Metabolomic Profiling of In Vivo Plasma Responses to DioxinAssociated Dietary Contaminant Exposure in Rats: Implications for Identification of Sources of Animal and Human Exposure

Dioxin contamination of the food chain typically occurs when cocktails of combustion residues or polychlorinated biphenyl (PCB) containing oils become incorporated into animal feed. These highly toxic compounds are bioaccumulative with small amounts posing a major health risk. The ability to identify animal exposure to these compounds prior to their entry into the food chain may be an invaluable tool to safeguard public health. Dioxin-like compounds act by a common mode of action and this suggests that markers or patterns of response may facilitate identification of exposed animals. However, secondary co-contaminating compounds present in typical dioxin sources may affect responses to compounds. This study has investigated for the first time the potential of a metabolomics platform to distinguish between animals exposed to different sources of dioxin contamination through their diet. Sprague-Dawley rats were given feed containing dioxin-like toxins from hospital incinerator soot, a common PCB oil standard and pure 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (normalized at 0.1 µg/kg TEQ) and acquired plasma was subsequently biochemically profiled using ultra high performance liquid chromatography (UPLC) quadropole time-of-flight-mass spectrometry (QTof-MS). An OPLS-DA model was generated from acquired metabolite fingerprints and validated which allowed classification of plasma from individual animals into the four dietary exposure study groups with a level of accuracy of 97-100%. A set of 24 ions of importance to the prediction model, and which had levels significantly altered between feeding groups, were positively identified as deriving from eight identifiable metabolites including lysophosphatidylcholine (16:0) and tyrosine. This study demonstrates the enormous potential of metabolomic-based profiling to provide a powerful and reliable tool for the detection of dioxin exposure in food-producing animals.

The use of handheld near-infrared reflectance spectroscopy (NIRS) for the proximate analysis of poultry feed and to detect melamine adulteration of soya bean meal

The use of handheld near infrared (NIR) instrumentation, as a tool for rapid analysis, has the potential to be used widely in the animal feed sector. A comparison was made between handheld NIR and benchtop instruments in terms of proximate analysis of poultry feed using off-the-shelf calibration models and including statistical analysis. Additionally, melamine adulterated soya bean products were used to develop qualitative and quantitative calibration models from the NIRS spectral data with excellent calibration models and prediction statistics obtained. With regards to the quantitative approach, the coefficients of determination (R2) were found to be 0.94-0.99 with the corresponding values for the root mean square error of calibration and prediction were found to be 0.081-0.215 % and 0.095-0.288 % respectively. In addition, cross validation was used to further validate the models with the root mean square error of cross validation found to be 0.101-0.212 %. Furthermore, by adopting a qualitative approach with the spectral data and applying Principal Component Analysis, it was possible to discriminate between adulterated and pure samples.

The effect of feeder type and change of feeder type on growing and finishing pig performance and behaviour

Reduced feed intake and hence lower growth rates commonly occur when the environment of the pig changes, e.g. at weaning and when pigs are moved from growing accommodation to finishing accommodation. It is hypothesised that if environmental factors, such as feeder type, remain the same in the weaning and finishing accommodation this 'growth check' may be reduced. A total of 640 pigs in 32 pens of 20 pigs per pen were used to investigate the effects of two feeder types and changing or not changing the feeder type at 10 weeks of age on growth performance and behaviour in the periods from 4 to 10, 10 to finish (22) and from 4 to finish (22) weeks of age, respectively. The two feeder types tested were a 'wet and dry' single-space (S) feeder and a 'dry' multi-space feeder (M).

Evidence of natural occurrence of the banned antibiotic chloramphenicol in herbs and grass

Chloramphenicol (CAP), a broad-spectrum antibiotic, was detected in several herb and grass samples from different geographic origins. Due to its suspected carcino-genicity and linkages with the development of aplastic anemia in humans, CAP is banned for use in food-producing animals in the European Union (EU) and many other countries. However, products of animal origin originating from Asian countries entering the European market are still found noncompliant (containing CAP) on a regular basis, even when there is no history of chloramphenicol use in these countries. A possible explanation for the continued detection of these residues is the natural occurrence of CAP in plant material which is used as animal feed, with the consequent transfer of the substance to the animal tissues. Approximately 110 samples were analyzed using liquid chromatography coupled with mass spectrometric detection. In 26 samples, the presence of CAP was confirmed using the criteria for banned substances defined by the EU. Among other plant materials, samples of the Artemisia family retrieved from Mongolia and from Utah, USA, and a therapeutic herb mixture obtained from local stores in the Netherlands proved to contain CAP at levels ranging from 0.1 to 450 mu g/kg. These findings may have a major impact in relation to international trade and safety to the consumer. The results of this study demonstrate that noncompliant findings in animal-derived food products may in part be due to the natural occurrence of chloramphenicol in plant material. This has implications for the application of current EU, USA, and other legislation and the interpretation of analytical results with respect to the consideration of CAP as a xenobiotic veterinary drug residue and the regulatory actions taken upon its detection in food.

