Growth characterization and predictive behavior of Eurotium species in a feedstuff matrix

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"ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:1 [ "total" => 0 ] "en" => array:14 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Debaryomyces hansenii F39A as biosorbent for textile dye removal" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:3 [ 0 => "en" 1 => "en" 2 => "es" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "257" "paginaFinal" => "265" ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Debaryomyces hansenii F39A como bioadsorbente para la remoción de colorantes textiles" ] ] "contieneResumen" => array:2 [ "en" => true "es" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 1112 "Ancho" => 1500 "Tamanyo" => 62472 ] ] "descripcion" => array:1 [ "en" => "Effect of the adsorbent dose on the biosorption capacity of Reactive Red 141 and Reactive Blue 19 (Co=0.0564mmol/l for RR-141 and Co=0.160mmol/l for RB-19, contact time=80min, rate=150rpm, T=20±2°C)." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Florencia Ruscasso, Brenda Bezus, Gabriela Garmendia, Silvana Vero, Gustavo Curutchet, Ivana Cavello, Sebastián Cavalitto" "autores" => array:7 [ 0 => array:2 [ "nombre" => "Florencia" "apellidos" => "Ruscasso" ] 1 => array:2 [ "nombre" => "Brenda" "apellidos" => "Bezus" ] 2 => array:2 [ "nombre" => "Gabriela" "apellidos" => "Garmendia" ] 3 => array:2 [ "nombre" => "Silvana" "apellidos" => "Vero" ] 4 => array:2 [ "nombre" => "Gustavo" "apellidos" => "Curutchet" ] 5 => array:2 [ "nombre" => "Ivana" "apellidos" => "Cavello" ] 6 => array:2 [ "nombre" => "Sebastián" "apellidos" => "Cavalitto" ] ] ] ] "resumen" => array:1 [ 0 => array:3 [ "titulo" => "Highlights" "clase" => "author-highlights" "resumen" => "<ul class="elsevierStyleList" id="lis0015"><li class="elsevierStyleListItem" id="lsti0005">•Debaryomyces hansenii F39A biomass can remove textile dyes from aqueous solutions.</li><li class="elsevierStyleListItem" id="lsti0010">•D. hansenii was able to adsorb from low to higher concentrations of dye.</li><li class="elsevierStyleListItem" id="lsti0015">•Kinetic and biosorption isotherms were modelled.</li><li class="elsevierStyleListItem" id="lsti0020">•D. hansenii could be an efficient and inexpensive biosorbent in wastewater treatment.</li></ul>" ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754120301218?idApp=UINPBA00004N" "url" => "/03257541/0000005300000003/v1_202109250622/S0325754120301218/v1_202109250622/en/main.assets" ] "itemAnterior" => array:19 [ "pii" => "S032575412030122X" "issn" => "03257541" "doi" => "10.1016/j.ram.2020.08.004" "estado" => "S300" "fechaPublicacion" => "2021-07-01" "aid" => "430" "copyright" => "Asociación Argentina de Microbiología" "documento" => "article" "crossmark" => 1 "licencia" => "http://creativecommons.org/licenses/by-nc-nd/4.0/" "subdocumento" => "fla" "cita" => "Rev Argent Microbiol. 2021;53:240-7" "abierto" => array:3 [ "ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:1 [ "total" => 0 ] "en" => array:13 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Anti-proliferative and apoptosis induction activities of curcumin on Leishmania major" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:2 [ 0 => "en" 1 => "es" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "240" "paginaFinal" => "247" ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Efectos de inducción antiproliferativa y apoptótica de la curcumina en Leishmania major" ] ] "contieneResumen" => 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Exponentially growing promastigotes were cultured in 96-well plates and treated at the indicated concentrations of curcumin for 48h. Cell death was analyzed using the MTT assay. The arrows indicate LC50. Error bars represent standard deviations of at least three different experiments." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Maha Elamin, Ebtsam Al-Olayan, Rewaida Abdel-Gaber, Ramy S. Yehia" "autores" => array:4 [ 0 => array:2 [ "nombre" => "Maha" "apellidos" => "Elamin" ] 1 => array:2 [ "nombre" => "Ebtsam" "apellidos" => "Al-Olayan" ] 2 => array:2 [ "nombre" => "Rewaida" "apellidos" => "Abdel-Gaber" ] 3 => array:2 [ "nombre" => "Ramy S." "apellidos" => "Yehia" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S032575412030122X?idApp=UINPBA00004N" "url" => "/03257541/0000005300000003/v1_202109250622/S032575412030122X/v1_202109250622/en/main.assets" ] "en" => array:21 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Growth characterization and predictive behavior of Eurotium species in a feedstuff matrix" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "248" "paginaFinal" => "256" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Mariana Greco, Graciela Pose, Alejandro Pardo" "autores" => array:3 [ 0 => array:4 [ "nombre" => "Mariana" "apellidos" => "Greco" "email" => array:1 [ 0 => "mariana.greco@gmail.com" ] "referencia" => array:3 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] 1 => array:3 [ "nombre" => "Graciela" "apellidos" => "Pose" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 2 => array:3 [ "nombre" => "Alejandro" "apellidos" => "Pardo" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, B1876BXD, Buenos Aires, Argentina" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina" "etiqueta" => "b" "identificador" => "aff0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Caracterización del crecimiento y comportamiento predictivo de especies de Eurotium en matriz de alimento balanceado" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 973 "Ancho" => 1520 "Tamanyo" => 77894 ] ] "descripcion" => array:1 [ "en" => "Growth/no growth interface of E. repens after 7, 15 and 40 days of incubation in a feed matrix. P (0.50): short dashed line: at 7 days; long dashed line: at 15 days; solid line: at 40 days; white square: growth at 7 days; gray square: growth at 15 days; black square: growth at 40 days; cross: no growth." ] ] ] "textoCompleto" => "IntroductionFeedlot practices were introduced in Argentina in the 90s mainly for economic reasons. Nowadays, this practice still continues and at the same time feedstuff production has globally increased. A recent feed survey indicates that worldwide feed production has exceeded a billion metric tons<a class="elsevierStyleCrossRef" href="#bib0255">4</a>; 10.6% of feed production in the Latin American region is represented by Argentina<a class="elsevierStyleCrossRef" href="#bib0405">34</a>. Particularly, more than 14 million tons were produced in 2015, while the most recent estimates suggest that around 25 million tons will be produced by 2020<a class="elsevierStyleCrossRef" href="#bib0370">27</a>. Animal feeds are intended for cattle, dairy cattle, porcine, poultry, aquaculture, rabbit and chinchilla, and more recently buffalo. Moreover, an important growth has been registered in the pet food industry in the last years. In addition to industrial feed production, small farmers also elaborate their own feed. However, in some circumstances, lack of quality assurance of raw material, improper technical production process or inappropriate storage conditions can lead to fungal spoilage development and mycotoxin production. In addition, sometimes due to lack of knowledge or negligence, despite the fact that visible fungal growth is observed, feeds are mixed with non-contaminated feed in order to dilute its contamination, thus economic losses are avoided. Therefore, feed quality decreases and becomes a risk for animals, and consequently for the complete food chain<a class="elsevierStyleCrossRef" href="#bib0420">37</a>.Animal feeds are composed of the necessary nutrients that animals need at every stage of their growth to reach a proper development and maximal growth in a profitable period of time for producers. Several parameters are involved and considered to define the quality of feed pellets such as formulation, durability, starch gelatinization and protein content, particle size, exposure to high pressure steam and proper cooling, among others<a class="elsevierStyleCrossRef" href="#bib0320">17</a>. In addition, the final product stability is regulated by aw. Therefore, aw is the most critical parameter to be taken into consideration since variations tend to increase fungal proliferation<a class="elsevierStyleCrossRefs" href="#bib0290">11,29,46</a>.Fungal contamination in raw materials and animal feed has been widely reported worldwide. In fact, most of them emphasize mycotoxigenic genera, mainly Alternaria, Aspergillus, Fusarium and Penicillium<a class="elsevierStyleCrossRefs" href="#bib0260">5,16,26,37,42</a>. In addition, based on the findings reported by Hassan et al.<a class="elsevierStyleCrossRef" href="#bib0365">26</a>, mixed cereals used as animal feeds are contaminated with high levels of mycotoxigenic fungi and mycotoxins. Moreover, several cases of human and animal intoxication and death caused by food and feedstuff contaminated with mycotoxins were also informed in the recent years<a class="elsevierStyleCrossRefs" href="#bib0325">18,21,33,38</a>.The presence and development of fungi cause changes in food and feed properties involving nutritional and organoleptic quality aspects. Due to their xerophilic physiology Eurotium species are able to grow at aw values as low as 0.65<a class="elsevierStyleCrossRefs" href="#bib0295">12,28</a>. Furthermore, these species are also able to produce a large variety of secondary metabolites<a class="elsevierStyleCrossRefs" href="#bib0285">10,22,32,43</a>. Therefore, it is of relevance to study the behavior of Eurotium species in order to prevent their growth in animal feed, and their potential production of secondary metabolites.In recent years mathematical modeling has gained importance since they are considered helpful tools applied to avoid fungal growth and increase food shelf-life.Previously, we have reported a high incidence of Eurotium species in raw materials and animal feedstuff and an initial study of the effect of aw and temperature on the growth of Eurotium species was performed on synthetic media<a class="elsevierStyleCrossRef" href="#bib0350">23</a>. However, considering the high incidence and magnitude of damages that this genus can cause, it is more accurate to study the effect of environmental parameters in a more realistic context thus, in this case in a feed matrix.Therefore, the aims of this work were (i) to study the effect of aw and temperature on the growth of Eurotium species in a feed matrix, and (ii) to obtain predictive mathematical models for each species: the cardinal model of Rosso for secondary kinetic growth, and the probabilistic model for growth/no-growth interface.Materials and methodsFungal isolatesFour Eurotium species previously isolated from rabbit and chinchilla feed were used in the present study. Eurotium amstelodami, E. chevalieri, E. repens and E. rubrum strains were previously identified by scanning electron microscopy and DNA sequencing and characterized by secondary metabolite profiling, and growth on synthetic media<a class="elsevierStyleCrossRefs" href="#bib0345">22,23</a>.Experimental design and data generationA full factorial design was used to study the growth of four Eurotium species in a feed matrix. The effect of nine aw and five temperatures were evaluated. The aw assayed were in the range 0.71–0.97 (0.71, 0.73, 0.75, 0.79, 0.83, 0.87, 0.91, 0.95, 0.97), and the incubation temperatures were 5, 15, 25, 30 and 37°C.Additional aw values were evaluated for some of the species: E. amstelodami at 0.77 and 0.81 aw at 15, 25, 30 and 37°C; E. repens at 0.85 and 0.89 aw and 5, 15, 25 and 30°C.The maximum aw that the feed could reach without showing excess of water was 0.97 aw corresponding to a moisture content of 60%. Above this value there was surrounding water.Feedstuff media and water activity adjustmentCommercial a package of rabbit and chinchilla feed was purchased from a local store. The total content was subdivided into 1kg samples, each of which was placed in plastic bags and the total number of bags were placed into the original package made of a double layer of wood paper, and then closed again. Immediately, the bag sample was sent to the Semi-Industrial Irradiation Plant (PISI) in the Ezeiza Atomic Center (Buenos Aires province, Argentina) where it was irradiated with 35kGy of gamma irradiation in order to achieve sterilization. Initial moisture content and aw were 11.06±0.43% (humid basis) and 0.687±0.014, respectively. Then, an adsorption isotherm was built. First, small samples (50-100g) were adjusted to the required aw, and then were scaled to 1kg sample size. Each kilogram of sample was put into a 3 liter glass jar previously sterilized in autoclave