Food Research International 130 (2020) 108912

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Food Research International

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Predictive modelling for the growth kinetics of Pseudomonas spp. on button T

mushroom (Agaricus bisporus) under isothermal and non-isothermal
conditions
Fatih Tarlaka, Murat Ozdemirb, , Mehmet Melikoglub
⁎

a
Department of Nutrition and Dietetics, Istanbul Gedik University, 34876 Kartal, Istanbul, Turkey
b
Department of Chemical Engineering, Gebze Technical University, 41400 Gebze, Kocaeli, Turkey

ARTICLE INFO ABSTRACT

Keywords: Baranyi model was fitted to experimental growth data of Pseudomonas spp. on the button mushrooms (Agaricus
Predictive microbiology bisporus) stored at different isothermal conditions (4, 12, 20 and 28 °C), and the kinetic growth parameters of
Microbiological change Pseudomonas spp. on the button mushrooms were obtained. The goodness of fit of the Baranyi model was
Dynamic condition evaluated by considering the root mean squared error (RMSE) and the adjusted coefficient of determination
Growth behaviour
(adjusted-R2). The Baranyi model gave RMSE values lower than 0.193 and adjusted-R2 values higher than 0.975
for all isothermal storage temperatures. The maximum specific growth rate (µmax) was described as a function of
temperature using secondary models namely, Ratkowsky and Arrhenius models. The Ratkowsky model described
the temperature dependence of µmax better than the Arrhenius model. Therefore, the differential form of the
Baranyi model was merged with the Ratkowsky model, and solved numerically using the fourth-order Runge-
Kutta method to predict the concentration of Pseudomonas spp. populations on button mushrooms under non-
isothermal conditions in which they are frequently subjected to during storage, delivery and retail marketing.
The validation performance of the dynamic model used was assessed by considering bias (Bf) and accuracy (Af)
factors which were found to be 0.998 and 1.016, respectively. The dynamic model developed also exhibited
quite small mean deviation (MD) and mean absolute deviation (MAD) values being −0.013 and 0.126 log CFU/
g, respectively. The modelling approach used in this work could be an alternative to traditional enumeration
techniques to determine the number of Pseudomonas spp. on mushrooms as a function of temperature and time.

1. Introduction Predictive microbiology can be considered as an important field in

Mushrooms have been considered as nutrient-rich food due to their
high protein and low cholesterol contents (Wani, Bodha, & Wani, 2010).
More than 2000 species of mushrooms exist in nature, but a few of those
are cultivated commercially in industrial scale (Valverde, Hernández-
Pérez, & Paredes-López, 2015). Among these species, the button mush-
room (Agaricus bisporus) is the most common edible mushroom all over
the world. Agaricus bisporus has no cuticle to protect it from physical
damage and microbial attack. Therefore, it can be easily contaminated
with microorganisms during growing and processing. Pseudomonas spp.
have been isolated as the most abundant bacterial genus, responsible for
the spoilage of Agaricus bisporus, and it was reported that Pseudomonas
spp. could range from 6.63 to 8.10 log CFU/g in mushrooms (Reyes,
Venturini, Oria, & Blanco, 2004; Simón & González-Fandos, 2010;
Venturini, Reyes, Rivera, Oria, & Blanco, 2011).
food microbiology in which it uses predictive models to describe the
microbial growth in different food products (Pérez-Rodríguez & Valero,
2013). Predictive models predict the growth of microorganisms quickly,
efficiently and in a cost-effective way as compared to traditional
methods of enumeration which are long-lasting, expensive and time-
consuming. The mathematical models used in predictive microbiology
are mainly categorised as primary, secondary and tertiary models
(Whiting, 1995). The primary models are the mathematical equations
that define the growth data as a function of time under a constant en-
vironmental condition. The Baranyi model (Baranyi & Roberts, 1994) is
the most extensively used primary model for describing microbial
growth data. The maximum specific growth rate, which is the one of the
most critical growth kinetic parameters, can be modelled using the
secondary models as a function of temperature, oxygen, pH and water
activity (aw). Temperature has the key role in affecting the growth

⁎
Corresponding author.
E-mail address: ozdemirm@gtu.edu.tr (M. Ozdemir).

