National Journal of Advanced Research

www.multidisciplinaryarticle.in
ISSN: 2455-216X
Received: 25-02-2024, Accepted: 19-03-2024, Published: 11-04-2024
Volume 10, Issue 1, 2024, Page No. 41-50

Artisanal fresh cheese quality improvement using hurdle technologies combination

Dely M1*, Zaouak A2, Ben Haj Koubair H1, Mnasser H1, Mankai M1
1
Department of Food Technology, University of Carthage, Higher Institute of Food Industries of Tunisia, ESIAT, Tunisia
2
Department of Nuclear Science and Technology, National Center for Nuclear Science and Technology, CNSTN, Tunisia

Abstract
The aim of this study was to compare the effect of combining electron beam irradiation with the activation of the
lactoperoxidase system as a decontamination tool. Microbiological, physico-chemical and organoleptic analyses were carried
out during storage of the cheeses at +4±1ºC for 18 days, and the shelf life was determined.
The use of electron beam irradiation combined with lactoperoxidase had a significant effect on microbiological quality
compared with control cheeses and those activated by lactoperoxidase alone throughout the shelf life.
However, the activation of lactoperoxidase alone had no significant impact on the development of these germs. With regard to
sensory analysis, no descriptor had a significant effect detected by the panel throughout the shelf life. On the other hand, these
conservation combination enabled the the shelf life to be extended.
The control cheese had a use-by date of 3.63 days and the cheese preserved with lactoperoxidase only 3.09 days. The
combination, low-dose electron beam irradiation with LPS extended the shelf life of cheese to 6.82 days, the medium dose
(1kGy) combined with lactoperoxidase had the shelf life of 10.92 days compared with 16.93 days for the highest dose
(1.5kGy) combined with lactoperoxidase.

Keywords: Microbiological quality, shelf life, electron accelerator, lactoperoxidase, organoleptic quality

Introduction ensuring microbiological safety (Predrag et al., 2020). The

The cheese market is booming due to the constant increase combination technique involves preserving food using
in its production and consumption (Emma et al., 2016). several agents/methods applied simultaneously or
Cheese is highly valued by consumers for its nutritional and sequentially (Peleg, 2020) [14].
organoleptic qualities as it is recognised as an important The combination of different technologies and/or the
supplier of nutrients, and is rich in many minerals, including adjustment of a food's pH, water activity (aW), oxygen
Ca, Mg, P and Zn, as well as vitamins A, D, E and K tension, etc., can be used to improve its preservation
(Emma et al., 2016). efficiency, which has been widely documented and has
Dairy products are the main source of dietary calcium in become common knowledge (Peleg, 2020) [14]. The latter is
many countries, including the US, UK and most of Northern widely used in industrialised as well as developing countries
Europe, providing a source that cannot be easily replaced by for efficient food preservation (Peng et al.,2015) [15].
other foods (Emma et al., 2016). Combination technology is an important approach that can
On the other hand, cheese has a fairly short shelf life due to be used to improve quality parameters during food
its multiple varieties of micro-organisms such as yeasts, processing and storage. Intelligent application of barriers
Pseudomonas sp, heterofermentative lactic bacteria, improves sensory characteristics, food chemical and
Clostridia, Bacillus sp, coliforms, Klebsiella pneumoniae, microbiological qualities (Peng et al., 2015) [15]. Moreover,
Penicillium and psychrophilic spoilage microflora (Zhu et more than 60 reported barriers are available, these can be
al., 2020) [22]. used in different combinations and concentrations for a wide
The application of heat treatments at very high temperatures range of foods. This versatility makes the application of the
can alter the composition of milk as it affects protein technology possible in modern and local food processing
structures and water-soluble vitamins, resulting in a (Peng et al., 2015) [15].
decrease in total fat and total solids and an increase in urea A particular example is the use of lactoperoxidase, a natural
(Bezie, 2019) [3]. enzyme considered to be an important element in the host's
Alternative processes are therefore needed to solve this natural defence system against bacterial infection. Indeed,
problem, mainly the use of combination technology (Peng et the LPO system can also be used to increase the stability of
al., 2015) [15]. milk storage at high ambient temperatures (Zarei et al.,
At the crossroads of major economic, environmental and 2016) [20].
public health issues, food preservation techniques today Non-thermal processes are implemented to inactivate
deserve the mobilisation of all stakeholders to ensure that spoilage micro-organisms and improve the nutritional,
technologies evolve towards solutions that are more sensory and microbial characteristics of the product; a good
respectful of both the environment and the consumer opportunity that presents itself is the use of irradiation
(Gontard et al., 2017) [7]. during cheese processing (Huo et al., 2013) [8].
This makes the application of barrier technology very Irradiation is the use of ionising radiation from radioactive
interesting as it can extend shelf life while preserving isotopes of cobalt or caesium or from accelerators producing
nutritional and sensory value. This technology works at controlled amounts of beta rays or X-rays on food (Huo et
lower temperatures and with shorter processing times, while al., 2013) [8].