at 121°C for 15min and dried in a drying oven at 105°C for 3h. Later, appropriate volumes of sterile distilled water were added in aseptic conditions, mixed and stored at 4°C for 7 days with periodic mixing. Moisture content was determined according to the AOAC Official Method<a class="elsevierStyleCrossRef" href="#bib0265">6</a> and aw was measured with LabSwift-aw Novasina. Each determination was done by triplicate, jars were vigorously shaken and samples were taken from different points. The feedstuff media were aseptically transferred into the Petri plates, a thin layer of feed was placed into them (10–15g), and previous to inoculation the plates were placed at each final incubation temperature for 24h in order to reach the incubation temperature<a class="elsevierStyleCrossRef" href="#bib0435">40</a>.Inoculum, inoculation and incubationHeavy sporulating cultures of each strain were obtained after seven days of growth at 25°C on Czapek yeast extract agar with 20% of sucrose (CY20S). Conidia were collected from the plate surface and suspended in 5ml of a water/glycerol solution adjusted to the required aw level and containing 0.05% v/v of Tween 80. Conidia concentration was determined with a Neubauer chamber and adjusted to a final concentration of 105conidia/ml. Petri plates containing the feed substrate were centrally inoculated with 1μl of the conidia suspension using a calibrated loop. Petri plates with the same aw were enclosed in polyethylene bags with plastic glasses containing water/glycerol solutions of the same aw in order to keep constant the relative humidity inside the bags and plates; water/glycerol solutions were renewed weekly. Incubation temperatures were 5, 15, 25, 30 and 37°C and the experiment had a duration of 40 days and a maximum of 90 days for the most unfavorable conditions. For each combination of aw*T*species ten plates were used. Additional non-inoculated plates with adjusted aw were placed into the corresponding bags to asses any aw deviation during the study.Growth assessmentPlates were observed twice a day with a binocular magnifier until growth was detected. Then, two perpendicular diameters of the colony were measured daily without opening the plates, until the colony reached the edge of the plate or until the end of the experiment.Growth data treatment and mathematical analysisData treatmentColony diameters were plotted against time, then linear regression was applied to obtain the growth rate (mm/day) as the slope of the line. All obtained data for each pair of (aw, T) conditions and replicates were transformed to obtain homogeneity of variance. All data were square root-transformed.Secondary modelSecondary models are used to describe the influence of environmental factors on fungal growth<a class="elsevierStyleCrossRef" href="#bib0430">39</a>. These models are of great importance and fundamentally very useful because they can be used to predict the microbiological shelf life of a food product<a class="elsevierStyleCrossRef" href="#bib0410">35</a>. In particular, in the Rosso cardinal model all of the studied parameters have a physiological meaning<a class="elsevierStyleCrossRef" href="#bib0330">19</a>. The data set obtained from this study was tested with other secondary models (data not shown), however the best fit was obtained with the Rosso cardinal model.Then, the influence of aw and temperature on fungal growth was studied. The Rosso cardinal model was applied to describe the effect of both environmental parameters on the radial growth rate. Each model performance was assessed by the root mean square error (RMSE) and coefficient of determination (R2).The equation describing the model is:<elsevierMultimedia ident="eq0005"></elsevierMultimedia>The equation was fitted using the nls function (nonlinear least squares) from the R Stats Package, in R (version 3.4.0).The parameter aw max was fixed at 0.97 since it was the maximum value that the feed can reach by addition of water without having free water/surrounding water. It is significant to mention that above 0.95 aw values there are notorious changes in feed appearance (pellet disintegration). Moreover, according to Sautour et al.<a class="elsevierStyleCrossRef" href="#bib0440">41</a>awmax should be set to one only for non-xerophilic fungi.Growth boundary modelProbabilistic models are used when the aim is to determine if growth can happen or not under specific conditions<a class="elsevierStyleCrossRef" href="#bib0330">19</a>. The information provided by these models is very useful because it allows to predict and avoid fungal (or bacterial) growth or even toxin production.<a class="elsevierStyleCrossRef" href="#bib0330">19</a> Then, this model instead of determining growth rate, determines whether or not growth occurs.The growth/no growth boundaries at different aw, temperature and time were built according to the linear logistic regression analysis. A value of 1 was assigned when growth was registered and 0 value when no growth occurred. A total of ten replicates were used.The equation for the logistic regression model is:<elsevierMultimedia ident="eq0010"></elsevierMultimedia>where P is the probability of growth (range 0–1), bi are the coefficients to be estimated, t (days) is the incubation time, aw is the water activity of the feed medium, and T is the incubation temperature (°C).The equation was fitted using the glm function (generalized linear models) from the R Stats Package, in R (version 3.4.0).The predicted growth/no growth boundaries interfaces for p=0.5 were calculated and plotted with Microsoft Excel 2007 Solver and Octave (GNU Project, Version 4.2.1).The predictive power/capacity and the goodness of fit of the logistic models were assessed by pseudo R2 indices (McFadden and Nagelkerke), percentage of concordance and accuracy<a class="elsevierStyleCrossRefs" href="#bib0250">3,14</a>.Performance of probability models at probability level of 0.50 was additionally evaluated by comparison with independent data obtained from studies carried out in analog media<a class="elsevierStyleCrossRefs" href="#bib0240">1,2,24,30,44</a>.ResultsGrowth description in feedGrowth of E. amstelodami, E. chevalieri, E. repens and E. rubrum was studied at different aw and temperatures.E. amstelodami grew in the range 0.77–0.97aw at 25 and 30°C. At 15°C the range was 0.81–0.97aw; and at 37°C the range was shorter, from 0.91 to 0.95 aw. The minimum growth rate was 0.19mm/day at 0.77aw and 25°C, while the maximum growth rate was 18.3mm/day at 0.91aw and 30°C. High growth rates were observed in the range 0.87–0.95 aw in the temperature interval of 15–37°C. Nevertheless, optimal aw and temperature were 0.91 and 30°C, respectively.E. chevalieri growth was observed in the range 0.79–0.97aw from 15 to 37°C and at 0.75aw at 30°C. The lowest growth rate was 0.13mm/day at 0.75aw and 30°C, while the highest growth rate was 23.6mm/day at 0.91aw and 30°C. High growth rates ranging from 10.5 to 15.8mm/day were also observed in the intervals of 0.83–0.91aw at 25–37°C temperature range.For E. repens, growth was registered from 0.91 to 0.97 aw at 5, 15, 25 and 30°C. Minimum growth rate was 0.29mm/day at 0.97aw and 5°C, whereas maximum growth occurred at 0.91aw and 25°C (20.16mm/day). At 15 and 30°C, high growth rates also occurred at both 0.91 and 0.95aw (10.2mm/day at 15°C and 16.7mm/day at 30°C; 9.3mm/day at 15°C and 6.3mm/day at 30°C, respectively).With regard to E. rubrum, growth was observed in the interval of 0.75–0.97aw. At 0.75aw only grew at 25°C, and growth at 5°C only occurred at 0.91aw. From 0.79 to 0.97aw growth took place at 15, 25 and 30°C. The optimal aw values ranged from 0.83 to 0.95 at 25 and 30°C with growth rates above 10mm/day, except at 0.95aw and 25°C. The maximum growth rate was 15.9mm/day (0.91aw at 25°C) and the minimum growth rate was 0.16mm/day (0.75aw and 25°C). No aw upper limit of growth was observed.It has been observed that the rate of growth in all species studied decreased at aw above 0.91 at all the temperatures tested.Secondary model<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a> shows the cardinal values of environmental factors obtained using the secondary cardinal model. The RMSE was calculated to assess the performance of the predictive models; in the calculation no-growth conditions next to the growth ones were included<a class="elsevierStyleCrossRef" href="#bib0390">31</a>. In general terms, the models showed a good fit with R2 ranging from 0.885 to 0.950. Nevertheless, the RMSE values were above 1, indicating a poor fit to experimental data. Despite the fact that all the RMSE values were higher than 1, there was high agreement between the parameters estimated by the model and the experimental data. For aw min, aw opt and aw max, the model provided values that fell within the experimental domain.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>For three out of four species there was agreement between the experimental aw opt and aw max and those estimated by the model (0.91 and 0.97, respectively). In addition, for Topt and Tmax, the model also showed high concordance with experimental data.With regard to Tmin for E. repens and E. rubrum, both species were able to grow at 5°C. The estimated value was below 5°C (2.78°C) for E. repens, whereas for E. rubrum, Tmin was estimated slightly above 5°C.With respect to optimal growth rate (μopt), there was high agreement for three out of four species. Nevertheless, there was a significant difference for E. repens, where the estimation of the model turned out to be twice as that of the experimental value.Growth/no-growth interface modelProbabilistic models were built for each of the Eurotium species at 7, 15 and 40 days. <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a> shows the coefficients obtained for each model.<elsevierMultimedia ident="tbl0010"></elsevierMultimedia>Developed models include linear, quadratic and interaction terms. There were two exceptions since terms were not significant (p-value>0.001): aw*T for E. chevalieri (p-value=0.0079), and aw2 for E. rubrum (p-value=0.452). Despite the fact that coefficients to assess predictive power and goodness of fit were accurate in both cases, the terms aw*T and aw2 were suppressed in each respective case in order to improve model fit and avoid overfitting (backward elimination of terms).<a class="elsevierStyleCrossRefs" href="#fig0005">Figures 1–4</a> show plots of probability of growth (p=0.50) for temperature and aw at 7, 15 and 40 days of incubation for each species. As it can be seen from the figures, growth occurred at almost all suitable conditions during the first 7 days. In general, a high percentage of logistic model agreement with the experimental data was found. The models showed a concordance percentage above 95 and accuracy above 0.929 (Table S1, supplementary material).<elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="fig0020"></elsevierMultimedia>Figures S1–S4 (supplementary material) show plot probability of growth vs. temperature for different values of aw and incubation times of 7, 15 and 40 days. These figures were also constructed by using the equations obtained by the model, and are complementary to <a class="elsevierStyleCrossRefs" href="#fig0005">Figures 1–4</a>.It is important to notice that the probability curves are almost centered at the experimental optimal temperature of growth for each species (except for E. repens, whose probability curve is centered at a lower temperature), and show how the probability of growth increases with time. It is worth noting that for E. chevalieri and E. rubrum this probability increment is notorious when comparing figures for 7 and 40 days respectively (Figs. S2 and S4).In addition, the validation of the models was also performed with external data from other authors<a class="elsevierStyleCrossRefs" href="#bib0240">1,2,24,30,44</a>. This comparison is shown in Table S2 (supplementary material), where the probability of growth of the models is shown on the “P model” column. Despite the fact that some disagreements were observed (8 false-positives and 5 false-negatives out of a total of 70 samples), the models predicted acceptably considering that the validation points were located at the domain-model boundaries.DiscussionGrowth and probabilistic models were developed for Eurotium species isolated from animal feed. It was found that in most of the evaluated conditions of aw and temperature, growth happened in a shorter period of time than in previously assayed synthetic media (MEA)<a class="elsevierStyleCrossRef" href="#bib0350">23</a>. Synthetic media are richer than natural feed and food, the latter provide a more real condition while synthetic media might induce an overestimated growth<a class="elsevierStyleCrossRef" href="#bib0270">7</a>. However, some discrepancies were found in the present study since in the feed matrix the temperatures required for growth at 0.75aw were 25 and 30°C (for E. rubrum and E. chevalieri, respectively), and no growth was detected at 15°C, while in synthetic media the lowest aw value at which growth occurred was 0.75 at 15 and 25°C. In addition, the temperature range of growth in feed was wider than in synthetic media, while the aw range of growth was more limited. Thus, we consider that the most appropriate choice is to perform the analysis in a culture media as close as possible to the original source from which molds were isolated. With regard to this topic, it was determined that the temperature and aw at which fungal growth occurred in animal feed were more in accordance with several authors<a class="elsevierStyleCrossRefs" href="#bib0240">1,13,15,47</a>. However, differences might be due to the nutritional composition of the substrates and geographical origin<a class="elsevierStyleCrossRefs" href="#bib0270">7,20,36</a>.In relation to the cardinal models, they provided accurate estimations for aw max and aw opt, μopt (except for E. repens), Tmax and Topt for all the four species studied (except Topt for E. chevalieri). On the other hand, the parameters aw min and Tmin were underestimated in all cases with the exception of aw min for E. chevalieri which was closer to the experimental observation.Rosso and Robinson<a class="elsevierStyleCrossRef" href="#bib0430">39</a> estimated E. amstelodami and E. chevalieri radial growth rate, aw min and aw opt from external data, which was obtained in a laboratory medium at 25°C and glycerol as substrate to control aw (among other solutes). These authors determined a radial growth rate of 2.52mm/day, 0.647 aw min and 0.974aw opt for E. amstelodami; while for E. chevalieri the radial growth rate was 2.664mm/day, 0.705aw min and 0.955aw opt. Once again, our findings differ widely since the radial growth rate in feed was about ten times higher than any informed value, aw min was higher than those and aw opt was lower than the values reported by the authors. However, Guynot et al.<a class="elsevierStyleCrossRef" href="#bib0360">25</a> reported a mean of maximum growth rates estimated by linear regression (at 0.90aw) comparable with the informed growth rates in the present study. In addition, Suhr and Nielsen<a class="elsevierStyleCrossRef" href="#bib0455">44</a> reported that E. repens was able to grow in rye bread medium at 0.80aw (controlled by glycerol) and exhibited visible growth before 30 days. Moreover, these authors reported that the shortest time for spoilage was observed at the highest aw (0.95) within a week<a class="elsevierStyleCrossRef" href="#bib0455">44</a>. In our study, E. repens was not able to grow below 0.91aw in animal feed and all the four species were able to grow at the highest aw (0.97) in less than a week's time.The RMSE and R2 were the parameters used to evaluate the goodness of fit and the fitting capacity of the models, respectively. The first parameter measures the deviation between observed and predicted values, while the second parameter indicates how useful the explanatory variables are in predicting the response variable<a class="elsevierStyleCrossRef" href="#bib0280">9</a>. A good fit is indicated by RMSE values close to zero, while a proper fitting capacity (R2) is indicated by values close to one.In the present work, despite the fact of having inoculated molds directly on the feed pellets, the obtained R2 values were very high (>0.885), comparable to values usually obtained from well-defined synthetic media<a class="elsevierStyleCrossRef" href="#bib0460">45</a>. Otherwise, RMSE values were too high to indicate a proper fit (2.139>RMSE>1.614). Similarly, Basak and Guha<a class="elsevierStyleCrossRef" href="#bib0275">8</a> informed an R2 range from 0.980 to 0.991 while RMSE ranged from 3.5 to 5.3 in a Penicillium expansum spore germination study.Probabilistic models developed for each Eurotium spp. showed agreement between observed and predicted data. This agreement was supported by the high values of the obtained coefficients. The pseudo R2 indices (McFadden and Nagelkerke) were accurate enough to indicate a good fit for each model. In addition, accuracy and concordance percentage for all developed models were also high, ranging from 95.58 to 97.81% and from 0.929 to 0.945, respectively. This agreement was also supported graphically, although probability models failed in some cases. Nevertheless, this kind of information can be usefully applied to prevent Eurotium spp. growth in animal feed during storage. As many of the conditions which favored growth occurred during the first 7 days, a safe storage from E. amstelodami growth will be achieved below 0.77aw regardless of the temperature (p of growth<0.10). To prevent E. chevalieri growth, aw above 0.79 (p of growth<0.10) should be avoided. In the case of E. repens aw should be kept below 0.91, and below 0.73aw for E. rubrum.In conclusion, the predictive models applied in the present study were able to satisfactorily describe and predict the effect of aw and temperature on growth and the probability of growth of Eurotium species in a feed matrix. Since these models were constructed in a real feed matrix they can be used as predictive tools to improve food safety, optimize food storage and prevent fungal spoilage. Thereby, important economic losses can be avoided and feed quality can be improved over time.To our knowledge this is the first report assessing the growth parameters of Eurotium species directly in animal feed.Conflict of interestThe authors declare that they have no conflicts of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:11 [ 0 => array:3 [ "identificador" => "xres1577109" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec1421245" "titulo" => "Keywords" ] 2 => array:3 [ "identificador" => "xres1577110" "titulo" => "Resumen" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0010" ] ] ] 3 => array:2 [ "identificador" => "xpalclavsec1421246" "titulo" => "Palabras clave" ] 4 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 5 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:6 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Fungal isolates" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Experimental design and data generation" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Feedstuff media and water activity adjustment" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Inoculum, inoculation and incubation" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Growth assessment" ] 5 => array:3 [ "identificador" => "sec0040" "titulo" => "Growth data treatment and mathematical analysis" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0045" "titulo" => "Data treatment" ] 1 => array:2 [ "identificador" => "sec0050" "titulo" => "Secondary model" ] 2 => array:2 [ "identificador" => "sec0055" "titulo" => "Growth boundary model" ] ] ] ] ] 6 => array:3 [ "identificador" => "sec0060" "titulo" => "Results" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0065" "titulo" => "Growth description in feed" ] 1 => array:2 [ "identificador" => "sec0070" "titulo" => "Secondary model" ] 2 => array:2 [ "identificador" => "sec0075" "titulo" => "Growth/no-growth interface model" ] ] ] 7 => array:2 [ "identificador" => "sec0080" "titulo" => "Discussion" ] 8 => array:2 [ "identificador" => "sec0085" "titulo" => "Conflict of interest" ] 9 => array:2 [ "identificador" => "xack557403" "titulo" => "Acknowledgments" ] 10 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2020-03-09" "fechaAceptado" => "2020-09-01" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec1421245" "palabras" => array:5 [ 0 => "Eurotium" 1 => "Feed matrix" 2 => "Environmental factors" 3 => "Water activity" 4 => "Modeling" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec1421246" "palabras" => array:5 [ 0 => "Eurotium" 1 => "Matriz alimento balanceado" 2 => "Factores ambientales" 3 => "Actividad de agua" 4 => "Modelado" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "Animal feeds are characterized by low water activity values. Nevertheless, fungal contamination with Eurotium species are quite common, causing nutritional depletion, spoilage and economic losses.The aim of this work was to assess Eurotium amstelodami, E. chevalieri, E. repens and E. rubrum growth in a feed matrix at different conditions of water activity (0.71–0.97) and temperature (5, 15, 25, 30 and 37°C).It was found that Eurotium species are able to grow in a wide range of water activity and temperature in a short period of time (7 days) and faster than in synthetic media. Rosso and probabilistic models were applied in order to determine the limiting and optimum growth conditions as well as growth probability at certain combinations of environmental factors. Both models provided an accurate fit to the cardinal parameters and good performance for growth/no growth cases.This is the first report assessing the growth parameters of Eurotium species directly in animal feed. Data obtained in the present study is useful to predict and avoid Eurotium species growth in animal feed." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "Los alimentos balanceados se caracterizan por tener bajos valores de actividad de agua. Sin embargo, la contaminación por hongos con especies de Eurotium es bastante común y causa agotamiento nutricional, deterioro y pérdidas económicas.El objetivo de este trabajo fue evaluar el crecimiento de Eurotium amstelodami, E. chevalieri, E. repens y E. rubrum en una matriz de alimento balanceado a diferentes condiciones de actividad de agua (0,71-0,97) y temperatura (5, 15, 25, 30 y 37°C).Se determinó que las especies de Eurotium pueden crecer en un amplio rango de actividades de agua y temperatura en un corto período de tiempo (7 días), y a mayor velocidad que en medio sintético. Se utilizaron los modelos de Rosso y probabilísticos para determinar las condiciones de crecimiento limitantes y óptimas, así como la probabilidad de crecimiento en ciertas combinaciones de factores ambientales. Ambos modelos proporcionaron un ajuste preciso a los parámetros cardinales y una buena performance para los casos de crecimiento/sin crecimiento.Este es el primer trabajo que evalúa los parámetros de crecimiento de las especies de Eurotium directamente en alimento balanceado. Los datos obtenidos en el presente estudio son útiles para predecir y evitar el crecimiento de especies de Eurotium en este tipo de alimentos." ] ] "apendice" => array:1 [ 0 => array:1 [ "seccion" => array:1 [ 0 => array:4 [ "apendice" => "The following are the supplementary data to this article:<elsevierMultimedia ident="upi0005"></elsevierMultimedia><elsevierMultimedia ident="upi0010"></elsevierMultimedia>" "etiqueta" => "Appendix A" "titulo" => "Supplementary data" "identificador" => "sec0095" ] ] ] ] "multimedia" => array:10 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 881 "Ancho" => 1575 "Tamanyo" => 74392 ] ] "descripcion" => array:1 [ "en" => "Growth/no growth interface of E. amstelodami after 7, 15 and 40 days of incubation in a feed matrix. P(0.50): short dashed line: at 7 days; long dashed line: at 15 days; solid line: at 40 days; white square: growth at 7 days; gray square: growth at 15 days; black square: growth at 40 days; cross: no growth." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 963 "Ancho" => 1461 "Tamanyo" => 72869 ] ] "descripcion" => array:1 [ "en" => "Growth/no growth interface of E. chevalieri after 7, 15 and 40 days of incubation in a feed matrix. P (0.50 short dashed line: at 7 days; long dashed line: at 15 days; solid line: at 40 days; white square: growth at 7 days; gray square: growth at 15 days; black square: growth at 40 days; cross: no growth." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 973 "Ancho" => 1520 "Tamanyo" => 77894 ] ] "descripcion" => array:1 [ "en" => "Growth/no growth interface of E. repens after 7, 15 and 40 days of incubation in a feed matrix. P (0.50): short dashed line: at 7 days; long dashed line: at 15 days; solid line: at 40 days; white square: growth at 7 days; gray square: growth at 15 days; black square: growth at 40 days; cross: no growth." