Determination of the useful life of food products by predictive models.

Diego Armando Ramírez Aguilar

Universidad Nacional Abierta y a Distancia - UNAD

216005_10: Desarrollo de nuevos productos

Ing. Elizabeth Hernández Alarcón

17 de abril de 2024

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 Goals

general objective

Carry out the simulation in the program proposed by the tutor, to determine various

growth curves, under mobile parameters such as water activity, pH and temperature, under a

certain deteriorating microorganism of the new worked product, in this way, assess the useful life

of the product. through the indirect Method.

specific objectives

• Learn to design growth curves in the proposed program.

• Identify the bacterial count from the proposed models based on the indicated time

(hours).

• Determine the shelf life of Bulgarian yogurt from the growth curves.

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 Determination of the useful life of food products by predictive
models.

Step 1. Registration and admission to the simulator ComBase

Figure 1. Registration to the ComBase simulator.

Figure 2. ComBase simulator login.

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Step 2. Identify the ComBase Simulator Tools

Figure 3. Recognition of the ComBase simulator.

Step 3. Enter the Broth Models link – Growth

Figure 4. Entering the Growth module of the ComBase simulator.

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 Step 4. Simulation for the microorganism involved in the deterioration of the

new food product.

Taking as reference the previous description of the simulator, perform the simulation for

the microorganism involved in the deterioration of the new food product, considering the

nature of it. For example, if the new product is for bakery, the one involved is probably

Bacillus Cereus; if it is a canned one, the Clostridium botulinum; if it is a fresh cheese

them involved could be Listeria monocytogenes.

It is necessary for the group to identify the microorganism that would be the main

responsible for the deterioration of the food, before proceeding to the simulation.

Selection of microorganisms

Microorganism involved in the deterioration of Bulgarian yogurt: molds and yeasts are

the main organisms responsible for the deterioration of yogurts and pasteurized products, which

is due to the fact that they are resistant to low pH, and are associated with the deterioration of

foods with high activity of water (Coronel, 2018).

Taking into account that the ComBase simulator only handles bacterial microorganisms,

and that yogurts are little attacked by pathogenic bacteria thanks to their pH and their probiotic

cultures, it was necessary to investigate which of the bacteria present can alter a yogurt-type

fermented drink.

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 According to the (FAO), poorly sanitized raw materials can contain pathogenic

microorganisms, such as Listeria monocytogenes, salmonella and Escherichia coli, therefore for

this indirect shelf life simulation exercise for Bulgarian yogurt, we will work with the e-coli

bacteria.

Figure 5. Selection of Echerichia coli Bacteria in ComBase simulator.

4.1 Identify the growth temperature range of the microorganism under study.

Consider the literature report on growth temperature.

microorganism Growth Temperature Optimum growth

Ranges temperature
Escherichia Coli 7ºC – 50ºC 37ºC
Note: Tomado de Hernández, Domínguez & Gonzaga (2015); OMS (2018).

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 Figure 6. Selection of optimal growth temperature for Echerichia coli (37ºC)

4.2 Establish the growth time of the microorganism in which the measurement will

be carried out, for example 25 hours.

microorganism Measurement time

Escherichia coli 24 hrs
Note: Own authorship

Figure 7. Growth time selection (24 hours)

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 4.3 Manipulate the aw - NaCl variables according to the ranges established by the

simulator and review the behavior of the growth curve (graph) and the data table (data

points). Don't forget that the level is 3 and indicate (Phys. State) the data that the

simulator defaults to.

Change of variables aw – NaCl (graph) Data table (data points)

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 Figure 8. Different ranges of aw - NaCl

Step 5. Behavior of microbial growth by manipulating the pH variable. Perform the same

procedure described above (Point 1).

Change of pH variables (graph) Data table (data points)

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 Figure 9. Different pH ranges

Step 6. According to the above, ask the following questions and argue in a way that

allows them to establish the criteria that must be considered to establish the useful life of the new

product:

6.1 Describe the nature of the new product and mention the reasons why the

microorganism was taken as a reference to develop the simulation.

Product Nature Selected

microorganism
Bulgarian Yogurt (Kefir) Dairy drink fermented by It is not common for
kefir granules, which are yogurt to be the target
colonies of of microbial attack by
polysaccharide matrix of bacteria, given its
microorganisms, which probiotic nature;
form a symbiosis however, molds and
between bacteria and yeasts can deteriorate
yeast. (Lopitz-Otsoa, the product, being the
Rementería, Elguezabal, most common in the
Garaizar, 2006). industry.
As no type of yeasts
and molds are found
in the list of
microorganisms in the
simulator, Echerichia
Coli is selected, as it
is a pathogenic
bacteria closely linked

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 to the quality of cow's
milk, which is
appropriate to work
with in the simulation,
if necessary. of
contamination of the
raw material during
milking, or poor
sanitation of the milk
in the plant during the
process
Note: Own authorship

6.2 Taking the data shown in the table (data point) and the behavior of the growth curve

(graph); identify the following aspects and conclude:

a. Under what parameters of aw - NaCl - pH; Does microbial kinetics take the microbial

growth curve in its 4 phases: initial (Lag) - Exponential - Stationary and death?

