SEMINARIO

1. Presentado por:
María Alejandra Nicholls Molina
Daniela Rios Carmona

2. Introduction
• AMS emphasis on learning activities that promote scientific thinking
and critical reasoning.
• Train students in all invaluable skills within the diagnostic laboratory.
• Many programs have done research projects to generate novel
experimental data in the form of ALUREs.
• Done because an introductory microbiology course for second year
students within a four year undergraduate science program.
• Prerequisite student knowledge.

3. Terminology
• Difference between community and hospital acquired
infections

4. ALURE
• Authentic Large-scale
Undergraduate Research
Experience is a method in which
the student himself drives a
research project designed to
generate novel experimental
data.
• ALURE was done for:
experience in technical
laboratory skills, critical
reasoning and problem solving
skills within diagnostic
microbiology

5. PCR
• Polymerase chain reaction is a tool used to focus in segments
of the DNA and copy it billions times over; is used mostly to
diagnose diseases, identify bacteria and viruses.
• In this study was used because they wanted to amplificate
mouth swab DNA by conducting mouth swabs for bacterial DNA
extraction

6. Relation
• Learning techniques (ALURE)
• Culture
• Extract genomic DNA
• 16S rRNA (sequencing)
• Identify Micro organisms on mouth swabs

7. General objective
• Develop confidence in undergraduate students in technical skills for
laboratory, scientific reasoning, and skills in problem solving within
diagnostic microbiology, by an ALURE in mapping the human oral
microbiome to learn about microbial identification and the relative
merits of culture dependent vs culture independent methods,
distinguishing the bacterial composition of healthy and diseased oral
cavities.

8. Materiales y Métodos
Sesión I: Creando habilidad
• Microscopía
• Cultivo
• Gram
• Platos de TSA
• Kit de tinción de gram
incubación de muestras
inoculadas a 37ºC durante 24
horas y almacenamiento a
4ºC hasta la sesión 2

9. Materiales y Métodos
Sesión II:muestreo de la
microbioma oral humana
• Extracción de DNAg de los hisopos
bucales
• PCR (amplificación de genes 16S
RNAr usando los primers: 803F;
803Fd; 1392wR) electroforesis
Realizar hisopos bucales para
la extracción de ADN
bacteriano (por PCR y
secuenciación de 16S RNAr) y
la inoculación de medios
selectivos y diferenciales de
cultivo (agar sangre, agar sal
manitol y placas de agar)

10. Materiales y Métodos
• 2 semanas requeridas para
optimizar la amplificación por
PCR de ADN de la muestra bucal,
la secuenciación de ADN, y la
agrupación en unidades
taxonómicas operacionales
(OTU). Imágenes electroforesis
en gel y tablas OTU deben estar
preparados para los estudiantes
de la sesión 4.
• incubación de muestras
inoculadas a 37ºC durante 24
horas y almacenamiento a 4ºC
hasta la sesión 3

11. Materiales y Métodos
Sesión III: identificación de
microbios orales (cultivo
dependientes)
• Identificación, que incluye kit de
coloración de gram,
microscopía, mechero, platos
de agar inoculados
Identificación de la
microbiota bucal de los
cultivos basada en tinción
de Gram, el crecimiento de
colonias en medios de agar
selectivo y diferencial,
pruebas bioquímicas y
pruebas inmunológicas

12. Materiales y Métodos
• 1-2 días (dependiendo del tamaño de clase) para preparar la
demostración de microorganismos, kits de pruebas bioquímicas e
inmunológicas.

13. Materiales y Métodos
Sesión IV: Identificación
de microbios orales
(cultivo independientes)
• Secuenciar
• Roche 454 GS-FLX Titanium
platform
• Acacia software
• OTU (97% de la información
agrupada)
• Base de datos de referencia
Greengenes
análisis e interpretación
de los datos recogidos a
través de identificación de
cultivo dependiente e
independiente de la
microbioma oral, a través
de cohorte de estudiantes

14. Materiales y Métodos
• 1-2 días para recopilar imágenes de electroforesis en gel y tablas
OTU para todos los estudiantes dentro de cohorte.

