[Psychosomatic, bonds and relational theory]. / Psychosomatique, liens et théorie relationnelle.

The term 'psychosomatic' conveys the idea of connections between the mind and the soma. These are also conceived between an organic pathology and a mental pathology, between the real and the imaginary, and oppose the separation of the body and mind. A bond is established through a relational situation, through words, and is based on values upheld by the caregivers such as humanity, individual commitment to the patient. Thereby created, it will found a relational psychosomatic dimension. This therapeutic approach consists in seeing the relationship rather than the division.

Modelling the effect of temperature, pH, water activity, and organic acids on the germination time of Penicillium camemberti and Penicillium roqueforti conidia.

In this study, the influence of environmental factors on the germination time of Penicillium camemberti and Penicillium roqueforti conidia was evaluated. To do so, the effects of i/temperature, pH, water activity, and ii/organic acids were determined using models based on i/cardinal values, and ii/minimum inhibitory concentration (MIC) respectively. Cardinal values for germination of conidia were not observed to be species dependent. Minimum temperatures were estimated to be below the freezing point, with an optimum of 26.9°C, and a maximum of 33.5°C. For both species, minimal and optimal aw values were found to be 0.83 and 0.99, respectively, while for pH these values corresponded to 2.9, and 5.6. MIC values could not be determined for lactic acid because conidia of both species germinated in up to 1M concentrations, the highest concentration tested. At pH5.6, P. camemberti (MIC=0.197M) was more sensitive to propionic acid than P. roqueforti (MIC=0.796M).

Validation of a predictive model for the growth of chalk yeasts on bread.

The present study focused on the effects of temperature, T, and water activity, aw, on the growth of Hyphopichia burtonii, Pichia anomala, and Saccharomycopsis fibuligera on Sabouraud Agar Medium. Cardinal values were estimated by means of cardinal models with inflection. All the yeasts were xerophilic, and they exhibited growth at 0.85 aw. The combined effects of T, aw, and pH on the growth of these species were described by the gamma-concept and validated on bread in the range of 15-25 °C, 0.91-0.97 aw, and pH 4.6-6.8. The optimum growth rates on bread were 2.88, 0.259, and 1.06 mm/day for H. burtonii, P. anomala, and S. fibuligera, respectively. The optimal growth rate of S. fibuligera on bread was about 2 fold that obtained on Sabouraud. Due to reproduction by budding, P. anomala exhibited low growth on Sabouraud and bread. However, this species is of major concern in the baker's industry because of the production of ethyl acetate in bread.

A model for the effect of pH on the growth of chalk yeasts.

Hyphopichia burtonii, Pichia anomala, and Saccharomycopsis fibuligera were isolated from spoiled packaged sliced bread. These chalk yeasts were characterized by a wide range of pH for which growth was almost optimum. Thus, the curve growth vs pH exhibited plateau and sharp profiles close to the minimum and the maximum pH. This study described a chalk yeast model (CYM) for the effect of pH derived from a new germination model for fungi (Dantigny, P., Nanguy, S., P.-M., Judet-Correia, D., and Bensoussan, M. 2011, International Journal of Food Microbiology, 146, 176-181). The CYM is asymmetric, versatile, based on parameters with biological significance, and compatible with the gamma concept. The CYM was compared to the cardinal pH model (CPM) which is widely used to describe the effect of pH on microbial growth. The CYM exhibited RMSE values two fold less than those obtained with the CPM for H. burtonii, and S. fibuligera for which plateaus were clearly observed. For P. anomala, the plateau was less obvious, but the RMSE value obtained with the CYM was similar to that found with the CPM. The CYM could extend its use to represent the effect of pH on mold growth.

Lag time for germination of Penicillium chrysogenum conidia is induced by temperature shifts.