https://doi.org/10.1016/j.foodres.2019.108912
Received 23 July 2019; Received in revised form 12 December 2019; Accepted 15 December 2019
Available online 18 December 2019
0963-9969/ © 2019 Elsevier Ltd. All rights reserved.
F. Tarlak, et al.
behaviour of microorganisms in foods. The Ratkowsky (Ratkowsky,
Olley, McMeekin, & Ball, 1982) and Arrhenius (Taoukis, Koutsoumanis,
& Nychas, 1999) models are frequently used secondary models to de-
termine temperature dependence of microbial growth. Tertiary models
are formed by combining primary and secondary models, and they use a
computer as an estimation tool, but these models suffer from the lack of
experimental information with regard to many specific foods including
mushrooms.
Wang et al. (2017) used the modified Gompertz model as the pri-
mary model to study the growth behaviour of Pseudomonas spp. on fresh
mushroom under isothermal conditions. Although the modified Gom-
pertz model represented the growth behaviour satisfactorily at constant
storage temperatures, the effect of changing storage temperatures on
the growth of Pseudomonas spp. on mushroom, which is the usual case
during the life cycle of the mushroom, was not evaluated. Tarlak,
Ozdemir, and Melikoglu (2018) used the modified Gompertz, logistic
and Baranyi models to describe the microbial growth data of Pseudo-
monas spp. on sliced mushroom (Agaricus bisporus) at different storage
temperatures. They found that the Baranyi model gave better goodness
of fit for describing the growth behaviour of Pseudomonas spp. on sliced
mushrooms than the modified Gompertz and logistic models.
The use of primary models under constant environmental conditions
provides a good description of microbial growth for static environ-
mental conditions. However, under real conditions, environmental
factors are not constant and stable during life cycle of a food product
(Zwietering, De Wit, Cuppers, & van't Riet, 1994). Therefore, dynamic
models are necessary to simulate changing environmental conditions by
considering a food product really subjects to (Pérez-Rodríguez &
Valero, 2013). As stated by Sardella, Gatt, and Valdramidis (2018),
there is a gap in the literature about the assessment of dynamic growth
models related to the growth of microorganisms on different foods.
Predictive models considering dynamically changed environmental
conditions have gained considerable attraction in predictive food mi-
crobiology in the last three years. Iannetti et al. (2017) separately in-
oculated Listeria monocytogenes and Yersinia enterocolitica to Italian style
fresh sausages and modelled the combined growth/death behaviours of
Listeria monocytogenes and Yersinia enterocolitica under dynamically
changed aw conditions at constant storage temperatures of 8, 12, 18 and
20 °C. Dolan, Meredith, Bolton, and Valdramidis (2019) coupled the gas
transfer through the packaging material with the microbial growth ki-
netics and estimated the growth kinetic parameters for Pseudomonas
and lactic acid bacteria in modified atmosphere packaged skinless
chicken fillets stored only at 2 °C. The most likely environmental factor
that varies and drastically affects the growth behaviour of micro-
organisms in foods is the temperature. Therefore, dynamic models
considering the effect of changing temperature are necessary to in-
vestigate and model the effect of varying temperature (non-isothermal
conditions) on microbial growth.
Tsironi et al. (2017) developed some predictive models including
the simulation of the growth behaviour of Pseudomonas spp. to estimate
the shelf-life of packed ready-to-eat salad under modified atmosphere
condition (3% O2, 10% CO2 and 87% N2). They described the growth
behaviour of Pseudomonas spp. under non-isothermal conditions based
on the effective temperature concept proposed by Giannakourou and
Taoukis (2003) where they considered a single value as the effective
temperature to represent dynamically changing storage temperatures.
Longhi et al. (2017) used optimal experimental design approach by
which primary and secondary model data could be simultaneously es-
timated in order to simulate the growth behaviour of Lactobacillus vir-
idescens in Man, Ragosa and Sharpe (MRS) culture medium under non-
isothermal storage conditions, but they did not validate their model