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 971 "Ancho" => 1534 "Tamanyo" => 74374 ] ] "descripcion" => array:1 [ "en" => "Growth/no growth interface of E. rubrum after 7, 15 and 40 days of incubation in a feed matrix. P (0.50): short dashed line: at 7 days; long dashed line: at 15 days; solid line at 40 days; white square: growth at 7 days; gray square: growth at 15 days; black square: growth at 40 days; cross: no growth." ] ] 4 => array:8 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at1" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Species \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Parameter \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Estimated value±SE \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Experimental data \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">R2 \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">RMSE \t\t\t\t\t\t\n \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. amstelodami</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">μopt (mm/day) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.76±0.912 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">18.3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.908 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.614 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.974±0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awopt \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.913±0.003 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.757±0.002 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.77 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmax (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">38.93±0.483 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Topt (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">30.41±0.597 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmin (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.09±0.436 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. chevalieri</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">μopt (mm/day) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24.88±6.450 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">23.6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.950 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.021 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.978±0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awopt \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.909±0.002 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.744±0.002 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.75 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmax (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37.20±0.402 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Topt (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">34.99±2.127 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmin (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.39±0.403 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. repens</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">μopt (mm/day) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.21±1.275 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">20.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.885 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.139 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.990±0.006 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awopt \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.928±0.010 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.897±0.002 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmax (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">36.96±0.249 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">30 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Topt (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24.44±0.603 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmin (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.78±1.810 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. rubrum</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">μopt (mm/day) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">14.50±0.605 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15.9 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.896 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.687 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmax \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.972±0.0006 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.97 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awopt \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.911±0.003 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">awmin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.727±0.002 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.75 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmax (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37.04±0.029 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Topt (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">26.17±0.497 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Tmin (°C) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.38±0.339 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab2703638.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Experimental and estimated cardinal values of water activity and temperature, growth rate and coefficients" ] ] 5 => array:8 [ "identificador" => "tbl0010" "etiqueta" => "Table 2" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at2" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "leyenda" => "p-value<0.001." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Species \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Variable \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Estimated value \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Standard Error \t\t\t\t\t\t\n \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="8" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. amstelodami</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Intercept \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−481.10000 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">14.66 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.25680 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.01026 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1010.00000 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">31.45 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.70700 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.10007 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−550.10000 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.43 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.03314 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.001079 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw*T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−1.16900 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.07698 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.00296 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.0001321 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. chevalieri</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Intercept \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−454.000 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.180 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.435 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.018 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">953.100 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">37.210 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.594 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.054 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−524.000 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">20.840 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.028 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.001 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.004 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.000 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="8" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. repens</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Intercept \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−3220 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">172 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.211 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.015 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">6840 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">365 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−1.63 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.312 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−3640 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">195 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.0355 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.00166 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw*T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.344 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.0017 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.00015 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">E. rubrum</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Intercept \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−51.7 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.208 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.1992 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.01027 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">39.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.091 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.114 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.0874 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">T2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.03568 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.00110 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">aw*T \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.613703 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.06746 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">t2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">−0.002345 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.00013 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab2703637.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Estimated parameters by secondary logistic regression models" ] ] 6 => array:5 [ "identificador" => "upi0005" "tipo" => "MULTIMEDIAECOMPONENTE" "mostrarFloat" => false "mostrarDisplay" => true "Ecomponente" => array:2 [ "fichero" => "mmc1.pdf" "ficheroTamanyo" => 95875 ] ] 7 => array:5 [ "identificador" => "upi0010" "tipo" => "MULTIMEDIAECOMPONENTE" "mostrarFloat" => false "mostrarDisplay" => true "Ecomponente" => array:2 [ "fichero" => "mmc2.pdf" "ficheroTamanyo" => 81422 ] ] 8 => array:5 [ "identificador" => "eq0005" "tipo" => "MULTIMEDIAFORMULA" "mostrarFloat" => false "mostrarDisplay" => true "Formula" => array:5 [ "Matematica" => "μmax0.5=(μopt*((aw−aw max)*(aw−aw min)2)/((aw opt−aw min)*((aw opt−aw min)*(aw−aw opt)−(aw opt−aw max)*(aw opt+aw min−2*aw)))*((T−Tmax)*(T−Tmin)2)/((Topt−Tmin)*((Topt−Tmin)*(T−Topt)−(Topt−Tmax)*(Topt+Tmin−2*T))))0.5" "Fichero" => "STRIPIN_si1.jpeg" "Tamanyo" => 6482 "Alto" => 19 "Ancho" => 1025 ] ] 9 => array:5 [ "identificador" => "eq0010" "tipo" => "MULTIMEDIAFORMULA" "mostrarFloat" => false "mostrarDisplay" => true "Formula" => array:5 [ "Matematica" => "Logit P=ln(P/(1−P))=b0+b1aw+b2T+b3t+b4aw2+b5T2+¦b6awT+b7t2" "Fichero" => "STRIPIN_si2.jpeg" "Tamanyo" => 2566 "Alto" => 13 "Ancho" => 326 ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0015" "bibliografiaReferencia" => array:47 [ 0 => array:3 [ "identificador" => "bib0240" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Water activity and temperature effects on germination and growth of Eurotium amstelodami, E. chevalieri and E. herbariorum isolates from bakery products" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "M. Abellana" 1 => "J. Benedí" 2 => "V. Sanchis" 3 => "A.J. Ramos" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1046/j.1365-2672.1999.00828.x" "Revista" => array:6 [ "tituloSerie" => "J Appl Microbiol" "fecha" => "1999" "volumen" => "87" "paginaInicial" => "371" "paginaFinal" => "380" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/10540239" "web" => "Medline" ] ] ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bib0245" "etiqueta" => "2" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Water activity and temperature effects on growth of Eurotium amstelodami, E. chevalieri and E. herbariorum on a sponge cake analogue" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "M. Abellana" 1 => "X. Magrí" 2 => "V. Sanchis" 3 => "A.J. 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Caracterización del crecimiento y comportamiento predictivo de especies de Eurotium en matriz de alimento balanceado

Mariana Grecoa,b,

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mariana.greco@gmail.com

Corresponding author.