16. Resultados

20. Explicación e interpretación de los datos
Calificación Justificación
Reprueba No muestra una correcta comprensión
de la diferencia entre los métodos de
identificación
Pasa Falta profundidad y detalles al
describir los datos obtenidos
Pasa con alto nivel Además de la correcta descripción
utiliza citas de otras bibliografías
Tabla 5. Criterios de evaluación

21. Uso de fuentes bibliográficas para evaluar
críticamente los resultados
Tabla 6. Criterios de evaluación
Calificación Justificación
Reprueba Incorrecta comparación entre el
estudio citado y el propio
experimento
Pasa Comparación válida, omite el tamaño
de la muestra y los datos
demográficos de los voluntarios
Pasa con alto nivel Comparación válida, profunda
discusión de los datos demográficos
de la población

22. Conclusions
Author Citation Agreed or disagreed
Merkel, S. “Furthermore, a majority of students
received a High Pass for
the criteria relating to explanation and
interpretation of results
and using sources for critical evaluation
in both 2012 and 2013,
which represent core competencies that
emphasize scientific
thinking and deeper conceptual
understanding and align with
learning objectives 4 and 5”

23. Author Citation Agreed or disagreed
Hanauer, D.I, D.Jacob-
Sera, M. L.Pedulla, S.
G.Cresawn, R.W.
Hendrix, and
G.F.Hatfull.
Lord, T., and T.
Orkwiszewski.
Wang, J. T. H., M. A.
Schembri, M.
Ramakrishna,
E. Sagulenko, and J. A.
Fuerst.
Weaver, G. C., C. B.
Russell, and D. J.
Wink.
“The positive impact
of this ALURE on student
development of research
skills is
consistent with previous
reports in chemistry,
biochemistry, and
microbiology (12, 19, 26,
27), and this project has
been able to
provide another
documented case of
integrative research
experiences
that are adaptable for
large undergraduate
classes”

24. Author Citation Agreed or disagreed
Petrosino, J. F., S.
Highlander, R. A. Luna, R.
A. Gibbs,
and J. Versalovic.
“A similar experimental
approach could be adopted
in mapping the microbiome
across different parts of the
human body, in line with
the
holistic approach adopted
by the Human Microbiome
Project the culture-
dependent tests and
diagnostic
standard operating
procedures would need to
be adjusted
accordingly depending on
common resident
microbiota at
the respective body sites”

25. Author Citation Agreed or disagreed
Kroes, I., P. W. Lepp, and
D. A. Relman.
“If next-generation
sequencing technology is
not available
for potential adopters, 16S
rRNA amplicons can be
ligated into
plasmids before direct
plasmid sequencing of the
PCR fragments, which is
feasible for smaller class
sizes”

26. Conclusion #1
This type of study will help the future of science
itself because it will provide to our society more
prepared professionals, capable of critical
reasoning, and more experience in the
laboratory field, and so that more and more
undergraduate students will make a difference in
science research since such a young age.

27. Conclusion #2
The results that are shown prove
that the majority of the
undergraduate students have
learned and achieved the main
objectives of the study, and they
didn’t do it poorly as it is perfectly
clear that most of the student
population has obtained a rating
that exceeds excellence.

28. Conclusion #3
Also this type of studies that helps students
improve in their laboratory techniques will
help us to have every time less mistakes
and have the right results so we can give
more accurate diagnosis and treatments for
community and hospital-acquired infections.

29. Conclusion #4
Most of us are not conscious of the amount
of bacteria and microorganisms that
naturally live in our body, with this study we
were able to identify the different types of
microbiome with a large rate of techniques.

30. Alejandra Nicholls Molina

31. Daniela Rios Carmona