In vitro fermentation characteristics of diets with different forage/concentrate ratios: comparison of rumen and faecal inocula

BACKGROUND: The aim of this trial was to evaluate the replacement of rumen fluid with faeces as inoculum in studying the in vitro fermentation characteristics of diets for ruminants using the in vitro gas production technique. Six iso‐protein diets with different forage/concentrate ratios were incubated with rumen fluid (RI) or faeces (FI) collected from sheep. RESULTS: Most of the fermentation parameters were influenced by diet and inoculum (P < 0.01). With both inocula, organic matter degradability (dOM), cumulative gas production (OMCV) and maximum fermentation rate (R_max) increased as the amount of concentrate in the diet increased. R_max was lower with FI vs RI (P < 0.01); dOM was higher with FI vs RI and the diet × inoculum interaction was significant. As expected, with both inocula, R_max increased as the neutral detergent fibre content of the diet decreased. Significant correlations were obtained using both inocula between OMCV/dOM and gas/volatile fatty acid (VFA), while the correlation VFA/dOM was significant only with FI. The microbial biomass yield calculated by stoichiometric analysis for all diets was higher with FI vs RI. With FI the organic matter used for microbial growth showed an overall decreasing trend as the amount of concentrate in the diet increased. CONCLUSION: The results indicate that both faeces and rumen fluid from sheep have the potential to be used as inoculum for the in vitro gas production technique. Copyright © 2011 Society of Chemical Industry     

Quality control of Aspergillus flavus and A. parasiticus agar and comparison with dichloran 18% glycerol agar: a collaborative study

AFPA culture medium, which is used for recognition of Aspergillus flavus and A. parasiticus, has been validated in a collaborative study including nine laboratories located in Australia, Brazil, Denmark, The Netherlands, Sweden and United Kingdom. Three freeze-dried fungal mixtures, containing A. flavus/A. parasiticus and background fungi, were produced and checked for homogeneity. The coefficients of variance were low, ranging from 0.81% to 1.09% for total fungal counts and between 2.50% and 2.72% for counts of A. flavus/A. parasiticus. The laboratories analysed the contents of two vials of each mixture on commercial A. flavus and A. parasiticus agar (AFPA), in-house-made AFPA, and on a standard media, dichloran 18% glycerol agar (DG18). Reproducibility values for counts of A. flavus/A. parasiticus indicated no differences between the commercial AFPA and the in-house-made AFPA. Variation between laboratories was low, indicating that the medium was effective in use. Reproducibility values for DG18 were higher. There were no differences in counts of A. flavus/A. parasiticus on AFPA and DG18. However, DG18 gave slightly higher total fungal counts compared to AFPA.     

Rheology and breadmaking performance of rice-buckwheat batters supplemented with hydrocolloids

The aim of this work was to investigate the effects of hydrocolloid addition on rheological properties and breadmaking performance of rice-buckwheat batter at different water levels. Xanthan gum (XG) and propylene glycol alginate (PGA) were added to rice-buckwheat blend (60:40) at levels of 0.5–1.5%. Batter rheological properties were investigated using dynamic measurements in the linear viscoelastic range (frequency sweep and time cure tests). The addition of both hydrocolloids significantly enhanced the storage modulus (G′) of batter. XG exerted greater effect on G′ than PGA. Different effects on starch gelatinisation were observed for the two hydrocolloids. PGA breads showed higher improvement in terms of increased specific volume (V_s), decreased crumb firmness and crumb structure than XG breads. Different technological behaviours were explained on the basis of batter rheological properties.     

Effect of coffee physical structure on volatile release

The mechanisms of volatile release from soluble coffee powders with different roasting degrees were studied. The presence of volatiles in the headspace during coffee humidification was analyzed by gas chromatography. Small amounts of volatiles were observed at low water activities (a_w), independently from the roasting degree; as the a_w increased headspace volatiles rose and then decreased as the moisture further increased. The changes in a_w and volatiles went along with the changes of coffee structure from a free-flowing powder to a sticky viscous fluid. The mechanism of volatiles release was controlled by both kinetic and thermodynamic factors. The former prevailed at low a_w in glassy systems; while the latter became important at high a_w when, due to increased mobility, equilibrium conditions were approached. Modified state diagrams were used to predict the critical temperature and a_w at which structural collapse and volatile release occurred. As far as equilibrium condition was achieved, coffee volatiles were partially re-adsorbed in the liquid phase.     

Aqueous photochemical reaction kinetics and transformations of fluoxetine

Fluoxetine (FLX) was shown to be photoreactive in sunlit surface waters. FLX degraded in deionized water when exposed to simulated sunlight with a half-life of 55.2+/-3.6 h(-1). Photodegradation products were identified using high performance liquid chromatography-UV (HPLC-UV) and liquid chromatography tandem mass spectrometry (LC-MS-MS) using electrospray (ES) ionization. Defluorination of the trifluoromethyl group in FLX and in fluometuron and flutalanil,two other compounds containing this functional group, is suggested to be a common direct photolysis pathway for trifluoromethylated compounds. Products resulting from O-dealkylation of FLX were also observed. The rate of degradation was faster in synthetic field water where .OH was the likely dominant oxidant in the system. The bimolecular rate constant for the reaction between FLX and .OH was measured as (8.4+/-0.5) x 10(9) and (9.6 +/-0.8) x 10(9) M(-1) s(-1) using two different methods of competition kinetics. Indirect photodegradation reactions could lead to the production of hydroxylated and O-dealkylated compounds. Although direct photolysis could potentially limitthe persistence of FLX in surface waters, its degradation by indirect photolysis would proceed faster. Thus, this latter process could be important in the elimination of FLX in surface waters.     

