Universidad Nacional Abierta y a Distancia

Vicerrectoría Académica y de Investigación

Guide for the Development of the Practical Component of the Course: Phase 4
- Development of the simulated practical component of the learning strategy.

1. Description of the Practical Component

Type of Practical Component: Virtual Lab
Type of Activity: Collaborative
Evaluation Moment: Intermediate Unit 2
Highest Activity Score: 100 points
The activity starts on: Friday, May 29, The activity ends on: Thursday, June
2020 11, 2020
With this activity, the following learning outcomes are expected:

Analyze the behavior of pathogenic microorganisms and their implications on the

quality of the new product, through indirect methods that relate to predictive
microbiology, to estimate the shelf life of the product.
The materials required for the development of your Practical Component are
the following:

Computer
Internet
Software ComBase
Username and password
The activities to be developed in the Practical Component are:

The name of the practice: Determination of the useful life of food products by
predictive models.

To determine the useful life of food products can be done by the direct method or by
indirect methods. For the development of the practice is to work with one of the
indirect methods that correspond to the microbiological type predictive models that
are supported by mathematical equations that use information from a database to
predict the growth of bacteria under defined conditions.

So, it is necessary to review the following concepts: water activity - aw and

concentration of NaCl, pH of the food product and storage temperature. It is required
the previous knowledge about the bacterial growth.

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Therefore, it is going to use a free software program of predictive
microbiology called ComBase. It is a repository of online database that describes the
survival and growth of pathogenic microorganisms in different environmental
conditions and a set of predictive software based on these data

It was taken as reference Demo 1: bacterial growth. Predictive ComBase models of

the course: Microbial alteration and useful life of the food from the General
Foundation of Salamanca University.

I. Registration and admission to the simulator ComBase

1. Enter the address: https://www.combase.cc/index.php/en/

2. Click the button Login/Register

3. To access the simulator must be registered, click Sign Up

4. You will see a screen similar to the one in the image below. Fill out the
registration form and then click on the blue button to 'Sign up'
5. When sign in the main page of the simulator

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II. Review of the ComBase tools

1. ComBase Browser: is composed of

thousands of microbial growth curves
and survival, which have been collated
in research institutes and publications.
2. ComBase Predictor: they are a
collection of tools based on the data
of ComBase to predict the growth or
inactivation of microorganisms.
3. Combase Predictive Models
4. DMFIT: Online DMFit and DMFit Excel
5. Resources
6. Help

III. Preliminary Simulation. To use the tool ComBase Predictor,

specifically the growth model

In the column on the left side of the

screen select ComBase Predictor

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ComBase Predictor is a set of growth
models and models of thermal death for
predicting the response of various
microorganisms to various
environmental factors. There are
different types of models: 1. Models of
growth; 2. Thermal inactivation
models; 3. Non-thermal inactivation
models

ComBase Predictor is a highly effective

tool for the design, comparison, and
validation of predictive models.
Effective in predictive microbiology.

Select the Growth Model

The main page of ComBase Predictor Growth Model can be seen in the image at
the bottom, and allows you to:

 Make predictions with a static temperature (over time) or under conditions

of fluctuating temperature in time.

 Makeup to four simultaneous predictions.

Where:

1. Indicates the prediction model selected.

2. The area to set temperature is static or dynamic. The Dynamic modality is to
enter different temperature ranges over time
3. Aqueous activity expressed as NaCl or as aw
4. Area to choose the type of microorganism.
5. Area to enter the parameters of the simulation: temperature, pH,
concentration of NaCl or aw and initial level of microorganisms.
6. Area of the results of the prediction expressed by the kinetic constants of
growth: maximum rate of growth and doubling time.
7. Area of results obtained graphically or numerically (table). To move from one
to another, you must activate the link Chart or data points.
8. Area to extend or reduce the time of prediction. Enter the number of hours in
the box or click the + or - as the case may be.

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IV. ComBase Growth Model: Make a prediction

1. Select Category: Growth Model

2. Set the mode:

 In Temperature: select static mode.
 In aqueous activity: select NaCl

3. Select microorganism:

In the areas to secure the microorganism select in the

drop-down menu: Brochothrix thermosphacta is a
microorganism responsible for the deterioration of meat
and fish products.

