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Johan Debevere

Publications and source records attributed to Johan Debevere.

At least 19 recordsLinked to original sources

Computer aided boar semen motility analysis for cereulide detection in different food matrices.

Computer Aided Semen Analysis (CASA) study of the boar semen motility has been demonstrated to be an appropriate assay for detection of cereulide (Bacillus cereus emetic toxin). Application of the boar semen bio-assay to detect cereulide directly in foods requires investigation of potential interference of food components, preservatives and other microbial and chemical food contaminants with the bio-assay. Current study provides evidence that none of included Staphylococcus aureus enterotoxins A, B, C and D nor B. cereus Hemolysin BL (HBL) and non-hemolytic enterotoxin (NHE) and three mycotoxins (Sterigmatocystin, Fumonisin B1 and Patulin) exhibited a toxic impact on semen progressive motility. Aflatoxin M1, M3 and zearalenone impaired semen motility only at concentrations (0.004 mg ml(-1), 0.1 mg ml(-1) and 10 mg ml(-1), respectively) much higher than those found in foods and those permitted by legislation, in comparison to cereulide which induces motility cease at concentrations lower than 20 ng ml(-1). Ten commonly used preservatives, namely potassium sorbate, sodium benzoate, (DL) malic acid, citric acid, (L+) tartaric acid, acetic acid, (DL) lactic acid, (L+) ascorbic acid, sodium chloride and sucrose induced no cease in spermatozoa motility even at preservative concentrations higher than permitted by legislation. Dioxins, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and acrylamide had no acute effect on spermatozoa motility at concentrations of 500 and 10,000 mg ml(-1), respectively. Robustness of computer aided boar semen motility analysis, tested with 14 different foods inoculated with cereulide producing B. cereus, showed distinct cereulide production in seven samples (although B. cereus growth to counts higher than 8 log CFU g(-1) was noted in 11 samples), in amounts close to those reported in foodborne outbreaks. Test evaluation in 33 samples suspected to hold cereulide showed actual cereulide presence in ten samples and no interference of food matrix with the assay.

Animals↗

Growth kinetics of cultures from single spores of Aspergillus flavus and Fusarium verticillioides on yellow dent corn meal.

A dilution protocol originally developed for the isolation of single bacterial cells was modified to suite the specificities of fungal growth. The modified protocol was used to study the growth kinetics of single spores of Aspergillus flavus and Fusarium verticillioides on yellow dent corn meal. Both a(w) and temperature significantly influenced the distributions of the colony growth rates and lag phases and the rate at which individual spores of both isolates completed the lag period. An interaction between a(w) and temperature was noted on the spread of the distributions of these growth parameters. The histograms of the single spore colony growth rates and lag phases generally became wider the more compromising the conditions for growth became, indicating a greater variation in growth ability at these conditions. The rate at which the single spores passed through the lag phases generally decreased with decrease in temperature and/or a(w), with an interaction again noted between these two factors on the rate. These results show the potential range and variability in growth of individual fungal spores at the lowest inoculum level possible.

Aspergillus flavus↗

The efficacy of electrolysed oxidising water for inactivating spoilage microorganisms in process water and on minimally processed vegetables.

The efficacy of Electrolysed Oxidising Water (EOW) for inactivating spoilage microorganisms in process water and on minimally processed vegetables was investigated. The direct effect of EOW on three important spoilage bacteria namely; Pseudomonas fluorescens, Pantoea agglomerans or Rahnella aquatilis was determined by inoculating tap water or "artificial process water" with approximately 8 log CFU/ml pure culture and electrolysing the resultant solutions. The three bacteria were each reduced to undetectable levels at low (0.5 A) and relatively higher levels (1.0 A) of current in tap water and "artificial process water", respectively. The residual effect of EOW on P. fluorescens, P. agglomerans or R. aquatilis was determined by incubating at room temperature 1 ml (approximately 9 log CFU/ml) pure culture suspensions in 9 ml of EOW-T (EOW produced from tap water), EOW-A (EOW produced from "artificial process water" supplemented with approximately 60.7 mg Cl(-)/l and 39.3 mg Na(+)/l) or deionised water (control) for 0, 15, 45 or 90 min. The bactericidal activity of both EOW-T and EOW-A increased with the concentration of free oxidants and incubation period and the three bacteria were completely reduced at free oxidants-incubation period combinations of 3.88 mg/l-45 min and 5.1 mg/l-90 min in EOW-T and EOW-A, respectively. Two types of industrial vegetable process water; salad-mix and soup process water, which had each a total psychrotrophic count of approximately 8 log CFU/ml were then electrolysed. Without any NaCl addition, only 1.2 and 2.1 log reductions of the psychrotrophs in soup and salad-mix process water was attained respectively. Supplementation of the process water with approximately 60.7 mg Cl(-)/l and 39.3 mg Na(+)/l afterwards resulted in complete reduction of the psychrotrophic count in both process waters, but soup process water required relatively higher levels of current compared to salad-mix water. Finally, fresh-cut lettuce was washed in EOW-T containing 3.62 mg free oxidants/l, EOW-IP (EOW produced from industrial process water) containing 2.8 mg free oxidants/l or tap water (control) for 1 or 5 min. Washing the vegetables for 1 min in EOW-T resulted in 1.9, 1.2, and 1.3 log reductions of psychrotrophs, lactic acid bacteria and Enterobacteriacae, respectively, which increased to 3.3, 2.6, and 1.9 log reductions after washing for 5 min instead. EOW-IP tested in this work had no bactericidal effect on the microflora of fresh-cut lettuce. Electrolysis could therefore be used to decontaminate process water for vegetable pre-washing and to sanitise tap water for final rinsing of vegetables, respectively.

