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Biomedical subjects

M L Delignette-Muller

Publications and source records attributed to M L Delignette-Muller.

At least 19 recordsLinked to original sources

Growth and survival of non-O157:H7 Shiga-toxin-producing Escherichia coli in cow manure.

AIMS: The main objective of this study was to evaluate the behaviour of non-O157:H7 Shiga-toxin-producing Escherichia coli (STEC) strains in cow manure. METHODS AND RESULTS: A mixture of eight green-fluorescent-protein-labelled STEC strains was inoculated around 10(6)-10(7) CFU g(-1) into four manure heaps. Two heaps were regularly turned and the two others remained unturned. STEC counts and physical parameters (temperature, pH, moisture content and oxido-reduction potential) were monitored for 1000 manure samples. The highest mean pH values were obtained near the surface at the base of all manure heaps. At the surface, the moisture content decreased from 76.5% to 42% in turned heaps. Temperatures reached 65 degrees C near the main body of all manure heaps, and only 35 degrees C near the superficial parts located at the base of them. These two sites (the centre and the base) were associated with D values for the STEC counts of 0.48 and 2.39 days, respectively. We were able to detect STEC strains during 42 days in turned manure heaps and during at least 90 days in unturned ones. CONCLUSIONS: These results emphasize the long-term survival of non-O157:H7 STEC in cow manure. SIGNIFICANCE AND IMPACT OF THE STUDY: Good management practices (e.g. turning) should be respected in order to minimize the risk of environmental contamination by STEC.

Animals↗

Effect of temperature, water-phase salt and phenolic contents on Listeria monocytogenes growth rates on cold-smoked salmon and evaluation of secondary models.

Salting and smoking are ancient processes for fish preservation. The effects of salt and phenolic smoke compounds on the growth rate of L. monocytogenes in cold-smoked salmon were investigated through physico-chemical analyses, challenge tests on surface of cold-smoked salmon at 4 degrees C and 8 degrees C, and a survey of the literature. Estimated growth rates were compared to predictions of existing secondary models, taking into account the effects of temperature, water phase salt content, phenolic content, and additional factors (e.g. pH, lactate, dissolved CO2). The secondary model proposed by Devlieghere et al. [Devlieghere, F., Geeraerd, A.H., Versyck, K.J., Vandewaetere, B., van Impe, J., Debevere, J., 2001. Growth of Listeria monocytogenes in modified atmosphere packed cooked meat products: a predictive model. Food Microbiology 18, 53-66.] and modified by Giménez and Dalgaard [Giménez, B., Dalgaard, P., 2004. Modelling and predicting the simultaneous growth of Listeria monocytogenes and spoilage micro-organisms in cold-smoked salmon. Journal of Applied Microbiology 96, 96-109.] appears appropriate. However, further research is needed to understand all effects affecting growth of L. monocytogenes in cold-smoked salmon and to obtain fully validated predictive models for use in quantitative risk assessment.

Animals↗

Use of Bayesian modelling in risk assessment: application to growth of Listeria monocytogenes and food flora in cold-smoked salmon.

An attempt to use a Bayesian approach to model variability and uncertainty separately in microbial growth in a risk assessment is presented. It was conducted within the framework of a French project aiming at assessing the exposure to Listeria monocytogenes in cold-smoked salmon. The chosen model describes the effect of time and temperature on bacterial growth. A Bayesian approach close to the one proposed by Pouillot et al. [Int. J. Food Microbiol. 81 (2003) 87] is used to estimate the variability and uncertainty of growth parameters from both literature data and data experimentally acquired during the project. Variability between strains and between products is taken into account. The growth of the food flora of cold-smoked salmon is also modelled by the same method. The results obtained for both models are used to predict the simultaneous growth of L. monocytogenes and food flora in cold-smoked salmon with a competitive model, expressing variability and uncertainty through a second-order Monte Carlo simulation.

Animals↗

Prevalence of Listeria monocytogenes in 13 dried sausage processing plants and their products.

The aims of the present study were: (i) to investigate the occurrence of Listeria monocytogenes in dried sausage processing plants on surfaces before and during processing, (ii) to study the contamination in meat and sausages at different stages of maturation, (iii) to assess the distribution of L. monocytogenes in the different plants and products studied. Thirteen dried sausage processing plants were sampled at two different times of the working day. The studies were repeated twice to evaluate the persistence of the pathogen. A total of 1029 samples were collected. Among swabbed samples, 15% were positive before the beginning of the working day and 47.3% during working day. Results showed that effectiveness of cleaning and disinfecting operations could be linked with the complexity of processing lines and machines used. The presence of L. monocytogenes in mixed meat amounted to 71.6% of the collected samples. A decrease of the contamination rate in dry sausage was noted, particularly during the drying stage. Nevertheless 3 sausages studied presented a low contamination rate (<3 cfu/g) when ready for consumption. A total of 996 strains of L. monocytogenes were characterised by biochemical tests and serotyping. A majority of isolates were 1/2a (49.5%), 1/2c (19.5%) and 1/2b (13%) strains. A high heterogeneity of serotypes was observed in all plants, raw meat and in sausages during maturation.