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Food irradiation has been identified as a safe technology on in the presence of phenolphthalein at 1% (w/v), (1 mL of
the food process and used to disinfect and preserve food, NaOH (N/9) corresponds to 0.01 g of lactic acid per cent)
including extending shelf life (Huo et al., 2013) [8]. (Fedala et al., 2020) [6].

Materials and methods Determination of fat

Cheese manufacture The method is based on the acid-butyrometric method of
The cheese was produced from raw cow’s milk. Van Gulik. It is based on the dissociation of cheese proteins
Subsequently, the standardized milk was pasteurized at by the addition of sulfuric acid and separation of the fat by
85°C/10 min, cooled to 32 C, and inoculated by adding centrifugation in a Van Gulik butyrometer.
0.125 g/l of mesophilic starter culture (Barukcic et al., In a Van – Gulik butyrometer, put 3g of cheese, add sulfuric
2020) [2]. Fermentation (acid coagulation) of the milk was acid so that it covers the cheese mass, making the proteins
carried out at 30 C for about 16 h until curd was formed and dissociate in a water bath. After complete dissociation, fill
pH of about 4.6 was reached (Barukcic et al., 2020) [2]. the graduated rod with sulfuric acid and add 1 ml of isoamyl
At this point, the fermentation process was interrupted by alcohol (Fedala et al., 2020) [6]. The separation of fat is done
rapid cooling. The resulting curd was cut by a sterile knife by centrifugation (Fedala et al., 2020) [6].
and heated to a maximum of 40°C while continuing The fat expressed in g/100g of cheese is obtained by direct
agitation to allow separation of the whey. Transferred to reading on the butyrometer scale (Fedala et al., 2020) [6]
molds in cold storage (15-16°C) and drained (Barukcic et
al., 2020) [2]. Determination of total dry extract
The cheese yield is calculated according to the following The total dry extract (TDE) is determined using an oven set
formula: at 103±2°C. The test samples are measured to the nearest 1
mg and the dry matter is expressed as a percentage by
weight by the remainder after drying (Fedala et al., 2020) [6].
Two grams of cheese are weighed after homogenization of
the cheese paste, and the dry matter is expressed as weight
Y : means the cheese yield in %. percentage by the remainder after drying (Fedala et al.,
Wc : means the weight of cheese obtained in kilograms. 2020) [6].
W : means the weight of the milk in kilograms.

LPS activation
The sample was subjected to refrigeration combined with TDE: total dry extract
activation of the LP system by addition of sodium E1: weight in grams of the capsule and the test sample.
thiocyanate (NaSCN) as a source of thiocyanate (SCN) to a E2: weight in grams of the capsule and residue after drying
final concentration of 14 mg⁄L (Boulares et al., 2011) [4]. and cooling.
After 1 min of thorough mixing of the milk, 30 mg ⁄L of Mc: weight in grams of the empty capsule.
sodium percarbonate (2Na2CO33H2O) was added as a
source of hydrogen peroxide (H2O2) as recommended by the Determination of the dry defatted extract
International Dairy Federation (IDF1988). It is the result of the difference between the total dry extract
and the fat content and
Technique of combining LPS with irradiation determined as follows:
This technique consists in activating the milk with LPS
which will be used for the manufacture of cheese, which
will be subsequently irradiated. The cheeses were irradiated
in the National Center for Nuclear Science and Technology TDE : total dry extract
(CNSTN) of Sidi Thabet of Tunisia using a gas pedal 10 ESD: dry extract defatted
MeV electron beam at doses of 0.5; 1; 1.5 kGy respectively MG: Fatty matter
(Huo et al., 2013) [8].
Measurement of moisture
The samples were distributed as follows The moisture is calculated by the following formula:
Samples Control LPS LPS+ 0,5 kGy LPS+ 1 kGy LPS+ 1,5 kGy
All the tests are spread over three periods: t0, t9 and t18.