using the growth data obtained from real foods. da Silva et al. (2017)
used the growth data obtained from the MRS medium in order to pre-
dict the growth behaviour of Lactobacillus viridescens in vacuum-packed
sliced ham under non-isothermal storage conditions. This model lacks
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Food Research International 130 (2020) 108912
the capability of accurately predicting the growth behaviour of micro-
organisms on real foods under dynamic conditions.
The growth behaviour of Pseudomonas spp., which are one of the
most abundant microorganisms widely isolated from food products,
have been studied under non-isothermal conditions using dynamic
models for some foods including fish (Koutsoumanis, 2001), poultry
(Gospavic, Kreyenschmidt, Bruckner, Popov, & Haque, 2008), pork
(Bruckner, Albrecht, Petersen, & Kreyenschmidt, 2013), shrimp
(Dabadé et al., 2015), chicken breast (Lytou, Panagou, & Nychas,
2016). Recently, Manthou et al. (2019) have attempted to develop a
dynamic model for the growth behaviour of Pseudomonas spp. on oyster
mushroom only at three different temperature conditions. The pre-
dictive capability of the model could be further improved by increasing
the number of storage temperatures.
The main objective of the present work was to quantitatively de-
scribe the growth behaviour of existing Pseudomonas spp. on button
mushrooms under non-isothermal conditions using dynamic modelling
approach. The novelty of this work from the existing dynamic model-
ling approaches used for predicting the growth of Pseudomonas spp. was
to improve the predictive capability of the dynamic model by analysing
the growth behaviour of Pseudomonas spp. at a wider temperature range
with more temperature measurements with respect to the changing
environmental conditions in which mushrooms really subject to.
2. Materials and methods
2.1. Sample preparation and microbiological analysis
White button mushrooms (Agaricus bisporus) were obtained from
MUPA Agriculture and Industry Inc. (Izmit, Kocaeli, Turkey).
Mushrooms were picked at the closed cap stage (cap diameter
3.5–4.5 cm) and immediately shipped to the laboratory at 4 °C which is
the industrial practice for storage and delivery of the mushrooms.
Damaged, bruised and shrivelled mushrooms were discarded, and the
mushrooms were not subjected to any treatments. Whole mushrooms
were placed in polystyrene trays (200 g/tray) with the dimension of
22.5 × 13.5 × 3 cm. The trays were not overwrapped with any
packaging material and placed in a temperature/humidity controlled
climate chambers (Binder MKFT 115, Binder GmbH, Tuttlingen,
Germany) at different isothermal conditions (4, 12, 20 and 28 °C) and
95% relative humidity (RH). The temperature and humidity variations
in the climate chambers were less than ± 0.2 °C and ± 2.5% RH, re-
spectively. For non-isothermal conditions, the trays were subjected to
periodically changing dynamic temperatures (24 h at 4 °C and 12 h at
10 °C) in the same climate chambers. The temperature in the climate
chambers were recorded using a data logger (Testo 174, Testo SE & Co.
KGaA, Lenzkirch, Germany) in every 15 min. Microbiological data were
collected under conditions in which mushrooms are usually subjected
to during storage, delivery and retail marketing.
Twenty-five grams of mushrooms were aseptically weighed and
homogenized using a stomacher (Interscience, Bag Mixer 400VW, USA)
at high speed for 2 min with 225 ml of sterile peptone (0.1%, w/v)
water (Oxoid, Basingstoke, UK) for each temperature with the corre-
sponding sampling frequency. Appropriate serial decimal dilutions
were made in serial dilution tubes by taking 1 ml of sample with 9 ml of
0.1% (w/v) sterile peptone water. Pseudomonas spp. were enumerated
in King's B medium (King, Ward, & Raney, 1954), following incubation
at 25 °C for 48 h. Total mushroom trays of 297 corresponding to three
independent batches were analysed for Pseudomonas spp. The initial
concentration of Pseudomonas spp. on mushrooms was determined as
soon as the mushrooms reached the laboratory, and Pseudomonas spp.
counts during storage were enumerated in three different trays at each
storage temperature for each sampling point for a maximum duration of
240 h (10 days). Results were expressed as the average ± standard
