Recently, residues of 4-acetaminophen (paracetamol) have been found in urine of slaughter animals. To investigate its possible origin, a 7 week animal experiment was performed with 5 groups of 4 calves fed different amounts of roughage, and treated or not with 4-acetaminophen. Group 1 (control): new-born calves were fed, after the first days of colostrum, solely dairy based calf milk replacer (CMR). The animals from the other groups were 2-3 weeks of age at the start of the experiment and were fed the same CMR. Group 2 was administered oral Pracetam (R) 30 mg acetaminophen/kg Body Weight (BW) in the CMR, 5 days a week at alternate weeks, with maximum roughage. Group 3 was fed legal minimum roughage. Group 4 received the maximum roughage. Group 5 was administered human paracetamol pills 30 mg/kg BW once a week at weeks 2-6 with maximum roughage. The animals were housed in separate pens on slatted rubber floors without bedding. The controls were housed in a different room than the other animals. Urine was sampled once in the acclimatization period and for 5 days after treatment. At week 2, 4 and 6 at respectively 1, 2 and 3 days after treatment, one animal from groups 2 and 5 was sacrificed. An additional animal from group 4 was sacrificed at week 4, the rest of the animals at day 43. Urine, muscle, liver and kidney were analysed for residues of 4-acetaminophen and its metabolites. It appeared that controls and the low and high roughage animals had almost no 4-acetaminophen (<3 mu g/L) in the urine and low levels of metabolites, whereas high roughage had at slaughter higher levels of metabolites than low roughage. Pracetam (R) treated animals showed high levels 4-acetaminophen (up to 130.000 mu g/L) and metabolites during treatment, and lower levels (2800 mu g/L) at slaughter. In the paracetamol treated animals 4-acetaminophen and metabolites were detectable during treatment (up to 130.000 mu g/L) and until 7 days after treatment (110 mu g/L). 4-Acetaminophen and metabolites were only present in tissues of treated animals, during treatment (11-14.800 mu g/L) and a few days after treatment (7-55 mu g/L). A striking effect was the decrease in concentration of 4-acetaminophen and metabolites in time during treatment in the tissues. Liver enzyme induction may provide a possible explanation for this.

In 2005, a study was carried out with chicks fed either organic feed or conventional feed. The aim of the trial was to see whether there was a difference in health between organic and non-organic fed chickens, as a stepping stone to a study in pigs and ultimately in humans. Thus, the final goal was to see whether organic food has positive health effects on humans. In the study, it appeared that animals fed organic feed showed a stronger immune response after a challenge than animals not fed organic feed. However, the researchers found that no firm conclusions could be drawn about the health status of organically fed animals. Based on these results, it was decided not to commission a follow-up study because the differences would be too small. The current knowledge and insights may give new reasons for follow-up research. For this purpose, a report was published with a summary of the original research and subsequent publications, as well as results from literature since 2005 on the health effects of organic feed for animals and organic products for humans. After the 2005 study, no comparable animal studies were carried out with organic and conventional feed. It is precisely in the case of animals that only the feed can differ, which can provide valuable insight into the effects of using only organic nutrition. Research has been done on the differences in composition between conventional and organic products. Human cohort studies have also been conducted on the effects of organic food on health parameters and the occurrence of various types of cancer. Although most researchers are very cautious about concluding the health effects of organic food, there are several relevant findings on differences in the composition of products, most of which are related to the organic production method. These include no synthetic chemical crop protection agents and fewer biocides as compared to conventional production, more antioxidants, and phenols and fewer antibiotic-resistant germs than conventional food. Moreover, milk has a more beneficial fatty acid pattern. There are some indications that eating organic food lowers the risk of developing certain conditions, such as allergies, metabolic syndrome and obesity, and certain cancers.