, Graciela Posea,b, Alejandro Pardoa,b

a Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, B1876BXD, Buenos Aires, Argentina

b Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina

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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Feedlot practices were introduced in Argentina in the 90s mainly for economic reasons&#46; Nowadays&#44; this practice still continues and at the same time feedstuff production has globally increased&#46; A recent feed survey indicates that worldwide feed production has exceeded a billion metric tons<a class="elsevierStyleCrossRef" href="#bib0255"><span class="elsevierStyleSup">4</span></a>&#59; 10&#46;6&#37; of feed production in the Latin American region is represented by Argentina<a class="elsevierStyleCrossRef" href="#bib0405"><span class="elsevierStyleSup">34</span></a>&#46; Particularly&#44; more than 14 million tons were produced in 2015&#44; while the most recent estimates suggest that around 25 million tons will be produced by 2020<a class="elsevierStyleCrossRef" href="#bib0370"><span class="elsevierStyleSup">27</span></a>&#46; Animal feeds are intended for cattle&#44; dairy cattle&#44; porcine&#44; poultry&#44; aquaculture&#44; rabbit and chinchilla&#44; and more recently buffalo&#46; Moreover&#44; an important growth has been registered in the pet food industry in the last years&#46; In addition to industrial feed production&#44; small farmers also elaborate their own feed&#46; However&#44; in some circumstances&#44; lack of quality assurance of raw material&#44; improper technical production process or inappropriate storage conditions can lead to fungal spoilage development and mycotoxin production&#46; In addition&#44; sometimes due to lack of knowledge or negligence&#44; despite the fact that visible fungal growth is observed&#44; feeds are mixed with non-contaminated feed in order to dilute its contamination&#44; thus economic losses are avoided&#46; Therefore&#44; feed quality decreases and becomes a risk for animals&#44; and consequently for the complete food chain<a class="elsevierStyleCrossRef" href="#bib0420"><span class="elsevierStyleSup">37</span></a>&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall">Animal feeds are composed of the necessary nutrients that animals need at every stage of their growth to reach a proper development and maximal growth in a profitable period of time for producers&#46; Several parameters are involved and considered to define the quality of feed pellets such as formulation&#44; durability&#44; starch gelatinization and protein content&#44; particle size&#44; exposure to high pressure steam and proper cooling&#44; among others<a class="elsevierStyleCrossRef" href="#bib0320"><span class="elsevierStyleSup">17</span></a>&#46; In addition&#44; the final product stability is regulated by <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#46; Therefore&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> is the most critical parameter to be taken into consideration since variations tend to increase fungal proliferation<a class="elsevierStyleCrossRefs" href="#bib0290"><span class="elsevierStyleSup">11&#44;29&#44;46</span></a>&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">Fungal contamination in raw materials and animal feed has been widely reported worldwide&#46; In fact&#44; most of them emphasize mycotoxigenic genera&#44; mainly <span class="elsevierStyleItalic">Alternaria</span>&#44; <span class="elsevierStyleItalic">Aspergillus</span>&#44; <span class="elsevierStyleItalic">Fusarium</span> and <span class="elsevierStyleItalic">Penicillium</span><a class="elsevierStyleCrossRefs" href="#bib0260"><span class="elsevierStyleSup">5&#44;16&#44;26&#44;37&#44;42</span></a>&#46; In addition&#44; based on the findings reported by Hassan et al&#46;<a class="elsevierStyleCrossRef" href="#bib0365"><span class="elsevierStyleSup">26</span></a>&#44; mixed cereals used as animal feeds are contaminated with high levels of mycotoxigenic fungi and mycotoxins&#46; Moreover&#44; several cases of human and animal intoxication and death caused by food and feedstuff contaminated with mycotoxins were also informed in the recent years<a class="elsevierStyleCrossRefs" href="#bib0325"><span class="elsevierStyleSup">18&#44;21&#44;33&#44;38</span></a>&#46;</p><p id="par0020" class="elsevierStylePara elsevierViewall">The presence and development of fungi cause changes in food and feed properties involving nutritional and organoleptic quality aspects&#46; Due to their xerophilic physiology <span class="elsevierStyleItalic">Eurotium</span> species are able to grow at <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> values as low as 0&#46;65<a class="elsevierStyleCrossRefs" href="#bib0295"><span class="elsevierStyleSup">12&#44;28</span></a>&#46; Furthermore&#44; these species are also able to produce a large variety of secondary metabolites<a class="elsevierStyleCrossRefs" href="#bib0285"><span class="elsevierStyleSup">10&#44;22&#44;32&#44;43</span></a>&#46; Therefore&#44; it is of relevance to study the behavior of <span class="elsevierStyleItalic">Eurotium</span> species in order to prevent their growth in animal feed&#44; and their potential production of secondary metabolites&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">In recent years mathematical modeling has gained importance since they are considered helpful tools applied to avoid fungal growth and increase food shelf-life&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">Previously&#44; we have reported a high incidence of <span class="elsevierStyleItalic">Eurotium</span> species in raw materials and animal feedstuff and an initial study of the effect of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature on the growth of <span class="elsevierStyleItalic">Eurotium</span> species was performed on synthetic media<a class="elsevierStyleCrossRef" href="#bib0350"><span class="elsevierStyleSup">23</span></a>&#46; However&#44; considering the high incidence and magnitude of damages that this genus can cause&#44; it is more accurate to study the effect of environmental parameters in a more realistic context thus&#44; in this case in a feed matrix&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">Therefore&#44; the aims of this work were &#40;i&#41; to study the effect of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature on the growth of <span class="elsevierStyleItalic">Eurotium</span> species in a feed matrix&#44; and &#40;ii&#41; to obtain predictive mathematical models for each species&#58; the cardinal model of Rosso for secondary kinetic growth&#44; and the probabilistic model for growth&#47;no-growth interface&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Fungal isolates</span><p id="par0040" class="elsevierStylePara elsevierViewall">Four <span class="elsevierStyleItalic">Eurotium</span> species previously isolated from rabbit and chinchilla feed were used in the present study&#46; <span class="elsevierStyleItalic">Eurotium amstelodami</span>&#44; <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#44; <span class="elsevierStyleItalic">E&#46; repens</span> and <span class="elsevierStyleItalic">E&#46; rubrum</span> strains were previously identified by scanning electron microscopy and DNA sequencing and characterized by secondary metabolite profiling&#44; and growth on synthetic media<a class="elsevierStyleCrossRefs" href="#bib0345"><span class="elsevierStyleSup">22&#44;23</span></a>&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Experimental design and data generation</span><p id="par0045" class="elsevierStylePara elsevierViewall">A full factorial design was used to study the growth of four <span class="elsevierStyleItalic">Eurotium</span> species in a feed matrix&#46; The effect of nine <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and five temperatures were evaluated&#46; The <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> assayed were in the range 0&#46;71&#8211;0&#46;97 &#40;0&#46;71&#44; 0&#46;73&#44; 0&#46;75&#44; 0&#46;79&#44; 0&#46;83&#44; 0&#46;87&#44; 0&#46;91&#44; 0&#46;95&#44; 0&#46;97&#41;&#44; and the incubation temperatures were 5&#44; 15&#44; 25&#44; 30 and 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Additional <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> values were evaluated for some of the species&#58; <span class="elsevierStyleItalic">E&#46; amstelodami</span> at 0&#46;77 and 0&#46;81 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 15&#44; 25&#44; 30 and 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#59; <span class="elsevierStyleItalic">E&#46; repens</span> at 0&#46;85 and 0&#46;89 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 5&#44; 15&#44; 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">The maximum <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> that the feed could reach without showing excess of water was 0&#46;97 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> corresponding to a moisture content of 60&#37;&#46; Above this value there was surrounding water&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Feedstuff media and water activity adjustment</span><p id="par0060" class="elsevierStylePara elsevierViewall">Commercial a package of rabbit and chinchilla feed was purchased from a local store&#46; The total content was subdivided into 1<span class="elsevierStyleHsp" style=""></span>kg samples&#44; each of which was placed in plastic bags and the total number of bags were placed into the original package made of a double layer of wood paper&#44; and then closed again&#46; Immediately&#44; the bag sample was sent to the Semi-Industrial Irradiation Plant &#40;PISI&#41; in the Ezeiza Atomic Center &#40;Buenos Aires province&#44; Argentina&#41; where it was irradiated with 35kGy of gamma irradiation in order to achieve sterilization&#46; Initial moisture content and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> were 11&#46;06<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;43&#37; &#40;humid basis&#41; and 0&#46;687<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;014&#44; respectively&#46; Then&#44; an adsorption isotherm was built&#46; First&#44; small samples &#40;50-100<span class="elsevierStyleHsp" style=""></span>g&#41; were adjusted to the required <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#44; and then were scaled to 1<span class="elsevierStyleHsp" style=""></span>kg sample size&#46; Each kilogram of sample was put into a 3 liter glass jar previously sterilized in autoclave at 121<span class="elsevierStyleHsp" style=""></span>&#176;C for 15<span class="elsevierStyleHsp" style=""></span>min and dried in a drying oven at 105<span class="elsevierStyleHsp" style=""></span>&#176;C for 3<span class="elsevierStyleHsp" style=""></span>h&#46; Later&#44; appropriate volumes of sterile distilled water were added in aseptic conditions&#44; mixed and stored at 4<span class="elsevierStyleHsp" style=""></span>&#176;C for 7 days with periodic mixing&#46; Moisture content was determined according to the AOAC Official Method<a class="elsevierStyleCrossRef" href="#bib0265"><span class="elsevierStyleSup">6</span></a> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> was measured with LabSwift-aw Novasina&#46; Each determination was done by triplicate&#44; jars were vigorously shaken and samples were taken from different points&#46; The feedstuff media were aseptically transferred into the Petri plates&#44; a thin layer of feed was placed into them &#40;10&#8211;15<span class="elsevierStyleHsp" style=""></span>g&#41;&#44; and previous to inoculation the plates were placed at each final incubation temperature for 24<span class="elsevierStyleHsp" style=""></span>h in order to reach the incubation temperature<a class="elsevierStyleCrossRef" href="#bib0435"><span class="elsevierStyleSup">40</span></a>&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Inoculum&#44; inoculation and incubation</span><p id="par0065" class="elsevierStylePara elsevierViewall">Heavy sporulating cultures of each strain were obtained after seven days of growth at 25<span class="elsevierStyleHsp" style=""></span>&#176;C on Czapek yeast extract agar with 20&#37; of sucrose &#40;CY20S&#41;&#46; Conidia were collected from the plate surface and suspended in 5<span class="elsevierStyleHsp" style=""></span>ml of a water&#47;glycerol solution adjusted to the required <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> level and containing 0&#46;05&#37; v&#47;v of Tween 80&#46; Conidia concentration was determined with a Neubauer chamber and adjusted to a final concentration of 10<span class="elsevierStyleSup">5</span><span class="elsevierStyleHsp" style=""></span>conidia&#47;ml&#46; Petri plates containing the feed substrate were centrally inoculated with 1<span class="elsevierStyleHsp" style=""></span>&#956;l of the conidia suspension using a calibrated loop&#46; Petri plates with the same <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> were enclosed in polyethylene bags with plastic glasses containing water&#47;glycerol solutions of the same <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> in order to keep constant the relative humidity inside the bags and plates&#59; water&#47;glycerol solutions were renewed weekly&#46; Incubation temperatures were 5&#44; 15&#44; 25&#44; 30 and 37<span class="elsevierStyleHsp" style=""></span>&#176;C and the experiment had a duration of 40 days and a maximum of 90 