Microbial Parameter of aw-NaCl

growth
curve in 4
phases
(initial,
exponential,
stationary
and death
In no
growth
curve of
Echerichia
coli are the
4 complete
phases
presented,
however, in
this curve
the lag,
exponential
and
aw activity - NaCl
stationary

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 phases are
clearly
presented.
pH parameter
In no
growth
curve are
the 4
complete
phases
presented,
however, in
this curve
the latency,
exponential
and
stationary pH activity
phase can be
evidenced.
Note: Own authorship

b. What is the microbial growth reached at the end of the time established for the

simulation?

For aw – NaCl – pH at 24 hours:

8.7 (Log UFC /g)

c. In what time range, is the stationary phase of the microbial growth curve presented

and what is the microbial count expressed in Conc (Log10 cells / g)?

Time range stationary phase time

(hours):20.2

Conc count (Log10 cells/g):

8.47

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 Note: Own authorship

Step 7. Analyze how the behavior of microbial growth is when it develops at the

optimum temperature ideal for its development over time. To do this, take the microbial count

data expressed in Conc (Log10 cells / g) in the initial phase (latency) of growth and compare

them with those reported in the stationary phase. If the temperature of the experiment is lowered

to 5°C, how is the behavior of the graph and the data in the table?

At ideal temperature (37ºC), the lag phase

lasts approximately 3 hours, and then enters
the exponential phase almost 24 hours into
the experiment.
In the growth curves, the stationary phase
cannot be clearly seen (only in 1 curve),
which leads to the conclusion that microbial
growth lasts more than 24 hours in all
experiments, in order to be able to make a
complete curve.

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Reduction to 5ºC (the simulator only allows
the t to be lowered to 10ºC)
Figure 10. Microbial growth at 37ºC and 10ºC
By reducing the temperature to 5ºC, it is
evident that the bacteria enter a constant
steady state during the 24 hours studied, with
very low microbial growth, reflecting the
importance in this simulation of keeping the
product stored and refrigerated at least
between 2ºC and 10ºC, to offer optimal
microbiological and organoleptic quality.
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 5. Conclude on the answers to the questions and establish the importance of controlling

the parameters that affect the development of microorganisms that can cause deterioration of the

new food product and therefore by controlling these parameters, it is possible to predict their

expected lifespan.

According to the simulations carried out previously, it was evident that the maximum

growth of Echerichia coli at different temperatures and water activity is (Log10 cells/g): 8.47 for

24, which would indicate that in 2 days it would double, passing the limits indicated by the

Codex Alimentarius, which indicates that within fermented milks, during the shelf life, the sum

of indicator microorganisms must be at a minimum value of 10.7 cfu/g (Coronel, 2018).

The product under the previous conditions is prone to deterioration due to physical

factors, such as temperature, affected amino acid levels in the product and the appearance of

volatile gases due to the presence of Echerichia coli.

In order to guarantee the survival of the beneficial bacteria, and the organoleptic and

microbiological characteristics of the yogurt, it must be guaranteed that it is preserved at a low

temperature (between 5 and 10ºC), since according to the simulation carried out at this

temperature, the curve. The growth rate is satisfactory for the useful life of the product

(persistent stationary phase), and it can be preserved for much longer.

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 conclusions

• We learned to design growth curves in the ComBase program, where the most relevant

thing was to distinguish the abrupt behavior that the Echecrichia coli bacteria has at low

temperatures.

• It allowed the bacterial count to be identified from the proposed models based on the

indicated time (hours), resulting in high counts in 24 hours, which automatically makes the

product susceptible to rapid deterioration.

• The shelf life of Bulgarian yogurt is determined from the growth curves, reaching the

conclusion that storing the fermented drink between 5 and 10ºC manages to partially inactivate

the bacteria, entering a stationary phase for a long time.

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 bibliographic references

Coronel Feijo M.A (2018). Estudio de las características fisicoquímicas y sensoriales de

yogurt enriquecido con quinua. Tesis Doctoral, Universidad de Extremadura.

https://dehesa.unex.es/bitstream/10662/9273/1/TDUEX_2019_Coronel_Feijo.pdf

Fernando Lopitz-Otsoa, Aitor Rementeria, Natalia Elguezabal, Javier Garaizar. (2006).

Kefir: una comunidad simbiótica de bacterias y levaduras con propiedades saludables. Revista

Iberoamericana de Micologìa. Volumen 23, Pages 67-74, ISSN 1130-1406.

https://doi.org/10.1016/S1130-1406(06)70016-X.

Hernandez-Jimenez.S.J, Dominguez-Toro.E.L, Gonzaga-Gutierrez.L (2015).Magnetic

field influence in E.Coli and S.cerevisiae growth and the ability of pseudomoanas sp and bacillius

sp to be industrial usage.Universidad del valle.

http://www.scielo.org.co/pdf/rcien/v19n1/v19n1a08.pdf

Organizacion mundial de la salud OMS (2018).E.Coli – Datos y cifras.

https://www.who.int/es/news-room/fact-sheets/detail/e-

coli#:~:text=E.%20coli%20productora%20de%20toxina%20Shiga%20puede%20crecer%20a%2

0temperaturas,)%20m%C3%ADnima%20de%200%2C95.

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