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Azole resistance in Aspergillus fumigatus (ARAf) is becoming a worldwide health threat due to increasing occurrence in the environment. However, environmental surveillance programs are not commonly in place and are lacking in Belgium. Since no data on the occurrence of ARAf and the presence of hotspots for the selection of azole resistance is available in Belgium, a first study on the prevalence of ARAf in the environment was conducted. A total of 232 air and compost or soil samples were taken from two composting facilities, and from horticultural and agricultural crops. The azole susceptibility pattern was determined using the EUCAST method (E. Def. 9.4), and the cyp51A gene and its promotor region were sequenced in A. fumigatus isolates with phenotypic azole resistance. Six pan-azole-resistant A. fumigatus isolates were identified, originating from compost and horticultural crops. Four isolates carried the TR34/L98H mutation, and one isolate carried the TR46/Y121F/T289A mutation. However, we did not observe any ARAf isolates from agricultural crops. In conclusion, this study reported the first TR34/L98H and TR46/Y121F/T289A mutation isolated from a composting facility and horticulture in Belgium. The implementation of standardization in environmental surveillance of A. fumigatus on a European level would be beneficial in order to identify hotspots.

The ubiquitous fungus Aspergillus fumigatus can cause serious invasive diseases in humans. Due to the extensive use of environmental azoles, an increase of clinical infections with azole-resistant A. fumigatus is seen. This pilot study aimed to estimate the prevalence of azole-resistant A. fumigatus in environmental reservoirs in Belgium.

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Choline is a vitamin-like essential nutrient, important throughout one's lifespan. Therefore, choline salts are added to infant formula, supplements and functional foods. However, if choline is present in a natural form, e.g. bound to phospholipids, it may be more efficiently absorbed. The study's aim was to evaluate if choline uptake is improved after consumption of an egg yolk phospholipid drink, containing 3 g of phospholipid bound choline, compared to a control drink with 3 g of choline bitartrate. We performed a randomized, double blind, cross-over trial with 18 participants. Plasma choline, betaine and dimethylglycine concentrations were determined before and up to six hours after consumption of the drinks. The plasma choline response, as determined by the incremental area under the curve, was four times higher after consumption of the egg yolk phospholipid drink compared with the control drink (p < 0.01). Similar outcomes were also observed for choline's main metabolites, betaine (p < 0.01) and dimethylglycine (p = 0.01). Consumption of natural choline from egg yolk phospholipids improved choline absorption compared to consumption of chemically produced choline bitartrate. This information is of relevance for the food industry, instead of adding choline-salts, adding choline from egg yolk phospholipids can improve choline uptake and positively impact health.

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In recent years, the begonia mite (Polyphagotarsonemus lotus) has become an important threat to different ornamental cultures in warm greenhouses. At present there are no professional plant protection products registered in Belgium for the control of mites of the Tarsonemidae family. In a screening trial, we evaluated the efficacy of a range of different acaricides: abamectin, milbemectin, pyridaben, spirodiclofen. Based on the results of the screening trial several products were selected for a full efficacy trial following EPPO guidelines. The best control results were obtained with two products from the avermectine group: abamectin and milbemectin. As growers currently have to rely solely on the use of natural enemies there is a strong need for practical evaluation of efficacies of the various predatory mite species (Amblyseius swirskii, A. cucumeris, A. andersoni) used in biological mite control. In a series of experiments, we screened the use of different species of predatory mites. The first efficacy trials on heavily infested plants at different rates of dosage and under different circumstances (temperature, dose rate, application technique) were started in May 2008. In these experiments Amblyseius swirskii showed good efficacy. But temperature was the limiting factor: the predatory mite needed a minimal temperature of 18 degrees C to obtain good results. Further research is necessary to search for predatory mites that can be used in winter conditions (lower temperatures, less light).

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Hydrogen peroxides are commonly used in greenhouses for cleaning purposes and disinfection of irrigation water systems, i.e., to prevent clogging by duckweed (Lemna minor), algae and other (micro)organisms. This use contains a potential risk of involuntary contact to the plants, e.g., to roots through irrigation or to the plant leaves through accidental droplets (spraying mist). To help growers to maximize disinfection with minimal risks, the efficacy and plant safety of a variety of commercial available peroxide formulations were compared, i.e., pure peroxide products, peroxide products with additives: Ag, performic acid, peracetic acid and sorbitol. Starting from pure (clean and without fertilizers) irrigation water the peroxides with Ag-stabilisers were most stable and most effective for algae prevention. In screenings for the curative effect on algae, duckweed and bacteria the best results were obtained with peroxide formulations with performic acid. In plant safety tests on potted Ficus benjamina, sprays and irrigations above the plants gave no toxicity till 500 ppm a.i.; irrigations below the plants didn't show toxicity but the plant growth was reduced with weekly applications of 2000 ppm a.i. On the contrary several applications were risky on herbaceous plants, sometimes even with very low dosages (12.5 ppm peroxide).

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The effectiveness of high-temperature, short holding time (HTST) pasteurization and homogenization with respect to inactivation of Mycobacterium avium subsp. paratuberculosis was evaluated quantitatively. This allowed a detailed determination of inactivation kinetics. High concentrations of feces from cows with clinical symptoms of Johne's disease were used to contaminate raw milk in order to realistically mimic possible incidents most closely. Final M. avium subsp. paratuberculosis concentrations varying from 10(2) to 3.5 x 10(5) cells per ml raw milk were used. Heat treatments including industrial HTST were simulated on a pilot scale with 22 different time-temperature combinations, including 60 to 90 degrees C at holding (mean residence) times of 6 to 15 s. Following 72 degrees C and a holding time of 6 s, 70 degrees C for 10 and 15 s, or under more stringent conditions, no viable M. avium subsp. paratuberculosis cells were recovered, resulting in >4.2- to >7.1-fold reductions, depending on the original inoculum concentrations. Inactivation kinetic modeling of 69 quantitative data points yielded an E(a) of 305,635 J/mol and an lnk(0) of 107.2, corresponding to a D value of 1.2 s at 72 degrees C and a Z value of 7.7 degrees C. Homogenization did not significantly affect the inactivation. The conclusion can be drawn that HTST pasteurization conditions equal to 15 s at > or =72 degrees C result in a more-than-sevenfold reduction of M. avium subsp. paratuberculosis.

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In nutrition research the number of human in vivo experiments is limited because of the many restrictions and the high costs of testing in humans. Up to now predictive computer models aiming to enhance research have been rare or too complex, with many nonmeasurable adjustable parameters. This study aimed to develop a basic physicochemical computer model for a first quantitative interpretation of results obtained from in vivo intestinal experiments with bacteria. This new modeling approach is validated with results obtained from gut infection studies in vivo. The design of the model is described, and its ability to reproduce experimental data is evaluated. The model predictions are compared with new experimental data. The phenomena that take place in the gastrointestinal tract are summarized by model constants for growth, adherence, and release of bacteria. Although the model is far from describing all details and many processes in the intestine are combined, the model calculation results lead to reasonable conclusions and interesting hypotheses. One of these hypotheses concluded from the model outcomes is that Escherichia coli bacteria have a much lower intestinal growth rate in humans than in rats. Extra laboratory validation experiments proved the reliability of this hypothesis predicted by the model. In addition, the known protective effect of dietary calcium and detrimental effect of clindamycin on the growth and adherence of Salmonella bacteria could be quantified. From these results it is clear that the model enhances the interpretation of in vivo gastrointestinal experiments and will facilitate research trajectories towards new functional foods that improve resistance to pathogenic bacteria in humans.

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