Furan in coffee: pilot studies on formation during roasting and losses during production steps and consumer handling.

The occurrence of furan in some food products has already been known for a few decades, and it has been reconfirmed in more recent investigations that furan is present in a variety of foodstuffs. This list of products includes roasted coffee, which has been shown to generate furan as a result of the heat treatment at roasting which is applied to achieve the desired aroma and flavour profile of a roasted coffee. The objective of this study is to provide data to allow a better understanding of the available data of furan in coffee, the kinetics of furan generated during roasting, and to estimate the reduction of furan levels afterwards due to subsequent processing steps and consumer handling. Finally, the study is meant as a contribution to establish exposure data on the basis of scientific data at the stage of coffee consumption. This paper shows that the formation of furan during roasting is dependent on roasting conditions and is, therefore, directly linked to achieving targeted flavour profiles. Furthermore, it is demonstrated that modifications in process conditions potentially to reduce furan levels may have the opposite effect on other undesired reaction products of the roasting chemistry such as, for example, acrylamide. Due to the high volatility of furan, any subsequent processing step or consumer handling has an impact on the level of furan. As a guidance from this study and in consideration of the identified losses of each process and handling step on the basis of the trial conditions, it is estimated that only approximately 10% of the initially generated furan during roasting gets into the cup of coffee for consumption.

Effect of administration of Lactobacillus crispatus, Clostridium lactatifermentans and dietary lactose on the development of the normal microflora and volatile fatty acids in the caeca of broiler chicks.

1. Lactobacillus crispatus and Clostridium lactatifermentans, both isolated from the caeca of chickens, grown together in an in vitro model system are able to ferment lactose to acetate and propionate. In this study, the capabilities of these organisms were studied in vivo. 2. The effect on concentrations of volatile fatty acids and lactate, together with the development of some bacterial groups in the caeca of chicks, was studied after oral inoculation with L. crispatus and C. lactatifermentans together with dietary lactose. For this purpose, chicks were divided into 4 groups: (i) control group, (ii) dietary lactose, (iii) L. crispatus and C. lactatifermentans, and (iv) dietary lactose together with L. crispitaus and C. lactatifermentans. 3. In general, concentrations of (undissociated) volatile fatty acids in the caeca were not significantly different in broilers receiving both bacteria and dietary lactose compared with control broilers. Concentrations of lactate in the caeca of 14-d-old broilers treated with any of the three treatments were significantly higher than in the caeca of control broilers. 4. This indicates that L. crispatus or other lactate-producing organisms were responsible for the elevated concentrations of lactic acid. Clostridium lactatifermentans has probably not colonised the caeca sufficiently to ferment this lactate further to acetate and propionate. 5. Numbers of Enterobacteriaceae and enterococci in the caeca of broilers receiving both bacteria and dietary lactose were not different from control broilers. 6. We conclude from these results that under the conditions applied in this study a mixture of L. crispatus and C. lactatifermentans with dietary lactose was able to increase lactate concentrations but was unable to increase concentrates of acetate and propionate in the caeca of broiler chicks.

Spatial and temporal variation of the intestinal bacterial community in commercially raised broiler chickens during growth.

The objective of this study was to determine whether host, compartment, or environmental specific factors play an important role in the establishment of the intestinal microflora in broiler chickens during growth. This objective was addressed using a 16S rDNA approach. PCR-amplicons from the V6 to V8 regions of the 16S rDNA of intestinal samples were separated by denaturing gradient gel electrophoresis (DGGE). The number of bands in all intestinal compartments increased when broilers grew older, indicating that the dominant bacterial community becomes more complex when chickens age. Each chicken had a unique banding pattern for all locations in the intestinal tract, irrespective of the age of chickens. This suggests that host-related factors affect the establishment of the dominant bacterial community. Banding patterns of intestinal compartments within one chicken were different from each other for broilers older than 4 days, except for both ceca which were highly similar. In 4-day-old broilers, banding patterns from crop, duodenum, and ileum were very similar. We conclude that (unknown) host specific factors play an important role in the development of the intestinal bacterial community in each broiler chicken. Furthermore, compartment-specific factors play an important role in the bacterial development of each intestinal compartment within one chicken.