error for each sampling point in terms of log CFU/g.
F. Tarlak, et al.
2.2. Modelling
2.2.1. Primary model
The Baranyi model (Baranyi & Roberts, 1994) was used to de-
termine the kinetic growth parameters of Pseudomonas spp. on button
mushrooms stored at isothermal conditions:
e µmax F(t) 1
y(t) = y0 + µmax F(t) ln 1 +
e(ymax y0) (1)
−1
where µmax is the maximum specific growth rate (h ), t is the time (h),
y(t) is the concentration of microbial population at time t (ln CFU/g), y0
is the initial concentration of microbial population (ln CFU/g) and ymax
is the maximum concentration of microbial population (ln CFU/g).
For the Baranyi model, the parameter F(t) is defined by Eq. (2):
1 vt µ max
F(t)t + ln(e +e e( vt µ max ) )
v (2)
λ is the lag phase duration (h), and v is rate of increase of the limiting
substrate, assumed to be equal to µmax (Juneja, Melendres, Huang,
Subbiah, & Thippareddi, 2009).
All growth parameters and their standard errors were calculated by
considering the average values of the independent observations using
the NonLinearModel.fit command which uses Levenberg-Marquardt
algorithm in the Matlab 8.3.0.532 (R2014a) software (MathWorks Inc.,
Natick, MA, USA). After the model fitting, conversion from ln CFU/g to
log CFU/g was performed for the parameters y0 and ymax.
Experimentally determined minimum and maximum microbial popu-
lations were chosen as the starting value for y0 and ymax, respectively.
The starting value of µmax was calculated by taking the maximum value
of first derivative of the natural logarithm of the concentration of mi-
crobial population as a function of time. After several successive
iterations in the nonlinear procedure, estimated growth kinetic para-
meters were obtained.
2.2.2. Secondary models
The Ratkowsky (Eq. (3)) and Arrhenius (Eq. (4)) models were used
to determine the relationship between the temperature and maximum
specific growth rate obtained from the Baranyi model
µmax = b1 (T T0) (3)
Ea
µ max = b2 exp
R (4)
where T is the temperature (°C), T0 is the theoretical minimum tem-
perature for microbial growth (°C), µmax is the maximum specific
growth rate (h−1), b1 and b2 are the regression coefficients, Ea is the
activation energy (J/mol), R is the universal gas constant (8.314 J/mol
K) and θ is the absolute temperature (K). Secondary model parameters
were obtained using the NonLinearModel.fit command in Matlab
software.
2.2.3. Evaluation of the goodness of fit of the primary and secondary
models
The goodness of fit of the primary and secondary models was
evaluated by considering the root mean square error (RMSE) and the
adjusted coefficient of determination
(adjusted-R2) values using Eqs. (5) and (6), respectively:
n
(observedi fittedi ) 2
RMSE =
i=1
n s (5)
n 1 SSE
adjusted-R2 = 1
n s SST (6)
where observedi is the observed value, fittedi is the fitted value, n is the
number of total observations, s is the number of parameters of the
model, SSE is the sum of squares of errors and SST is the total sum of
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Food Research International 130 (2020) 108912
squares.
Temperature data were correlated with µmax for the Ratkowsy
model while temperature data were correlated with µmax for the
Arrhenius model. Therefore, the RMSE values were calculated based on
µmax for both secondary models. The model with the lower RMSE and
the higher adjusted-R2 was selected as the appropriate secondary model
to describe the temperature dependence of µmax.
2.2.4. Dynamic model
The prediction of the microbial growth under non-isothermal con-
ditions was carried out using Eqs. (7) and (8) as described by Baranyi
and Roberts (1994):
dy 1
= µmax (T(t))[1 e(y(t) ymax) ]
dt 1+e Q(t) (7)
dQ
= µmax (T(t))
dt (8)
The initial conditions to solve Eqs. (7) and (8) are y(0) = y0 (the
initial population concentration, ln CFU/g) and Q(0) = lnq0, respec-
tively. Q(t) is the natural logarithm of q(t), a dimensionless variable
related to physiological state of the cells. µmax and λ values were esti-
mated for each isothermal condition using the Baranyi model. For each
isothermal condition, h0 values were then calculated from the equation,
μmax × λ = h0. The average h0 was used to obtain the initial q0 in the
differential form of the Baranyi model using Eq. (9) (Baranyi & Roberts,