In the environment, fungal conidia are subject to transient conditions. In particular, temperature is varying according to day/night periods. All predictive models for germination assume that fungal spores can adapt instantaneously to changes of temperature. The only study that supports this assumption (Gougouli and Koutsoumanis, 2012, Modelling germination of fungal spores at constant and fluctuating temperature conditions. International Journal of Food Microbiology, 152: 153-161) was carried out on Penicillium expansum and Aspergillus niger conidia that, in most cases, already produced germ tubes. In contrast, the present study focuses on temperature shifts applied during the first stages of germination (i.e., before the apparition of the germ tubes). Firstly, germination times were determined in steady state conditions at 10, 15, 20 and 25 °C. Secondly, temperature shifts (e.g., up-shifts and down-shifts) were applied at 1/4, 1/2, and 3/4 of germination times, with 5, 10 and 15 °C magnitudes. Experiments were carried out in triplicate on Penicillium chrysogenum conidia on Potato Dextrose Agar medium according to a full factorial design. Statistical analysis of the results clearly demonstrated that the assumption of instantaneous adaptation of the conidia should be rejected. Temperature shifts during germination led to an induced lag time or an extended germination time as compared to the experiments conducted ay steady state. The induced lag time was maximized when the amplitude of the shift was equal to 10 °C. Interaction between the instant and the direction of the shift was highlighted. A negative lag time was observed for a 15 °C down-shift applied at 1/4 of the germination time. This result suggested that at optimal temperature the rate of germination decreased with time, and that the variation of this rate with time depended on temperature.

Effect of inoculum size and water activity on the time to visible growth of Penicillium chrysogenum colony.

In order to assess the effect of the inoculum size on the time to visible growth for Penicillium chrysogenum, the correlation described by González et al. (González, H.H.L., Resnik, S.L., Vaamonde, G., 1987. Influence of inoculum size on growth rate and lag phase of fungi isolate from Argentine corn. International Journal of Food Microbiology 4, 111-117) was compared to the model introduced by Gougouli et al. (Gougouli, M., Kalantzi, K., Beletsiotis, E., Koutsoumanis, K.P., 2011. Development and application of predictive models for fungal growth as tools to improve quality control in yogurt production. Food Microbiology 28, 1453-1462). Based on the regression coefficient, the latter model performed better than the former one to fit the data obtained for P. chrysogenum grown on Potato Dextrose Agar at 25 °C. Inoculum sizes in the range 10(1)-10(5) spores were tested at 0.930, 0.950, 0.970, and 0.995 aw. By extrapolation of the straight line, the model of Gougouli et al. (2011) provided accurate estimations of the time to visible growth for a single spore inoculum, tvg (N=1). In order to avoid experiments at reduced water activities, the influence of water activity on the model parameters, and on the ratio tvg (N=1) over the germination time was assessed.

Effect of storage conditions (relative humidity, duration, and temperature) on the germination time of Aspergillus carbonarius and Penicillium chrysogenum.

Fungal conidia are disseminated, often in the air, for a certain period of time, prior to contaminating food products. The objective of this study was to examine the effects of the relative humidity, RH (%), time (day), and temperature (°C) during this period of time, called "storage", on the germination time, τ (h), of Aspergillus carbonarius and Penicillium chrysogenum. A Doehlert design was used in the range, 20-100% RH, 2-28 days, and 5-25 °C. As compared to un-stored conidia, the germination time of conidia stored at 60% RH, 15 days, 5 °C was increased by 23 and 28%, for A. carbonarius and P. chrysogenum, respectively. Stored conidia exhibited a minimum τ value at 60% RH, and 100% RH for A. carbonarius and P. chrysogenum, respectively. For these species, τ was minimum for 2 days of storage. The effect of temperature was RH dependent for A. carbonarius. The germination time of stored conidia was clearly greater than that of fresh conidia obtained in the laboratory. This result should be taken into account to determine the mould free shelf-life of food products.

A new model for germination of fungi.

The objective of this study was to design a germination model dedicated to fungi. The percentage of germinated spores, P(%), depended on the maximum percentage of germination P(max) (%), the germination time, τ (h) and a design parameter, d (-) according to : [formula in text]. The model was capable to fit satisfactorily either apparent symmetric and asymmetric shapes of germination curves. The accuracy of τ determined by using the logistic or the present model was at least twice that obtained by the Gompertz equation. In contrast to the logistic model, the new model is by essence asymmetric. Therefore, its use is consistent with skewed distributions of the individual germination times that were observed experimentally in many cases.

Impact of water activity of diverse media on spore germination of Aspergillus and Penicillium species.

