potential as a trap crop for M. chitwoodi. Copyright © 2011 Society of Chemical Industry     

Stable carbon isotope composition of perirenal adipose tissue fatty acids from Engadine sheep grazing either mountain or lowland pasture

To provide further insights into ruminant lipid digestion and metabolism, and into cis-9,trans-11 18:2 synthesis, 12 growing Engadine lambs grazing either mountain pasture (2,250 m above sea level; n = 6) or lowland pasture (400 m above sea level; n = 6) were studied. Both pastures consisted exclusively of C(3) plants. Before the experiment, all animals grazed a common pasture for 6 wk. Grasses and perirenal adipose tissues of the sheep were analyzed for fatty acids by gas chromatography. Stable C-isotope ratios (δ(13)C values in ‰ vs. the Vienna Pee Dee Belemnite standard) were determined in the composite samples by elemental analysis-isotope ratio mass spectrometry. The δ(13)C of the individual fatty acids were measured by gas chromatography-combustion-isotope ratio mass spectrometry. The δ(13)C value of the entire mountain pasture grass was -27.5‰ (SD 0.31), whereas that of the lowland pasture grass was -30.0‰ (SD 0.07). This difference was reflected in the perirenal adipose tissues of the corresponding sheep (P < 0.05), even though the δ(13)C values were less in the animals than in the grass. The δ(13)C values for cis-9 16:1 and cis-9 18:1 in perirenal fat differed between mountain and lowland lambs (P < 0.05). The 16:0 in the adipose tissue was enriched in (13)C by 5‰ compared with the dietary 16:0, likely as a result of partly endogenous synthesis. The δ(13)C values of cis-9,trans-11 18:2 (cis-9,trans-11 CLA) in the adipose tissue were smaller than those of its dietary precursors, cis-9,cis-12 18:2 and cis-9,cis-12,cis-15 18:3; conversely, the δ(13)C values of trans-11 18:1 were not, suggesting that large proportions of perirenal cis-9,trans-11 18:2 were of endogenous origin and discrimination against (13)C occurred during Δ(9)-desaturation. The same discrimination was indicated by the isotopic shift between 16:0 and cis-9 16:1 in the mountain grazing group. Furthermore, the δ(13)C values of cis-9,trans-11 18:2 were smaller relative to the precursor fatty acids in the mountain lambs compared with the lowland group. This result suggests a reduced extent of biohydrogenation in lambs grazing on mountain grass in comparison with those grazing on lowland grass. This was supported by the smaller cis-9,trans-11 18:2 concentrations in total fatty acids found in the adipose tissues of the lowland lambs (P < 0.001). The results of this study demonstrate that natural differences between δ(13)C values of swards from different pastures and the adipose tissue fatty acids could be used as tracers in studies of lipid metabolism in ruminants.     

Pig producer attitude towards surgical castration of piglets without anaesthesia versus alternative strategies

A group of 160 Flemish pig farmers (n=160) were surveyed about their attitude towards surgical castration with anaesthesia (SCA) and without (SCN), immunocastration (IC), sperm sexing (SS) and the production of entire males (EM). The farmers' general preference was recorded as SCN>SS>SCA=IC>EM. The farmers' self-reported knowledge of the strategies was associated with their preference for SCA (negatively) and SS (positively). Herd size was correlated with farmers' preference for SCA (negatively) and SS (positively). Farmers perceived surgical castration without anaesthesia as the most favourable strategy in terms of farm profitability, animal performance and effectiveness against boar taint, but they expected the lowest consumer acceptance for this strategy. Surgical castration with anaesthesia was ranked the least favourable in terms of labour conditions. Production of entire males was viewed as the least profitable and the least effective strategy. Sperm sexing was positively perceived, particularly in terms of labour conditions, animal welfare, effectiveness and expected consumer acceptance. The farmers' opinion was quite homogeneous, especially regarding surgical castration without anaesthesia and production of entire males. Increasing our understanding of the farmers' perceptions will hopefully facilitate communication about this intricate issue.