See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/348448028

Modelling viability of Listeria monocytogenes in paneer

Article  in  Food Microbiology · January 2021

DOI: 10.1016/j.fm.2021.103738

CITATIONS READS

2 32

5 authors, including:

Dipon Sarkar John P Bowman

Tasmanian Institute of Agriculture University of Tasmania
5 PUBLICATIONS   19 CITATIONS    622 PUBLICATIONS   13,748 CITATIONS   

SEE PROFILE SEE PROFILE

Bing Wang David Ratkowsky

University of Nebraska at Lincoln University of Tasmania
42 PUBLICATIONS   455 CITATIONS    318 PUBLICATIONS   11,749 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Mitigating the Risk of Campylobacter spp. in Broiler Chicken Supply Systems Using Quantitative Microbial Risk assessment and Cost-effectiveness Approaches View
project

Investigating opportunities to influence gut microbiota in farmed Atlantic Salmon View project

All content following this page was uploaded by Dipon Sarkar on 08 April 2021.

The user has requested enhancement of the downloaded file.

 Food Microbiology 97 (2021) 103738

Contents lists available at ScienceDirect

Food Microbiology
journal homepage: www.elsevier.com/locate/fm

Modelling viability of Listeria monocytogenes in paneer

Dipon Sarkar a, David A. Ratkowsky a, Bing Wang b, John P. Bowman a, Mark L. Tamplin a, *
a
Centre of Food Safety & Innovation, University of Tasmania, Private Bag 54, Sandy Bay, Tasmania, 7005, Australia
b
Department of Food Science and Technology, University of Nebraska-Lincoln, 1901 N 21st St, Lincoln, NE, 68588, United States

A R T I C L E I N F O A B S T R A C T

Keywords: Paneer is a fresh, soft ready-to-eat cheese that is susceptible to Listeria monocytogenes contamination, exemplified
Soft cheese by product recalls in Australia, Canada, and the USA. Previous research demonstrates that L. monocytogenes
Listeriosis grows in paneer, however there are no paneer-specific predictive models that quantify the effect of environ­
Predictive microbiology
mental conditions on L. monocytogenes viability. This study measured the viability of a five-strain cocktail of
Growth kinetics
Food safety
L. monocytogenes in freshly prepared paneer incubated at 4–40 ◦ C. Growth rates were fitted with the extended
Ratkowsky square root model, with growth rates ranging from 0.014 to 0.352 log10 CFU/h. In comparison with
published models, only the ComBase L. monocytogenes broth model acceptably predicted growth (Bf = 1.01, Af =
1.12) versus the developed model. The influence of paneer pH (5.0–6.0) and storage temperature (41–45 ◦ C) on
L. monocytogenes growth at the upper temperature growth boundary was described using a logistic model. These
models provide quantitative tools to improve the safety of paneer processing conditions, shelf-life estimation,
food safety management plans, and risk assessment.

1. Introduction
Milk and dairy products are the major contributors of foodborne
disease in India (Kristkova et al., 2017), a country with the world’s
lowest per capita consumption of meat (http://www.fao.
org/faostat/en/#data/FBS). Among Indian dairy foods, paneer is
widely consumed, reflected in an estimated annual domestic production
of 110 million tonnes (USDA, 2015) and in increasing popularity in
many other countries. Traditionally in India, paneer production has
been mainly produced in homes and by an unorganised dairy sector
consisting of small- and medium-scale local enterprises (Khan and Pal,
2011; USDA, 2015; Varalakshmi and Leysen, 2020). The potential for
Listeria monocytogenes contamination is significant, considering the
numerous small-scale producers that account for a large portion of
paneer production, many of which use raw milk as the starting
component (Bhilegaonkar et al., 1997; Nahidul-Islam et al., 2018; Soni
et al., 2013; Vaishnavi et al., 2001).
Paneer is a non-ripened, non-melting, soft cheese produced by
curdling milk with acidifying agents, such as citric acid or vinegar (Khan
and Pal, 2011). Production involves heating milk to 80–90 ◦ C, cooling,
coagulating with acid, draining whey, and pressing curd (Kumar et al.,
2014). Paneer is not subjected to post-packaging heat treatment but
instead is cut into blocks before packaging, increasing the opportunity
for pathogen contamination. The pH of the final product ranges from 5.5
to 6.0, with a moisture content of <70% and 0.5% titratable acidity (%
lactic acid), characteristics that readily support viability of bacterial
pathogens, such as L. monocytogenes. However, there are few quantita­
tive risk management tools to aid the design of paneer food safety plans.
Listeriosis outbreaks linked to cheese consumption have occurred in
various countries (Martinez-Rios and Dalgaard, 2018; Gaulin et al.,
2012; Gould et al., 2014; Koch et al., 2010). Paneer has already been
linked to food recalls due to L. monocytogenes contamination in Australia
(Product Safety Australia, 2014), Canada (Food Poisoning bulletin.,
2015), and the USA (CDC, 2015). In India, there is a relatively high risk
of listeriosis due to inadequate regulation of small-scale processors, milk
quality, cold chain management, food handling, and hygienic conditions
(Chawla and Chawla, 2009). While L. monocytogenes has been shown to
grow in paneer via different modes of contamination, such as during
coagulum preparation, water immersion, and/or packaging (Vara­
lakshmi and Leyson, 2020), L. monocytogenes growth kinetics have not
been reported.
Effective food safety management relies on science-based evidence
to estimate the fate of hazards over a series of processing and handling
operations. In this regard, predictive microbiology provides tools used

* Corresponding author. Tasmanian Institute of Agriculture, Private Bag 98, University of Tasmania, Hobart, Tasmania, 7001, Australia.
E-mail addresses: dipon.sakar@utas.edu.au (D. Sarkar), d.ratkowsky@utas.edu.au (D.A. Ratkowsky), bing.wang@unl.edu (B. Wang), john.bowman@utas.edu.au
(J.P. Bowman), mark.tamplin@utas.edu.au (M.L. Tamplin).

https://doi.org/10.1016/j.fm.2021.103738
Received 28 October 2020; Received in revised form 27 December 2020; Accepted 7 January 2021
Available online 12 January 2021
0740-0020/© 2021 Elsevier Ltd. All rights reserved.
D. Sarkar et al. Food Microbiology 97 (2021) 103738

