Fasciola hepatica - monitoring the milky way? The use of tank milk for liver fluke monitoring in dairy herds as base for treatment strategies

In this study 595 lactating cows originating from 31 carinthian farms were investigated in accordance of liver fluke infection using individual and tank milk as well as individual blood and faecal samples. Two commercial ELISAs were used to test the milk and blood serum, and the results were compared with coproscopy and a commercial copro-antigen ELISA. Receiver operating characteristics (ROC) and two-graph operating characteristics (TG ROC) of tank milk results were conducted based on the individual milk to determine the minimum reliable in-herd antibody prevalence for the predominant condition in the investigation area. In 17.8% of the examined individuals located in 64.5% of the farms eggs were detected by coproscopy. The copro-antigen ELISA delivered 13.4% positive individuals from 54.8% of the farms. The milk ELISAs showed 42.7% (Euroclone) and 44.2% (Pourquier) positive cows on 90.3% of the farms. The blood samples were positive in 43% (Euroclone) and 45.2% (Pourquier) of the individuals from 90.3% to 96.8% of the herds, respectively. Based on the milk and the blood an average in-herd prevalence of 30-45% can be assumed. The serum and milk samples delivered correlating results with kappa values between 0.94 and 0.97, whereas the coproscopy and copro-antigen ELISA did not correlate well with the ELISA results. The two different ELISA tests highly correlated on individual and on herd level. Both showed a reliable minimum in-herd prevalence of ～20%, meaning that one fifth of the individuals in a herd have to be positive to obtain a positive bulk tank milk result. In the investigated area a higher in-herd prevalence is expected, therefore the tank milk is useful as a monitoring tool and can be used as a basis for intervention strategies.     

Optimisation of amino sugar quantification by HPLC in soil and plant hydrolysates

Amino sugars are increasingly used as indicators for the accumulation of microbial residues in soil and plant material. A reverse-phase high-performance liquid chromatography method was improved for the simultaneous determination of muramic acid, mannosamine, glucosamine and galactosamine in soil and plant hydrolysates via ortho-phthaldialdehyde (OPA) pre-column derivatisation and fluorescence detection. The retention time was reduced, and the separation of muramic acid and mannosamine was optimised by modifying the mobile phase. The effects of excitation wavelength, OPA reaction time, tetrahydrofuran concentration and pH value of the mobile phase on the amino sugar separation were tested. Quantification limits were in the range of 0.13 to 0.90 μg ml⁻¹. No interferences exist from amino acids or other primary amines, occurring in soil and plant hydrolysates.     

The impact of extraction methodologies on the toxicity of sediments in the zebrafish (Danio rerio) embryo test

Purpose

Traditionally, methods for sediment extractions are characterised using chemical analyses. However, in order to evaluate sediment extracts with regard to biological effects and, thus, bioaccessibility, extraction methods have to be compared to effect data obtained from experiments with in situ exposure scenarios, i.e., sediment contact tests. This study compares four extraction methods and sediment contact test data from a previous project with respect to predictive power in the fish embryo test with zebrafish (Danio rerio).

Materials and methods

A natural and an artificial sediment spiked with a mixture of six organic pollutants (2,4-dinitrophenol, diuron, fluoranthene, nonylphenol, parathion and pentachlorophenol) were extracted using (a) membrane dialysis extraction (MDE), (b) a Soxhlet procedure, (c) hydroxypropyl-??-cyclodextrin (HPCD) or (d) Tenax^®-TA. Whereas the former two are regarded being exhaustive with respect to non-covalently bound contaminants, the latter two are considered to predict bioaccessibility. Resulting extracts were tested in the fish embryo assay with D. rerio for embryotoxic and teratogenic potential.

Results and discussion

Mortalities caused by organic extracts from Soxhlet extraction and MDE were high. However, HPCD extracts turned out to be at least as effective as extracts obtained with these two methods. One possible reason might be short ageing time of the spiked sediments. Only Tenax^®-TA extracts gave results comparable to the sediment contact assay for natural sediment, but revealed low reproducibility. Significant differences between natural and artificial sediment were found for extracts obtained with techniques using native (i.e., non-freeze-dried) sediments, i.e., HPCD and Tenax^®-TA. In contrast, MDE and Soxhlet extracts used freeze-dried sediment and did not differentiate between natural and artificial sediment. Therefore, freeze-drying has likely altered and equalised sediment properties that influence accessibility, such as composition of bacterial communities and organic matter quality.

Conclusions

Four extraction methods were successfully characterised with respect to their stringency and predictiveness for bioaccessibility. MDE was confirmed as an alternative to Soxhlet extraction. High mortalities induced by HPCD extracts underline the need to include ageing into consideration when assessing sediments. Although Tenax^®-TA may basically be used to predict bioaccessibility in the fish embryo test, the high variability observed warrants further investigation of the relation between effect and extractability. Apparently, freeze-drying can severely affect sediment properties, potentially eliminating individual properties of natural sediments.     