days for the most unfavorable conditions&#46; For each combination of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#42;<span class="elsevierStyleItalic">T</span>&#42;species ten plates were used&#46; Additional non-inoculated plates with adjusted <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> were placed into the corresponding bags to asses any <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> deviation during the study&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Growth assessment</span><p id="par0070" class="elsevierStylePara elsevierViewall">Plates were observed twice a day with a binocular magnifier until growth was detected&#46; Then&#44; two perpendicular diameters of the colony were measured daily without opening the plates&#44; until the colony reached the edge of the plate or until the end of the experiment&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Growth data treatment and mathematical analysis</span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Data treatment</span><p id="par0075" class="elsevierStylePara elsevierViewall">Colony diameters were plotted against time&#44; then linear regression was applied to obtain the growth rate &#40;mm&#47;day&#41; as the slope of the line&#46; All obtained data for each pair of &#40;<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#44; <span class="elsevierStyleItalic">T</span>&#41; conditions and replicates were transformed to obtain homogeneity of variance&#46; All data were square root-transformed&#46;</p></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Secondary model</span><p id="par0080" class="elsevierStylePara elsevierViewall">Secondary models are used to describe the influence of environmental factors on fungal growth<a class="elsevierStyleCrossRef" href="#bib0430"><span class="elsevierStyleSup">39</span></a>&#46; These models are of great importance and fundamentally very useful because they can be used to predict the microbiological shelf life of a food product<a class="elsevierStyleCrossRef" href="#bib0410"><span class="elsevierStyleSup">35</span></a>&#46; In particular&#44; in the Rosso cardinal model all of the studied parameters have a physiological meaning<a class="elsevierStyleCrossRef" href="#bib0330"><span class="elsevierStyleSup">19</span></a>&#46; The data set obtained from this study was tested with other secondary models &#40;data not shown&#41;&#44; however the best fit was obtained with the Rosso cardinal model&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Then&#44; the influence of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature on fungal growth was studied&#46; The Rosso cardinal model was applied to describe the effect of both environmental parameters on the radial growth rate&#46; Each model performance was assessed by the root mean square error &#40;RMSE&#41; and coefficient of determination &#40;<span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span>&#41;&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">The equation describing the model is&#58;<elsevierMultimedia ident="eq0005"></elsevierMultimedia></p><p id="par0095" class="elsevierStylePara elsevierViewall">The equation was fitted using the nls function &#40;nonlinear least squares&#41; from the R Stats Package&#44; in R &#40;version 3&#46;4&#46;0&#41;&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">The parameter <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> max</span> was fixed at 0&#46;97 since it was the maximum value that the feed can reach by addition of water without having free water&#47;surrounding water&#46; It is significant to mention that above 0&#46;95 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> values there are notorious changes in feed appearance &#40;pellet disintegration&#41;&#46; Moreover&#44; according to Sautour et al&#46;<a class="elsevierStyleCrossRef" href="#bib0440"><span class="elsevierStyleSup">41</span></a><span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span><span class="elsevierStyleInf">max</span> should be set to one only for non-xerophilic fungi&#46;</p></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Growth boundary model</span><p id="par0105" class="elsevierStylePara elsevierViewall">Probabilistic models are used when the aim is to determine if growth can happen or not under specific conditions<a class="elsevierStyleCrossRef" href="#bib0330"><span class="elsevierStyleSup">19</span></a>&#46; The information provided by these models is very useful because it allows to predict and avoid fungal &#40;or bacterial&#41; growth or even toxin production&#46;<a class="elsevierStyleCrossRef" href="#bib0330"><span class="elsevierStyleSup">19</span></a> Then&#44; this model instead of determining growth rate&#44; determines whether or not growth occurs&#46;</p><p id="par0110" class="elsevierStylePara elsevierViewall">The growth&#47;no growth boundaries at different <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#44; temperature and time were built according to the linear logistic regression analysis&#46; A value of 1 was assigned when growth was registered and 0 value when no growth occurred&#46; A total of ten replicates were used&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">The equation for the logistic regression model is&#58;<elsevierMultimedia ident="eq0010"></elsevierMultimedia>where <span class="elsevierStyleItalic">P</span> is the probability of growth &#40;range 0&#8211;1&#41;&#44; <span class="elsevierStyleItalic">b</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">i</span></span> are the coefficients to be estimated&#44; t &#40;days&#41; is the incubation time&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> is the water activity of the feed medium&#44; and T is the incubation temperature &#40;&#176;C&#41;&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">The equation was fitted using the glm function &#40;generalized linear models&#41; from the R Stats Package&#44; in R &#40;version 3&#46;4&#46;0&#41;&#46;</p><p id="par0125" class="elsevierStylePara elsevierViewall">The predicted growth&#47;no growth boundaries interfaces for <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;5 were calculated and plotted with Microsoft Excel 2007 Solver and Octave &#40;GNU Project&#44; Version 4&#46;2&#46;1&#41;&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">The predictive power&#47;capacity and the goodness of fit of the logistic models were assessed by pseudo <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> indices &#40;McFadden and Nagelkerke&#41;&#44; percentage of concordance and accuracy<a class="elsevierStyleCrossRefs" href="#bib0250"><span class="elsevierStyleSup">3&#44;14</span></a>&#46;</p><p id="par0135" class="elsevierStylePara elsevierViewall">Performance of probability models at probability level of 0&#46;50 was additionally evaluated by comparison with independent data obtained from studies carried out in analog media<a class="elsevierStyleCrossRefs" href="#bib0240"><span class="elsevierStyleSup">1&#44;2&#44;24&#44;30&#44;44</span></a>&#46;</p></span></span></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Results</span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Growth description in feed</span><p id="par0140" class="elsevierStylePara elsevierViewall">Growth of <span class="elsevierStyleItalic">E&#46; amstelodami</span>&#44; <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#44; <span class="elsevierStyleItalic">E&#46; repens</span> and <span class="elsevierStyleItalic">E&#46; rubrum</span> was studied at different <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperatures&#46;</p><p id="par0145" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">E&#46; amstelodami</span> grew in the range 0&#46;77&#8211;0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; At 15<span class="elsevierStyleHsp" style=""></span>&#176;C the range was 0&#46;81&#8211;0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#59; and at 37<span class="elsevierStyleHsp" style=""></span>&#176;C the range was shorter&#44; from 0&#46;91 to 0&#46;95 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#46; The minimum growth rate was 0&#46;19<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 0&#46;77<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; while the maximum growth rate was 18&#46;3<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; High growth rates were observed in the range 0&#46;87&#8211;0&#46;95 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> in the temperature interval of 15&#8211;37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; Nevertheless&#44; optimal <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature were 0&#46;91 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; respectively&#46;</p><p id="par0150" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">E&#46; chevalieri</span> growth was observed in the range 0&#46;79&#8211;0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> from 15 to 37<span class="elsevierStyleHsp" style=""></span>&#176;C and at 0&#46;75<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The lowest growth rate was 0&#46;13<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 0&#46;75<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; while the highest growth rate was 23&#46;6<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; High growth rates ranging from 10&#46;5 to 15&#46;8<span class="elsevierStyleHsp" style=""></span>mm&#47;day were also observed in the intervals of 0&#46;83&#8211;0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 25&#8211;37<span class="elsevierStyleHsp" style=""></span>&#176;C temperature range&#46;</p><p id="par0155" class="elsevierStylePara elsevierViewall">For <span class="elsevierStyleItalic">E&#46; repens</span>&#44; growth was registered from 0&#46;91 to 0&#46;97 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 5&#44; 15&#44; 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; Minimum growth rate was 0&#46;29<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 5<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; whereas maximum growth occurred at 0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 25<span class="elsevierStyleHsp" style=""></span>&#176;C &#40;20&#46;16<span class="elsevierStyleHsp" style=""></span>mm&#47;day&#41;&#46; At 15 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; high growth rates also occurred at both 0&#46;91 and 0&#46;95<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> &#40;10&#46;2<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 16&#46;7<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#59; 9&#46;3<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 15<span class="elsevierStyleHsp" style=""></span>&#176;C and 6&#46;3<span class="elsevierStyleHsp" style=""></span>mm&#47;day at 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; respectively&#41;&#46;</p><p id="par0160" class="elsevierStylePara elsevierViewall">With regard to <span class="elsevierStyleItalic">E&#46; rubrum</span>&#44; growth was observed in the interval of 0&#46;75&#8211;0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#46; At 0&#46;75<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> only grew at 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; and growth at 5<span class="elsevierStyleHsp" style=""></span>&#176;C only occurred at 0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#46; From 0&#46;79 to 0&#46;97<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> growth took place at 15&#44; 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The optimal <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> values ranged from 0&#46;83 to 0&#46;95 at 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C with growth rates above 10<span class="elsevierStyleHsp" style=""></span>mm&#47;day&#44; except at 0&#46;95<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The maximum growth rate was 15&#46;9<span class="elsevierStyleHsp" style=""></span>mm&#47;day &#40;0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#41; and the minimum growth rate was 0&#46;16<span class="elsevierStyleHsp" style=""></span>mm&#47;day &#40;0&#46;75<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#41;&#46; No <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> upper limit of growth was observed&#46;</p><p id="par0165" class="elsevierStylePara elsevierViewall">It has been observed that the rate of growth in all species studied decreased at <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> above 0&#46;91 at all the temperatures tested&#46;</p></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Secondary model</span><p id="par0170" class="elsevierStylePara elsevierViewall"><a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a> shows the cardinal values of environmental factors obtained using the secondary cardinal model&#46; The RMSE was calculated to assess the performance of the predictive models&#59; in the calculation no-growth conditions next to the growth ones were included<a class="elsevierStyleCrossRef" href="#bib0390"><span class="elsevierStyleSup">31</span></a>&#46; In general terms&#44; the models showed a good fit with <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> ranging from 0&#46;885 to 0&#46;950&#46; Nevertheless&#44; the RMSE values were above 1&#44; indicating a poor fit to experimental data&#46; Despite the fact that all the RMSE values were higher than 1&#44; there was high agreement between the