Livestock is an important and integral component of agriculture production in Yemen and contributes 28% of the total agricultural production income. Research in the field of Yemeni ethnoveterinary medicine is limited to a few studies. Therefore, our work aims to substantiate scientifically the ethnoveterinary use of some documented plant species based on a literature review of their bioactivities and toxicological properties. Searching the scientific literature has revealed various pharmacological activities that may support the claimed healing activities of 11 out of 14 plant species for some of their ethnoveterinary utilization. This comprises the use of Aloe spp. latex for constipation, worms, boils, and wounds; Boswellia sacra underbark for wounds and its oleo-gum resin for mastitis; Soqotraen Boswellia species as an insect repellent; Cissus rotundifolia for stomach pain; Cyphostemma digitatum as an appetite stimulant; Psiadia punctulate for bone fracture; Pulicaria undulata as an insect repellent; combinations of Aristolochia bracteolate with Sorghum bicolor grains for bloating; Rumex nervosus and salt for eye pimples; and Trigonella foenum-graecum seeds with Hordeum vulgare grains for constipation. Some plants were found to demonstrate various toxic effects in in vivo and in vitro experimental studies. The local administration of Calotropis procera latex was also reported to induce an intense inflammatory response. It can be concluded that our work has provided valuable scientific information on the biological and toxic activities of some Yemeni ethnoveterinary remedies that could be utilized for the benefit of farmers to ration the use of these remedies and avoiding their toxicity.

Antibiotics are used to control infectious diseases in both animals and humans. They can be life-saving compounds but excessive use in animal husbandry leads to the development of antibiotic resistance which can impact the public health. Since similar antibiotics are used in both animal and human healthcare, it is important to reduce the use of antibiotics in production animals. In the Netherlands policies have been developed aiming for a decrease of antibiotic usage in animals, and alternatives to antibiotics are investigated. Currently, a one-on-one relationship between farmer and veterinarian is successfully implemented and (national) registration of antibiotic usage is mandatory. Unfortunately, after a 70% decrease in antibiotic usage since 2009, this decrease is now stagnating in most sectors. Innovative strategies are required to facilitate a further reduction. One promising option is a focus on farm management and natural alternatives to antibiotics. The Dutch government has invested in the spread of knowledge of natural remedies and good animal management to support animal health via so called Barnbooks for farmers and veterinarians. Another option is the analysis of on-farm antibiotic use to prevent unregistered applications. New (bio)analytical strategies to monitor the correct and complete registration of antibiotic usage have been developed and trial-tested in the Netherlands. Such strategies support a risk-based monitoring and allow effective selection of high-risk (high antibiotic use or illegal antibiotic) users. Both effective monitoring and the availability and knowledge of alternatives is a prerequisite to achieve a further significant decrease in antibiotic veterinary usage.

Microplastics (MPs) are considered an emerging issue as environmental pollutants and a potential health threat. This review will focus on recently published data on concentrations in food, possible effects, and monitoring methods. Some data are available on concentrations in seafood (fish, bivalves, and shrimps), water, sugar, salt, and honey, but are lacking for other foods. Bottled water is a considerable source with numbers varying between 2600 and 6300 MPs per liter. Particle size distributions have revealed an abundance of particles smaller than 25 µm, which are considered to have the highest probability to pass the intestinal border and to enter the systemic circulation of mammals. Some studies with mice and zebrafish with short- or medium-term exposure (up to 42 days) have revealed diverse results with respect to both the type and extent of effects. Most notable modifications have been observed in gut microbiota, lipid metabolism, and oxidative stress. The principal elements of MP monitoring in food are sample preparation, detection, and identification. Identified data gaps include a lack of occurrence data in plant- and animal-derived food, a need for more data on possible effects of different types of microplastics, a lack of in silico models, a lack of harmonized monitoring methods, and a further development of quality assurance.

Wound healing is a complex process of integrated and overlapping phases resulting in the restoration of structural and functional integrity of injured tissues. Wounds and burn wounds represent a significant burden on the patients and health care professionals worldwide. There is still an obvious lack of suitable wound healing drugs since most of the available products can have some side effects and limitations. Traditional medicine has been practiced by the indigenous people of Yemen from antiquity until today. However, many of herbal and non-herbal products, used for wounds treatment, have not been the subject of any scientific investigation. Hence, the aim of this study was to document and scientifically evaluate the claimed wound healing efficiency of twelve herbal and non-herbal remedies (nine plants, one mineral (potassium alum), one marine product (cuttlefish bone) and one animal product (honey) used in Sana’a by indigenous people to treat wounds and burns. Searching electronic databases has indicated various experimental studies demonstrating four distinct pharmacological activities (anti-inflammatory, antioxidant, antimicrobial activities and promoting various phases of wound healing) displayed by the raw materials, extracts, chemical groups and some isolated compounds of the studied herbal and non-herbal remedies. Moreover, it has been found that the studied remedies do not produce serious side effects and can be considered generally safe upon appropriate topical usage. This study provides scientific data that support the wound healing efficiency of the studied remedies and thus lend some scientific justification for their traditional use in Sana’a for the treatment of wounds and burns. These herbal and non-herbal remedies can be considered as leads for future materials for wound healing and therefore further experimental and clinical studies are required to validate their effectiveness and safety.

