Detection of ruminant meat and bone meals in animal feed by real-time polymerase chain reaction: result of an interlaboratory study

The commercialization of animal feeds infected by prions proved to be the main cause of transmission of bovine spongiform encephalopathy (BSE). Therefore, feed bans were enforced, initially for ruminant feeds, and later for all feeds for farmed animals. The development and validation of analytical methods for the species-specific detection of animal proteins in animal feed has been indicated in the TSE (Transmissible Spongiform Encephalopathies) Roadmap (European Commission. The TSE (Transmissible Spongiform Encephalopathy) roadmap. URL: http://europa.eu.int/comm/food/food/biosafety/bse/roadmap_en.pdf, 2005) as the main condition for lifting the extended feed ban. Methods based on polymerase chain reaction (PCR) seem to be a promising solution for this aim. The main objective of this study was to determine the applicability of four different real-time PCR methods, developed by three National expert laboratories from the European Union (EU), for the detection and identification of cattle or ruminant species in typical compound feeds, fortified with meat and bone meals (MBM) from different animal species at different concentration levels. The MBM samples utilized in this study have been treated using the sterilization condition mandatory within the European Union (steam pressure sterilization at 133 degrees C, 3 bar, and 20 min), which is an additional challenge to the PCR methods evaluated in this study. The results indicate that the three labs applying their PCR methods were able to detect 0.1% of cattle MBM, either alone or in mixtures with different materials such as fishmeal, which demonstrates the improvement made by this technique, especially when compared with results from former interlaboratory studies.     

Analyses of the impact of changes in atmospheric deposition and climate on forest growth in European monitoring plots: A stand growth approach

During the last 15 years a number of studies have shown increasing forest growth in central Europe, rather than a decline as was expected due to negative effects of air pollution. We have here used data from intensive monitoring plots spread over Europe for a five year period in order to examine the influence of environmental factors on forest growth. Evaluations focussed on the influence of nitrogen, sulphur and acid deposition, temperatures, precipitation and on a drought index calculated as deviation from the long-term mean. The study included the main tree species Norway spruce, Scots pine, common beech as well as European and sessile oak and was based on data from 363 plots. As many other factors besides nitrogen and temperature influence tree growth, expected stem volume increments were modelled using site productivity, stand age and a stand density index. Relative volume increment was then calculated as actual increment in % of expected increment. The site productivity, assumed to be given by site conditions and past environmental conditions, was either taken from expert estimates or computed from site index curves from northern, central and southern Europe. The model explained between 18% and 39% of the variance with site productivity being positively related and age negatively related to actual increment. The various models and statistical approaches were fairly consistent, and indicated a fertilizing effect of nitrogen deposition, with slightly above one percent increase in volume increment per kg of nitrogen deposition per ha and year. This was most clear for spruce and pine, and most pronounced for plots having soil C/N ratios above 25. Also, we found a positive relationship between relative increment and summer temperature, i.e. May–August mean temperature deviation from the 1961–1990 means. The cause–effect relationship here is, however, less certain. Other influences were uncertain. Possibly, sulphur and acid deposition have effects on growth, but these effects are obscured by, and outweighed by the positive effect of nitrogen deposition, because of collinearity between these variables. Drought effects were uncertain also, and one reason for this might be large uncertainties in the precipitation data: precipitation measured on some 50% of the plots correlated poorly with the precipitation data obtained from Europe-wide databases. The major finding of this study was a positive relationship between higher than normal volume increment on one hand and nitrogen deposition on the other hand.     

Trace elements bioavailability to <Emphasis Type="Italic">Triticum aestivum</Emphasis> and <Emphasis Type="Italic">Dendrobaena veneta</Emphasis> in a multielement-contaminated agricultural soil amended with drinking water treatment residues

Purpose

The in situ stabilization of multielement-contaminated agricultural soils has limited effectiveness when using common single amendments. This study examined the use of drinking water treatment residues (WTR), based on (hydr)oxides of Fe, Al, or Mn, as a cost-effective solution to optimize the immobilization of metals (Cd, Pb, Zn) and As.

Materials and methods

Trace elements (TE) bioavailability was assessed under semi-controlled conditions in a pot study cultivating winter wheat (Triticum aestivum L. cv. Tiger) until maturity. An Fe-based WTR and a Mn-based WTR, applied at rates of 0.5 and 1% (m/m), were related to effects of lime marl (LM) application. Additionally, a bioassay with earthworms (Dendrobaena veneta) was conducted. Both bioassays were compared with measurements of NH₄NO₃-soluble, diffusive gradients in thin film (DGT)-available and soil solution TE concentrations, representing well-established surrogates for mimicking the bioavailable element fractions in soil.

Results and discussion

The application of the Fe-based WTR reduced As accumulation in vegetative wheat tissues (by up to 75%) and earthworms (by up to 41%), which corresponded with the findings from soil chemical analyses and improved plant growth and earthworm body weight. However, As concentrations in cereal grains were not affected, Cd or Pb accumulation by wheat was not mitigated, and Zn uptake was enhanced. By contrast, the Mn-based WTR effected the greatest reduction in Pb uptake, and lowered Cd transfer to wheat grain (by up to 25%). Neither the NH₄NO₃-soluble nor DGT-available concentrations matched with Cd and Zn accumulation in plants or earthworms, indicating interferences due to competition for binding sites according to the biotic ligand model.