Physicochemical analysis Rheological study

pH Hardness (N), of cheese was determined by texturometer
The pH is measured with a pH meter (KDD002) to 0.01 analyzer type TVT 6700. Cheese samples were cut into 5 *
units of accuracy. For milk the determination is made on 10 5 cm cubes and stored at a room temperature before
mL of the sample. For cheese, the electrode of the pH meter measurements (Barukcic et al., 2020) [2].
is placed directly in the cheese (Fedala et al., 2020) [6] Then the prepared samples were subjected to double
compression at a traverse speed of 1 mm/s and a penetration
Titratable acidity distance of 40 mm up and down with 10 s between the two
This is an acid-base titration, lactic acid is neutralized by a cycles (Barukcic et al.,2020) [2].
solution of sodium hydroxide NaOH in the presence of
phenolphthalein as a colored indicator ((Fedala et al., 2020) Sensory analysis
[6]
. Once the production is finished, a hedonic test is carried out
For milk the acidity is expressed in degree Dornic (°D). The for the two cheeses. The purpose of this test is to compare
titration is performed by an alkaline solution (NaOH, N/9) the overall hedonic appreciation of the different cheeses by

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focusing on the individual feelings related to the pleasure or milk constituents during draining (Vignola et al., 2002) [19].
displeasure caused by the food (Fedala et al., 2020) [6]. It requires a high level of dry extract and more specifically
Consumer acceptance was determined using a 9-point scale. protein (casein) and high fat concentrations (Vignola et al.,
The results of the sensory analysis are represented by a 2002) [19].
radar graph. It allows to highlight the data around a According to our results, we obtained a Cheese yield Y=
reference value (Fedala et al., 2020) [6]. 10.7%.

Microbiological analysis Physicochemical analyses

The cheese was aseptically removed from the package and Effect of Hurdle technology on pH
the surface was cut with a sterile knife. The cheese slices No significant difference (P > 0.05) in pH was observed
were weighed into a sterile beaker with 100 ml of sterile between the control and the LPS-activated cheese. Seifu et
distilled water. The cheese was grated and homogenized al, (2004) [17] stated that activation of cheese by LPS had no
thoroughly (Fedala et al., 2020) [6] effect on pH. Similarly, no significant difference in pH was
Serial dilutions of the homogenates were placed on observed between the controls and the cheeses activated by
appropriate media in Petri dishes and analyzed immediately LPS throughout the ripening period (Boulares et al., 2011)
(Fedala et al., 2020). [6] The numbers of total coliforms, total [4]
.
aerobic mesophilic flora, yeasts and molds were monitored Fig 1 Shows the effect of ebeam treatment (0.5, 1 and 1.5
on consecutive days during storage (Fedala et al,. 2020) [6] kGy) and LPS activation on cheese pH at (0 d) t0, (9 d) t9
The media and incubation conditions used were as follows: and (18 d) t18 of storage at 4±1°C.
Coliforms: spread on violet red bile agar (VRBA), with a In fact, we noted that the pH of the control cheeses at t9 and
double cover layer of the same medium, incubated at 30° C t18 and those activated by LPS decreased significantly
for 24 to 48 hours (Fedala et al,. 2020) [6] compared to the other samples that underwent the combined
Total aerobic mesophilic flora: spread on PCA (Plate Count technique.
Agar) and incubation is done at 30°C for 48 hours (Fedala et This significant decrease can be attributed to the inhibition
al,. 2020) [6] of the microbial flora and particularly the lactic acid bacteria
Yeasts and molds: spread on Sabouraud dextrose agar, responsible for the acidification of the cheese.
incubated at 25° C for 5 to 7 days (Fedala et al,. 2020) [6] For the products treated with ebeam combined with LPS and
for the low dose of 0.5 kGy, the pH fell from 4.51 ± 0.01 at
The determination of shelf life
t0 to 4.47 ± 0.01 at t18, while for the medium dose of 1 kGy
The general equation that describes the loss of quality of a
the pH fell from 4.49 ± 0.01 at t0 to 4.45 ± 0.01 at t18.
food is applicable for any factor A is as follows:
Moreover, the highest dose, 1.5 kGy, reduced the pH of the
cheeses studied from 4.48 ± 0.01 at t0 to 4.43 ± 0.01 at t18.
This decrease in pH can be explained by the inhibition of
the microbial flora and in particular the lactic bacteria
r : rate of the degradation reaction = rate of formation of A ; responsible for the acidification of the cheese spread
K : reaction rate constant or apparent rate ; (Agherghour et al., 2015) [1].
A : concentration of the factor to be followed; The pH is a quality index that determines the ability of food
n : order of the degradation reaction. to be preserved. It is one of the main obstacles that
microbial flora must overcome in order to proliferate
Statistical analysis (Agherghour et al., 2015) [1].
All experiments were repeated at least three times. The It is therefore important to measure the pH in order to
results were subjected to a one-factor analysis of variance determine the stability of the food with respect to
with a significance level of 95% using Excel for the radar microorganisms, which are pathogenic to humans rarely
presented for the sensory analysis and by SPSS software develop at an acid pH, below 4 (Agherghour et al., 2015) [1].
using the one-factor ANOVA test. Most microorganisms grow at pH close to neutral, whereas
moulds grow at acid pH (Agherghour et al., 2015) [1].
Results and disscusion Our results disagree with those of Kim (2010) [9] who
Cheese yield studied the effect of ebeam with different doses of 1, 3 and 5
Cheese yield is of great interest in the cheese industry kGy on the pH of cheese and reported that electron beam
because it reflects the overall quantitative distribution of irradiation also had no significant effect on pH.