The effects of water activity (a(w)) of diverse media i/ culture medium for sporogenesis, a(w sp) ii/ liquid spore suspension medium, a(w su) and iii/ medium for germination, a(w ge), on the germination time t(G) of Aspergillus carbonarius, Aspergillus flavus, Penicillium chrysogenum and Penicillium expansum were assessed according to a screening matrix at 0.95 and 0.99 a(w). It was shown that i/ reduced t(G)s were obtained at 0.95 a(w sp) except for P. expansum ii/ a significant effect of a(w su) on t(G) was demonstrated for A. carbonarius, P. chrysogenum and P. expansum iii/ the most important factor for controlling the germination time was the medium for germination except for A. carbonarius (a(w su)). In accordance with the fact that fungal spores can swell as soon as they are suspended in an aqueous solution it is recommended to re-suspend fungal spores in a solution at the same water activity as that of subsequent germination studies.

Validation of a predictive model for the growth of Botrytis cinerea and Penicillium expansum on grape berries.

The objective of this study was to develop and to validate a model for predicting the combined effect of temperature and a(w) on the radial growth rate, mu, of Botrytis cinerea and Penicillium expansum on grape berries. The proposed strategy was based on the gamma-concept developed previously [Zwietering, M.H., Wijtzes, T., de Wit, J.C., van't Riet, K. 1992. A decision support system for prediction of the microbial spoilage in foods. Journal of Food Protection. 12, 973-979]: mu=mu(opt).gamma(T).gamma(a(w)), where the gamma functions were cardinal models with inflection (CMI), mu(opt) the radial growth rate on grape berries. Firstly, the cardinal temperatures and a(w)'s were estimated independently from experiments carried out on Potato Dextrose Agar. Secondly, the gamma concept was validated i/ on a synthetic grape juice medium (SGJ) and ii/ on a grape juice agar (GJA). Accuracy and bias factors were closer to 1 with the latter analogue, thus suggesting that GJA should be preferred to SGJ. Thirdly, an experimental protocol taken into account the isotropic nature of fungal growth was developed for estimating mu(opt) on grape berries. This study demonstrated that CMI's can be validated on agri-food products over a wide range of temperature and a(w) using the described methodology.

Distributions of the growth rate of the germ tubes and germination time of Penicillium chrysogenum conidia depend on water activity.

The effects of water activities for sporulation (a(wsp)) and germination (a(wge)) on the distributions of the growth rate of the germ tubes (mu) and the germination time (t(G)) of Penicillium chrysogenum conidia were determined by monitoring the length of the same germ tubes throughout the experiments automatically. No relationship between the individual t(G)'s and mu's could be established. Irrespective of the water activity for germination, mu was greater and t(G) was less for conidia produced at 0.95a(wsp) than that at 0.99a(wsp). At 0.99 a(wge) the mean and the standard deviation of t(G) were smaller than those obtained at 0.95a(wge). At 0.99a(wge), normal distributions for mu and t(G) were exhibited, but not at 0.95a(wge). The cumulative frequencies were used to reconstruct the germination curves. Great differences in the percentage of spores capable of germination (P(G)) and in the mean germination times between conidia produced at 0.95a(wsp) and at 0.99a(wsp) were clearly exhibited at 0.95a(wge), thus demonstrating the paramount influence of sporulation conditions on germination kinetics.

Inactivation of conidia of Penicillium chrysogenum, P. digitatum and P. italicum by ethanol solutions and vapours.

A fractional factorial design, 2(5-1) experiments, was used for assessing the influence of 5 factors: water activity, aw [0.7, 0.9], temperature, T ( degrees C) [10, 30], mode of application, A [liquid, vapour], ethanol concentration, E (% w/w) [5, 10] and time, t (d) [1, 4] on the inactivation of spores of Penicillium chrysogenum, P. digitatum and P. italicum. Survival was determined by germination at optimal conditions within 3d. The experimental response was log (N 0/Nt), where N 0 and Nt (spore ml(-1)) the concentrations of viable spores at t=0 and t respectively. By a decreasing order of sensitivity to ethanol, moulds were ranked as followed: P. digitatum, P. italicum and P. chrysogenum. A greater inactivation for P. digitatum, P. italicum, that were the most sensitive moulds to ethanol, was obtained by applying vapour rather than ethanol solution. The order of significance of the main factors depended upon the mould. The key factor for explaining inactivation of P. chrysogenum was water activity. But, temperature was the main factor for explaining inactivation of P. digitatum and P. italicum. In the more drastic conditions, (i.e., 0.7 aw, 30 degrees C, 10% w/w ethanol), all spores were inactivated by applying liquid solution for 4d.

The logarithmic transformation should be avoided for stabilising the variance of mould growth rate.