by food companies and regulatory bodies to quantify microbial behav­ 2.3. Isothermal growth studies
iour under different environmental conditions. This information assists
in the design and decision-making components of food safety plans Each freshly prepared paneer batch was cut into 5-g blocks using a
(Plaza-Rodríguez et al., 2015; Tamplin, 2005). For L. monocytogenes, sterilized chopping board and knife. Twenty microliters of ambient
Codex and food regulatory guidelines encourage risk-based approaches temperature (~25 ◦ C) L. monocytogenes cocktail was inoculated onto the
that integrate predictive models (FAO/WHO, 2007). surface of each block (~25 ◦ C), producing an initial concentration of
Several mathematical models have been developed for ~103 CFU/g. Controls were inoculated with 20 μL sterile 0.1% peptone
L. monocytogenes in fresh and soft cheese, including queso fresco water. Four blocks of inoculated paneer were placed in a single sterile
(Thomas et al., 2019), queso blanco (Uhlich et al., 2006), cottage cheese Petri dish with the inoculation side facing upward; four negative control
(Østergaard et al., 2014), soft blue-white cheese (Rosshaug et al., 2012), blocks were placed in a separate dish. Petri dishes were placed in a
minas fresh cheese (Cadavez et al., 2019) and fresh ricotta (Tirloni et al., sealed UV-sterilized polypropylene container (17.5 cm × 12 cm x 5 cm)
2019). Models developed for other soft cheeses may not be applicable to containing a sponge moistened in sterile distilled water to maintain
paneer due to differences in organic acid levels and inhibitory effects of humidity and then incubated at 4, 10, 15, 20, 25, 30, 35 and 40 ◦ C. Two
starter cultures on L. monocytogenes growth. To address this knowledge biologically independent replicates were analysed at each temperature.
gap, L. monocytogenes viability in paneer was measured from 4 to 45 ◦ C,
a temperature range that typically occurs during paneer production and 2.3.1. Sampling and enumeration
storage. The resulting predictive models were compared to others in the The sampling frequency for each temperature was based on pre­
published literature. dictions by the ComBase broth growth model for L. monocytogenes/
innocua (https://browser.combase.cc/ComBase_Predictor.aspx?model
2. Materials and methods =1), with sampling frequency ranging from every 48 h at 4 ◦ C to
every 2 h at 40 ◦ C. At each sampling time, three paneer blocks (inocu­
2.1. Bacterial strains and inoculum lated and controls) from a single Petri dish were combined, diluted 10-
fold in sterile 0.1% peptone water that had been pre-chilled to 4 ◦ C, and
Five strains of L. monocytogenes (Table 1; Shabala et al., 2008), homogenized for 1 min with a Colworth Stomacher (model 400, U.K.)
previously stored at − 80 ◦ C, were cultured on Brain Heart Infusion (BHI) The homogenate was serially diluted in sterile 0.1% peptone water, and
agar (Amyl Media, Dandendong, Vic., Australia) at 37 ◦ C for 24 h. Each then 100 μL of appropriate dilutions plated in duplicate on Palcam agar
strain was then cultured overnight in 5 mL BHI broth at 37 ◦ C until the (Oxoid CM0877B, U.K.) containing Palcam Selective Supplement (Oxoid
OD600 was approximately 1.0 (109 CFU/mL). Next, each of the five SR0150E, U.K.); agar plates were incubated at 37 ◦ C for 48 h to
strains were serially diluted in 0.1% peptone water to 106 CFU/mL, and enumerate L. monocytogenes. The fourth block of inoculated paneer was
then mixed in equal volumes to obtain 106 total CFU/mL for the inoc­ used to measure pH at each sampling time. The mean CFU for both
ulum cocktail. replicates were counted and converted to log10 CFU/g.
For the three negative control paneer blocks, 100 μL of diluted
2.2. Paneer production sample was plated on Standard Plate Count Agar (PCA; Oxoid, CM0463,
U.K.) and incubated at 25 ◦ C for 48 h to enumerate Total Viable Count
Paneer was produced in the laboratory on the day of experimentation (TVC), and also on Palcam agar to measure potential background levels
following procedures described by the National Dairy Development of L. monocytogenes. The minimum detection limit was 1.98 log10 CFU/g.
Board, India (Aneja et al., 2002). In brief, pasteurized non-homogenized The fourth negative control block of paneer was used to measure pH.
cow milk (~3 L) was heated on an induction cooktop to 85 ◦ C and
maintained at that temperature for 5 min. The milk was then cooled to 2.4. Growth/no-growth experiments
70 ◦ C and coagulated with 2% (w/v %; approximately 0.37 mM undis­
sociated citric acid) food-grade citric acid (McKenzie, Australia). The Growth/no-growth (G/NG) potential of the L. monocytogenes cocktail
coagulum was collected in cheese cloth-lined hoops, and excess moisture was measured in broth from 41 to 45 ◦ C, over a pH range of 5.0–6.0. The
drained and pressed-out. The subsequent paneer batch (700–750 g) was temperature range selected was based on the notional maximum growth
removed from the hoops and air-dried, after which pH (Orion Star A215, limit (Tmax) as estimated by the developed paneer model (Ratkowsky
Thermofisher, Vic., Australia) and water activity were measured et al., 1983), and also considering relevant temperatures used in paneer
(Aqualab CX-2, Decagon Devices, Pullman, WA, USA). production and published studies (Membré et al., 2004; Nunes Silva
et al., 2020). A modified factorial design was used to initially test 5 × 5
combination of pH and temperature, with additional temperatures and
pH later tested to ascertain the G/NG boundary with greater accuracy.
The pH of 50 mL of BHI broth in a 250 mL Erlenmeyer flask fitted with a
Table 1
screw cap was adjusted with 2% food-grade citric acid. pH was
Characteristics of L. monocytogenes strains, including reported minimum pH and
measured after sterilization to confirm that it was the same as the batch
maximum NaCl (%, wt/vol) for growth in BHI broth at 20 ◦ C (Shabala et al.,
2008).
of paneer. The water activity of paneer batches ranged from 0.980 to
0.990 and BHI was 0.987; due to this similarity, BHI water activity was
Strain Source Serotype Minimum pH Maximum NaCl (%)
not adjusted. The BHI broth was inoculated with 100 μL of
FW03/0034 fooda 1/2a 4.2 12.1 L. monocytogenes cocktail to produce an initial concentration of ~104
FW03/0032 fooda 3a 4.1 11.2 CFU/mL, and then flasks incubated at the specified temperatures. After
F2365 foodb 4b nre nr
Silliker 204231-1 foodc 1/2c 4.1 11.2
12 h incubation (i.e. a length of time sufficient for L. monocytogenes to
102-231-s-7-566 foodd 3b 4.2 11.2 reach exponential phase at 40 ◦ C), samples were diluted in 0.1% peptone
a water and plated on BHI to enumerate L. monocytogenes. An increase ≥1
Melbourne Microbiological Diagnostic Unit Public Health Laboratory, North
log10 CFU/mL, which is equivalent to three replication cycles, was
Melbourne, Vic., Australia.
b
Linnan et al., 1988. defined as growth. At each test condition, two biologically independent
c
Silliker Australia Pty Ltd, now Mérieux NutriSciences, Regents Park, NSW, replicates were tested; each replicate was assigned a score of 1 if growth
Australia. occurred or 0 if growth was not observed. The data obtained from these
d
Holah et al., 2004. experiments were used to develop a G/NG boundary model.
e
Not reported. The predictions from the G/NG boundary model developed in broth