Fig 1: Effect of ebeam irradiation on the pH of fresh cheese (Control; LPS; 0.5kGy +LPS; 1 kGy+LPS; 1.5kGy+LPS)

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Effect of Hurdle technology on acidity control and a sample activated only with LPS. We can see
The acidity developed in cheese results from the that there was a significant increase in acidity in the cheeses
transformation of lactose into lactic acid. It is measured by that had undergone electron beam irradiation combined with
titration (Gadi et al., 2020). The low titratable acidity values LPS compared with the other control cheeses and those
recorded in our results reflect weak lactic fermentation in activated by LPS only at t0.
the cheese samples, which depends on the casein, mineral However, the increase in acidity in cheeses irradiated with
salt and ion content. It also depends on the hygienic electron beams combined with LPS compared with control
conditions during milking, the total microbial flora and its cheeses and cheeses irradiated with LPS alone was not
metabolic activity and the handling of the milk (Gadi et al., significant at t9 and t18.
2020). This increase can be explained by the deacidifying effect of
certain germs such as psychotropics and moulds and the
Fig 2 Shows the effect of cheese acidity at t0, t9 and t18 of resulting degradation or inhibition of the microbial flora and
storage at 4±1°C after a combination of ebeam treatment in particular the lactic acid bacteria responsible for the
with 0.5; 1 and 1.5 kGy and LPS activation compared to a acidification of the cheese (Leksir, 2018).

Fig 2: Effect of ebeam irradiation on titratable acidity of fresh cheese (Control ;

LPS ; 0.5kGy +LPS ; 1 kGy+LPS; 1.5kGy+LPS)

Effect of Hurdle technology on total dry extract compared with a control and a sample activated only with
The level of dry extract varies from one type of cheese to LPS.
another. This difference is due to the use of salt and the Graphical analysis shows an increase in dry extract content
draining time. The cheese's high dry matter content gives it as a function of dose, from a minimum of 35.73 g/kg
a relatively firm consistency. (Control) to a maximum of 36.93 g/kg (1.5 kGy).
Fig 3 shows the effect on the total dry extract of the cheese This increase was significant at t0, t9 and t18 and could be
at t0, t9 and t18 of storage at 4±1°C after a combination of due to the phenomenon of radiolysis of the water, which
ebeam treatment with 0.5, 1 and 1.5 kGy and LPS activation increased the percentage of dry extract.

Fig 3: Effect of ebeam irradiation on the total dry extract of fresh cheese (Control; LPS;
0.5kGy +LPS; 1 kGy + LPS; 1.5kGy+LPS)

Effect of Hurdle technology on fat content acids. Milk fat plays an essential role in the development of
Fat content and dry matter are very important in cheese both the taste and texture of cheese (Vignola, 2002) [19].
production because milk lipids are characterised by the Cheese texture depends on its fat content. In fact, the water
presence of relatively short-chain fatty acids which can be content and the proportions of long polysaturated fatty acids
absorbed by a simpler mechanism than long-chain fatty in the milk determine the texture of the dough: extra hard,