Conclusions

The results obtained in this study suggest that a bioassay with key species prior to field application should be mandatory when designing in situ stabilization options. The application of WTR to an agricultural soil strongly affected TE bioavailability to plants and earthworms. Low application rates tended to improve biomass production of biota. Higher application rates involved risks (e.g., P fixation, TE inputs), and none of the amendments tested could immobilize all targeted elements.

     

Assessment of computed tomography derived cricoid cartilage and tracheal dimensions to evaluate degree of cricoid narrowing in brachycephalic dogs

The aims of this observational, analytical, retrospective study were to (i) obtain computed tomographic (CT) cricoid dimensions (height, width, and transverse‐sectional area), (ii) compare the cricoid dimensions between brachycephalic and mesaticephalic breeds, and (iii) compare cricoid cartilage dimensions between dogs without and affected with brachycephalic airway syndrome. The study is important to help to further evaluate and understand the anatomical components of brachycephalic airway syndrome. Measurements were performed in 147 brachycephalic and 59 mesaticephalic dogs. The cricoid cartilage was found to be significantly more oval in Pugs and French Bulldogs compared to mesaticephalic breeds. The cricoid cartilage transverse‐sectional area was smallest for the Pug and, after adjusting for weight, significantly smaller for Pugs (P < 0.001), Boston Terriers (P = 0.001), and French Bulldogs (P < 0.001) compared to Jack Russell Terriers. The tracheal transverse‐sectional area at C4 of English Bulldogs was significantly smaller than for Jack Russell Terriers (P = 0.005) and Labradors (P < 0.001). The cricoid cartilage transverse‐sectional area:weight ratio was significantly lower in brachycephalic breeds compared to mesaticephalic breeds (P < 0.001). The cricoid cartilage:trachea at C4 transverse‐sectional area for brachycephalic dogs was significantly larger than for mesaticephalic dogs (<0.001), demonstrating that the trachea was the narrowest part of the airway. No significant differences were found for cricoid dimensions between dogs affected with and without brachycephalic airway syndrome. However, large individual variation was found among the brachycephalic breeds and further studies investigating the relationship between cricoid cartilage size, laryngeal collapse, concurrent tracheal hypoplasia, and/or severity of brachycephalic airway syndrome are warranted.     

Effects of granivorous rodents on direct seeding of oak and beech in relation to site preparation and sowing date

Planting bare-rooted seedlings is a rather expensive method to regenerate oak (Quercus robur) and beech (Fagus sylvatica). Direct seeding could reduce artificial regeneration costs substantially, but the method currently suffers from a number of problems, one of which is seed consumption by rodents. In the present study clear-cut areas in southern Sweden were prepared using one of four different mechanical site preparation methods – disc trenching, patch scarification, topsoil removal or mounding – then manually sown with acorns and beech nuts. Acorns were sown in May and July 2006, and both acorns and beech nuts were sown in May 2007. Three years following sowing, best seedling establishment from viable acorns, 60–70%, was achieved after sowing in mounds in May, and the subsequent survival reached 90–100%. After the first growing season for beech, both seedling establishment and subsequent survival was lower than for oak, which coincides with a higher number of lost beech nuts at the excavation to investigate the fate of seeds. Granivorous rodents (Apodemus flavicollis, A. sylvaticus, Mus musculus, and Myodes glareolus) were live-trapped more frequently in the vicinity of slash piles compared to other micro-site features, such as uprooted stumps, snags and stones. In 2006, no connection was found between number of rodents captured and proportion of lost seeds, while in 2007 the two were in good accordance. No clear relationship was found between number of captured rodents, vegetation cover class and mechanical site preparation method. More rodents were captured in 2007 than in 2006, and there was an increase in captures from spring until late summer/autumn for both years, which was mirrored in the seedling establishment result from the three different times of sowing. Based on experiences from the present study the recommendation for successful direct seeding of oak on clear-cuts is a combination of mounding site preparation, removal of slash and sowing in May rather than July. However, the results indicate that successful direct seeding of beech is more difficult to achieve.     

Marennine,Promising Blue Pigments from a Widespread Haslea Diatom Species Complex

In diatoms, the main photosynthetic pigments are chlorophylls a and c, fucoxanthin, diadinoxanthin and diatoxanthin. The marine pennate diatom Haslea ostrearia has long been known for producing, in addition to these generic pigments, a water-soluble blue pigment, marennine. This pigment, responsible for the greening of oysters in western France, presents different biological activities: allelopathic, antioxidant, antibacterial, antiviral, and growth-inhibiting. A method to extract and purify marennine has been developed, but its chemical structure could hitherto not be resolved. For decades, H. ostrearia was the only organism known to produce marennine, and can be found worldwide. Our knowledge about H. ostrearia-like diatom biodiversity has recently been extended with the discovery of several new species of blue diatoms, the recently described H. karadagensis, H. silbo sp. inedit. and H. provincialis sp. inedit. These blue diatoms produce different marennine-like pigments, which belong to the same chemical family and present similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms thus present a commercial potential for aquaculture, cosmetics, food and health industries.