4. Select Conditions:
The introduction of data can be done only by entering the figures or by moving
the cursor
 With regard to the parameters of the Phys. state, it must be preserved by
the resulting from the program by default.
 As the program uses the American numeric notation, decimals should be
written with point (.) instead of a comma (,).
4.1 Init. Level, write the figure of 3 or verify
that the value.
The initial level is the number of
microorganisms per gram expressed as log
UFC/g. Therefore, a 3 indicates log UFC/g=3
which is equivalent to 1000 UFC/g.

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4.2 Phys.state. The box is left empty or clears
the existing figure. In this way, the program
automatically completes the most suitable
figure.

The Phys.state is a dimension between 0 and

1 related to the Lag phase or phase of latency
of the micro-organism. A 1 means that the
micro-organism will multiply immediately
without lag phase.
4.3 Temperature: In the check Temp (°C)
write the value of 20 or verify that the value
exists.

4.4 pH. In the pH box, write the figure of 7 or

verify that the value.

4.5 NaCl. In the NaCl box (%), twrite the

figure of 0.6 or verify that the value.

4.6 Time (h). In the box that appears below

the chart Time (h) enters 25 hours.

5. Run the prediction

5.1 Is required to press the Enter or Return key after changing a parameter.

5.2 To run the model with the data entered or selected are generated the results
of the prediction:

5.3 The results of the prediction are expressed by the kinetic constants of
growth: maximum rate of growth (Log. conc/h) and doubling time (h) in the
highlighted area in red.

5.4 The area of results obtained graphically or numerically. To pass from one to
the other, activate the button Chart or data points.
6. Results of the prediction

6.1 Results in graph: To run the model under the conditions laid down and during
the hours fixed, is the prediction by following the formulas of the model in a
graphical way expressing the log UFC/g for the duration set, which for example is
of 25 hours, and results in the following graph:
 6.2 Results in Table: at the top of the graph are two push buttons to view the
results graphically or numerically. To view the results in the form of a table click
in the data points, which displays a table with the concentration of micro-
organisms in Conc (log10 UFC/g) at regular intervals of time, as noted below:

6.3 Kinetic results: Below the area where you enter the conditions appear two
(2) measures that derermine bacterial growth for these conditions, which are:

Maximum speed or rate of growth Max.rate (log.conc/h)

Doubling Time Dbl.time(Hours), time in hours in which it doubles the population.

V. Simulation for the pathogenic microorganism involved in the deterioration

of the new food product

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

simulation for the pathogenic 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.
1. Behavior of microbial growth taking into account variables such as aw and NaCl
1.1 Identify the growth temperature range of the study microorganism. Take
into account the literature report on the temperature of growth.
1.2 Establish the growth time for the microorganism in which the measurement
is to be performed. For example, 25h.
1.3 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).
1.4 Do not forget that the level is 3 and state (Phys. State) the data that the
simulator throws is kept by default.

2. Behavior of microbial growth by manipulating the pH variable. Perform the

same procedure described above (Point 1).

3. According to the above, ask the following questions and argue in a way that
allows them to establish the criteria that must be taken into account to establish
the useful life of the new product:
3.1 Describe the nature of the new product and mention the reasons why the
microorganism was taken as a reference to develop the simulation.
3.2 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?
b. What is the microbial growth reached at the end of the time established for
the simulation?
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)?

4. 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?

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.
Keep in mind for the development of the Practical Component:

In the Initial Information Environment, you must:

Make judicious reading of the guide for the development of the practical
component. Make predictions in the ComBase simulator.
Keep in mind the rubric of evaluation, in each of its criteria.
Check the agenda for the delivery date of the activity.
Review the care schedule via Skype for clarification of Doubts. Participate in
sessions of Skype.
Keep in mind the programming of web conferences

In the Learning Environment, you must:

Consult the bibliographic references on shelf life in Chapter 6.

In the Evaluation Environment, you must:

Delivery of the group report according to the structure requested in the guide for
the development of the practical component.
Individual Work Evidences:
No individual evidence is required in this activity.