The prediction of intake potential and organic matter digestibility of grass silages by near infrared spectroscopy analysis of undried samples

A study was undertaken to examine a range of sample preparation and near infrared reflectance spectroscopy (NIPS) methodologies, using undried samples, for predicting organic matter digestibility (OMD g kg(-1)) and ad libitum intake (g kg(-1) W-0.75) of grass silages. A total of eight sample preparation/NIRS scanning methods were examined involving three extents of silage comminution, two liquid extracts and scanning via either external probe (1100-2200 nm) or internal cell (1100-2500 nm). The spectral data (log 1/R) for each of the eight methods were examined by three regression techniques each with a range of data transformations. The 136 silages used in the study were obtained from farms across Northern Ireland, over a two year period, and had in vivo OMD (sheep) and ad libitum intake (cattle) determined under uniform conditions. In the comparisons of the eight sample preparation/scanning methods, and the differing mathematical treatments of the spectral data, the sample population was divided into calibration (n = 91) and validation (n = 45) sets. The standard error of performance (SEP) on the validation set was used in comparisons of prediction accuracy. Across all 8 sample preparation/scanning methods, the modified partial least squares (MPLS) technique, generally minimized SEP's for both OMD and intake. The accuracy of prediction also increased with degree of comminution of the forage and with scanning by internal cell rather than external probe. The system providing the lowest SEP used the MPLS regression technique on spectra from the finely milled material scanned through the internal cell. This resulted in SEP and R-2 (variance accounted for in validation set) values of 24 (g/kg OM) and 0.88 (OMD) and 5.37 (g/kg W-0.75) and 0.77 (intake) respectively. These data indicate that with appropriate techniques NIRS scanning of undried samples of grass silage can produce predictions of intake and digestibility with accuracies similar to those achieved previously using NIRS with dried samples. (C) 1998 Elsevier Science B.V.

The application of Near-Infrared Reflectance Spectroscopy (NIRS) to detect melamine adulteration of soya bean meal

Soya bean products are used widely in the animal feed industry as a protein based feed ingredient and
have been found to be adulterated with melamine. This was highlighted in the Chinese scandal of
2008. Dehulled soya (GM and non-GM), soya hulls and toasted soya were contaminated with melamine
and spectra were generated using Near Infrared Reflectance Spectroscopy (NIRS). By applying chemometrics
to the spectral data, excellent calibration models and prediction statistics were obtained. The coefficients
of determination (R2) were found to be 0.89–0.99 depending on the mathematical algorithm used,
the data pre-processing applied and the sample type used. The corresponding values for the root mean
square error of calibration and prediction were found to be 0.081–0.276% and 0.134–0.368%, respectively,
again depending on the chemometric treatment applied to the data and sample type. In addition, adopting
a qualitative approach with the spectral data and applying PCA, it was possible to discriminate
between the four samples types and also, by generation of Cooman’s plots, possible to distinguish
between adulterated and non-adulterated samples.

Determination of geographical origin of distillers dried grains and solubles using isotope ratio mass spectrometry

In recent years distillers dried grains and solubles (DDGS), co-products of the bio-ethanol and beverages industries, have become globally traded commodity for the animal feed sector. As such it is becoming increasingly important to be able to trace the geographical origin of commodities in case of a contamination incident or authenticity issue arise. In this study, 137 DDGS samples from a range of different geographical origins (China, USA, Canada and European Union) were collected and analyzed. Isotope ratio mass spectrometry (IRMS) was used to analyze the DDGS for ²H/¹H, ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O and ³⁴S/³²S isotope ratios which can vary depending on geographical origin and processing. Univariate and multivariate statistical techniques were employed to investigate the feasibility of using the IRMS data to determine botanical and geographical origin of the DDGS. The results indicated that this commodity could be differentiated according to their place of origin by the analysis of stable isotopes of hydrogen, carbon, nitrogen and oxygen but not with sulfur. By adding data to the models produced in this study, potentially an isotope databank could be set up for traceability procedures for DDGS, similar to the one established already for wine which will help in feed and food security issues arising worldwide.

Effect of two different rice dehusking procedures on total arsenic concentration in rice

Pollution of subterranean water by arsenic (As) in Asia has resulted in the worst chemical disaster in human history. For populations living on subsistence rice diets, As contamination of rice grain contributes greatly to dietary As exposure. The main objectives of this study were to compare two dehusking processes: (a) wet process (soaking of rice, boiling and mechanical hulling) and (b) dry process (mechanical hulling), and recommend the method leading to a lower As content in commercial rice. In general, hulling of paddy rice (373 mu g As kg(-1)) significantly decreased As content in rice grain (311 mu g As kg(-1)). The final As concentrations in boiled rice (final product of the wet process) and atab rice (dry process) were 332 and 290 mu g kg(-1). Thus, the dry method is recommended for dehusking paddy rice if not As-free water is available. However, villagers can reduce the As content in the wet system by discarding the soaking water and using new water for the light boiling. Finally, it is not recommended to use rice husk for feeding animals because the As concentration is very high, approximately 1,000 mu g As kg(-1).

Determination of the Mycotoxin Content in Distiller's Dried Grain with Solubles Using a Multianalyte UHPLC-MS/MS Method

There are more than 300 potential mycotoxins that can contaminate food and feed and cause adverse effects in humans and animals. The data on the co-occurrence of mycotoxins in novel animal feed materials, such as distiller's dried grain with solubles (DDGS), are limited. Thus, a UHPLC-MS/MS method for the quantitation of 77 mycotoxins and other fungal metabolites was used to analyze 169 DDGS samples produced from wheat, maize, and barley and 61 grain samples. All DDGS samples analyzed were contaminated with 13-34 different mycotoxins. Fumonisins were present in all 52 maize DDGS samples (81.0-6890 μg/kg for fumonisin B1), and deoxynivalenol was present in all 99 wheat DDGS samples (39.3-1120 μg/kg). A number of co-occurring mycotoxins were also identified. Due to the high co-occurrence of mycotoxins, routine screening of the animal feed ingredients is highly recommended to allow the highlighted risks to be effectively managed.