Radial growth rate, micro (mm d(-1)) was evaluated by plotting the radius of the colony, r (mm) versus time (d) for Alternaria alternata, Aspergillus flavus, Cladosporium cladosporioides, Mucor racemosus, Rhizopus oryzae and Trichoderma harzianum at different T and a(w). For each of the 12 data sets, an analysis of variance of the raw growth rate data was performed. It was observed from the P-values that all square-root transformed values of micro were non-significant at the significance level alpha=0.05, whereas for untransformed values of micro, three of the 12 values were significant and for logarithmically transformed micro, nine of the 12 values were significant at the significance level. Therefore the logarithmic transformation that is used for bacteria should be avoided for these moulds except for A. flavus. In contrast the square-root transformation appeared suitable for stabilising the variance of mould growth rate in all cases.

Standardisation of methods for assessing mould germination: a workshop report.

The first workshop on predictive mycology was held in Marseille, France, 2--4 February 2005 under the auspices of the French Microbiological society. The purpose of the workshop was to list the different techniques and definitions used by scientists for assessing mould germination and to evaluate the influence of the different techniques on the experimental results. Recommendations were made when a large consensus was obtained. In order to facilitate the study of germination, alternative methods to microscopic examination were examined.

Modeling the effect of ethanol vapor on the germination time of Penicillium chrysogenum.

The influence of ethanol vapor on germination of Penicillium chrysogenum was determined on yeast nitrogen base plus glucose agar medium at 25 degrees C. Ethanol vapors were generated by 0 to 6% (wt/wt) ethanol solutions at the bottom of hermetically closed petri dishes. The logistic equation was used to describe the data as the percentage of germination versus time and to estimate the germination time. The effect of ethanol concentration on germination time was described by a new reparameterized equation, resulting in an estimated limiting ethanol concentration of 4.3%. Up to 3% ethanol, all spores germinated, and the germination time increased with increasing ethanol concentration. At 3.5 and 4%, some spores formed abnormal germ tubes and others were inhibited at the swelling stage. The inhibiting effect of ethanol was reversible under these experimental conditions.

Basis of predictive mycology.

For over 20 years, predictive microbiology focused on food-pathogenic bacteria. Few studies concerned modelling fungal development. On one hand, most of food mycologists are not familiar with modelling techniques; on the other hand, people involved in modelling are developing tools dedicated to bacteria. Therefore, there is a tendency to extend the use of models that were developed for bacteria to moulds. However, some mould specificities should be taken into account. The use of specific models for predicting germination and growth of fungi was advocated previously []. This paper provides a short review of fungal modelling studies.

Modelling the effect of ethanol on growth rate of food spoilage moulds.

The effect of ethanol (E) on the radial growth rate (mu) of food spoilage moulds (Aspergillus candidus, Aspergillus flavus, Aspergillus niger, Cladosporium cladosporioides, Eurotium herbariorum, Mucor circinelloides, Mucor racemosus, Paecilomyces variotii, Penicillium chrysogenum, Penicillium digitatum, Rhizopus oryzae and Trichoderma harzianum) was assessed in Potato Dextrose Agar (PDA) medium at a(w) 0.99, 25 degrees C. In order to model this effect, the Monod type equation described previously by Houtsma et al. (Houtsma, P.C., Kusters, B.J.M., de Wit, J.C., Rombouts, F.M., Zwietering, M.H., 1994. Modelling growth rates of Listeria monocytogenes as a function of lactate concentration. Int. J. Food. Microbiol. 24, 113-123.) was re-parameterised: mu = mu(opt)[K(E(max)-E)/K E(max)-2KE+E(max)E]; E(max) (%, wt/wt): ethanol concentration at which no growth occurs, K (%, wt/wt): ethanol concentration at which mu = mu(opt)/2, mu(opt) (mm day(-1)): growth rate at 0% ethanol. The model was capable of describing curves, mu vs. E, with either a concave shape (KE(max)/2) with a good accuracy (root mean square error (RMSE) < or = 0.136) with the notable exception of R. oryzae and T. harzianum. After growth rate data were square-root transformed to stabilise the variance, E(max) was estimated in the range 3% to 5% for all moulds with the exception of T. harzianum (E(max) 2.14%) and P. variotii (E(max) 6.43%). Ethanol would appear an effective additional barrier to inhibit fungal growth in food products and would represent an interesting alternative to the use of preservatives.