2
D. Sarkar et al. Food Microbiology 97 (2021) 103738

were evaluated by conducting G/NG analysis in freshly prepared paneer

following the methodologies described in section 2.3.1. The pH of the √ μmax = a(T − Tmin ){1 − exp[b(T − Tmax )]} (5)
paneer was not adjusted during the experiment, and instead the innate
where a and b are model fitting parameters with units of log10 CFU/h/◦ C
pH was measured. The incubation temperatures were the same as for
and ◦ C− 1, respectively, T is temperature (◦ C), and Tmax and Tmin notional
BHI broth G/NG studies. After 12 h of incubation, L. monocytogenes was
growth limits (◦ C), also model fitting parameters.
enumerated following the procedures in section 2.4. An increase ≥1
The model was fitted using the MATLAB (Version R2017a, Math­
log10 CFU/mL was defined as growth.
works, Natick, MA) nonlinear least squares curve-fitting toolbox with
the trust-region algorithm. Root mean square error (RMSE), sum of
2.5. Model and data comparisons squared errors of prediction (SSE), and coefficient of determination (R2)
were checked for model goodness-of-fit.
Comparisons were made between the developed model and popular
online model platforms, published L. monocytogenes models and data for 2.6.3. Growth/no-growth model
different cheeses (Table 4). These comparisons were made using indices For the G/NG experiments, a logistic regression model (Equation (6))
of model performance, bias (Bf) and accuracy factor (Af), described by was used to predict the probability of growth under tested conditions,
Ross (1996), where, based on the method of Ratkowsky and Ross (1995),
( ) ( )
P
∑ predicted Logit(P) = Ln =f (6)
log
observed 1− P
Bf = 10 n (1)
P is the probability of growth being modelled and f is the function
∑
( )⃒
⃒ defined by Equation (7),
predicted ⃒
|log
⃒
(7)
observed
f = bo + b1 .T + b2 .pH + b3 .T 2 + b4 .pH 2 + b5 .T.pH
Af = 10 n (2)

A Bf or Af value of 1.0 indicates unbiased and accurate predictions
(Oscar, 2005; Ross, 1996). For Bf > 1.0, Af should be < 1.1.5; whereas
for Bf ≤ 1.0, Af should be < 1.30 (Oscar, 2005). For ComBase, Pathogen
Modeling Program (PMP) and the Food Safety and Spoilage Predictor
(FSSP) models, inputs for pH and aw were adjusted to approximate
experimental paneer; comparisons were made only in a common tem­
perature range between the model and this study. For the ComBase
broth model, inputs were 4–40 ◦ C, pH 5.5, aw 0.999; for PMP (aerobic
and anaerobic), 4–35 ◦ C, pH 5.5, NaCl 0.5%, 0 ppm NaNO2; for FSSP,
4–25 ◦ C, NaCl 0.6%, pH 5.6, 0% CO2, 0 mg/kg nitrite; 0 ppm phenol.
Comparisons with published models for cheese were limited to those in
which L. monocytogenes growth was modelled only as a function of
temperature, similar to the present study.
where b0, b1, b2, b3, b4 and b5 are model fitting parameters. The data
were fitted using SAS PROC LOGISTIC (SAS Institute, Cary, N.C., USA)
with the Fisher Scoring method used for optimization of the maximum
likelihood of the parameters. A forward stepwise selection of the logistic
procedure was used to determine the significant terms in Eq. (7). Model-
fitting was evaluated using chi-square tests, the Hosmer–Lemeshow
goodness-of-fit test, correct prediction percentage, and concordant rate
(Skandamis et al., 2007; Astoreca et al., 2010). The Hosmer–Lemeshow
goodness-of-fit test was used to decide whether there was a lack of fit,
with a small value of the statistic indicating that the model fits the data
well. For the model output, P = 0.5 was considered the G/NG boundary.
3. Results

The pH of each of the 16 freshly prepared paneer batches was rela­

2.6. Data analyses tively similar, ranging from 5.42 to 5.61 with an average of 5.51. Water
activity for all batches was 0.999. Background L. monocytogenes was not
2.6.1. Primary models detected in any negative control sample.
The primary model of Baranyi and Roberts (1994) was fitted to ki­
netic data using the DMFit version 3.5 Excel add-in, downloaded from 3.1. Primary models for isothermal growth kinetics
ComBase (Baranyi and Tamplin, 2004) (https://browser.combase.
cc/DMFit_Excel.aspx). The model is described as: L. monocytogenes replicated in paneer from 4 to 40 ◦ C (Fig. 1,
( ) Table 2). μmax increased from an average of 0.014 log10 CFU/h at 4 ◦ C, to
eμmax F(t) − 1
y(t) = y0 + μmax F(t) − ln 1 + (ymax − y ) (3) a maximum of 0.352 log10 CFU/h in paneer. At 40 ◦ C, average μmax
e
decreased to 0.178 log10 CFU/h. The highest average MPD was 8.08
0

log10 CFU/g (30 ◦ C) and the lowest MPD was 7.49 log10 CFU/g (10 ◦ C).
where y0 and y(t) are the microbial population concentration (log10
The highest average LT (71.2 h) was detected at 4 ◦ C for paneer and
CFU/h) at time 0 and t h, respectively, μmax is the maximum growth rate
generally decreased with increasing temperature (Table 2). LT showed
(log10 CFU/h), and ymax is the maximum cell population (MPD, log10
high variability between replicates, differing by as much as 148% (i.e.
CFU/g). The parameter F(t) is defined as:
difference between replicates divided by mean). Due to these reasons LT
( )
1 was not modelled in this study. The kinetic profiles of TVC in negative
F(t) = t + ln e− vt + e− μmax λ + e(− vt− μmax λ) (4)
v control samples showed similar trends as L. monocytogenes in paneer.

where λ is the lag phase duration (LT, h) and v is rate of increase of the
limiting substrate, which is assumed to be equal to μmax . Using the
model, kinetic parameters including LT, μmax, and MPD were deter­
mined for L. monocytogenes growth in paneer. Primary model-fitting was
evaluated using goodness-of-fit statistics for coefficient-of-
determination (R2), and standard error of fitting (se).
3.2. Secondary growth model for isothermal studies
The extended Ratkowsky model was fitted to the observed square
root of μmax as a function of temperature (Fig. 2, Table 3), showing a
high goodness-of-fit statistic (RMSE = 0.0348, SSE = 0.0145, R2 =
0.9486).

2.6.2. Secondary growth model

The extended Ratkowsky model (Ratkowsky et al., 1983) was fitted
to the square root of μmax as a function of temperature (Equation (1)),

3
D. Sarkar et al. Food Microbiology 97 (2021) 103738

Fig. 1. Growth of L. monocytogenes in paneer from 4 to 40 ◦ C, for one of the two trials. Blue symbols, L. monocytogenes log10 CFU/g; yellow symbol, TVC in negative
control; red line, primary model fitted to L. monocytogenes kinetics. (For interpretation of the references to colour in this figure legend, the reader is referred to the
Web version of this article.)

Table 2
L. monocytogenes growth kinetics in paneer based on primary model fit.
Temperature pH LTa μmaxb MPDc sed R2
(◦ C) (log10 CFU/ (log10
(h)
h) CFU/g)