1994):
q0 =
1
e h0 1 (9)
Because µmax is a function of both temperature and time, µmax values
estimated by the appropriate secondary model were put into the dif-
ferential form of the Baranyi model. The temperature data recorded
with the data logger were used to solve Eqs. (7) and (8). Results were
obtained using ode45 command which implements the fourth-order
Runge-Kutta method in the Matlab software (Velugoti et al., 2011).
The predicted concentration of microbial population using dynamic
modelling was compared with the observed growth data under non-
isothermal storage conditions for the model validation assessed by
considering the bias (Bf) and accuracy (Af) factors (Ross, 1996; Wang
et al., 2017) given in Eqs. (10) and (11), respectively:
n log(y
i=1 predicted /yobserved)
Bf = 10 n (10)
n |log(y
i=1 predicted /yobserved)|
Af = 10 n (11)
where ypredicted is the predicted concentration of microbial population
(log CFU/g), yobserved is the observed concentration of microbial po-
pulation (log CFU/g) and n is the number of total observations. The Bf is
a measure of average variation between the observed and predicted
values. The Af measures the average difference between the observed
and predicted values by disregarding whether the difference is positive
or negative. A value of 1 for Bf and Af indicates that there is a perfect
agreement between all the observed and predicted values. The mean
deviation (MD) and the mean absolute deviation (MAD) between the
observed and predicted concentrations of microbial populations were
also calculated to assess the prediction performance of the dynamic
model as suggested by Le Marc et al. (2008).
3. Results and discussion
Experimental growth data of Pseudomonas spp. on button mushroom
obtained at different isothermal storage temperatures (4, 12, 20 and
28 °C) are given in Fig. 1. The growth curves for Pseudomonas spp. on
button mushrooms stored at different temperatures exhibited a sigmoid
trend which conforms to the general growth behaviour of Pseudomonas
spp. for fish (Koutsoumanis, 2001), poultry (Gospavic et al., 2008),
F. Tarlak, et al.
Fig. 1. Pseudomonas spp. growth as fitted by the Baranyi model for button
mushroom stored at 4, 12, 20 and 28 °C.
pork (Bruckner et al., 2013), shrimp (Dabadé et al., 2015), chicken
breast (Lytou et al., 2016) and ready to eat fresh vegetable salad
(Tsironi et al., 2017). The initial concentration of Pseudomonas spp. on
the button mushrooms harvested at different times was found to be
7.05 ± 0.14 log CFU/g which is in good agreement with the initial
Pseudomonas spp. counts found in mushrooms where Pseudomonas spp.
counts ranged from 6.63 to 8.10 log CFU/g (Reyes et al., 2004; Simón &
González-Fandos, 2010; Venturini et al., 2011). High initial counts of
Pseudomonas spp. can be attributed to the absence of cuticle to protect
Agaricus bisporus from physical damage and microbial attack where it is
easily contaminated with microorganisms especially Pseudomonas spp.
during growth, harvesting and processing.
µmax and λ are the most important critical parameters to describe the
growth behaviour of microorganisms on food, and temperature has a key
role in affecting directly both of these growth parameters (Huang, 2008).
The kinetic parameters including µmax and λ belonging to Pseudomonas
spp. on the button mushrooms derived from the Baranyi model for each
storage temperature are presented in Table 1. The µmax increased from
0.043 to 0.202 h−1 with increasing the storage temperature from 4 to
28 °C while an opposite trend was observed for λ which decreased from
44.1 to 20 h as the storage temperature increased from 4 to 28 °C. The
increase in µmax and decrease in λ mean that Pseudomonas spp. on button
mushrooms could grow faster and adapt themselves to growth conditions
easier when the temperature increased from 4 to 28 °C. Maximum con-
centrations of Pseudomonas spp. on button mushrooms during this study
were 8.65 ± 0.16, 10.21 ± 0.18, 10.78 ± 0.16 and 10.62 ± 0.15
log CFU/g at the storage temperatures of 4, 12, 20 and 28 °C, respectively
(Table 1). The difference in the maximum and initial populations of
Pseudomonas spp. on button mushrooms stored at 4 °C was smaller
compared to the difference for the storage temperatures of 12, 20 and