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semi-soft, soft, and so on. For example, with more than 60% cheeses and those activated by LPS and those irradiated
fat and less than 51% water, you get an extra-hard cheese. with 0.5 and 1 kGy combined with LPS at t0.
Too high a fat content can lead to problems with draining On the other hand, all the other samples showed a
and coagulation (Vignola, 2002) [19]. significant increase compared with the controls and those
Fig 4 shows the variation in the fat content of cheeses activated by LPS at t9 and t18.
treated with the first combination and control cheeses over a This increase could be due to oxidation of the lipids by the
storage period of 18 days at 4±1°C. irradiation process, producing OH radicals formed mainly as
The highest dose 1.5 kGy combined with LPS activation a result of radiolysis of water.
had a significant effect on fat content compared to all other This was approved by Hyun et al, (2010) who showed that
samples throughout the storage period. electron beam irradiation induced higher values of
However, there was a slight increase in fat content as a thiobarbituric acid reactive substances (TBARS) in samples
function of dose, but it did not exceed 2% in the control irradiated at 5 kGy than in other samples.

Fig 4 : effect of ebeam irradiation on the fat content of fresh cheese (Control; LPS;
0.5kGy +LPS; 1 kGy+LPS; 1.5kGy+LPS)

Effect of Hurdle technology on defatted dry extract with the control cheeses and those activated by LPS
Figure 5 shows the variation in fat-free dry matter of treated throughout the shelf life.
and untreated cheeses stored at 4±1°C. It can be seen that However, there was a significant variation in the defatted
the defatted dry extract values increased in proportion to the dry matter of cheeses irradiated with the highest dose (1.5
technique applied, rising from 5.40, 4.53 and 4.17 g/kg at t0. kGy) combined with LPS compared with control cheeses
The three doses applied did not have a significant effect and those activated by LPS throughout t9.
(p>0.05) on the fat-free dry extract of the cheeses compared

Fig 5: Effect of ebeam irradiation on the defatted dry extract of fresh cheese (Control; LPS;
0.5kGy +LPS; 1 kGy+LPS; 1.5kGy+LPS)

Effect of Hurdle technology on humidity moisture content is less than 58%), or a fairly moist, and
For the moisture parameter, if the values are outside the therefore fragile, dough if the moisture content% is greater
standards, there is a risk of having a fairly hard dough (if the than 60%. The TSE content is linked to the moisture

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content%, so outside the range given in table (40-42), there activated by LPS. On the other hand, no significant variation
is a risk of varying the moisture content% and therefore in humidity was observed in the LPS-activated cheese
ultimately contributing to the hardness or brittleness of the compared with the controls at t0.
dough. On the other hand, there was a significant difference
Fig 6 shows that the control cheese samples had an average between the cheeses that had been subjected to the different
moisture content of 64.27% and the LPS-activated cheese doses with LPS in terms of moisture compared to the
had a value of 64.83%, while the irradiated cheese samples controls and those activated only by LPS, and no significant
had values of 62.47%, 61.83% and 60.03% respectively at difference between the cheeses activated by LPS and the
t0. controls at t9 and t18. Thus the moisture content of the
In fact, there was a significant difference between the irradiated cheese samples was significantly lower than that
control and irradiated groups combined with LPS, since of the controls and those activated only by LPS.
there was a significant decrease in the cheeses with the This decrease can be attributed to the effect of gamma
highest dose combined with LPS compared with all the irradiation on the capacity of cheese proteins to retain water.
other samples at t0. Almost similar results were reported by Gosh et al, (1999).
Moreover, the other two doses combined with LPS had a
significant effect compared with the controls and those

Fig 6 : Effect of ebeam irradiation on the moisture of fresh cheese (Control; LPS; 0.5 kGy
+LPS;1 kGy+LPS; 1.5kGy+LPS)