4 5.42 65.8 0.013 7.83 0.17 0.99

(25.03)e (0.001)e
4 5.57 76.6 0.015 7.8 0.16 0.99
(21.32) (0.001)
10 5.56 ndf 0.035 7.16 0.39 0.94
(0.005)
10 5.61 30.4 0.064 7.82 0.22 0.99
(6.25) (0.009)
15 5.53 nd 0.064 8.17 0.32 0.97
(0.007)
15 5.42 nd 0.079 7.95 0.23 0.99
(0.004)
20 5.37 0.44 0.135 7.46 0.28 0.98
(0.31) (0.014)
20 5.56 nd 0.102 8.34 0.53 0.93
(0.013)
25 5.42 nd 0.207 7.63 0.46 0.9
(0.015)
25 5.49 nd 0.208 7.91 0.23 0.99 Fig. 2. Secondary model of L. monocytogenes growth in paneer (red solid line)
(0.015) with 95% confidence interval of the prediction (dotted line) and mean observed
30 5.57 <0.01 0.18 8.36 0.32 0.95 rates (blue circles) in paneer. Error bars are equal to one standard deviation.
(2.11) (0.024) (For interpretation of the references to colour in this figure legend, the reader is
30 5.46 <0.01 0.221 7.8 0.19 0.98 referred to the Web version of this article.)
(1.54) (0.016)
35 5.58 2.57 0.371 8.01 0.15 0.99
(0.85) (0.036)
35 5.42 1.26 0.332 7.64 0.24 0.99 Table 3
(1.01) (0.046) Coefficients for paneer secondary models with 95% confidence intervals.
40 5.58 <0.01 0.173 7.25 0.2 0.98 a b Tmin Tmax RMSE R2
(1.51) (0.012)
40 5.53 <0.01 0.183 7.83 0.24 0.98 0.01382 0.3885 − 5.109 42.91 0.03482 0.9486
(1.6) (0.018) (0.01095, (− 0.2317, (− 9.781, (38.59,
0.0167) 1.009)a − 0.4371) 47.24)
a
Lag time.
a
b
Growth rate. 95% confidence intervals in parentheses.
c
Maximum population density.
d
Standard error-of-fit. 3.3. Comparison of the paneer growth model with published data and
e
Standard error of parameter. L. monocytogenes growth models
f
Not detected <0.01 h.
The temperature range of the ComBase L. monocytogenes broth model

4
D. Sarkar et al. Food Microbiology 97 (2021) 103738

was the same as that of the present studies (Table 4). Adjusting ComBase
model inputs for pH and aw to 5.5 and 0.999, respectively, ComBase
under- and over-predicted L. monocytogenes μmax, showing an inflection
point at ~20 ◦ C (Fig. 3), and an acceptable performance (Bf = 1.01, Af =
1.12) compared to the developed paneer model. The PMP
L. monocytogenes aerobic and anaerobic broth models (4–35 ◦ C, pH 5.5,
NaCl 0.5%) were unacceptable, demonstrating marked over-predictions
for temperatures >15 ◦ C by the aerobic model (Bf = 0.84, Af = 1.22) and
for temperatures 4–30 ◦ C for the anaerobic model (Bf = 0.88, Af = 1.13).
Likewise, the FSSP L. monocytogenes broth model (4–25 ◦ C, NaCl 0.6%,
pH 5.6) produced unacceptable over-predictions (Bf = 0.81, Af = 1.24).
Published models for L. monocytogenes growth in queso fresco,
ricotta, camembert, and blue cheeses had narrow predictions ranges and
markedly under-predicted growth in paneer (Fig. 3, Table 4). In
contrast, a model for camembert cheese over-predicted growth (Lobacz
et al., 2013). When compared to independent data for other cheeses,
growth in Greek whey cheeses (myzithra, anthotyros, and manouri) was
accurately predicted by the paneer model, whereas growth in gusto
cheeses with different spices was markedly over-predicted.
3.4. Upper temperature growth/no-growth boundary
For all pH levels, growth (i.e. ≥ 1 log10 cfu/mL) in broth or paneer
did not occur at 43, 44, or 45 ◦ C (Fig. 4). At 41, 41.5, and 42 ◦ C, growth
in broth occurred at pH ≥ 5.28, ≥5.4, and ≥5.74. For paneer (Table 5;
Fig. 4), growth occurred in all samples at 41 ◦ C but growth was not
detected at 43 ◦ C, as observed in broth. At 42 ◦ C, growth occurred in
paneer at pH ≥ 5.66; at pH 5.61, one of the two replicates displayed
growth.
The developed G/NG model, based upon minimizing the maximum
likelihood, had inputs for pH and temperature but the higher order
terms (T*pH, T2 and pH2) were not significant (P > 0.05). It is described
as,
f = 199.9 − 6.92∗T + 16.14∗pH (8)
where f is the logit of the probability of growth and T = temperature
(◦ C).
With P ≥ 0.5 as the criterion for growth, 90.5% of the observations
were estimated with a high concordance rate of 98.25%. The Hosmer-
Lemeshow goodness-of-fit statistic gave a non-significant p-value (χ2
= 5.32, df = 8, p = 0.72), indicating that the model fit the data well. This
is also depicted in Fig. 4, with only four misclassifications between the
observed and estimated values at the probability level P = 0.5. The
paneer validation experiment showed that the model predicted 93.34%
of observations correctly, with only two misclassifications, indicating
that the developed model can be successfully used for estimating G/NG
outcomes in paneer.
4. Discussion
A key tenet of food safety risk management is understanding how
hazards quantitively change as a function of food processing and
handling. In this regard, relatively little information is available about
pathogen behaviour in paneer, even though each year this product is
consumed by millions of individuals worldwide. In response to this in­
formation gap, this study describes and models viability kinetics of
L. monocytogenes in paneer over a wide range of temperature relative to
production and storage.
These experiments describe L. monocytogenes growth in fresh paneer
over a temperature range of 4–45 ◦ C, in which the average μmax ranges
from 0.014 to 0.352 log10 CFU/h. The rate observed at 4 ◦ C is similar to
water-contaminated paneer but is approximately 1.6 times greater than
that for surface-contaminated paneer (Varalakshmi and Leysen, 2020).
Over the range of 4–40 ◦ C, the growth rate profile in paneer was similar
to that reported for other matrices, displaying a maximum rate at
~37 ◦ C (Membré et al., 2004; Buazzi et al., 1992; Duh and Schaffner,
1993). The growth kinetics of TVC in negative control samples did not
suggest that background microbiota would inhibit growth of L. mono­
cytogenes in paneer through the ‘’Jameson effect’’ (Jameson, 1962). Lag
time was not detected in all replicates, however when detected the
highest value occurred at 4 ◦ C and then decreased with increasing
temperature. The inoculum was cultured at 37 ◦ C and then maintained
at ~25◦ before inoculation, pre-culture conditions that may have
contributed to the observed variation in LT (Hudson, 1993; Dufrenne
et al., 1997).
L. monocytogenes growth has been well characterized and, in some
cases, modelled in traditional and niche market cheeses. For example,
growth kinetics have been measured in minas frescal (Cabral et al.,
2019), feta (Andritsos et al., 2019), gouda (Wemmenhove et al., 2013,
2016), fresh ricotta (Tirloni et al., 2019), queso fresco (Thomas et al.,
2019), queso blanco (Ulhich et al., 2006), camembert (Lobacz et al.,
2013) and blue cheeses (Lobacz et al., 2013). When compared to

Table 4
Comparison of the developed paneer model to published models and data.
Model/Reference na Matrix Minimum temperatureb Maximum temperatureb Bf Af

Online software
ComBasec 8 broth 4 40 1.01 1.12
PMP (aerobic)d 7 broth 4 35 0.84 1.22
PMP (anaerobic) 7 broth 4 35 0.88 1.13
FSSPe 5 broth 4 25 0.81 1.24
Published models
Ulhich et al. (2006) 5 queso blanco 4 25 1.15 1.19
Lobacz et al. (2013) 5 blue cheese 3 15 0.76 1.3
Lobacz et al. (2013) 5 camembert cheese 3 15 1.55 1.55
Tirloni et al. (2019) (model #1) 4 fresh ricotta 4.1 20 1.91 1.91
Thomas et al. (2019) 6 queso fresco 5 30 1.53 1.53
Published growth data for cheese
Papageorgiou et al. (1996) 3 myzithra 5 22 1 1.03
Papageorgiou et al. (1996) 3 anthotyros 5 22 0.98 1.06
Papageorgiou et al. (1996) 3 manouri 5 22 0.99 1.06
Lobacz et al. (2016) 5 gusta cheese with red pepper 3 15 0.31 3.13
Lobacz et al. (2016) 5 gusta cheese with garlic pepper 3 15 0.36 2.77
Lobacz et al. (2016) 5 gusta cheese with mixed herbs 3 15 0.36 2.71
a
Number of observations.
b
Maximum and minimum temperature of the models or data (◦ C).
c
Baranyi and Tamplin (2004).
d
USDA, 2013.
e
DTU Food, National Food Institute, version 4.0.

