Quantifying fecal shedding of Mycobacterium avium ssp. paratuberculosis from calves after experimental infection and exposure

Johne's disease, a chronic enteritis caused by Mycobacterium avium ssp. paratuberculosis (MAP), causes large economic losses to the dairy industry worldwide. Fecal shedding of MAP contaminates the environment, feed, and water and contributes to new infections on farm, yet there is limited knowledge regarding mechanisms of shedding, extent of intermittent shedding, and numbers of MAP bacteria shed. The objectives were to (1) compare (in an experimental setting) the frequency at which intermittent shedding occurred and the quantity of MAP shed among pen mates that were inoculated or contact-exposed (CE); and (2) determine whether an association existed between inoculation dose and quantity of MAP shed. In the first experiment, 32 newborn Holstein-Friesian bull calves were allocated to pens in groups of 4, whereby 2 calves were inoculated with a moderate dose (MD; 5 × 10⁸ cfu) of MAP and 2 calves acted as CE. Calves were group-housed for 3 mo, fecal samples were collected and cultured, and culture-positive samples were quantified. In the second experiment, 6 calves were inoculated with either a low (LD) or high (HD) dose of MAP (1 × 10⁸ or 1 × 10¹⁰ cfu, respectively), and fecal samples were collected for 3 mo and cultured for detection of MAP. The amount of MAP was quantified using direct extraction (DE) of DNA from fecal samples and F57-specific quantitative PCR. In experiment 1, the average amount of MAP in all culture-positive samples did not differ between MD and CE calves. In experiment 2, when comparing inoculation doses, LD calves had the lowest proportion of MAP-positive culture samples and HD had the highest, but no difference was detected in the average quantity of MAP shed. This study provided new information in regards to Johne's disease research and control regarding shedding from various inoculation doses and from CE animals; these data should inform future trials and control programs.     

Detection of noroviruses in shellfish in the Netherlands

Shellfish from oyster farms in the Netherlands and imported from other European countries were examined for viral contamination. A method that allows sequence matching between noroviruses from human cases and shellfish was used. The samples of shellfish (n = 42) were analyzed using a semi-nested RT-PCR that had been optimized for detection of norovirus in shellfish (SR primer sets). In addition, a different genome region was targeted using a second primer set which is routinely used for diagnosis of norovirus infection in humans (JV12Y/JV13I). To improve the detection limit for this RT-PCR a semi-nested test format was developed (NV primer sets). One of 21 oyster samples (4.8%) from Dutch farms was norovirus positive, whereas norovirus was detected in 1 out of 8 oyster samples (12.5%) and 5 out of 13 mussel samples (38.5%) collected directly after importation in the Netherlands. RNA from samples associated with an outbreak of gastro-enteritis in the Netherlands in 2001 was re-analyzed using the NV primer sets. At least one identical sequence (142/142 nt) was found in three fecal and in two oyster samples related to this outbreak. Further surveillance of norovirus by detection and typing of viruses from patients with gastroenteritis and shellfish is warranted to clarify the causes of future outbreaks.     

Beer quality screening by FT-IR spectrometry: Impact of measurement strategies, data pre-processings and variable selection algorithms

In the presented research three measurement strategies of Fourier transform infrared (FT-IR) spectrometry (horizontal- and micro-attenuated total reflection (HATR and μATR, respectively) and a novel high throughput transmission (HTT)) in combination with extensive chemometric approach have been applied for analysis of beer. Fifty Belgian beer samples were analyzed: lager and white beers, dark and blonde ales, lambic and Trappist beers. Partial least square (PLS) regression was used for prediction of different physicochemical parameters of beer based on mid-IR data, modified with different spectral preprocessing and variable selection algorithms for higher prediction performance. It was found that HATR and μATR have the same prediction capability with a slight advantage of μATR and excellent correlations between spectra and reference values were demonstrated for original (R² 0.95), apparent (R² 0.89) and real (R² 0.95) extracts, real and apparent degrees of fermentation (R² 0.91 and 0.89), alcohol content (R² 0.94) and energetic value (R² 0.95). Good results (R² 0.83) were obtained from the HTT experiments for prediction of real extract and allow measurement of large sample numbers in short time (1 sample/minute) with further potential increase in a sample throughput and has the possibility for automation. The obtained results indicate that mid-IR spectrometry in combination with advanced pre-processing and variable selection methods is a valuable analytical tool for high throughput measurement of beer quality attributes and thus has potential for use in product development and quality control.     