Animals↗

Fate of Listeria monocytogenes in experimentally contaminated French sausages.

Listeria monocytogenes has been recognized as one of the most important foodborne pathogens dealt with by the food. The bacterium has been found in every part along the pork processing industry from the slaughterhouse to the cutting room and the delicatessen factories. During the fermentation and drying of sausages, L. monocytogenes tends to decrease substantially. However, despite the various hurdles in the dry sausage manufacturing process, L. monocytogenes is able to survive and is detected in the final products. The present study has evaluated growth and survival of eight different L. monocytogenes strains (originating from sausage, sausage industry environment and from clinical cases of listeriosis) in experimentally inoculated French sausages with 10(4) cfu g(-1). This study points out the fact that the decrease of L. monocytogenes contamination rate during the manufacturing process of sausages is strain dependent (p < 0.001) and mainly due to the drying and maturation step than to the fermentation itself. Whatever the strains studied, almost no decrease of the contamination rate was noted during the fermentation step. However hurdle-adapted strains (those isolated from sausages or sausage industry environment) were more difficult to cure from sausages (decrease by 1.5 log10) than non-adapted strains (decrease by 3 log10) at the end of the drying period (day 35), when sausages were ready for consumption. These sausages became safe only at the best before date. As a consequence, L. monocytogenes and more particularly those "adapted" strains might represent a very important issue for hygienists since these strains originating from sausages or production environment themselves are likely to contaminate sausages during manufacturing and remain in the final products. However, the high inoculum levels used in the study (10(4) cfu g(-1)) are not representative of the natural contamination of L. monocytogenes commonly encountered in the raw material for sausages. If such contamination happened to be inferior to 100 cfu g(-1), then the manufacturing process used in this study would be able to produce "safe" sausages according to the European regulation requiring the absence of L. monocytogenes in 25 g of food with a tolerance of below 100 cfu g(-1) at the best before date.

Animals↗

Modelling the effect of a temperature shift on the lag phase duration of Listeria monocytogenes.

The aim of this work is to study and model the effect of a temperature shift on h(0), the product of the growth rate by the lag phase duration (mulambda). Our work is based on the data of Whiting and Bagi [Int. J. Food Microbiol. 73 (2002) 291], who studied the influence of both the pre-incubation temperature (T(prior)) and the growth temperature (T(growth)) on lambda values of Listeria monocytogenes. We introduce a new model to describe the evolution of the parameter h(0) as a function of T(prior) and T(growth), and compare it to Whiting and Bagi's published polynomial model that describes the influence of T(prior) and T(growth) on lambda independently of mu. For exponential as well as stationary phase cells, h(0) increases almost linearly with the magnitude of the temperature shift. A simple linear model of h(0) turns out to be more suitable to predict lambda values than a polynomial model of lambda.

Food Microbiology↗

Effect of injecting collagenase into the uterine artery during a caesarean section on the placental separation of cows induced to calve with dexamethasone.

A caesarean section was performed on 30 cows before normal term and 16 to 20 hours after the induction of parturition with dexamethasone. During the surgical procedure, 20,000 U of bacterial collagenase was injected into the uterine artery of 15 of the cows. The average periods of retention of the fetal membranes were 40 hours in the treated cows and 114 hours in the control cows (P<0.001). At 36 hours after the surgery six of the treated cows (40 per cent) but all 15 of the control cows had retained fetal membranes. The collagenase-treated cows showed no abnormal clinical signs during the 10 days after the operation.

Animals↗

Modeling the lag time of Listeria monocytogenes from viable count enumeration and optical density data.

The following two factors significantly influence estimates of the maximum specific growth rate ( micro (max)) and the lag-phase duration (lambda): (i) the technique used to monitor bacterial growth and (ii) the model fitted to estimate parameters. In this study, nine strains of Listeria monocytogenes were monitored simultaneously by optical density (OD) analysis and by viable count enumeration (VCE) analysis. Four usual growth models were fitted to our data, and estimates of growth parameters were compared from one model to another and from one monitoring technique to another. Our results show that growth parameter estimates depended on the model used to fit data, whereas there were no systematic variations in the estimates of micro (max) and lambda when the estimates were based on OD data instead of VCE data. By studying the evolution of OD and VCE simultaneously, we found that while log OD/VCE remained constant for some of our experiments, a visible linear increase occurred during the lag phase for other experiments. We developed a global model that fits both OD and VCE data. This model enabled us to detect for some of our strains an increase in OD during the lag phase. If not taken into account, this phenomenon may lead to an underestimate of lambda.