5
D. Sarkar et al. Food Microbiology 97 (2021) 103738

Fig. 3. Comparison of predictions from developed paneer model with other models and data.

Fig. 4. Growth/no-growth data for L. monocytogenes cocktail tested under different conditions of pH and temperature in BHI broth and paneer, along with the
developed model.

commonly used L. monocytogenes broth-based models in public software over-predicted rate above ~20 ◦ C. Both the FSSP and PMP models
packages, the ComBase L. monocytogenes/innocua model more accu­ markedly over-predicted growth rate compared to the paneer model.
rately predicted growth rate (Bf = 1.01, Af = 1.12) in paneer, however Such differences might be attributed to variations in food matrix
the ComBase model under-predicted growth rate below ~20 ◦ C and microstructure, which is known to influence bacterial growth (Noriega

6
D. Sarkar et al. Food Microbiology 97 (2021) 103738

Table 5 is relevant to paneer processing. This includes the upper temperature

Parameter estimates with performance statistics for the model. boundary that affects L. monocytogenes viability as milk cools following
Parameter Estimated value Standard error Pr > ChiSq the initial heating process. Additionally, prepared paneer is often held
on hot plates in restaurants and catering services. Therefore, the
Intercept 199.9 59.04 0.0007
T − 6.92 1.94 0.0004 developed G/NG model may assist in the design of safe holding tem­
pH 16.14 4.43 0.0003 peratures implemented by the paneer industry.
Hosmer-Lemeshow 5.32 (df = 8, p = 0.72)
Following the approach of other G/NG boundary studies (Skandamis
% concordant 98.25 et al., 2007; Belessi et al., 2011) that used low citric acid concentrations,
% correct predictions 90.5 pH was described as a function of citric acid. Growth boundary/inter­
face models for L. monocytogenes have been reported by several authors
(Tienungoon et al., 2000; Gysemans et al., 2007; Bolton and Frank,
et al., 2008; Skandamis and Jeanson, 2015). Therefore, these results
1999; Koutsoumanis et al., 2004; Vermeulen et al., 2007, 2009). Valero
highlight potential drawbacks when using broth-based models to inform
et al. (2006) and Koutsoumanis et al. (2004) studied growth up to 30 ◦ C
risk assessment. Models developed for fermented cheeses such as
and Tienungoon et al. (2000) up to 36 ◦ C, in combination with factors
camembert and blue, and for queso fresco and queso blanco,
including pH and aw. The majority of G/NG studies have been conducted
under-predicted growth in paneer (Lobacz et al., 2013).
at refrigeration or near-refrigeration temperatures, and/or a combina­
Under-prediction might be due to inhibition of L. monocytogenes by
tion of pH, aw, and organic acid. Duh and Schaffner (1993) and Membré
mesophilic starter cultures and/or salt used during processing (Leggett
et al. (2004) studied growth at 40 and 42 ◦ C, respectively. Of note, the
et al., 2012). Other reasons could include potential inhibiting effect of
extensive Microbial Response Viewer database (Koseki, 2009) has few
bacteriocins and other secondary metabolites produced in fermented
reports about L. monocytogenes survival at temperatures above Tmax
products. A similar pattern of growth under-estimation was observed by
(Polese et al., 2011).
Tirloni et al. (2019) when they compared predictions between fer­
For the upper temperature boundary studies, a synergistic effect of
mented cheese models and fresh ricotta. Although fresh ricotta is also an
pH and temperature was observed, as a small pH shift from 5.74 to 5.72
acid coagulated fresh cheese, model #1 reported by Tirloni et al. (2019)
at 42 ◦ C resulted in no growth of L. monocytogenes. Similarly, Tie­
underpredicts L. monocytogenes growth in paneer. This could possibly be
nungoon et al. (2000) observed an abrupt transition in G/NG for
attributed to higher concentrations of lactic acid and NaCl in ricotta
L. monocytogenes when pH changed 0.1–0.2 pH units. At 30 ◦ C, a mini­
versus paneer.
mum pH of 4.39 in broth (with HCl as acidulant) supports
Food Standards Australia and New Zealand regulations state that
L. monocytogenes growth (George et al., 1988); we observed a minimum
L. monocytogenes must be absent in five 25-g samples of ready-to-eat
pH (using citric acid as acidulant) of 5.72 at 42 ◦ C. In this regard, we
products at the time of manufacture or should not exceed 2 log10
used a mixed culture of L. monocytogenes to describe a worst-case sce­
CFU/g at the time of consumption (FSANZ, 2014). Assuming hypo­
nario; Gysemans et al. (2007) reported that the boundary of a multiple
thetical scenarios in which a fresh batch of paneer is contaminated with
strain culture may be a composite line consisting of the growth limits of
an initial load of 1 CFU/g (0 log10 CFU/g) of L. monocytogenes and then
the individual strains under the most stringent conditions.
stored at 4 or 7 ◦ C, mild abuse temperatures common for domestic re­
In conclusion, this research quantifies the behaviour of
frigerators (Evans and Redmond, 2015), or 22 ◦ C, the paneer model
L. monocytogenes in fresh paneer over the temperature range of 4–45 ◦ C
estimates L. monocytogenes would reach 2 log CFU/g in 126.2 h, 60.93 h,
and identifies the limitations of other published models in successfully
and 14.25 h, respectively. This indicates that an opened package of
predicting the growth of the pathogen in paneer. Overall, the need to
paneer should be consumed within 5 d when stored at 4 ◦ C, 2 d at 7 ◦ C,
monitor the temperature of the product during manufacturing and
and 14 h at 22 ◦ C. These are important considerations in terms of food
storage of the product is highlighted in this study, and the developed
safety management plans considering that large quantities of paneer are
predictive model can be used in foods safety management systems to
produced at home or by the unorganised dairy sector.
reduce the risk of L. monocytogenes growth.
Collating information available on the physicochemical character­
istics of paneer produced and sold in Australia (personal communica­
Declaration of competing interest
tion, Dr. Gabrielle Cook, Dairy Food Safety Victoria), pH, aw, and
moisture content of commercial samples in Victoria, Australia ranged
None.
from 5.45 to 5.76, 0.98 to 0.99, and 48.9–59%, respectively. The
maximum sodium chloride concentration and lactic acid concentration
Acknowledgements
was 0.1% and 1.4 mM, respectively. pH can be a possible hurdle, but the
pH range of commercial samples is well within the growth range of
This work was supported by the Centre of Food Safety and Innova­
L. monocytogenes. Organic acids in their undissociated forms inhibit the
tion in the Tasmanian Institute of Agriculture. This research did not
growth of L. monocytogenes in food products. Among commonly used
receive any specific grant from funding agencies in the public, com­
organic acids in food formulation, lactic acid and citric acid exhibit the
mercial, or not-for-profit sectors.
highest and lowest antilisterial activity, respectively, when used in a
The authors generously thank Caroline Claye for technical assistance
model cheese system (Engstrom et al., 2020). Wemmenhove et al.
and Dr. Gabrielle Cook, Dairy Food Safety Victoria for assisting with
(2016) reported that for six different strains of L. monocytogenes tested in
data and valuable discussions.
broth media, the average minimum inhibitory concentration (MIC) for
undissociated citric acid for all strains tested was 3.8 mM, at pH 5.2–5.6.
References
However, for fresh ricotta, Tirloni et al. (2019) reported the MIC for
undissociated citric acid and undissociated lactic acid required to pre­ Andritsos, N.D., Kallitsis, T., Roukas, D., 2019. Growth potential of Listeria monocytogenes
vent L. monocytogenes growth was 24 mM and 5.1–14.7 mM, respec­ in ready-to-eat Feta cheese-based sauce stored at 4◦ C. J. Food Saf. 39, e12599
tively. The maximum lactic acid concentration in commercial paneer is https://doi.org/10.1111/jfs.12599.
Aneja, R.P., Mathur, B.N., Chandan, R.C., Banerjee, A.K., 2002. Technology of Indian
very low (1.4 mM), which is common for fresh cheeses that have no Milk Products. Dairy India Publications, Delhi, India.
added lactate, thus making refrigeration the most important Astoreca, A., Barberis, C., Magnoli, C., Dalcero, A., 2010. Aspergillus carbonarius growth
post-processing hurdle for processed paneer, similar to other fresh and ochratoxin A production on irradiated dried grapes under different water
activity and temperature conditions. World Mycotoxin J. 3, 175–182. https://doi.
cheeses (ILSI Research Foundation and Risk Science Institute, 2005). org/10.3920/WMJ2009.1164.
Optimally, G/NG models should encompass a temperature range that