Colony Count, Microbial↗

Influence of experimental parameters on the fluorescence response and recovery of the high-performance liquid chromatography analysis of fumonisin B1.

The effect of mobile phase pH, column temperature, extraction method and derivatisation time on the fluorescence response and recovery of fumonisin B1 in corn was investigated. Column temperature and mobile phase pH were negatively and positively correlated with the fluorescence response, respectively. Use of an Ultraturrax blender for extraction resulted in higher FB1 recoveries compared to a rotary shaker. In contrast to other reports, maximum fluorescence response occurred after a derivatisation time of 8 min. These results reflect on the absolute requirement for standardisation of the aforementioned parameters.

Chromatography, High Pressure Liquid↗

Development of a real-time NASBA assay for the detection of Campylobacter jejuni cells.

The objectives of this study were the development of a real-time NASBA assay for the detection of Campylobacter jejuni mRNA and the evaluation of its potential to determine the viability of the detected C. jejuni cells. A set of specific primers and probes was chosen to amplify the mRNA of the tuf-gene and the GTPase-gene. Only the tuf-assay was able to detect as low as 10(2) cells per NASBA reaction and was specific for Campylobacter. However, as the assay was able to detect dead cells, it cannot be used to demonstrate the viability of C. jejuni cells. The tuf-gene mRNA is no good viability indicator due to its stability.

Base Sequence↗

Immunoquantitative real-time PCR for detection and quantification of Staphylococcus aureus enterotoxin B in foods.

A real-time immunoquantitative PCR (iqPCR) method for detection of Staphylococcus aureus enterotoxin B (SEB) was developed and evaluated using both pure cultures and foods. The assay consisted of immunocapture of SEB and real-time PCR amplification of the DNA probe linked to the detection antibody. iqPCR was compared to an in-house enzyme-linked immunosorbent assay (ELISA) using the same couple of capture-detection antibodies and to commercial kits for detection of S. aureus enterotoxins (SE). The iqPCR was approximately 1,000 times more sensitive (<10 pg ml(-1)) than the in-house ELISA and had a dynamic range of approximately 10 pg ml(-1) to approximately 30,000 pg ml(-1). iqPCR was not inhibited by any of the foods tested and was able to detect SEB present in these foods. No cross-reactivity with SE other than SEB was observed. Application of iqPCR for detection of SEB in cultures of S. aureus revealed the onset of SEB production after 4 h of incubation at 22, 37, and 42 degrees C, which was in the first half of the exponential growth phase. The total amounts of SEB produced by the two strains tested were larger at 42 degrees C than at 37 degrees C and were strain dependent.

Bacterial Toxins↗

Influence of type of food on the kinetics and overall production of Bacillus cereus emetic toxin.

Potato puree and penne pasta were inoculated with cereulide producing B. cereus 5964a and B. cereus NS117. Static incubation at 28 degrees C proved these two foods to be a better substrate for higher cereulide production (4,080 ng/g in puree and 3,200 ng/g in penne were produced by B. cereus 5964a during 48 h of incubation) compared with boiled rice (2,000 ng/g). This difference occurred despite B. cereus counts of more than 10(8) CFU/g in all three products. Aeration of cultures had a negative effect on cereulide production, causing concentrations more than 10-fold lower than in some statically incubated samples. Cereulide production remained undetectable in shaken milk, whereas it reached 1,140 ng/ml in statically incubated milk. At 12 and 22 degrees C, presence of background flora was also a determinative factor. A total B. cereus count of more than 106 CFU/ml did not necessarily lead to uniform cereulide production and was also dependent on the B. cereus strain involved. In this study, we confirm that a number of factors play a crucial role in the determination of the extent to which, if at all, cereulide will be produced. Among those, type of the food, temperature, pH, and whether additional aeration (via incubation on an orbital shaker) is induced had an important role. An important effect was also induced by the cereulide-producing strain involved.