Risk of cross-contamination due to the use of antimicrobial medicated feed throughout the trail of feed from the feed mill to the farm

The cross-contamination of non-medicated feed with residues of antimicrobials causes an animal and public health concern associated with the potential for the selection and dissemination of resistance in commensal bacteria and potentially zoonotic bacteria. To identify the extent of this situation, we built a risk model that provides a way to estimate the percentage of cross-contaminated feed in total and at the different levels at which cross-contamination may occur (i.e. the feed mill, the transport truck, the farm), for different levels of antimicrobial medicated feed produced in a country per year. The model, estimated that when antimicrobial medicated feed represents a hypothetical x_i = 2% of the total feed produced in a country per year, then 5.5% (95% CI = 3.4%; 11.4%) of the total feed produced in a year could be cross-contaminated with different levels of antimicrobials due to practices related to medicated feed. In detail, 1.80% (95% CI = 0.2%; 7.7%) of the total feed produced in such a country would be cross-contaminated due to antimicrobial carryover occurring at the feed mill level, 1.83% (95% CI = 1.3%; 2.0%) at the transport truck level and 1.84% (95% CI = 1.2%; 2.0%) at the farm level. The model also demonstrated that even in cases where antimicrobial medicated feed would be produced in end-of-line mixers or a fine dosing system on trucks, the risk of cross-contamination would not be negligible; the percentage of cross-contaminated feed produced in a country (where x_i = 2%) per year would be 3.7% (95% CI = 2.9%; 4.0%) and 2.4% (95% CI = 1.6%; 2.7%), respectively. It is hard to reduce the risk to zero as it is the result of factors occurring at different levels. Thus, the use of antimicrobial medicated feed should be avoided as much as possible to reduce selection pressure.     

Effect of water and monoglyceride concentration on the behaviour of monoglyceride containing fat systems

The effect of water and monoglycerides on the structural behaviour of a selected fat system was investigated to explore their role in structure development of products like margarine and sponge cake. Firstly, the monoglyceride and triglyceride system was selected based on solid fat content (SFC) determination. This system was further investigated by differential scanning calorimetry (DSC) and polarized light microscopy (PLM). Secondly, the role of water concentration in this system was investigated by SFC-determination and hardness measurements at different water concentrations. These investigations demonstrate clearly the important role of water, especially at lower concentrations. Finally, the influence of monoglycerides on droplet size distribution in margarines and on textural properties of sponge cakes was investigated. These investigations reveal a larger droplet size of monoglyceride containing margarines and a more dense and harder texture of sponge cakes containing monoglycerides.     

A study on the use of empirical models to predict the formation of acrylamide in potato crisps

The formation of acrylamide in potato crisps was fitted by empirical mathematical models. Potato slices were fried under the same experimental conditions for different times. Besides the content of precursors in the raw potato slices, acrylamide and water content in the potato crisps were quantified after predetermined times (2-6 min). The temperature developments in the surrounding oil and outer cell layer of the potato slices were monitored, giving more insight in the frying process and making future comparisons between studies possible. The pattern found for the formation of acrylamide, which was similar to earlier studies, was fitted to three empirical models. Statistical methods were used to compare the performance of the models, with the "Logistic-Exponential" and "Empirical" model performing equally well. The obtained model parameters were in the range of earlier reported studies, although this comparison is not unequivocal as the experimental conditions differed between studies. The precision of parameter estimates was problematic; this should be improved by better experimental design. Nevertheless, the approach of this study will make it possible to truly compare acrylamide formation patterns and model parameters in the future, with the ability to develop a tool to predict acrylamide formation in potato crisps.     

Nocturnal tidal barrier management improves glass eel migration in times of drought and salinization risk

Adjusted tidal barrier management (ATBM, setting tidal doors ajar during tidal rise) is currently applied in Belgium to improve glass eel passage through tidal gates. However, salt intrusion in the receiving waters due to upwelling and accumulation of saline groundwater as a result of intensive drought put severe pressure on the unrestrained use of this cost-efficient mitigation measure. We evaluated the efficacy of a very restricted ATBM at a tidal barrier on a small canalized waterway, located 3.5 km from the sea: one out of seven gate doors was left 20 cm ajar for ca. 30 min when the water levels on both sides of the tidal barrier were approximately equal. During 20 inflow events differing in tidal flow (i.e., flood or ebb tide) and daytime (i.e., day or night) in March/April 2019, migrating glass eels were caught with a fyke net fixed on the temporally opened gate. In total, 12,853 glass eels were captured, almost all during flood tides at night (96%), with a maximum catch of 3,827 individuals per inflow event. These results show that even small windows of opportunity can help glass eels pass tidal barriers.     

Catalytic One‐Pot Oxetane to Carbamate Conversions: Formal Synthesis of Drug Relevant Molecules

Oxetanes are versatile building blocks in drug‐related synthesis to induce property‐modulating effects. Whereas related oxiranes are widely used in coupling chemistry with carbon dioxide (CO₂) to afford value‐added commodity chemicals, oxetane/CO₂ couplings remain extremely limited despite the recent advances in the synthesis of these four‐membered heterocycles. Here we report an effective one‐pot three‐component reaction (3CR) strategy for the coupling of (substituted) oxetanes, amines and CO₂ to afford a variety of functionalized carbamates with excellent chemoselectivity and good yields. The process is mediated by an aluminium‐based catalyst under relatively mild conditions and the developed catalytic methodology can be applied to the formal synthesis of two pharmaceutically relevant carbamates with the 3CR being a key step.