7
D. Sarkar et al.
Baranyi, J., Roberts, T.A., 1994. A dynamic approach to predicting bacterial growth in
food. Int. J. Food Microbiol. 23, 277–294. https://doi.org/10.1016/0168-1605(94)
90157-0.
Baranyi, J., Tamplin, M.L., 2004. ComBase: a common database on microbial responses
to food environments. J. Food Protect. 67, 1967–1971. https://doi.org/10.4315/
0362-028X-67.9.1967.
Belessi, C.E.A., Gounadaki, A.S., Schvartzman, S., Jordan, K., Skandamis, P.N., 2011.
Evaluation of growth/no growth interface of Listeria monocytogenes growing on
stainless steel surfaces, detached from biofilms or in suspension, in response to pH
and NaCl. Int. J. Food Microbiol. 145, S53–S60. https://doi.org/10.1016/j.
ijfoodmicro.2010.10.031.
Bhilegaonkar, K.N., Kulshrestha, S.B., Kapoor, K.N., Kumar, A., Agarwal, R.K., Singh, B.
R., 1997. Isolation of Listeria monocytogenes from milk. J. Food Sci. Technol. 34,
248–250.
Bolton, L.F., Frank, J.F., 1999. Defining the growth/no-growth interface for Listeria
monocytogenes in mexican-style cheese based on salt, ph, and moisture content.
J. Food Protect. 62, 601–609. https://doi.org/10.4315/0362-028X-62.6.601.
Buazzi, M.M., Johnson, M.E., Marth, E.H., 1992. Survival of Listeria monocytogenes during
the manufacture and ripening of Swiss cheese. J. Dairy Sci. 75, 380–386. https://doi.
org/10.3168/jds.S0022-0302(92)77772-8.
Cabral, G.J., Valencia, G.A., Carciofi, B.A.M., Monteiro, A.R., 2019. Modeling microbial
growth in Minas Frescal cheese under modified atmosphere packaging. J. Food
Process. Preserv. 43 https://doi.org/10.1111/jfpp.14024.
Cadavez, V.A.P., Campagnollo, F.B., Silva, R.A., Duffner, C.M., Schaffner, D.W.,
Sant’Ana, A.S., Gonzales-Barron, U., 2019. A comparison of dynamic tertiary and
competition models for describing the fate of Listeria monocytogenes in Minas fresh
cheese during refrigerated storage. Food Microbiol. 79, 48–60. https://doi.org/
10.1016/j.fm.2018.11.004.
CDC, 2015. Recall & Advice to Consumers, Restaurants, and Retailers (Final Update).
Multistate outbreak of Listeriosis linked to soft cheeses. https://www.cdc.gov/liste
ria/outbreaks/soft-cheeses-09-15/advice-consumers.html. (Accessed 27 January
2020).
Chawla, A., Chawla, N., 2009. Milk and Dairy Products in India – Production,
Consumption and Exports. Hindustan Studies & Services Ltd.
Dufrenne, J., Delfgou, E., Ritmeester, W., Notermans, S., 1997. The effect of previous
growth conditions on the lag phase time of some foodborne pathogenic micro-
organisms. Int. J. Food Microbiol. 34, 89–94. https://doi.org/10.1016/S0168-1605
(96)01170-1.
Duh, Y.-H., Schaffner, D.W., 1993. Modeling the effect of temperature on the growth rate
and lag time of Listeria innocua and Listeria monocytogenes. J. Food Protect. 56,
205–210. https://doi.org/10.4315/0362-028x-56.3.205.
Engstrom, S.K., Cheng, C., Seman, D., Glass, K.A., 2020. Growth of Listeria monocytogenes
in a model high-moisture cheese based on pH, moisture, and acid type. J. Food
Protect. 83, 1335–1344. https://doi.org/10.4315/jfp-20-069.
Evans, E.W., Redmond, E.C., 2015. Analysis of older adults’ domestic kitchen storage
practices in the United Kingdom: identification of risk factors associated with
listeriosis. J. Food Protect. 78, 738–745. https://doi.org/10.4315/0362-028X.JFP-
14-527.
FAO/WHO, 2007. Working principles for risk analysis for food safety for application by
governments. URL http://www.fao.org/3/a-a1550t.pdf. (accessed 1.23.20).
Food Poisoning bulletin, 2015. International recalls of Listeria contaminated cheese. URL.
https://foodpoisoningbulletin.com/2015/international-recalls-of-listeria-co
ntaminated-cheese/.
Food Standards Australia and New Zealand (FSANZ), 2014. Guidance on the Application
of Microbiological Criteria for Listeria Monocytogenes in RTE Food.
Gaulin, C., Ramsay, D., Bekal, S., 2012. Widespread listeriosis outbreak attributable to
pasteurized cheese, which led to extensive cross-contamination affecting cheese
retailers, Quebec, Canada, 2008. J. Food Protect. 75, 71–78. https://doi.org/
10.4315/0362-028X.JFP-11-236.
George, S.M., Lund, B.M., Brocklehurst, T.F., 1988. The effect of pH and temperature on
initiation of growth of Listeria monocytogenes. Lett. Appl. Microbiol. 6, 153–156.
https://doi.org/10.1111/j.1472-765X.1988.tb01237.x.
Gould, L.H., Mungai, E., Casey, B.B., 2014. Outbreaks attributed to cheese: differences
between outbreaks caused by unpasteurized and pasteurized dairy products, United
States, 1998–2011. Foodborne Pathol. Dis. 11, 545–551. https://doi.org/10.1177/
0003122413519445.
Gysemans, K.P.M., Bernaerts, K., Vermeulen, A., Geeraerd, A.H., Debevere, J.,
Devlieghere, F., Van Impe, J.F., 2007. Exploring the performance of logistic
regression model types on growth/no growth data of Listeria monocytogenes. Int. J.
Food Microbiol. 114, 316–331. https://doi.org/10.1016/j.ijfoodmicro.2006.09.026.
Holah, J.T., Bird, J., Hall, K.E., 2004. The microbial ecology of high-risk, chilled food
factories; evidence for persistent Listeria spp. and Escherichia coli strains. J. Appl.
Microbiol. 97 (1), 68–77. https://doi.org/10.1111/j.1365-2672.2004.02272.x.
Hudson, J.A., 1993. Effect of pre-incubation temperature on the lag time of Aeromonas
hydrophila. Lett. Appl. Microbiol. 16, 274–276. https://doi.org/10.1111/j.1472-
765X.1993.tb01417.x.
ILSI Research Foundation, Risk Science Institute, 2005. Achieving continuous
improvement in reductions in foodborne listeriosis–a risk-based approach. J. Food