Rheological study Calcium from the core then migrates to the surface to re
Texture is an essential factor in consumer acceptance of a establish the equilibrium, causing the micelles to destabilise
product. The term 'firmness' is commonly used to describe a and the cheese to soften as a result McSweeney, (2004) [11].
parameter assessed by means of empirical mechanical tests The changes observed in the texture of the untreated and
and considered as an attribute that must be maintained treated samples could be linked to this phenomenon.
during storage and processing. However, the three doses applied in combination with LPS
Table 1 shows the variation in firmness of treated and did not have a significant effect on cheese firmness at t0, t9
untreated cheeses stored at 4±1°C. and t18.
It can be seen that the average firmness value of the cheeses
decreases directly following irradiation and over time. Table 1: Effect of ebeam irradiation on the firmness of fresh
In fact, non-irradiated cheeses went from 0.28 to 0.2 N after cheese (Control ; LPS ; 0.5kGy +LPS ; 1 kGy+LPS; 1.5kGy+LPS)
18 days of refrigerated storage.
For cheeses activated only by LPS, firmness fell from 0.277 Time Doses T0 T9 T18
Control 0,28±0,0 0,247±0,005 0,2±0,01
to 0.21 N after 18 days of refrigerated storage.
LPS 0,277±0,01 0,243±0,01 0,21±0,00
As for the cheeses treated with the low dose combined with
0,5kGy 0,273±0,01 0,24± 0,00 0,207±0,01
LPS, their firmness fell from 0.27 to 0.21 N. For the 1 kGy 0,27±0,0 0,237± 0,00 0,2±0,0
medium dose (1 kGy) combined with LPS, the average 1,5 kGy 0,265±0,01 0,233± 0,01 0,187±0,01
firmness value rose from 0.27 to 0.20 N.
For the 1.5 kGy dose with LPS, a decrease was observed, Microbiological quality
reaching 0.26 N after 18 days of conservation. The results of the evolution of the microbial flora of the
This decrease in texture over time could be due to the cheeses at (t0) and after 9 and 18 days of storage at 4±1°C.
migration of calcium in response to the pH gradient and the The germs sought and counted in our work are considered to
activities of fungal enzymes and rennet. Several authors, be indicators of the overall quality of the finished product
Spinnler et al, (2004) [18] have reported that the change in and reflect compliance or non-compliance with good
texture is due to this phenomenon, as calcium phosphate hygiene practices (Gadi et al., 2020).
precipitates at the generally high pH of cheese rinds.

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Table 2: Effect of combining ebeam with LPS activation on microbiological quality

Total coliforms (UFC/g) Total mesophilic aerobes (UFC/g) Yeasts and molds (UFC/g)
t0 : 0,53±0,09 t0 : 3,00±0,09 t0 : 1,48±0,00
Témoin t9 : 1,41±0,03 t9 : 4,34±0,01 t9 : 3,88±0,02
t18 : 2,87±0,03 t18 : 4,52±0,02 t18 : 3,97±0,01
t0 : 0,34±0,11 t0 : 2,93±0,02 t0 : 1,52±0,04
LPS t9 : 1,34±0,03 t9 : 4,27±0,01 t9 : 3,85±0,01
t18 : 2,87±0,00 t18 : 4,45±0,01 t18 : 3,95±0,02
LPS t0 : 0,00±0,00 t0 : 0,59±0,05 t0 : 1,04±0,07
+ t9 : 0,95±0,08 t9 : 4,19±0,01 t9 : 2,33±0,00
0,5kGy t18 : 1,13±0,07 t18 : 4,27±0,02 t18 : 3,69±0,02
LPS t0 : 0,00±0,00 t0 : 0,07±0,13 t0 : 0,88±0,1
+ t9 : 0,81±0,1 t9 : 3,99±0,01 t9 : 2,21±0,02
1 kGy t18 : 0,95±0,03 t18 : 4,18±0,01 t18 : 2,45±0,03
LPS t0 : 0,00±0,00 t0 :0,00±0,00 t0 : 0,62±0,17
+ t9 : 0,46±0,15 t9 : 1,39±0,03 t9 : 1,68±0,14
1,5 kGy t18 : 0,78±0,17 t18 : 1,47±0,02 t18 : 2,10±0,00