A new method for detecting low levels of growth promoters in bovine meat was developed with the following goal: easy, fast and sensitive analysis of a wide range of compounds, with reduced consumption of chemicals and disposables. Several classes of growth promoters were included, i.e. resorcylic acid lactones (RALs) and steroids, the latter including corticosteroids and gestagens. For sample treatment, 0.5 g of homogenised bovine meat was simultaneously disrupted and extracted in a bead-ruptor machine. The organic extraction solvent was further processed by solid-phase extraction (SPE) clean-up using 96-Well Oasis® HLB Plates. Six SPE washing steps were applied to remove matrix compounds after which the growth promoters were eluted and analysed using UHPLC-MS/MS. To achieve lower detection levels and to reduce LC-solvent consumption, a narrow-bore column with an internal diameter of 1 mm was used, instead of the conventional 2.1 mm. During analysis, the mass spectrometer was operated in negative and positive ionisation mode (ion switching). The newly developed method was validated according to the Commission Decision 2002/657. The results demonstrate that the method meets the criteria as established in this Commission Decision. The precision of the method for exogenous steroids varies between 85 and 115%, the CCα for the compounds ranges from 0.1–0.9 μg kg −1 and the expanded measurement uncertainty was lower than 36%. Compared to our current in-house methods with analysis times of 2 days for a maximum of 24 samples, the new method offers improved sample throughput (96 samples in less than 24 h) and lower detection limits.

Objective was to assess the use by veterinarians of the Guides for farmers to keep their animals healthy with natural products. These booklets are published by the Dutch government to help in reducing the use of antibiotics in livestock by veterinarians. A questionnaire according to the iDiaLo og methodology, was distributed among all approved veterinarians that take part in a quality assurance program for a specific species. This Idialoog methodology is a proven research technique that allows for a high response and delivers relevant information on the primary question for the target group. Additional information on use of natural products and the role of the veterinarian was collected. All responses (answers and opinions) are anonymised and grouped. The participants receive this as feedback via email (figure 1). Within iDiaLoog, all participants are equal and can participate at any time that suits them best. Because of its near-real-time feedback properties it is attractive for participants to join and thus ensuring a high response rate. iDiaLoog is a simple, effective and efficient manner to gather and share relevant and actual information about specific topics. This provides practitioners with insight of their peer group.

To investigate the potential transfer of pyrrolizidine alkaloids (PAs), laying hens were fed for 14 days with diets containing 0.5% of dried common ragwort, common groundsel, narrow-leaved ragwort or viper's bugloss, or 0.1% of common heliotrope. This resulted in total PA levels in feed of respectively 5.5, 11.1, 53.1, 5.9 and 21.7 mg kg-1, with varying composition. PAs were transferred to eggs, in particular yolk, with steady-state levels of respectively 12, 21, 216, 2 and 36 µg kg-1. Overall transfer rates for the sum of PAs were estimated between 0.02% and 0.23%, depending on the type of PAs in the feed. In animals slaughtered shortly after the last exposure, levels in meat were slightly lower than those in eggs, levels in livers somewhat higher. When switched to clean feed, levels in eggs gradually decreased, but after 14 days were still above detection limits in the hens exposed to higher PA levels. Similar was the case for meat and especially kidneys and livers. It is concluded that the intake of PA containing herbs by laying hens may result in levels in eggs and meat that could be of concern for consumers, and as such should be avoided.

Food producing animals are exposed to biologically active plant compounds through feed and roughages, presenting a potential risk to the animal but also consumers of food of animal origin. To evaluate to which plant compounds of concern dairy cows in the Netherlands are exposed, a ranking filter model was developed, combining information on abundance of plant species in vegetation plots in the Netherlands (183,905 plots of three different vegetation types) with plant-compound combinations (700), and with consumption data of fresh grass, grass silage and corn silage by cattle. The most abundant plant genera are those producing cyanogenic glycosides, coumarins and benzofuranocoumarins, being predominantly fodder plants (alfalfa, clover and some grasses) considered to be safe. Highest exposures were estimated for plant genera producing piperidine alkaloids (horsetail), furanocoumarins (parsley and relatives), pyrrolizidine alkaloids (Symphytum, Senecio, Leucanthemum, Eupatorium) and essential oils. The current results allow to prioritise future scientific research on these compounds.