Protect. 68, 1932–1994.
Jameson, J.E., 1962. A discussion of the dynamics of Salmonella enrichment. J. Hyg. 60,
193–207.
Khan, S.U., Pal, M.A., 2011. Paneer production: a review. J. Food Sci. Technol. 48,
645–660. https://doi.org/10.1007/s13197-011-0247-x.
Koch, J., Dworak, R., Prager, R., Becker, B., Brockmann, S., Wicke, A., Wichmann-
Schauer, H., Hof, H., Werber, D., Stark, K., 2010. Large listeriosis outbreak linked to
8
Food Microbiology 97 (2021) 103738
cheese made from pasteurized milk, Germany, 2006-2007. Foodb. Pathog. Dis. 7,
1581–1584. https://doi.org/10.1089/fpd.2010.0631.
Koseki, S., 2009. Microbial Responses Viewer (MRV): a new ComBase-derived database
of microbial responses to food environments. Int. J. Food Microbiol. 134, 75–82.
https://doi.org/10.1016/j.ijfoodmicro.2008.12.019.
Koutsoumanis, K.P., Kendall, P.A., Sofos, J.N., 2004. A comparative study on growth
limits of Listeria monocytogenes as affected by temperature, pH and aw when grown
in suspension or on a solid surface. Food Microbiol. 21, 415–422. https://doi.org/
10.1016/j.fm.2003.11.003.
Kristkova, S.Z., Grace, D., Kuiper, M., 2017. The Economics of Food Safety in India – a
Rapid Assessment. Wageningen University & Research, Netherlands, 2017.
Kumar, S., Rai, D.C., Niranjan, K., Bhat, Z.F., 2014. Paneer—an Indian soft cheese
variant: a review. J. Food Sci. Technol. 51, 821–831. https://doi.org/10.1007/
s13197-011-0567-x.
Leggett, L.N., Tomasula, P.M., Van Hekken, D.L., Porto-fett, A.C.S., Shoyer, B., Renye, J.
A., Luchansky, J.B., Farkye, N., 2012. Effect of storage at 4 and 10c on the growth of
Listeria monocytogenes in and on queso fresco. J. Food Saf. 32, 236–245. https://doi.
org/10.1111/j.1745-4565.2012.00373.x.
Linnan, M.J., Mascola, L., Lou, X.D., Goulet, V., May, S., Salminen, C., Hird, D.W.,
Yonekura, M.L., Hayes, P., Weaver, R., 1988. Epidemic listeriosis associated with
Mexican-style cheese. N. Engl. J. Med. 13, 823–828.
Lobacz, A., Kowalik, J., Tarczynska, A., 2013. Modeling the growth of Listeria
monocytogenes in mold-ripened cheeses. J. Dairy Sci. 96, 3449–3460. https://doi.
org/10.3168/jds.2012-5964.
Lobacz, A., Zulewska, J., Kowalik, J., 2016. The analysis of the behaviour of Listeria
monocytogenes in fresh cheeses with various spices during storage. Procedia Food
Sci 7, 80–84. https://doi.org/10.1016/j.profoo.2016.02.092.
Martinez-Rios, V., Dalgaard, P., 2018. Prevalence of Listeria monocytogenes in European
cheeses: a systematic review and meta-analysis. Food Contr. 84, 205–214. https://
doi.org/10.1016/j.foodcont.2017.07.020.
Membré, J.-M., Kubaczka, M., Dubois, J., Chèné, C., 2004. Temperature effect on Listeria
monocytogenes growth in the event of contamination of cooked pork products.
J. Food Protect. 67, 463–469. https://doi.org/10.4315/0362-028x-67.3.463.
Nahidul-Islam, S.M., Kuda, T., Takahashi, H., Kimura, B., 2018. Bacterial and fungal
microbiota in traditional Bangladeshi fermented milk products analysed by culture-
dependent and culture-independent methods. Food Res. Int. 111, 431–437. https://
doi.org/10.1016/J.FOODRES.2018.05.048.
National Food Institute (DTU Food). Food spoilage & safety predictor. URL. http://fssp.
food.dtu.dk/ (accessed 10.10.19).
Noriega, E., Laca, A., Díaz, M., 2008. Modelling of diffusion-limited growth to predict
Listeria distribution in structured model foods. J. Food Eng. 87, 247–256. https://
doi.org/10.1016/j.jfoodeng.2007.11.035.
Nunes Silva, B., Cadavez, V., António Teixeira, J., Ellouze, M., Gonzales-Barron, U.,
2020. Cardinal parameter meta-regression models describing Listeria monocytogenes
growth in broth. Food Res. Int. 136, 109476. https://doi.org/10.1016/j.
foodres.2020.109476.
Oscar, T.E., 2005. Validation of lag time and growth rate models for Salmonella
Typhimurium: acceptable prediction zone method. J. Food Sci. 70, M129–M137.
https://doi.org/10.1111/j.1365-2621.2005.tb07103.x.
Østergaard, N.B., Eklöw, A., Dalgaard, P., 2014. Modelling the effect of lactic acid
bacteria from starter- and aroma culture on growth of Listeria monocytogenes in
cottage cheese. Int. J. Food Microbiol. 188, 15–25. https://doi.org/10.1016/j.
ijfoodmicro.2014.07.012.
Papageorgiou, D.K., Bori, M., Mantis, A., 1996. Growth of Listeria monocytogenes in the
whey cheeses Myzithra, Anthotyros, and Manouri during storage at 5, 12, and 22◦ C.
J. Food Prot. 59, 1193–1199. https://doi.org/10.4315/0362-028X-59.11.1193.
Plaza-Rodríguez, C., Thoens, C., Falenski, A., Weiser, A.A., Appel, B., Kaesbohrer, A.,
Filter, M., 2015. A strategy to establish food safety model repositories. Int. J. Food
Microbiol. 204, 81–90. https://doi.org/10.1016/j.ijfoodmicro.2015.03.010.
Polese, P., Del Torre, M., Spaziani, M., Stecchini, M.L., 2011. A simplified approach for
modelling the bacterial growth/no growth boundary. Food Microbiol. 28, 384–391.
https://doi.org/10.1016/j.fm.2010.09.011.
Product Safety Australia, 2014. Omar Cheese Pty Ltd—Village Paneer and Indya Paneer
Cheese. https://www.productsafety.gov.au/recall/omar-cheese-pty-ltd-village
-paneer-and-indya-paneer-cheese?source=recalls. (Accessed 27 January 2020).
Ratkowsky, D.A., Lowry, R.K., McMeekin, T.A., Stokes, A.N., Chandler, R.E., 1983.
Model for bacterial culture growth rate throughout the entire biokinetic temperature
range. J. Bacteriol. 154, 1222–1226.
Ratkowsky, D.A., Ross, T., 1995. Modelling the bacterial growth/no growth interface.