parameters estimated by the model and the experimental data&#46; For <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span>&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> max</span>&#44; the model provided values that fell within the experimental domain&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0175" class="elsevierStylePara elsevierViewall">For three out of four species there was agreement between the experimental <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> max</span> and those estimated by the model &#40;0&#46;91 and 0&#46;97&#44; respectively&#41;&#46; In addition&#44; for <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">opt</span> and <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">max</span>&#44; the model also showed high concordance with experimental data&#46;</p><p id="par0180" class="elsevierStylePara elsevierViewall">With regard to <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">min</span> for <span class="elsevierStyleItalic">E&#46; repens</span> and <span class="elsevierStyleItalic">E&#46; rubrum</span>&#44; both species were able to grow at 5<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The estimated value was below 5<span class="elsevierStyleHsp" style=""></span>&#176;C &#40;2&#46;78<span class="elsevierStyleHsp" style=""></span>&#176;C&#41; for <span class="elsevierStyleItalic">E&#46; repens</span>&#44; whereas for <span class="elsevierStyleItalic">E&#46; rubrum</span>&#44; <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">min</span> was estimated slightly above 5<span class="elsevierStyleHsp" style=""></span>&#176;C&#46;</p><p id="par0185" class="elsevierStylePara elsevierViewall">With respect to optimal growth rate &#40;<span class="elsevierStyleItalic">&#956;</span><span class="elsevierStyleInf">opt</span>&#41;&#44; there was high agreement for three out of four species&#46; Nevertheless&#44; there was a significant difference for <span class="elsevierStyleItalic">E&#46; repens</span>&#44; where the estimation of the model turned out to be twice as that of the experimental value&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Growth&#47;no-growth interface model</span><p id="par0190" class="elsevierStylePara elsevierViewall">Probabilistic models were built for each of the <span class="elsevierStyleItalic">Eurotium</span> species at 7&#44; 15 and 40 days&#46; <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a> shows the coefficients obtained for each model&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><p id="par0195" class="elsevierStylePara elsevierViewall">Developed models include linear&#44; quadratic and interaction terms&#46; There were two exceptions since terms were not significant &#40;<span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#58; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#42;<span class="elsevierStyleItalic">T</span> for <span class="elsevierStyleItalic">E&#46; chevalieri</span> &#40;<span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;0079&#41;&#44; and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span><span class="elsevierStyleSup">2</span> for <span class="elsevierStyleItalic">E&#46; rubrum</span> &#40;<span class="elsevierStyleItalic">p</span>-value<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;452&#41;&#46; Despite the fact that coefficients to assess predictive power and goodness of fit were accurate in both cases&#44; the terms <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#42;<span class="elsevierStyleItalic">T</span> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span><span class="elsevierStyleSup">2</span> were suppressed in each respective case in order to improve model fit and avoid overfitting &#40;backward elimination of terms&#41;&#46;</p><p id="par0200" class="elsevierStylePara elsevierViewall"><a class="elsevierStyleCrossRefs" href="#fig0005">Figures 1&#8211;4</a> show plots of probability of growth &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;50&#41; for temperature and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at 7&#44; 15 and 40 days of incubation for each species&#46; As it can be seen from the figures&#44; growth occurred at almost all suitable conditions during the first 7 days&#46; In general&#44; a high percentage of logistic model agreement with the experimental data was found&#46; The models showed a concordance percentage above 95 and accuracy above 0&#46;929 &#40;Table S1&#44; supplementary material&#41;&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="fig0020"></elsevierMultimedia><p id="par0205" class="elsevierStylePara elsevierViewall">Figures S1&#8211;S4 &#40;supplementary material&#41; show plot probability of growth vs&#46; temperature for different values of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and incubation times of 7&#44; 15 and 40 days&#46; These figures were also constructed by using the equations obtained by the model&#44; and are complementary to <a class="elsevierStyleCrossRefs" href="#fig0005">Figures 1&#8211;4</a>&#46;</p><p id="par0210" class="elsevierStylePara elsevierViewall">It is important to notice that the probability curves are almost centered at the experimental optimal temperature of growth for each species &#40;except for <span class="elsevierStyleItalic">E&#46; repens</span>&#44; whose probability curve is centered at a lower temperature&#41;&#44; and show how the probability of growth increases with time&#46; It is worth noting that for <span class="elsevierStyleItalic">E&#46; chevalieri</span> and <span class="elsevierStyleItalic">E&#46; rubrum</span> this probability increment is notorious when comparing figures for 7 and 40 days respectively &#40;Figs&#46; S2 and S4&#41;&#46;</p><p id="par0215" class="elsevierStylePara elsevierViewall">In addition&#44; the validation of the models was also performed with external data from other authors<a class="elsevierStyleCrossRefs" href="#bib0240"><span class="elsevierStyleSup">1&#44;2&#44;24&#44;30&#44;44</span></a>&#46; This comparison is shown in Table S2 &#40;supplementary material&#41;&#44; where the probability of growth of the models is shown on the &#8220;<span class="elsevierStyleItalic">P</span> model&#8221; column&#46; Despite the fact that some disagreements were observed &#40;8 false-positives and 5 false-negatives out of a total of 70 samples&#41;&#44; the models predicted acceptably considering that the validation points were located at the domain-model boundaries&#46;</p></span></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Discussion</span><p id="par0220" class="elsevierStylePara elsevierViewall">Growth and probabilistic models were developed for <span class="elsevierStyleItalic">Eurotium</span> species isolated from animal feed&#46; It was found that in most of the evaluated conditions of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature&#44; growth happened in a shorter period of time than in previously assayed synthetic media &#40;MEA&#41;<a class="elsevierStyleCrossRef" href="#bib0350"><span class="elsevierStyleSup">23</span></a>&#46; Synthetic media are richer than natural feed and food&#44; the latter provide a more real condition while synthetic media might induce an overestimated growth<a class="elsevierStyleCrossRef" href="#bib0270"><span class="elsevierStyleSup">7</span></a>&#46; However&#44; some discrepancies were found in the present study since in the feed matrix the temperatures required for growth at 0&#46;75<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> were 25 and 30<span class="elsevierStyleHsp" style=""></span>&#176;C &#40;for <span class="elsevierStyleItalic">E&#46; rubrum</span> and <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#44; respectively&#41;&#44; and no growth was detected at 15<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; while in synthetic media the lowest <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> value at which growth occurred was 0&#46;75 at 15 and 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; In addition&#44; the temperature range of growth in feed was wider than in synthetic media&#44; while the <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> range of growth was more limited&#46; Thus&#44; we consider that the most appropriate choice is to perform the analysis in a culture media as close as possible to the original source from which molds were isolated&#46; With regard to this topic&#44; it was determined that the temperature and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> at which fungal growth occurred in animal feed were more in accordance with several authors<a class="elsevierStyleCrossRefs" href="#bib0240"><span class="elsevierStyleSup">1&#44;13&#44;15&#44;47</span></a>&#46; However&#44; differences might be due to the nutritional composition of the substrates and geographical origin<a class="elsevierStyleCrossRefs" href="#bib0270"><span class="elsevierStyleSup">7&#44;20&#44;36</span></a>&#46;</p><p id="par0225" class="elsevierStylePara elsevierViewall">In relation to the cardinal models&#44; they provided accurate estimations for <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> max</span> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span>&#44; <span class="elsevierStyleItalic">&#956;</span><span class="elsevierStyleInf">opt</span> &#40;except for <span class="elsevierStyleItalic">E&#46; repens</span>&#41;&#44; <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">max</span> and <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">opt</span> for all the four species studied &#40;except <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">opt</span> for <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#41;&#46; On the other hand&#44; the parameters <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> and <span class="elsevierStyleItalic">T</span><span class="elsevierStyleInf">min</span> were underestimated in all cases with the exception of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> for <span class="elsevierStyleItalic">E&#46; chevalieri</span> which was closer to the experimental observation&#46;</p><p id="par0230" class="elsevierStylePara elsevierViewall">Rosso and Robinson<a class="elsevierStyleCrossRef" href="#bib0430"><span class="elsevierStyleSup">39</span></a> estimated <span class="elsevierStyleItalic">E&#46; amstelodami</span> and <span class="elsevierStyleItalic">E&#46; chevalieri</span> radial growth rate&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span> from external data&#44; which was obtained in a laboratory medium at 25<span class="elsevierStyleHsp" style=""></span>&#176;C and glycerol as substrate to control <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> &#40;among other solutes&#41;&#46; These authors determined a radial growth rate of 2&#46;52<span class="elsevierStyleHsp" style=""></span>mm&#47;day&#44; 0&#46;647 <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> and 0&#46;974<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span> for <span class="elsevierStyleItalic">E&#46; amstelodami</span>&#59; while for <span class="elsevierStyleItalic">E&#46; chevalieri</span> the radial growth rate was 2&#46;664<span class="elsevierStyleHsp" style=""></span>mm&#47;day&#44; 0&#46;705<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> and 0&#46;955<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span>&#46; Once again&#44; our findings differ widely since the radial growth rate in feed was about ten times higher than any informed value&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> min</span> was higher than those and <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span> opt</span> was lower than the values reported by the authors&#46; However&#44; Guynot et al&#46;<a class="elsevierStyleCrossRef" href="#bib0360"><span class="elsevierStyleSup">25</span></a> reported a mean of maximum growth rates estimated by linear regression &#40;at 0&#46;90<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span>&#41; comparable with the informed growth rates in the present study&#46; In addition&#44; Suhr and Nielsen<a class="elsevierStyleCrossRef" href="#bib0455"><span class="elsevierStyleSup">44</span></a> reported that <span class="elsevierStyleItalic">E&#46; repens</span> was able to grow in rye bread medium at 0&#46;80<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> &#40;controlled by glycerol&#41; and exhibited visible growth before 30 days&#46; Moreover&#44; these authors reported that the shortest time for spoilage was observed at the highest <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> &#40;0&#46;95&#41; within a week<a class="elsevierStyleCrossRef" href="#bib0455"><span class="elsevierStyleSup">44</span></a>&#46; In our study&#44; <span class="elsevierStyleItalic">E&#46; repens</span> was not able to grow below 0&#46;91<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> in animal feed and all the four species were able to grow at the highest <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> &#40;0&#46;97&#41; in less than a week&#39;s time&#46;</p><p id="par0235" class="elsevierStylePara elsevierViewall">The RMSE and <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> were the parameters used to evaluate the goodness of fit and the fitting capacity of the models&#44; respectively&#46; The first parameter measures the deviation between observed and predicted values&#44; while the second parameter indicates how useful the explanatory variables are in predicting the response