Monitoring of transmission of Salmonella enterica serovars in pigs using bacteriological and serological detection methods.

The standard method to detect Salmonella positive pigs is bacteriological examination of the faeces, but in recent years the use of Salmonella-ELISA's have become available to screen pigs for serological evidence of infection. This study was conducted to monitor the transmission of five different Salmonella enterica serovars (S. Typhimurium, S. Brandenburg, S. Panama, S. Livingstone, and S. Goldcoast) in fattening pigs and to test the feasibility of Salmonella-ELISA, using seeder pigs as a mode of transmission. Serovar dependence in transmission was observed. The Salmonella-ELISA proved to be useful to detect S. Typhimurium and S. Brandenburg in herds but was of limited value to demonstrate S. Livingstone, S. Goldcoast, and S. Panama.

Survival of Escherichia coli O157:H7 ATCC 43895 in a model apple juice medium with different concentrations of proline and caffeic acid.

The effects of proline and caffeic acid on the survival of Shiga toxin-producing Escherichia coli (STEC) O157:H7 strain ATCC 43895 in a model apple juice medium were studied. It is hypothesized that the inhibitory effect of caffeic acid may explain why almost all outbreaks of STEC O157:H7 infections linked to apple juice or cider have occurred in October or November.

Inhibition of a glucose-limited sequencing fed-batch culture of Salmonella enterica serovar enteritidis by volatile fatty acids representative of the ceca of broiler chickens.

The effects of concentrations of volatile fatty acids on an anaerobic, glucose-limited, and pH-controlled growing culture of Salmonella enterica serovar Enteritidis were studied. Suddenly increasing volatile fatty acids to the concentrations representative of the ceca of 15-day-old broiler chickens caused washout of serovar Enteritidis. In contrast, a sudden increase to the volatile fatty acid concentrations representative of the ceca of younger broiler chickens caused a reduction in the biomass but not washout. Gradually increasing volatile fatty acids caused a gradual decrease in the biomass of serovar Enteritidis. We conclude that the concentrations of volatile fatty acids present in the ceca of broilers with a mature microflora can cause washout of serovar Enteritidis in an in vitro system mimicking cecal ecophysiology.

Development of the Specific and Random Amplification (SARA)-PCR for both species identification of enterococci and detection of the vanA gene.

A rapid and reliable polymerase chain reaction (Specific and Random Amplification (SARA)-PCR) has been developed to identify enterococcal species and to detect the vanA gene in one single reaction. This technique was based on the use of the primer set previously designed to amplify a part of the vanA gene (Dutka-Malen et al., J. Clin. Microbiol., 33 (1995) 24-27). In the chosen low stringency conditions complex patterns were obtained, with a sharp band of approximately 700 bp in cases where the vanA gene was present. Discrimination at the species and strain level was achieved by applying the SARA-PCR assay to a collection of 55 enterococcal isolates and type strains. Simultaneously the vanA gene was detected in all strains showing high resistance to vancomycin.

Role of volatile fatty acids in development of the cecal microflora in broiler chickens during growth.

It is known that volatile fatty acids can inhibit growth of species of the family Enterobacteriaceae in vitro. However, whether these volatile fatty acids affect bacterial populations in the ceca of chickens is unknown. Therefore, a study was conducted to investigate if changes in volatile fatty acids in ceca of broiler chickens during growth affect bacterial populations. Results showed that members of the Enterobacteriaceae and enterococci are present in large numbers in 3-day-old broilers and start to decrease when broilers grow older. Lactobacilli are present in large numbers as well in 3-day-old broilers, but they remain stable during the growth of broilers. Acetate, butyrate, and propionate increase from undetectable levels in 1-day-old broilers to high concentrations in 15-day-old broilers, after which they stabilize. Significant negative correlations could be calculated between numbers of Enterobacteriaceae and concentrations of undissociated acetate, propionate, and butyrate. Furthermore, pure cultures of Enterobacteriaceae isolated from the ceca were grown in the presence of volatile fatty acids. Growth rates and maximal optical density decreased when these strains grew in the presence of increasing volatile fatty acid concentrations. It is concluded that volatile fatty acids are responsible for the reduction in numbers of Enterobacteriaceae in the ceca of broiler chickens during growth.