Colony Count, Microbial↗

Optimal growth temperature of O157 and non-O157 Escherichia coli strains.

AIMS: There are several biological characteristics that differ between Escherichia coli O157:H7, a dangerous food-borne pathogen, and the other serotypes of E. coli. METHODS AND RESULTS: The optimal growth temperatures (T(opt)) were determined for 32 E. coli strains, whether each strain belonged to the O157 serotype or not. The mean values of T(opt) for the O157 and non-O157 groups were 40.2 and 41.2 degrees C, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This difference is statistically significant (P=0.0002) but has no biological implication.

Animals↗

Biological variability and exposure assessment.

Predictive models are now commonly used for exposure assessment, with growth parameters defined for each microbial species. In this study, we tried to take into account microbial growth variability among strains of a single species. Bacillus cereus in pasteurized milk was chosen to illustrate the influence of the biological variability on the outcome of exposure assessment. Each parameter of the exposure assessment (growth parameters, shelf-life conditions) was characterized by a probability distribution describing variability and/or uncertainty. The impact of the intra-species variability on the result of the exposure assessment was then quantified and discussed. Two simple domestic shelf life conditions were tested. The results confirm that the biological variability has a great impact on the accuracy of the result and should not be systematically neglected.

Animals↗

Determination of parotid urea secretion in sheep by means of ultrasonic flow probes and a multifactorial regression analysis.

For determination of the dynamics of parotid urea secretion in conscious sheep, a previously standardized transit time ultrasonic flow metering system was used to measure bilateral parotid flow. Six ewes fed for ad libitum consumption were prepared under halothane anesthesia with ultrasonic probes around both parotid ducts; these ducts were also cannulated orally. After probe encapsulation (8 d), parotid flows were recorded during 24 h, and samples of saliva and blood for urea determination were obtained hourly. Jaw movements were recorded by means of a submandibular balloon to monitor feeding behavior. Urea concentration in parotid saliva was 60 to 74% of that in plasma (a positive linear correlation existed) and was poorly influenced by the parotid flow. The amount of urea secreted with parotid saliva was directly related to the salivation rate. To calculate the urea secretion in parotid saliva, a multiple linear regression model was developed from computer-calculated parotid flows over 1-min periods and plasma urea concentration. The model was accurate because the plot of calculated vs measured values was not significantly different from the line of identity. The daily parotid urea N varied from .35 to 1.02 g among ewes. The higher urea secretion rate found during rumination and eating (1.32+/-.42 and .98 +/-.33 mg/min, respectively) vs. during rest (.60+/-.39 mg/ min, P<.05) was due to higher salivation rates (5.17 +/-1.46, 3.56+/-.90, and 2.04+/-.52 mL/min, respectively, P<.05) rather than to changes in saliva urea concentrations (saliva:plasma urea ratio = .65+/-.04, .67+/-.04, and .68+/-.03, respectively). Of the daily parotid urea output, 40.8% was secreted during rest. The contribution of parotid urea N to the ruminal N pool was relatively small (1.2 to 3.7% of the N intake, which was 23.0 to 33.6 g/d). These techniques allowed direct and precise measurements of parotid urea secretion without disturbing the animal or altering the physiological regulation of salivary secretion.

Animals↗

Characterization of unexpected growth of Escherichia coli O157:H7 by modeling.

Modeling of batch kinetics in minimal synthetic medium was used to characterize Escherichia coli O157:H7 growth, which appeared to be different from the exponential growth expected in minimal synthetic medium and observed for E. coli K-12. The turbidimetric kinetics of 14 of the 15 O157:H7 strains tested (93%) were nonexponential, whereas 25 of the 36 other E. coli strains tested (70%) exhibited exponential kinetics. Moreover, the anomaly was almost corrected when the minimal medium was supplemented with methionine. These observations were confirmed with two reference strains by using plate count monitoring. In mixed cultures, E. coli K-12 had a positive effect on E. coli O157:H7 and corrected its growth anomaly. This demonstrated that commensalism occurred, as the growth curve for E. coli K-12 was not affected. The interaction could be explained by an exchange of methionine, as the effect of E. coli K-12 on E. coli O157:H7 appeared to be similar to the effect of methionine.

Animals↗

Relation between the generation time and the lag time of bacterial growth kinetics.