Table 1
Estimated kinetic growth parameters for the Baranyi model.
Temperature (°C) y0a (log CFU/g) ymaxa (log CFU/g)
4 7.08 ± 0.06 8.65 ± 0.05
12 7.08 ± 0.06 10.14 ± 0.05
20 7.06 ± 0.12 10.71 ± 0.13
28 7.05 ± 0.07 10.64 ± 0.07
a
Estimated growth parameters ± standard errors.
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Food Research International 130 (2020) 108912
28 °C. This means that the growth potential of Pseudomonas spp. on
button mushrooms stored at 4 °C was lower than the storage tempera-
tures of 12, 20 and 28 °C. The growth potential of Pseudomonas spp. on
oyster mushrooms stored at 4 °C was also found lower than the growth
potentials of Pseudomonas spp. stored at higher temperatures as reported
by Manthou et al. (2019).
The Baranyi model yielded RMSE values lower than 0.193 and ad-
justed-R2 values higher than 0.975 for all storage temperatures. The
goodness of fit of the Baranyi model indicated that the Baranyi model
could be reliably used to describe the growth behaviour of Pseudomonas
spp. on button mushrooms in the temperature range of 4 and 28 °C
under isothermal conditions (Table 1).
The µmax derived from the Baranyi model was modelled as a func-
tion of temperature using the Ratkowsky (Fig. 2a) and Arrhenius
(Fig. 2b) models. For the Ratkowsky model, the estimated value of T0
for the growth of Pseudomonas spp. on button mushrooms was found to
be −17.5 °C (Table 2). Similar results were also reported for Pseudo-
monas spp. on mushrooms in which T0 values for the growth of Pseu-
domonas spp. were found to be −20.4 (Wang et al., 2017) and −12.5 °C
(Tarlak et al., 2018) for whole and sliced button mushrooms, respec-
tively. However, it needs to be highlighted that T0 represents only the
theoretical minimum temperature, and estimated T0 can be con-
siderably lower than the real minimum temperature (Ross & Dalgaard,
2004; Lianou, Moschonas, Nychas, & Panagou, 2018). For the Arrhenius
model, the estimated value of Ea for the growth of Pseudomonas spp. on
button mushrooms was 39.5 kJ/mol (Table 2). Bruckner et al. (2013)
also used the Arrhenius model to determine the value of Ea for the
growth of Pseudomonas spp. in poultry and pork where the Ea values
were found to be 103.0 kJ/mol and 69.5 kJ/mol, respectively. Steeper
slope obtained from the Arrhenius model for the poultry and pork
means that Pseudomonas spp. growth in poultry and pork is more sen-
sitive to small changes in temperature compared to the Pseudomonas
spp. growth on button mushrooms.
The RMSE and adjusted-R2 values calculated for the Ratkowsky and
Arrhenius models are tabulated in Table 2. The RMSE values were
found to be 7.83 × 10−5 and 8.40 × 10−3 for the Ratkowsky and
Arrhenius models, respectively, while the adjusted-R2 values were
0.993 and 0.984, respectively. The lower RMSE and higher adjusted-R2
values indicated that the Ratkowsky model could satisfactorily describe
the relationship between the storage temperature of mushrooms and
the maximum specific growth rate of Pseudomonas spp. Therefore,
Ratkowsky model was selected to model the temperature dependence of
the maximum specific growth rate in dynamic modelling predictions.
The growth behaviour of Pseudomonas spp. has been extensively
investigated since the food spoilage has been directly linked to the
concentration of Pseudomonas spp. populations (Raposo, Pèrez, de
Faria, Ferrús, & Carrascosa, 2017). Because µmax is one of the most
important parameters characterizing microbial growth behaviour, µmax
values of Pseudomonas spp. were determined for some foods including
fish (Koutsoumanis, 2001), poultry (Dominguez & Schaffner, 2007),
pork (Koutsoumanis, Stamatiou, Drosinos, & Nychas, 2008) and beef
(Zhang et al., 2011) at different temperatures. The results collected
from these studies and our results are compared in Table 3. At the same
temperature, µmax values of Pseudomonas spp. on button mushroom are
considerably lower than the µmax values obtained for Pseudomonas spp.
in fish, poultry, pork and beef. This is probably due to the difference in
µmaxa (h−1) λa (h) RMSE adjusted-R2
0.043 ± 0.007 44.1 ± 10.4 0.103 0.975
0.091 ± 0.006 33.2 ± 3.6 0.193 0.977
0.134 ± 0.015 23.5 ± 4.7 0.146 0.990
0.202 ± 0.013 20.0 ± 1.9 0.122 0.993
F. Tarlak, et al. Food Research International 130 (2020) 108912