Collaborative Evidences:
The collaborative evidence to be submitted is:

The structure of the report should be as follows:

- Home with student data

- Screenshots which demonstrates the use of the simulator: numerals I to III. Each
image must have a header and an explanation.
- Analyze and conclude with arguments the obtained data in the numeral IV.
ComBase Growth Model: Make a prediction.
- Results of the simulation performed for the pathogenic microorganism involved in
the deterioration of the new food product. Numeral V.
 2. General Guidelines for the Development of Evidences

For Collaborative evidence, consider the following:

1. All members of the group must participate with their contributions in the
development of the activity.

2. In each group a single member will be chosen to submit the requested product in
the environment indicated by the teacher.

3. Before delivering the requested product, students should check that it meets all
the requirements mentioned in this activity guide.

4. Only the members of the group that participated with contributions during the
time assigned for the activity should be included as authors of the submitted
product.
Please keep in mind that all individual or collaborative written products must comply
with the spelling rules and presentation conditions defined in this activity guide.
Regarding the use of references, consider that the product of this activity must
comply with APA Format.
In any case, make sure you comply with the rules and avoid academic plagiarism.
You can review your written products using the Turnitin tool found on the virtual
campus.

Under the Academic Code of Conduct, the actions that infringe the academic order,
among others, are the following: paragraph e) "Plagiarism is to present as your own
work all or part of a written report, task or document of invention carried out by
another person. It also implies the use of citations or lack of references, or it
includes citations where there is no match between these and the reference" and
paragraph f) "To reproduce, or copy for profit, educational resources or results of
research products, which have rights reserved for the University ". (Acuerdo 029 -
13 de diciembre de 2013, Artículo 99)

The academic penalties students will face are:

a) In case of academic fraud demonstrated in the academic work or evaluation, the
score obtained will be zero (0.0) without any disciplinary measures being derived.
b) In case of proven plagiarism in academic work of any nature, the score obtained
will be zero (0.0), without any disciplinary measures being derived.
 3. Evaluation Rubric

Type of Activity: Collaborative

Evaluation Moment: Intermediate Unit 2
Highest Activity Score: 100 points
First Evaluation High Level: Evidence through a screenshot of the step-by-
Criterion: step implementation and the results of the prediction of
each one of the conditions raised.
Procedural Criteria:
Presentation of If your work is at this level, you can get between 10
simulation supports points and 20 points
in the ComBase
Software. Numerals Average Level: Evidence in part by a screenshot of the
I and II step- by-step implementation and the results of the
prediction of each one of the conditions raised.
This criterion
represents 20 If your work is at this level, you can get between 4
points of the total points and 9 points
of 100 points of
the activity. Low level: Do not evidence that you have completed the
simulation on the prediction of each one of the conditions
raised.

If your work is at this level, you can get between 0

points and 3 points
Second Evaluation High Level: Analyze and conclude with clear arguments the
Criterion: data obtained in the prediction of numeral IV.

Content Criteria: If your work is at this level, you can get between 15
Implementation and points and 30 points
results of the
prediction are Average Level: Analyze and conclude with moderately clear
selected. Numeral IV arguments the data obtained in the prediction of numeral IV.

This criterion If your work is at this level, you can get between 5
represents 30 points and 14 points
points of the total
of 100 points of Low level: Do not show with arguments the prediction of
the activity. the numeral IV or not present.
 If your work is at this level, you can get between 0
points and 3 points
Third Evaluation High Level: Analyze with scientific criterion the behavior of
Criterion: m.o pathogens in food and the incidence of these in the
shelf life of the new product, but do not take as reference
Results of the the results of the simulation and/or analyze that present are
simulation not properly grounded.
performed for the
pathogenic If your work is at this level, you can get between 30
microorganism points and 50 points
involved in the
deterioration of the Average Level: Analyze with scientific criterion the
new food product. behavior of m.o pathogens in food and the incidence of
Numeral V. these in the shelf life of the new product, but do not take as
reference the results of the simulation and/or analyzes that
This criterion present are not properly grounded.
represents 50
points of the total If your work is at this level, you can get between 15
of 100 points of points and 29 points
the activity.
Low level: Do not present results or critical analyzes of the
behavior of m.o pathogens in food and the incidence of
these in the shelf life of the new product.

If your work is at this level, you can get between 0

points and 14 points