In predictive microbiology, the relation between the lag time (Lag) and the generation time (Tg) is commonly assumed to be proportional, as long as the pre-incubation environmental conditions remain constant. This relation was statistically examined in nine published datasets. For every dataset, it was roughly proportional. However, a more advanced study showed that the ratio Lag/Tg was not totally independent of the environmental conditions. In particular, a significant negative effect of the pH on this ratio was observed in five of the nine datasets. For modeling the environmental dependence of microbial growth parameters, some authors independently deal with Lag and Tg. Other authors only model the environmental dependence of Tg, assuming Lag/Tg to be constant. These two modeling methods were statistically compared for the nine datasets under study. Results differed from one dataset to another. For some, the model developed with a constant ratio Lag/Tg sufficed to describe the data, whereas for the others, an independent modeling of Lag and Tg was more satisfactory.

Bacteria↗

Behaviour and enterotoxin production by Staphylococcus aureus during the manufacture and ripening of raw goats' milk lactic cheeses.

To study the possible presence of staphylococcal enterotoxin A in raw goats' milk lactic cheese, milk was inoculated with an enterotoxigenic Staphylococcus aureus strain to a final concentration of 4, 5 and 6 log(cfu/ml). Cheese was prepared following industrial specifications and ripened for 42 d. Detection of the enterotoxins was by the Vidas Staph enterotoxin test (BioMérieux) and by an indirect double-sandwich ELISA technique using anti-enterotoxin monoclonal antibodies. Staphylococcal counts declined markedly after draining, and by the end of ripening they had disappeared from some cheeses. In contrast, aerobic mesophilic organisms grew well. The level of staphylococcal enterotoxin A recovered varied from 1 to 2.5 ng/g cheese made with an initial population of 10(5) or 10(6) cfu/ml. Only traces of enterotoxin A (0.5 ng/g) were detected in cheeses made with the lowest Staph. aureus inoculum used in this study. Enterotoxin A was also detected in cheeses from which Staph. aureus had disappeared.

Animals↗

Application of a modified disc diffusion technique to antimicrobial susceptibility testing of Vibrio anguillarum and Aeromonas salmonicida clinical isolates.

Two techniques for antimicrobial susceptibility testing of Vibrio anguillarum and Aeromonas salmonicida strains were compared. The first method was the reference test that determines Minimal Inhibitory Concentrations (MIC); the second was a modified diffusion test that measures the Inhibitory Concentrations in Diffusion (ICD) by carrying out the diffusion test with five discs of differing contents. ICD measurement was not applicable for the susceptibility testing of oxytetracycline and sulfadimethoxine. On the other hand, a good correlation between the MICs and the ICDs was observed for oxolinic acid, sarafloxacin, chloramphenicol and trimethoprim. Moreover, the ICD values were close to those obtained for the MIC values. A. salmonicida resistant strains were detected by ICD determination. Thus, ICD could be used instead of MIC for oxolinic acid, sarafloxacin, trimethoprim and chloramphenicol susceptibility testings. The ICD technique is easy to carry out and is not dependent on the growth characteristics of bacteria.

Aeromonas↗

An economic approach to the MIC.

The determination of the Inhibitory Concentration in Diffusion (ICD) is proposed as an alternative to the agar dilution Minimal Inhibitory Concentration (MIC) that is time-consuming and cumbersome for routine use. Based on the technique of the disk diffusion test, it consists in calculating a continuous variable, the ICD, corresponding to the antibiotic concentration in the agar at the edge of the inhibition zone. Six antibiotics were tested (ampicillin, cefotaxime, erythromycin, gentamicin, nalidixic acid and rifampicin) each against 17 to 51 strains of enterobacteria, Staphylococcus aureus and Enterococcus spp. and six other antibiotics (cefsulodin, ceftazidime, imipenem, piperacillin, ticarcillin and tobramycin), against 13 to 25 strains of Pseudomonas aeruginosa. A total of 284 antibiotic-strain combinations were tested. Three different antibiotic charges were obtained for each antibiotic by cutting commercial disks in two and four equal pieces. The ICD was calculated for each strain from the size of inhibition zones around a full disk, a half and a quarter of a disk. Concurrently, the MIC was performed, using a conventional agar dilution method. There was a good correlation between the two methods and reproducibility for the ICD proved to be correct. This reliable technique is very efficient both in terms of laboratory time and cost of materials and could be proposed for widespread use in clinical laboratories.

Ampicillin↗

Accuracy of microbial growth predictions with square root and polynomial models.

The results of growth predictions using square root and polynomial models published in 14 papers were studied. Errors on quantities of practical interest such as lag time, generation time or the time required to reach a given increase in number of cells, are analyzed. The distribution of these errors was examined with the perspective of the practical use of predictive models in food industry. Highly unsafe predictions and significant average errors were observed in some cases. A good knowledge of predictive models accuracy seems essential for their efficient and safe use, for example to predict the shelf life of a product. Yet, authors generally gave no pragmatic information on such things as the average relative error or the range of errors on predicted variables. Problems of robustness of models when tested in different conditions were noticed, which corroborates the necessity of a systematic validation of models on new data.

Bacteria↗