variable<a class="elsevierStyleCrossRef" href="#bib0280"><span class="elsevierStyleSup">9</span></a>&#46; A good fit is indicated by RMSE values close to zero&#44; while a proper fitting capacity &#40;<span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span>&#41; is indicated by values close to one&#46;</p><p id="par0240" class="elsevierStylePara elsevierViewall">In the present work&#44; despite the fact of having inoculated molds directly on the feed pellets&#44; the obtained <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> values were very high &#40;&#62;0&#46;885&#41;&#44; comparable to values usually obtained from well-defined synthetic media<a class="elsevierStyleCrossRef" href="#bib0460"><span class="elsevierStyleSup">45</span></a>&#46; Otherwise&#44; RMSE values were too high to indicate a proper fit &#40;2&#46;139<span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>RMSE<span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>1&#46;614&#41;&#46; Similarly&#44; Basak and Guha<a class="elsevierStyleCrossRef" href="#bib0275"><span class="elsevierStyleSup">8</span></a> informed an <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> range from 0&#46;980 to 0&#46;991 while RMSE ranged from 3&#46;5 to 5&#46;3 in a <span class="elsevierStyleItalic">Penicillium expansum</span> spore germination study&#46;</p><p id="par0245" class="elsevierStylePara elsevierViewall">Probabilistic models developed for each <span class="elsevierStyleItalic">Eurotium</span> spp&#46; showed agreement between observed and predicted data&#46; This agreement was supported by the high values of the obtained coefficients&#46; The pseudo <span class="elsevierStyleItalic">R</span><span class="elsevierStyleSup">2</span> indices &#40;McFadden and Nagelkerke&#41; were accurate enough to indicate a good fit for each model&#46; In addition&#44; accuracy and concordance percentage for all developed models were also high&#44; ranging from 95&#46;58 to 97&#46;81&#37; and from 0&#46;929 to 0&#46;945&#44; respectively&#46; This agreement was also supported graphically&#44; although probability models failed in some cases&#46; Nevertheless&#44; this kind of information can be usefully applied to prevent <span class="elsevierStyleItalic">Eurotium</span> spp&#46; growth in animal feed during storage&#46; As many of the conditions which favored growth occurred during the first 7 days&#44; a safe storage from <span class="elsevierStyleItalic">E&#46; amstelodami</span> growth will be achieved below 0&#46;77<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> regardless of the temperature &#40;<span class="elsevierStyleItalic">p</span> of growth<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;10&#41;&#46; To prevent <span class="elsevierStyleItalic">E&#46; chevalieri</span> growth&#44; <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> above 0&#46;79 &#40;<span class="elsevierStyleItalic">p</span> of growth<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;10&#41; should be avoided&#46; In the case of <span class="elsevierStyleItalic">E&#46; repens a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> should be kept below 0&#46;91&#44; and below 0&#46;73<span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> for <span class="elsevierStyleItalic">E&#46; rubrum</span>&#46;</p><p id="par0250" class="elsevierStylePara elsevierViewall">In conclusion&#44; the predictive models applied in the present study were able to satisfactorily describe and predict the effect of <span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span> and temperature on growth and the probability of growth of <span class="elsevierStyleItalic">Eurotium</span> species in a feed matrix&#46; Since these models were constructed in a real feed matrix they can be used as predictive tools to improve food safety&#44; optimize food storage and prevent fungal spoilage&#46; Thereby&#44; important economic losses can be avoided and feed quality can be improved over time&#46;</p><p id="par0255" class="elsevierStylePara elsevierViewall">To our knowledge this is the first report assessing the growth parameters of <span class="elsevierStyleItalic">Eurotium</span> species directly in animal feed&#46;</p></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Conflict of interest</span><p id="par0260" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest&#46;</p></span></span>"
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          "titulo" => "Introduction"
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          "titulo" => "Materials and methods"
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              "titulo" => "Feedstuff media and water activity adjustment"
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              "titulo" => "Inoculum&#44; inoculation and incubation"
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          "titulo" => "Results"
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              "identificador" => "sec0065"
              "titulo" => "Growth description in feed"
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            1 => array:2 [
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              "titulo" => "Secondary model"
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            2 => array:2 [
              "identificador" => "sec0075"
              "titulo" => "Growth&#47;no-growth interface model"
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          "titulo" => "Discussion"
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          "titulo" => "Acknowledgments"
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          "titulo" => "References"
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    "fechaRecibido" => "2020-03-09"
    "fechaAceptado" => "2020-09-01"
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            0 => "<span class="elsevierStyleItalic">Eurotium</span>"
            1 => "Feed matrix"
            2 => "Environmental factors"
            3 => "Water activity"
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            1 => "Matriz alimento balanceado"
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        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Animal feeds are characterized by low water activity values&#46; Nevertheless&#44; fungal contamination with <span class="elsevierStyleItalic">Eurotium</span> species are quite common&#44; causing nutritional depletion&#44; spoilage and economic losses&#46;</p><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">The aim of this work was to assess <span class="elsevierStyleItalic">Eurotium amstelodami</span>&#44; <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#44; <span class="elsevierStyleItalic">E&#46; repens</span> and <span class="elsevierStyleItalic">E&#46; rubrum</span> growth in a feed matrix at different conditions of water activity &#40;0&#46;71&#8211;0&#46;97&#41; and temperature &#40;5&#44; 15&#44; 25&#44; 30 and 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#41;&#46;</p><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">It was found that <span class="elsevierStyleItalic">Eurotium</span> species are able to grow in a wide range of water activity and temperature in a short period of time &#40;7 days&#41; and faster than in synthetic media&#46; Rosso and probabilistic models were applied in order to determine the limiting and optimum growth conditions as well as growth probability at certain combinations of environmental factors&#46; Both models provided an accurate fit to the cardinal parameters and good performance for growth&#47;no growth cases&#46;</p><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">This is the first report assessing the growth parameters of <span class="elsevierStyleItalic">Eurotium</span> species directly in animal feed&#46; Data obtained in the present study is useful to predict and avoid <span class="elsevierStyleItalic">Eurotium</span> species growth in animal feed&#46;</p></span>"
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        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Los alimentos balanceados se caracterizan por tener bajos valores de actividad de agua&#46; Sin embargo&#44; la contaminaci&#243;n por hongos con especies de <span class="elsevierStyleItalic">Eurotium</span> es bastante com&#250;n y causa agotamiento nutricional&#44; deterioro y p&#233;rdidas econ&#243;micas&#46;</p><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">El objetivo de este trabajo fue evaluar el crecimiento de <span class="elsevierStyleItalic">Eurotium amstelodami</span>&#44; <span class="elsevierStyleItalic">E&#46; chevalieri</span>&#44; <span class="elsevierStyleItalic">E&#46; repens</span> y <span class="elsevierStyleItalic">E&#46; rubrum</span> en una matriz de alimento balanceado a diferentes condiciones de actividad de agua &#40;0&#44;71-0&#44;97&#41; y temperatura &#40;5&#44; 15&#44; 25&#44; 30 y 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#41;&#46;</p><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Se determin&#243; que las especies de <span class="elsevierStyleItalic">Eurotium</span> pueden crecer en un amplio rango de actividades de agua y temperatura en un corto per&#237;odo de tiempo &#40;7 d&#237;as&#41;&#44; y a mayor velocidad que en medio sint&#233;tico&#46; Se utilizaron los modelos de Rosso y probabil&#237;sticos para determinar las condiciones de crecimiento limitantes y &#243;ptimas&#44; as&#237; como la probabilidad de crecimiento en ciertas combinaciones de factores ambientales&#46; Ambos modelos proporcionaron un ajuste preciso a los par&#225;metros cardinales y una buena <span class="elsevierStyleItalic">performance</span> para los casos de crecimiento&#47;sin crecimiento&#46;</p><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Este es el primer trabajo que eval&#250;a los par&#225;metros de crecimiento de las especies de <span class="elsevierStyleItalic">Eurotium</span> directamente en alimento balanceado&#46; Los datos obtenidos en el presente estudio son &#250;tiles para predecir y evitar el crecimiento de especies de <span class="elsevierStyleItalic">Eurotium</span> en este tipo de alimentos&#46;</p></span>"
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          "en" => "<p id="spar0075" class="elsevierStyleSimplePara elsevierViewall">Growth&#47;no growth interface of <span class="elsevierStyleItalic">E&#46; rubrum</span> after 7&#44; 15 and 40 days of incubation in a feed matrix&#46;</p> <p id="spar0080" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">P</span> &#40;0&#46;50&#41;&#58; short dashed line&#58; at 7 days&#59; long dashed line&#58; at 15 days&#59; solid line at 40 days&#59; white square&#58; growth at 7 days&#59; gray square&#58; growth at 15 days&#59; black square&#58; growth at 40 days&#59; cross&#58; no growth&#46;</p>"
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                  \t\t\t\t">37&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;402&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">37&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">34&#46;99<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;127&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">2&#46;139&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">30&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">24&#46;44<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;603&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">25&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">5&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t">14&#46;50<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;605&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">15&#46;9&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;896&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;687&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">a</span><span class="elsevierStyleInf"><span class="elsevierStyleItalic">w</span></span><span class="elsevierStyleInf">max</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;972<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;0006&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;97&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
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Article information

ISSN: 03257541
Original language: English

DOI:10.1016/j.ram.2020.09.006

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Year/Month	Html	Pdf	Total

2024 November	2	0	2
2024 October	22	21	43
2024 September	52	19	71
2024 August	28	16	44
2024 July	46	30	76
2024 June	26	6	32
2024 May	16	11	27
2024 April	27	17	44
2024 March	54	11	65
2024 February	44	7	51
2024 January	34	7	41
2023 December	30	5	35
2023 November	52	8	60
2023 October	46	20	66
2023 September	30	14	44
2023 August	38	13	51
2023 July	27	4	31
2023 June	46	3	49
2023 May	89	10	99
2023 April	77	3	80
2023 March	33	2	35
2023 February	39	5	44
2023 January	40	16	56
2022 December	13	30	43
2022 November	20	21	41
2022 October	29	15	44
2022 September	15	26	41
2022 August	17	17	34
2022 July	15	5	20
2022 June	22	14	36
2022 May	26	19	45
2022 April	34	15	49
2022 March	45	30	75
2022 February	25	19	44
2022 January	26	12	38
2021 December	33	21	54
2021 November	40	24	64
2021 October	61	31	92
2021 September	35	25	60
2021 August	23	17	40
2021 July	16	25	41
2021 June	6	10	16
2021 May	4	14	18
2021 April	9	21	30
2021 March	10	12	22
2021 February	6	6	12
2021 January	9	4	13
2020 December	0	3	3

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