In agriforensics, time of administration is often debated when illegal drug residues, such as clenbuterol, are found in frequently traded cattle. In this proof-of-concept work, the feasibility of obtaining retrospective timeline information from segmented calf tail hair analyses has been studied. First, an ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) hair analysis method was adapted to accommodate smaller sample sizes and in-house validated. Then, longitudinal 1 cm segments of calf tail hair were analyzed to obtain clenbuterol concentration profiles. The profiles found were in good agreement with calculated, theoretical positions of the clenbuterol residues along the hair. Following assessment of the average growth rate of calf tail hair, time of clenbuterol administration could be retrospectively determined from segmented hair analysis data. The data from the initial animal treatment study (n = 2) suggest that time of treatment can be retrospectively estimated with an error of 3-17 days.

The EUBerry project is not only building on technical research but also studies berry production and consumption from a social sciences perspective. This paper aims to describe the approach used in the EUBerry project about business economics as well as marketing. In addition some preliminary results are presented. The social sciences part of EUBerry consists of four parts: economic viability, market situation, marketing strategies and evaluation. The economic viability of new production methods will be calculated beforehand as well as at the end of the project. The ex-ante analysis about critical success factors, with regard to expected costs and returns, gives the conditions for the improvement and implementation of new developing methods and can be seen as research guide. In this paper preliminary results of the ex-ante analysis for blueberries will be presented. With respect to market description, the initial market situation of fresh berries in selected European countries will be described. This description will be based on desk research as well as interviews. These results are taken as a basis for the development, testing and evaluation of marketing strategies, adapted from health benefits of berries in order to increase consumption. These marketing strategies will be tested together with SMEs. An overview of this marketing "plan of action method" will be given.

Background: Synthetic Amorphous Silica (SAS) is commonly used in food and drugs. Recently, a consumer intake of silica from food was estimated at 9.4 mg/kg bw/day, of which 1.8 mg/kg bw/day was estimated to be in the nano-size range. Food products containing SAS have been shown to contain silica in the nanometer size range (i.e. 5 - 200 nm) up to 43% of the total silica content. Concerns have been raised about the possible adverse effects of chronic exposure to nanostructured silica.Methods: Rats were orally exposed to 100, 1000 or 2500 mg/kg bw/day of SAS, or to 100, 500 or 1000 mg/kg bw/day of NM-202 (a representative nanostructured silica for OECD testing) for 28 days, or to the highest dose of SAS or NM-202 for 84 days.Results: SAS and NM-202 were extensively characterized as pristine materials, but also in the feed matrix and gut content of the animals, and after in vitro digestion. The latter indicated that the intestinal content of the mid/high-dose groups had stronger gel-like properties than the low-dose groups, implying low gelation and high bioaccessibility of silica in the human intestine at realistic consumer exposure levels. Exposure to SAS or NM-202 did not result in clearly elevated tissue silica levels after 28-days of exposure. However, after 84-days of exposure to SAS, but not to NM-202, silica accumulated in the spleen. Biochemical and immunological markers in blood and isolated cells did not indicate toxicity, but histopathological analysis, showed an increased incidence of liver fibrosis after 84-days of exposure, which only reached significance in the NM-202 treated animals. This observation was accompanied by a moderate, but significant increase in the expression of fibrosis-related genes in liver samples.Conclusions: Although only few adverse effects were observed, additional studies are warranted to further evaluate the biological relevance of observed fibrosis in liver and possible accumulation of silica in the spleen in the NM-202 and SAS exposed animals respectively. In these studies, dose-effect relations should be studied at lower dosages, more representative of the current exposure of consumers, since only the highest dosages were used for the present 84-day exposure study.