Total coliforms in the target organism. It also depends on pH, temperature,

In fact, the number of total germs reflects the level of water activity (Aw), and the nature of the radiation used in
hygiene of the cheese samples. the process (Molins et al., 2001) [12].
Table 2 shows a total absence of total coliforms The results obtained stated that the use of the hurdle
immediately after treatment (t0) with all the doses combined technique significantly reduced the growth of total coliforms
with LPS. Even with LPS activation alone, there was a at low doses.
reduction in proliferation compared with the control.
For products treated with different doses combined with Mesophilic aerobes
LPS, we can see that the total coliforms were reactivated, The number of total aerobic mesophilic flora in cheese can
hence their increase over time. This increase in the cause premature swelling and the production of enterotoxins
irradiated products was still less than in the control and in cheese (Seifu et al., 2004) [17].
LPS-activated cheeses. The application of ebeam and the activation of LPS made it
The numbers in all the groups irradiated with LPS decreased possible to reduce mesophilic aerobes directly after
with increasing irradiation dose. treatment (t0) until a total absence of mesophilic aerobes for
In fact, at t0 there was a significant decrease at all irradiation the combination of LPS with the highest dose of 1.5 kGy, as
doses with electron beams combined with LPS compared shown in Table 2.
with the control and LPS-activated cheeses, and LPS Moreover, this combination of LPS with the highest dose
activation significantly reduced the evolution of total (1.5 kGy) at t0 resulted in a significant reduction compared
coliforms. to that of the control cheeses and those activated by LPS and
On the other hand, at t9 there was a significant decrease in those with a dose of 0.5 kGy combined with LPS.
the evolution of total coliforms in the cheese irradiated with With an initial control of 3.0 log CFU/g, there was a slight
the highest dose, 1.5 kGy combined with LPS, compared but non-significant reduction (p>0.05) for the cheese
with the control and activated by LPS and the other two activated only by LPS at 2.93 log CFU/g.
doses. It was also noted that irradiation of the cheeses with In fact, the reduction in this flora in cheeses made from milk
0.5 and 1 kGy combined with LPS resulted in a significant preserved by activation of the LPS system suggests that
decrease in the evolution of total coliforms compared with activation of this system in the milk before cheese
the control and LPS. manufacture could be of great practical importance (Seifu et
However, there was no significant increase in total al., 2004) [17].
coliforms in the LPS-activated cheese and the control. Moreover, Seifu et al, (2004) [17], found that preservation of
Similarly, at t18, the same results as those obtained at t9 were
cheese milk by the LPS system can be used to improve
observed, with the exception that there was no longer a
microbiological quality. At t9 and t18, there was a significant
significant decrease in the evolution of these germs in the
difference in all samples.
cheese irradiated with the highest dose and that at 1 kGy.
In fact, the 1.5 kGy dose combined with LPS was the best of However, during storage, aerobic mesophilic bacteria
the three tested against total coliforms, as it allowed total increased. In fact, after one week of refrigeration, there was
inhibition at t0 and caused the greatest reduction during an increase in aerobic mesophiles compared to t0. This
storage. increase noted at t9 can be explained as follows: the rays
Similar results were reported by Aly et al, (2012) who damaged and lysed the cells of the microorganisms at t0 and
showed that gamma irradiation at high doses which are 1;3 over time, and thanks to their biological systems and
and 5 kGy decreased total coliforms. resistances, this lysis was repaired.
This was probably due to the effect of the energy produced Our results are consistent with those of Hyun et al, (2010)
from the irradiation breaking the DNA bonds. Some bacteria who reported that at t0 the mesophilic load was absent at the
can repair DNA strand damage and resist the effect of highest dose of 3kGy.
irradiation. The effectiveness of the process depends on the Irradiation of 1 and 2 kGy reduced the microbial load by
organism's sensitivity to irradiation, the rate at which it can approximately 1 or 2 decimal units of the original microbial
repair damaged DNA, and in particular the amount of DNA load.