Effect of cattle faeces with different microbial biomass content on soil properties, gaseous emissions and plant growth

A study was carried out to investigate the effects of different diets for heifers, low- and high-yielding cows on the microbial composition of their faeces and subsequently the impacts of these faeces on CO₂ and N₂O emissions, N mineralisation and plant N uptake. A diet low in N and high in acid detergent fibre offered to heifers resulted in faeces dominated by fungi. These faeces were characterised by a low content in microbial biomass C and N and a high ergosterol concentration in comparison to the faeces of high-yielding cows. Added to soil, faeces of heifers led to lower emission and stronger N immobilisation during a 14-day incubation in comparison to the faeces of high-yielding cows. Total N₂O emission was significantly (P?<?0.05) correlated with faecal microbial biomass N. Rye grass yield and N uptake were lowest in the soil supplemented with faeces from heifers in a 62-day pot experiment. Plant N uptake was influenced by the faecal microbial biomass C/N ratio and the fungal C to bacterial C ratio. In conclusion, the faecal microbial biomass was affected to a high degree by the feeding regime and faecal microbial characteristics revealed higher impacts on plant N uptake than soil microbial properties.     

Breeding for organic agriculture: the example of winter wheat in Austria

Breeding for organic agriculture (BFOA) is a strategy for a commercial breeding company based on the exploitation of the frequently observed high correlation for many traits between conventional, low input (LI) and organic agriculture (ORG). Indirect selection under LI can be useful to roughly divide the germplasm into the genotypes better adapted to high input and those better adapted to LI or ORG conditions. BFOA is an evolving process, in which two methods are currently applied: early generations are either selected following the pedigree system under LI, or grown as bulk populations on ORG fields. In the latter case the system switches to LI after individual ear selection under ORG conditions. In both methods, the first replicated yield trial is grown parallel under ORG and LI. Subsequently, the genotypes are grouped into conventional or ORG advanced trial series. The BFOA strategy allows that the larger genetic variability of both the organic and conventional gene pool can be exploited in the selection for ORG. Hitherto, seven winter wheat varieties were released in Austria after exclusive organic VCU testing.     

Estimating N status of winter wheat using a handheld spectrometer in the North China Plain

Excessive nitrogen (N) fertilizer application is very common in the North China Plain. Diagnosis of in-season N status in crops is critical for precision N management in this area. Remote sensing, as a timely and nondestructive tool, could be an alternative to traditional plant testing for diagnosing crop N status. The objectives of this study were to determine which vegetation indices could be used to estimate N status in winter wheat (Triticum aestivum L.) under high N input conditions, develop models to predict winter wheat N uptake using spectral vegetation indices and validate the models with data from farmers’ fields. An N rate experiment and a variety-N experiment were conducted in Huimin, Shandong Province from 2005/2006 to 2006/2007 to develop the models. Positive linear relationships between simple ratio vegetation indices (red vegetation index, RVI and green vegetation index, GVI) and N uptake were observed independent of growth stages and varieties (R², 0.48–0.74). In contrast, the relationships between normalized difference vegetation indices (NDVI and GNDVI), red and green normalized difference vegetation index (RGNDI), and red and green ratio vegetation index (RGVI) were exponentially related to N uptake (R², 0.43–0.79). Subsequently, 69 farmers’ fields in four different villages were selected as datasets to validate the developed models. The results indicated that the prediction using RVI had the highest coefficient of determination (R², 0.60), the lowest root mean square error (RMSE, 39.7 kg N ha⁻¹) and relative error (RE, 30.5%) across different years, varieties and growth stages. We conclude that RVI can be used to estimate nitrogen status for winter wheat in over-fertilized farmers’ fields before heading.     

Spatially resolved quantification of agrochemicals on plant surfaces using energy dispersive X‐ray microanalysis

BACKGROUND: In the present study the principle of energy dispersive X‐ray microanalysis (EDX), i.e. the detection of elements based on their characteristic X‐rays, was used to localise and quantify organic and inorganic pesticides on enzymatically isolated fruit cuticles. Pesticides could be discriminated from the plant surface because of their distinctive elemental composition. RESULTS: Findings confirm the close relation between net intensity (NI) and area covered by the active ingredient (AI area). Using wide and narrow concentration ranges of glyphosate and glufosinate, respectively, results showed that quantification of AI requires the selection of appropriate regression equations while considering NI, peak‐to‐background (P/B) ratio, and AI area. The use of selected internal standards (ISs) such as Ca(NO₃)₂ improved the accuracy of the quantification slightly but led to the formation of particular, non‐typical microstructured deposits. The suitability of SEM‐EDX as a general technique to quantify pesticides was evaluated additionally on 14 agrochemicals applied at diluted or regular concentration. CONCLUSION: Among the pesticides tested, spatial localisation and quantification of AI amount could be done for inorganic copper and sulfur as well for the organic agrochemicals glyphosate, glufosinate, bromoxynil and mancozeb. Copyright © 2009 Society of Chemical Industry