Fig. 2. Maximum specific growth rate values obtained from the Baranyi model as a function of storage temperature using (a) Ratkowsky and (b) Arrhenius models.

Table 2
Estimated parameters for the secondary models.
Secondary models Parameters Estimate ± standard error RMSEa adjusted-R2

µmax = b1 (T T0) b1 9.90 × 10−3 ± 4.95 × 10−4 7.83 × 10−5 0.993

T0 (°C) −17.5 ± 1.7
b2 1.45 × 106 ± 2.03 × 105 8.40 × 10−3 0.984
µmax = b2 exp ( )
R
Ea
Ea (kJ/mol) 39.5 ± 3.4

a
RMSE values were calculated based on µmax.

Table 3
Comparison of µmax values for Pseudomonas spp. on fish, poultry, beef, pork and
mushroom.
Temperature (°C) Fisha Poultryb Porkc Beefd Mushroome

4 – – – 0.089 0.045
5 0.103 0.095 0.110 – 0.049
10 0.191 0.204 0.202 0.206 0.074
12 – 0.259 – – 0.085
15 0.269 0.353 0.323 0.290 0.104
20 – 0.544 – 0.413 0.137

µmax values (h−1) were obtained from aKoutsoumanis (2001), bDominguez and
Schaffner (2007), cKoutsoumanis et al. (2008), dZhang et al. (2011) and ecal-
culated with the Ratkowsky model in this work.

growth potential of Pseudomonas spp. on different food substrates.

The step of assessing of the dynamic model performance is crucial to
validate the reliability of the estimated growth parameters of
Pseudomonas spp. on button mushrooms. Therefore, the growth data of
Pseudomonas spp. on button mushrooms subjected to non-isothermal
storage conditions (24 h at 4 °C and 12 h at 10 °C) were compared with
the predicted growth data using the differential form of Baranyi model
(Fig. 3). The Bf and Af were found to be 0.998 and 1.016, respectively.
Both Bf and Af close to one indicated that the dynamic model used in
this work had a high capability to predict the concentration of Pseu-
domonas spp. on the button mushrooms stored at changing tempera-
tures with time. The MD and MAD values for Pseudomonas spp. popu-
lations were −0.013 and 0.126 log CFU/g, respectively. The MD value
of −0.013 log CFU/g showed that on average the dynamic model
overestimated 0.013 log CFU/g while the MAD value of 0.126 log CFU/
g indicated that on average the predicted values were 0.126 log CFU/g
different (either higher or lower) from the observed ones. Manthou
et al. (2019) reported a MD value of −0.10 log CFU/g and MAD value
Fig. 3. Observed (×) and predicted ( ) concentrations of Pseudomonas spp. on
button mushrooms under dynamic temperature conditions (24 h at 4 °C and
12 h at 10 °C). The dashed line (–) shows the changing temperature during
storage.
of 0.22 log CFU/g for the Pseudomonas spp. populations on oyster
mushroom. All these results showed that the models used in the current
study have better predictive capability than the dynamic model pre-
viously developed for the oyster mushroom. As a result, the modelling
approach used in this work could be an alternative to traditional enu-
meration techniques to determine the number of Pseudomonas spp. on
mushrooms.

5
F. Tarlak, et al.
4. Conclusions
The growth behaviour of Pseudomonas spp. on button mushrooms
under isothermal storage temperatures between 4 and 28 °C was de-
scribed by the Baranyi model. The µmax values obtained from the
Baranyi model were correlated with the temperature using the
Ratkowsky and Arrhenius models. High adjusted-R2 and low RMSE
values showed that the Ratkowsky model could be used to describe the
relationship between the storage temperature of mushrooms and the
maximum specific growth rate of Pseudomonas spp. The successfully
validated differential form of the Baranyi model merged with the
Ratkowsky model provided valuable information to evaluate and si-
mulate the growth behaviour of the Pseudomonas spp. on button
mushrooms under fluctuating temperature conditions in which mush-
rooms are usually subjected to during storage, delivery and retail
marketing. The predictive models used in this work have a high po-
tential to be used as a simulation tool for the mushroom processors to
follow the microbiological quality of the mushrooms before they reach
to the consumers.
CRediT authorship contribution statement
Fatih Tarlak: Software, Validation, Formal analysis, Investigation,
Writing - original draft. Murat Ozdemir: Conceptualization,
Methodology, Writing - review & editing, Supervision. Mehmet
Melikoglu: Resources, Data curation, Project administration, Funding
acquisition.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
This work was financially supported by the Gebze Technical
University through Scientific Research Projects (BAP 2014 A-25 and
BAP 2015 A-40). Dr. Fatih Tarlak would like to thank The Scientific and
Technological Research Council of Turkey (TUBITAK) for granting
Postdoctoral Research Fellowship (2219).
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