Regulation of Product Formation in Bacteroides xylanolyticus X5-1 by Interspecies Electron Transfer.

Bacteroides xylanolyticus X5-1 was grown in pure culture and in mixed culture with Methanospirillum hungatei JF-1 under xylose limitation in the chemostat. In the pure culture, ethanol, acetate, CO(2), and hydrogen were the products. In the mixed culture, acetate, CO(2), and presumably hydrogen were the only products formed by B. xylanolyticus X5-1. The biomass yield of B. xylanolyticus X5-1 increased because of cocultivation. In cell extracts of the pure culture, both NAD- and NADP-dependent acetaldehyde dehydrogenase and ethanol dehydrogenase activities were found. In cell extracts of the mixed culture, activities of these enzymes were not detected. Inhibition of methanogenesis in the mixed culture by the addition of bromoethanosulfonic acid (BES) resulted in an accumulation of H(2), ethanol, and formate. Immediately after the addition of BES, NAD-dependent acetaldehyde dehydrogenase and ethanol dehydrogenase activities were detected. After a short lag phase, a NADP-dependent ethanol dehydrogenase was also detectable. The induction of acetaldehyde dehydrogenase and ethanol dehydrogenase was inhibited by chloramphenicol, suggesting de novo synthesis of these enzymes. These results are consistent with a model in which the shift in product formation caused by interspecies electron transfer is regulated at the level of enzyme synthesis.

Xylose and Glucose Utilization by Bacteroides xylanolyticus X5-1 Cells Grown in Batch and Continuous Culture.

During cultivation on a mixture of xylose and glucose, Bacteroides xylanolyticus X5-1 showed neither diauxic growth nor a substrate preference. Xylose-limited continuous-culture cells were able to consume xylose and glucose both as single substrates and as mixed substrates without any lag phase. When glucose was the growth-limiting substrate, the microorganism was unable to consume xylose. However, in the presence of a small amount of glucose or pyruvate, xylose was utilized after a short lag phase. In glucose-limited cells, xylose isomerase was present at low activity but xylulose kinase activity could not be detected. On addition of a mixture of xylose and glucose, xylose isomerase was induced immediately and xylulose kinase was induced after about 30 min. The induction of the two enzymes was sensitive to chloramphenicol, showing de novo synthesis. Xylose uptake in glucose-grown cells was very low, but the uptake rate could be increased when incubated with a xylose-glucose mixture. The increase in the uptake rate was not affected by chloramphenicol, indicating that a constitutive uptake system had to be activated. The inability of B. xylanolyticus X5-1 cells undergoing glucose-limited continuous culture to induce the xylose catabolic pathway after the addition of only xylose probably was caused by energy limitation.

D-xylose catabolism in Bacteroides xylanolyticus X5-1.

The xylose metabolism of Bacteroides xylanolyticus X5-1 was studied by determining specific enzyme activities in cell free extracts, by following 13C-label distribution patterns in growing cultures and by mass balance calculations. Enzyme activities of the pentose phosphate pathway and the Embden-Meyerhof-Parnas pathway were sufficiently high to account for in vivo xylose fermentation to pyruvate via a combination of these two pathways. Pyruvate was mainly oxidized to acetyl-CoA, CO2 and a reduced cofactor (ferredoxin). Part of the pyruvate was converted to acetyl-CoA and formate by means of a pyruvate-formate lyase. Acetyl-CoA was either converted to acetate by a combined action of phosphotransacetylase and acetate kinase or reduced to ethanol by an acetaldehyde dehydrogenase and an ethanol dehydrogenase. The latter two enzymes displayed both a NADH- and a NADPH-linked activity. Cofactor regeneration proceeded via a reduction of intermediates of the metabolism (i.e. acetyl-CoA and acetaldehyde) and via proton reduction. According to the deduced pathway about 2.5 mol ATP are generated per mol of xylose degraded.