620 Tamayo LJ et al.

Didactic model of the chicken embryo

Revista Colombiana de Ciencias Pecuarias

Didactic model of the chicken embryo development using

modified Dawson’s diaphanization and staining technique¤

Modelo didáctico del desarrollo embrionario del pollo usando la técnica modificada de Dawson para
transparentación y tinción

Modelo didático do desenvolvimento embrionário do frango usando a técnica modificada de Dawson

para diafanização e coloração

Lynda J Tamayo Arango1,*, MV, Dr Sci Agr; Paula A Suárez Avendaño2, Est Zoot; Ana I Cano Valderrama2,
Est Zoot; Brayian A Cuartas Martínez2, Est Zoot; Sergio A Yepes Ciro2, Est Zoot; Carlos A Mejía Giraldo3,
Biol, MSc; Yasser Y Lenis Sanin4, MVZ, Esp, MSc

1
Faculty of Agrarian Sciences, University of Antioquia, BIOGENESIS, Carrera 75 N° 65-87. AA 1226. Medellín, Colombia.
2
Faculty of Agrarian Sciences, University of Antioquia. Carrera 75 N° 65-87. AA 1226. Medellín, Colombia.
3
Faculty of Medicine, University of Antioquia. Carrera 51d N° 62-29. Medellín, Colombia.
4
Faculty of Agrarian Sciences, University of Antioquia, CENTAURO, Carrera 75 N° 65-87. AA 1226. Medellín, Colombia.

(Received: 4 may, 2012; accepted 19 august, 2012)

Summary

Background: didactic models are a very useful tool for pedagogy in Agrarian Science careers (i.e. Veterinary
Medicine and Animal Science). Objective: the aim of this work was to create a didactic model of the chicken
embryo development using modified Dawson´s diaphanization and staining technique, which allows the centers
of ossification to be viewed. Methods: chick embryos from day 5 to day 21 were diaphanized with KOH, stained
with Alizarin Red, and stored in glycerol. Results: growth of primary ossification centers during embryonic
development was easily visualized. Conclusion: to our knowledge, this is the first literature report showing an
anatomical model of all embryonic stages of chicken development. Impact of this model in pedagogy must be
evaluated in future works.

Key words: anatomy, cartilage, embryology, organic system, pedagogy.

Resumen

Antecedentes: los modelos didácticos son una muy buena herramienta para el proceso de enseñanza-
aprendizaje en los cursos de Ciencias Agrarias (Medicina Veterinaria y Zootecnia, entre otros). Objetivo:

¤ To cite this article: Tamayo LJ, Suárez PA, Cano AI, Cuartas BA, Yepes SA, Mejía CA, Lenis YY. Didactic model of the chicken embryo development using
modified Dawson´s diaphanization and staining technique. Rev Colomb Cienc Pecu 2012; 25:620-624.
*
Corresponding author: Lynda J Tamayo Arango. University of Antioquia, Faculty of Agrarian Sciences, BIOGENESIS. Carrera 75 N° 65-87. AA 1226.
Medellín, Colombia. Email: [email protected]

Rev Colomb Cienc Pecu 2012; 25:620-624

 Tamayo LJ et al. Didactic model of the chicken embryo 621

hacer un modelo didáctico del desarrollo embrionario del pollo usando la técnica modificada de Dawson para
transparentación y tinción, que permite visualizar los centros de osificación. Métodos: embriones de pollo
del día 5 al 21 fueron transparentados con KOH, teñidos con Rojo de Alizarina y almacenados en glicerina.
Resultados: se pudo visualizar el crecimiento de los centros primarios de osificación durante el desarrollo
embrionario. Conclusión: este es el primer reporte en literatura que muestra un modelo anatómico de todas
las etapas del desarrollo embrionario del pollo. En futuras investigaciones se debe evaluar el impacto que
tiene este modelo en el proceso enseñanza-aprendizaje.

Palabras clave: anatomía, cartílago, embriología, pedagogía, sistemas orgánicos.

Resumo

Antecedentes: os modelos didáticos são uma boa ferramenta para o processo ensino-aprendizado nos cursos
de Ciências Agrárias (Medicina Veterinária, Zootecnia, entre outros). Objetivo: fazer um modelo didático do
desenvolvimento do frango usando a técnica modificada de Dawson para diafanização e coloração, que permite
visualizar os centros de ossificação. Métodos: Embriões de frango do dia 5 ao 21 foram transparentados com
KOH, corados com Vermelho de Alizarina e armazenados em glicerina. Resultados: foi possível visualizar o
crescimento dos centros primários de ossificação durante o desenvolvimento embrionário. Conclusão: este é
o primeiro reporte na literatura de um modelo anatômico do desenvolvimento embrionário do frango. Futuras
pesquisas devem ser focadas no impacto que tem este modelo no processo ensino-aprendizado.

Palavras chave: anatomia, cartilagem, embriologia, pedagogia, sistemas orgânicos.