Bacillus cereus↗

Influence of eggshell condensation on eggshell penetration and whole egg contamination with Salmonella enterica serovar enteritidis.

Shells of agar-filled and whole eggs were inoculated with 10(3) to 10(4) CFU of Salmonella enterica serovar Enteritidis per eggshell. The agar-filled eggs were used to study bacterial eggshell penetration, and the whole egg results were used to characterize contamination of the egg contents. In each group, half of the eggs were stored for 21 days at 20 degrees C and 60% relative humidity (RH), and the other half was stored for 24 h at 6 degrees C and then for 20 days at 20 degrees C. The latter conditions resulted in condensation on the eggshell for 30 min from the moment the eggs were placed in the 20 degrees C chamber. Taking into account the ages at which hens were studied (39, 53, and 67 weeks), an average of 62% of the eggshells with condensate were penetrated compared with 43% for the control group; this difference was significant (P < 0.01). No significant difference in whole egg contamination was found; 18% of the control eggs were contaminated compared with 22% of the condensate eggs. Whole egg contamination was significantly higher for eggs from the hens at an older age (67 weeks). This difference probably was not due to a higher penetration potential because differences were not observed for the corresponding agar-filled eggs. Condensation on the eggshell seemed to encourage bacterial penetration of the eggshell but had a smaller impact on whole egg contamination.

Age Factors↗

Dynamics of boar semen motility inhibition as a semi-quantitative measurement of Bacillus cereus emetic toxin (Cereulide).

Qualitative and quantitative application of a computer assisted sperm analyzer (CASA) for detection and quantification of cereulide was described. The plot of the decrease of the percentage of boar semen progressive motility (PMOT%) in function of time and the visual inspection of curves provided a qualitative comparison between different samples (curve slope corresponds to the amount of cereulide in the sample). If the change of PMOT% over a time required for achieving PMOT% drop to 10% (DeltaPMOT%/Deltatau) is plotted against the standard curve (obtained with known concentrations of valinomycin), a semi-quantitative estimation of the amount of cereulide in the sample is obtained. An optimized CASA method was applied to determine the production of cereulide under different conditions. No cereulide was found in aerated samples and in samples incubated at 12 degrees C. The amount of cereulide produced depended on the agar medium used, type of Bacillus cereus strain and the amount of oxygen present in the atmosphere.

Animals↗

Effect of water activity and temperature on growth and the relationship between fumonisin production and the radial growth of Fusarium verticillioides and Fusarium proliferatum on corn.

The two major fumonisin-producing Fusarium species are Fusarium verticillioides and Fusarium proliferatum. The growth and fumonisin production of these two isolates on corn was studied at water activities (a(w)) between 0.860 and 0.975 and at temperatures between 15 and 30 degrees C. Growth rates (g, mm/day) were obtained by linear regression during the linear phase of growth. In general, growth rates for both isolates increased significantly (P < 0.05) with increases in a(w) and temperature. Both fumonisin production and radial growth (mycelial development) for both isolates increased with a(w) at all temperatures investigated, but the effect of temperature on this relationship was not obvious. The effect of temperature on fumonisin production at high a(w) values optimal for growth was only marginal, whereas at lower a(w) values the effect of temperature was more pronounced, with more fumonisin production occurring at temperatures not optimal for growth. The optimum temperature for fumonisin production was between 15 and 25 degrees C. For F. proliferatum, the optimum temperature for growth at all a(w) values, 30 degrees C, resulted in the poorest fumonisin production. For both isolates, the slowest initial rate of fumonisin production was at 15 degrees C, the temperature at which the slowest growth rates were obtained.

Animals↗

Concepts and tools for predictive modeling of microbial dynamics.

Description of microbial cell (population) behavior as influenced by dynamically changing environmental conditions intrinsically needs dynamic mathematical models. In the past, major effort has been put into the modeling of microbial growth and inactivation within a constant environment (static models). In the early 1990s, differential equation models (dynamic models) were introduced in the field of predictive microbiology. Here, we present a general dynamic model-building concept describing microbial evolution under dynamic conditions. Starting from an elementary model building block, the model structure can be gradually complexified to incorporate increasing numbers of influencing factors. Based on two case studies, the fundamentals of both macroscopic (population) and microscopic (individual) modeling approaches are revisited. These illustrations deal with the modeling of (i) microbial lag under variable temperature conditions and (ii) interspecies microbial interactions mediated by lactic acid production (product inhibition). Current and future research trends should address the need for (i) more specific measurements at the cell and/or population level, (ii) measurements under dynamic conditions, and (iii) more comprehensive (mechanistically inspired) model structures. In the context of quantitative microbial risk assessment, complexity of the mathematical model must be kept under control. An important challenge for the future is determination of a satisfactory trade-off between predictive power and manageability of predictive microbiology models.

Bacteria↗