Lett. Appl. Microbiol. 20, 29–33. https://doi.org/10.1111/j.1472-765X.1995.
tb00400.x.
Ross, T., 1996. Indices for performance evaluation of predictive models in food
microbiology. J. Appl. Bacteriol. 501–508. https://doi.org/10.1111/j.1365-
2672.1996.tb03539.x.
Rosshaug, P.S., Detmer, A., Ingmer, H., Larsen, M.H., 2012. Modeling the growth of
Listeria monocytogenes in soft blue-white cheese. Appl. Environ. Microbiol. 78,
8508–8514. https://doi.org/10.1128/AEM.01865-12.
Shabala, L., Lee, S.H., Cannesson, P., Ross, T., 2008. Acid and NaCl limits to growth of
Listeria monocytogenes and influence of sequence of inimical acid and NaCl levels on
inactivation kinetics. J. Food Protect. 71, 1169–1177. https://doi.org/10.4315/
0362-028x-71.6.1169.
Skandamis, P.N., Jeanson, S., 2015. Colonial vs. planktonic type of growth: mathematical
modeling of microbial dynamics on surfaces and in liquid, semi-liquid and solid
foods. Front. Microbiol. 6, 1178. https://doi.org/10.3389/fmicb.2015.01178.
Skandamis, P.N., Stopforth, J.D., Kendall, P.A., Belk, K.E., Scanga, J.A., Smith, G.C.,
Sofos, J.N., 2007. Modeling the effect of inoculum size and acid adaptation on
D. Sarkar et al.
growth/no growth interface of Escherichia coli O157:H7. Int. J. Food Microbiol. 120,
237–249. https://doi.org/10.1016/j.ijfoodmicro.2007.08.028.
Soni, D.K., Singh, R.K., Singh, D.V., Dubey, S.K., 2013. Characterization of Listeria
monocytogenes isolated from Ganges water, human clinical and milk samples at
Varanasi, India. Infect. Genet. Evol. 14, 83–91. https://doi.org/10.1016/j.
meegid.2012.09.019.
Tamplin, M.L., 2005. Chapter 7: modeling pathogen behavior in foods. In: Fratamico, P.,
Smith, J. (Eds.), Foodborne Pathogens: Microbiology and Molecular Biology. Caister
Academic Press, Norfolk, UK, pp. 113–120.
Thomas, M., Tiwari, R., Mishra, A., 2019. Predictive model of Listeria monocytogenes
growth in queso fresco. J. Food Protect. 82, 2071–2079. https://doi.org/10.4315/
0362-028X.JFP-19-185.
Tienungoon, S., Ratkowsky, D.A., McMeekin, T.A., Ross, T., 2000. Growth limits of
Listeria monocytogenes as a function of temperature, pH, NaCl, and lactic acid. Appl.
Environ. Microbiol. 66, 4979–4987. https://doi.org/10.1128/AEM.66.11.4979-
4987.2000.
Tirloni, E., Stella, S., Bernardi, C., Dalgaard, P., Rosshaug, P.S., 2019. Predicting growth
of Listeria monocytogenes in fresh ricotta. J. Food Microbiol. 78, 123–133. https://
doi.org/10.1016/j.fm.2018.10.004.
Uhlich, G.A., Luchansky, J.B., Tamplin, M.L., Molina-corral, F.J., Anandan, S., Porto-
fett, A.C.S., 2006. Effect of storage temperature on the growth of Listeria
monocytogenes on Queso Blanco slices. J. Food Saf. 26, 202–214.
U.S. Department of Agriculture, Agricultural Research Service, 2013. USDA-ARS.
Pathogen modeling Program (PMP) online. https://pmp.errc.ars.usda.gov/default.
aspx/. Accessed 20.2.2020.
U.S. Department of Agriculture, Foreign Agricultural Services, 2015. Cheese Demand
Rising - New Market Opportunities.
Vaishnavi, C., Singh, S., Grover, R., Singh, K., 2001. Bacteriological analysis of paneer in
chandigarh. Indian J. Med. Microbiol. 19, 224–226.
Valero, A., Carrasco, E., Pérez-Rodriguez, F., García-Gimeno, R.M., Zurera, G., 2006.
Growth/no growth model of Listeria monocytogenes as a function of temperature, pH,
View publication stats
Food Microbiology 97 (2021) 103738
citric acid and ascorbic acid. Eur. Food Res. Technol. 224, 91–100. https://doi.org/
10.1007/s00217-006-0293-1.
Varalakshmi, S., Leysen, S., 2020. Evaluation of the effectiveness of different
preservation techniques on the inactivation of Listeria monocytogenes by using
challenge testing protocol in the fresh, soft cheese - paneer. LWT (Lebensm.-Wiss. &
Technol.) 125, 109359. https://doi.org/10.1016/j.lwt.2020.109359.
Vermeulen, A., Gysemans, K.P.M., Bernaerts, K., Geeraerd, A.H., Van Impe, J.F.,
Debevere, J., Devlieghere, F., 2007. Influence of pH, water activity and acetic acid
concentration on Listeria monocytogenes at 7 ◦ C: data collection for the development
of a growth/no growth model. Int. J. Food Microbiol. 114, 332–341. https://doi.org/
10.1016/j.ijfoodmicro.2006.09.023.
Vermeulen, A., Gysemans, K.P.M., Bernaerts, K., Geeraerd, A.H., Debevere, J.,
Devlieghere, F., Van Impe, J.F., 2009. Modelling the influence of the inoculation
level on the growth/no growth interface of Listeria monocytogenes as a function of pH,
aw and acetic acid. Int. J. Food Microbiol. 135, 83–89. https://doi.org/10.1016/j.
ijfoodmicro.2009.07.038.
Wemmenhove, E., Stampelou, I., van Hooijdonk, A.C.M., Zwietering, M.H., Wells-
Bennik, M.H.J., 2013. Fate of Listeria monocytogenes in Gouda microcheese: No
growth, and substantial inactivation after extended ripening times. Int. Dairy J. 32,
192–198. https://doi.org/10.1016/j.idairyj.2013.05.004.
Wemmenhove, Ellen, van Valenberg, H.J.F., Zwietering, M.H., van Hooijdonk, T.C.M.,
Wells-Bennik, M.H.J., 2016. Minimal inhibitory concentrations of undissociated
lactic, acetic, citric and propionic acid for Listeria monocytogenes under conditions
relevant to cheese. Food Microbiol. 58, 63–67. https://doi.org/10.1016/j.
fm.2016.03.012.
Wemmenhove, E., Wells-Bennik, M.H.J., Stara, A., van Hooijdonk, A.C.M.,
Zwietering, M.H., 2016. How NaCl and water content determine water activity
during ripening of Gouda cheese, and the predicted effect on inhibition of Listeria
monocytogenes. J. Dairy Sci. 99, 5192–5201. https://doi.org/10.3168/jds.2015-
10523.