Furfurals removal from roasted coffee powder by vacuum treatment

The possibility of removing 5-hydroxymethylfurfural (HMF) and furfural from roasted coffee by application of vacuum treatments was studied. In particular, different combinations of pressure and time were used. Results showed that the vacuum treatment was ineffective in removing HMF and furfural from anhydrous (commercial) coffee powder, while it proved effective if applied to previously hydrated samples. Besides, the hydration step alone was responsible for furfurals removal, although prolonged times were needed. By applying a hydration step up to a_w 0.7, followed by a vacuum treatment at 2.7 kPa and 60 °C for 10 min, 20% and 100% HMF and furfural removals were achieved, respectively. These differences in the percentages of HMF and furfural removal can be attributed to differences in the chemical and physical properties of the two molecules. However, the vacuum treatments caused a significant decrease in the coffee headspace total volatiles that in turn was responsible for a lower odour intensity of the samples. Therefore further studies are needed to reveal process conditions able to minimise the loss of sensory properties, making this technology a reliable strategy to mitigate the furfural contents in coffee.     

Evaluation of <Emphasis Type=Italic>flaA</Emphasis> Short Variable Region Sequencing,Multilocus Sequence Typing,and Fourier Transform Infrared Spectroscopy for Discrimination between <Emphasis Type=Italic>Campylobacter jejuni</Emphasis> Strains

Discriminatory and robust typing methods are needed to improve the understanding of the dynamics of food-borne Campylobacter infections and epidemiology in primary animal production. To evaluate the strain discriminatory potential of typing methods, flaA short variable region (SVR) sequencing and Fourier transform infrared (FTIR) spectroscopy were applied on a collection of 102 epidemiologically related and unrelated Campylobacter jejuni strains. Previous application of FTIR spectroscopy for subtyping of Campylobacter has been limited. A subset of isolates, initially discriminated by flaA SVR sequencing, were further subjected to multilocus sequence typing (MLST). It was found that flaA SVR sequencing had a slightly higher discriminatory power than FTIR spectroscopy, based on the Simpson diversity index. The clustering of strains indicated that FTIR spectroscopy is indeed a suitable method for discrimination of Campylobacter. The isolates were assigned to six clusters based on flaA SVR sequences and nine clusters based on the FTIR spectroscopy profiles. Furthermore, the cluster analysis of flaA SVR sequences, MLST, and FTIR spectroscopy profiles showed a high degree of congruence, assigning the isolates to similar cluster structures. In conclusion, FTIR spectroscopy can be applied for subtyping of Campylobacter, and the high discriminatory potential of both flaA SVR sequencing and FTIR spectroscopy render them suitable screening methods for large numbers of strains.     

Interlaboratory comparison of real-time PCR protocols for quantification of general fecal indicator bacteria

The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized protocol requires information on the reproducibility and sources of variation associated with qPCR methodology across laboratories. This study examines interlaboratory variability in the measurement of enterococci and Bacteroidales concentrations from standardized, spiked, and environmental sources of DNA using the Entero1a and GenBac3 qPCR methods, respectively. Comparisons are based on data generated from eight different research facilities. Special attention was placed on the influence of the DNA isolation step and effect of simplex and multiplex amplification approaches on interlaboratory variability. Results suggest that a crude lysate is sufficient for DNA isolation unless environmental samples contain substances that can inhibit qPCR amplification. No appreciable difference was observed between simplex and multiplex amplification approaches. Overall, interlaboratory variability levels remained low (<10% coefficient of variation) regardless of qPCR protocol.     

Bioactive compound content,antioxidant activity,deoxynivalenol and heavy metal contamination of pearled wheat fractions

Wheat kernels are rich in antioxidant compounds, that are mainly present in the outer bran layers and which are removed during milling. Unfortunately, several contaminants, e.g., mycotoxins and heavy metals, are also concentrated in the external layers. Pearling of 3 wheat varieties gave five fractions (each 5% of the original grain weight), starting from the outer layer until the inner kernel, designated as 0–5%, 5–10%, 10–15%, 15–20%, 20–25%, respectively. The remaining 75% of the inner kernel was also collected. Dietary fibre, free phenolic acids and total antioxidant activity decreased progressively from the external to the internal layers. However, the 5–10% fraction was richer in β-glucan content than the external one (0–5%). Heavy metals were only found in the most external fraction. Deoxynivalenol contamination decreased from the external to the internal layers: 64% of total contamination of kernel was found in the 0–5% and 5–10% fractions. The 10–15% kernel fraction offered the best compromise between high nutritional value and low contamination risk.     

A comparative study of the purification of betanin

Betanin is a natural pigment with antioxidant properties used as a food colourant. This work describes the spectrophotometric and chromatographic quantification of betanin (2S/15S) and its epimer isobetanin (2S/15R) in fresh beetroot juice, food-grade beetroot powder and betanin standard diluted in dextrin. Absorption spectra of all three samples were deconvoluted using a mixed three-function model. Food-grade beetroot powder has the largest amount of violet-red impurities, probably formed during processing. The purification of betanin from these complex matrices was carried out by seven different methods. Ion exchange chromatography was the most efficient method for the purification of betanin from all samples; however, fractions contain high amounts of salt. Reversed-phase HPLC as well as reversed-phase column chromatography also produced good results at a much faster rate. The longer retention time of isobetanin when compared to betanin in reversed-phase conditions has been investigated by means of quantum-mechanical methods.