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The use of the combination of electron beam irradiation at On the basis of the deterioration index reported, if the
low doses produced similar results to those obtained by the overall acceptability score was less than 5, the product
use of electron beam irradiation at high doses. would be considered a putrid food (Lainez et al., 2008) [10].
The figures represent the results of the sensory analysis of
Yeasts and moulds the cheeses after treatment (t0), 9 days after irradiation (t9),
Yeasts and moulds in cheese are considered to be spoilage 18 days after irradiation (t18) stored at 4°C.
organisms leading to flavour and texture deterioration, The "aroma" descriptor for the cheeses was scored as
including softening and discolouration (Aly et al., 2012). follows: the highest score was awarded for cheeses activated
For yeasts and moulds (table 2), there was a significant only by LPS at t0, scored at 5.58 out of 9, but decreased to
decrease in the microbial load following the combination of 3.83 towards the end of storage at t18.
ebeam with LPS. On the other hand, the lowest score was awarded to cheese
In fact, the lowest dose and the average doses of 0.5 and irradiated with electron beams in combination with LPS,
1kGy combined with LPS resulted in an average value for which totalled 4.83 out of 9 and fell over time to 4.17 at t18.
yeasts and moulds (1.45 and 0.81 log CFU/g respectively) For the other descriptors such as salty taste and acid taste,
compared with the control, which was initially 2.34 log the lowest scores were attributed to the control cheeses,
CFU/g and 2.31 log CFU/g for those activated only by LPS, which scored 5.33 and 5.08 out of 9 respectively at t0 and
and almost zero with the highest dose combined with LPS. decreased over time to 5.00 and 4.67 out of 9 at t9. The
No significant difference in the number of moulds was panel gave a score of 4.50 and 3.75 over 18 days of storage.
observed between the control and the LPS-activated cheeses In fact, the descriptors studied, such as colour, acidity,
at t0. texture and odour, and linking them to physicochemical
parameters, will be detailed below.
Moreover, these results are approved by Seifu et al, (2004)
[17] For the "texture" descriptor, the scores attributed by our
who demonstrated that LPS activation has no significant
panel to the 0.5, 1 and 1.5 kGy doses combined with LPS
effect on the evolution of yeasts and moulds.
had no significant effect (p>0.05) on cheese texture. This is
During storage at 4±1°C, the yeast and mould levels of confirmed by the analytical test measuring the firmness of
untreated (non-irradiated) raspberries increased over time to the cheeses, which found that the different doses combined
3.97 compared with those treated only with LPS, which had with LPS had no significant effect compared to the controls
an average value of 3.95 log CFU/g at t18. or to the cheeses activated only by LPS. However, the panel
Similarly, for products irradiated and combined with LPS, found that texture deteriorated significantly over time.
an increase in fungal growth was observed over time, but For the descriptor "colour", the scores attributed by our
this increase remained significantly lower than for the panel to doses of 0.5, 1 and 1.5 kGy combined with LPS had
control and that activated only by LPS at t9 and t18. no significant effect (p>0.05), but we note that time
Similar results were reported by Aly et al, (2012) who significantly affected colour at t18 only, whereas there was
showed that gamma irradiation at the highest dose of 5 kGy no significant difference between t0 and t9.
reduced the proliferation of these microorganisms. The Concerning the odour descriptor rated by the panel at t0, t9
doses used by these authors were 1, 3 and 5 kGy. and t18. The scores given are not significant (p>0.05). In
fact, there were no significant differences between all the
Sensory analysis samples. On the other hand, we note that time has a
The sensory analysis, hedonic test, was carried out directly significant effect throughout the shelf life. These two
after treatment at t0, t9 and t18 days of storage at 4°C on descriptors, colour and odour, were highly correlated
control cheeses, activated by LPS, having undergone (p<0.01).
irradiation at 0.5, 1 and 1.5 kGy combined with LPS. We On the other hand, there was no significant difference in
chose to carry out these analyses throughout the 18-day overall acceptability between all the samples, although it
storage period on the basis of the cheese quality assessed. was noted that shelf life affected overall acceptability
The characterisation of the sensory properties of the cheeses significantly at t18 only, whereas there was no significant
was analysed using the radar diagram constructed with the difference between t0 and t9. Moreover, overall acceptability
scores obtained for the different parameters evaluated by the was strongly correlated with colour, odour and texture
panellists. (p<0.01).

Fig 7 : Effect of ebeam irradiation with LPS on the organoleptic properties of fresh cheese.

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National Journal of Advanced Research www.multidisciplinaryarticle.in

Effect of hurdle technologies on the evolution of the the acceptability) for the the combination throughout the shelf
shelf life of cheese life.
Estimation of the use-by date using the accelerated aging The study of the microbiological quality of the cheese
test showed that irradiation with ebeam combined with LPS had
The spoilage kinetics of a foodstuff is a representation of the a significant effect on the microbiological quality (total
deterioration of a parameter A as a function of time and coliforms, mesophilic aerobes, yeasts and moulds)
temperature. These kinetics are generally of order zero or of compared with control cheeses and cheeses activated only
order one. by LPS.
The determination of the order of the spoilage reaction is In addition, the determination of the shelf life showed that
obtained by comparing the coefficient of determination R2 the use of these combination resulted in its extension of by
of the linear regression of the three kinetic models related to 3.63 days, compared with 3.09 days for the LPS cheese
a quality criterion A by drawing the graphs: alone.
Order zero: A = f(t) In fact the combination, low-dose electron beam irradiation
Order 1: Ln (A) = f(t) with LPS extended the shelf life of the cheese to 6.82 days,
Order 2: (1/H) = f(t) compared with 10.92 and 16.93 days respectively for 1 and
Determination of the use-by date by monitoring the 1.5 kGy combined with activation of the lactoperoxidase
evolution of yeasts and molds, indeed the microbial limit of system.
acceptability was estimated by fitting the experimental data
to the Gompertz equation modified by Corbo (Zantar et al., References
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