To investigate whether clenbuterol-treated calves could contaminate untreated pen mates, three animal experiments were performed. (1) One calf of a pen of five was treated with clenbuterol by injection (Ventipulmin injection, REG NL 2532, 2.5mL/100 kg) twice a day for 10 days. (2) In two pens, one animal was treated with clenbuterol via oral administration (Ventipulmin syrup, REG NL 2532, 4mL/125 kg) for 4 weeks. (3) In two pens, one animal was treated with clenbuterol via the milk (Ventipulmin, REG NL 2532, 2.5mL/100 kg body weight) twice a day for 10 days. Here, the animal was set apart during treatment, cleaned and put back into the group. Levels of clenbuterol were analysed in hair and urine with LC-MS/MS. Clenbuterol administered by injection could not be transferred from treated to untreated calves. In the second experiment, all pen mates were found positive for clenbuterol in the hair. This contamination was probably due to licking the mouth of the treated animal or saliva from the treated animal spoiling the floor. In the third experiment, no pen mates were found positive for clenbuterol in the hair. Clenbuterol was found in the urine and hair of only treated animals.

Precision cut liver slices (PCLSs) are widely used as a model to study hepatotoxicity. For culturing of PCLS diverse protocols are used which could affect slices viability and results. We aimed to identify the most optimal culture protocol for mouse PCLS. Slices were cultured for 24h under different concentrations of serum, glucose, insulin, and oxygen. Thereafter, slices viability was assessed by biochemical methods. Transcriptome analysis was performed to identify changes introduced by culture at different oxygen concentrations (20%, 40%, 60%, and 80% of oxygen). Medium composition did not affect the slices viability. Although metabolic competence was unaffected by oxygen concentrations, culturing at 80% of oxygen yielded slices with the best biochemical characteristics. The comparison of uncultured vs. cultured slices revealed 2524 genes to be differentially expressed. Genes involved in drug metabolism, peroxisomal and mitochondrial functions were down-regulated while several adaptive/stress response processes were up-regulated. Moreover, 80% of oxygen was the most favorable condition with respect to maintenance of expression of genes involved in drug and energy metabolism. The outcome of this study indicates that mouse PCLS are a valuable tool in research on hepatic functions and toxicity, particularly if they are cultured under a controlled oxygen concentration of 80%.

BACKGROUND:Unexpected cholestasis substantially contributes to drug failure in clinical trials. Current models used for safety assessment in drug development do not accurately predict cholestasis in humans. Therefore, it is of relevance to develop new screening models that allow identifying drugs with cholestatic properties.METHODS:We employed mouse precision cut liver slices (PCLS), which were incubated 24 h with two model cholestatic compounds: cyclosporin A (CsA) and chlorpromazine (CPZ). Subsequently, transcriptome analysis using DNA microarrays and q-PCR were performed to identify relevant biological processes and biomarkers. Additionally, histology was carried out and levels of triglycerides (TG) and bile acids (BA) were measured. To verify the ex vivo mouse data, these were compared with publically available human data relevant for cholestasis.RESULTS:Whole genome gene expression analysis showed that CsA up-regulated pathways related to NF-κB, ER stress and inflammation. Both CsA and CPZ down-regulated processes related to extracellular matrix (ECM) remodelling, BA homeostasis, Fxr signalling, and energy metabolism. The differential expression of a number of characteristic genes (e.g. Abcg5, Abcg8, Klf15, and Baat) could be confirmed by q-PCR. Histology revealed that CsA but not CPZ induced "ballooning" of hepatocytes. No effects on TG and BA levels were observed after incubation of PCLS with CsA and CPZ. A substantial number of processes altered in CsA- and CPZ-treated mouse PCLS ex vivo was also found to be affected in liver biopsies of cholestatic patients.CONCLUSION:The present study demonstrated that mouse PCLS can be used as a tool to identify mechanisms of action of cholestatic model compounds. The induction of general stress responses and down-regulated Fxr signalling could play a role in the development of drug induced cholestasis. Importantly, comparative data analysis showed that the ex vivo mouse findings are also relevant for human pathology. Moreover, this work provides a set of genes that are potentially useful to assess drugs for cholestatic properties.

ISHS XXVIII International Horticultural Congress on Science and Horticulture for People (IHC2010): International Symposium on the Challenge for a Sustainable Production, Protection and Consumption of Mediterranean Fruits and Nuts INTRODUCING NEW APPLE CULTIVARS THROUGH A COORDINATED APPROACH FROM CONSUMER TILL BREEDER