Introduction have been made since the first description of the

technique (Randle, 1969; Tipton and Burt, 1977;
Didactic models are tools used to improve Sanchez et al., 1996; Staples and Schnell, 1964;
analytic and argumentative capacity, imagination, and Jensh and Brent, 1966; Kawamura et al., 1990),
innovation skills in students in order to enrich their and it is considered as a very useful technique for
learning process in agrarian careers. The integrative studying the embryological development of the
course in Veterinary Medicine and Animal Science skeletal system. Essentially, the method consists
at the Faculty of Agrarian Sciences of the University of macerating the fetus with alkali, followed by
of Antioquia is titled Organic Systems. Aiming to staining it with a dye, which has a strong affinity for
improve the integration of anatomy, histology and calcium salts, such as Alizarin Red (a dye derived
physiology concepts, students have to develop a from anthraquinone). The resulting specimens allow
didactic model and present it in an exhibition during the visualization of the stained red bone beneath the
this course (Lenis and Tamayo, 2011). transparent soft tissues.

Avian embryo development is of great economic It is necessary for Animal Science and Veterinary
importance because it is during this time that the Medicine students to develop and have access
genetic basis of the line or breed to be produced is to didactic models for studying skeletal system
consolidated. Nowadays, it is very common to use ossification. The study of this important process
animal didactic models for improving pedagogy in allows the students to better understand some of
agrarian science careers (i.e. Veterinary Medicine the skeletal problems that animals can encounter,
and Animal Science). especially those derived from developmental
impairments. The aim of this work was to describe
Dawson´s technique is an anatomical technique the diaphanization and stain of the skeletal system
which diaphanizes soft tissues and stains embryo of chicken embryos aged 5 to 21 days for its use as
bones (Dawson, 1926). Several modifications a didactic model for teaching purposes.

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622 Tamayo LJ et al. Didactic model of the chicken embryo

Material and methods

Animals
Chicken embryos provided by Marruecos poultry
farm (Medellín, Colombia) from 5 to 21 days of
incubation were used. Day 5 was chosen because
skeletal system development (endochondral and
intramembranous ossification) starts at that day,
with small ossification centers in the skull, limb
buds and prevertebrae (Sawad et al., 2009). Each
embryo was separated from the attached structures
of the egg and placed in individual 250 mL bottles.

Specimen processing
In order to avoid excessive manipulation and
damage the embryos were not transferred from
the bottles. Taking into account the small size of
the specimens, it was not considered necessary to
eviscerate or skin the embryos. They were fixed in
10% formalin (Protokimica, Medellín, Colombia) Figure 1. 16-day-old chick embryo. Primary centers of ossification are
clearly seen in red at bone diaphysis, while cartilage is seen in white at
for eight days to attain an adequate fixation, then bone extremities.
dehydrated in 70% ethyl alcohol (Protokimica,
Medellín, Colombia) mixed with molecular
iodine (Carlo Erba, Val-de-Reuil, France) for two
days. Molecular iodine was used as a mordant to
potentiate fixation of Alizarin Red in bone calcium.
Next, specimens were transferred to a 2% KOH
solution (Protokimica, Medellín, Colombia) added
to 1% Alizarin Red (Merck, Parkville, Australia)
for 24 hours. The embryos were transferred to a 2%
KOH solution for eight days for clearing. At this
time, embryos were completely stained by Alizarin
Red, and no difference between muscle and bone
was observed. For an additional clearing, a 1:1:1 2%
KOH solution, ammonia (Protokimica, Medellín,
Colombia) and glycerol (Protokimica, Medellín,
Colombia) was used for eight days. Finally, glycerol
was used to store the specimens.

Results
Embryos were successfully diaphanized,
yielding the ossification centers in red (Figure 1). To
attain the main objective of this work—that is, the
process of forming a didactic model—all the bottles
were placed in a specially designed shelf including Figure 2. Final aspect of the didactic model displaying chicken development
photos of the fresh specimen in all the stages to stages from days 5 to 21. Fetuses were placed in individual bottles.
Photographs were placed in the central panel for additional explanations.
enhance the steps of the embryologic development
process, as shown in figure 2.

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 Tamayo LJ et al. Didactic model of the chicken embryo 623

The resulting didactic embryo model was as avoiding discoloration of the specimen with time
the final project of the Organic Systems course and guaranteeing that the specimen will withstand
at the Faculty of Agrarian Sciences-University of time while remaining in good condition. The
Antioquia (Colombia), exhibited during the First addition of ammonia makes specimens more
Undergrad Students Exhibition of Animal Didactic transparent than only KOH and has the advantage
Models (Lenis and Tamayo, 2011). of being less corrosive to the delicate tissues of the
fetus.
Discussion
Although diaphanization protocols are widely
To our knowledge, this is the first literature used for research, they are poorly used as didactic
report showing an anatomical model of all the models for the explanation of basic science topics
embryonic stages of chicken development. The in agrarian careers. With this model we obtained a
modified Dawson´s technique stained the calcified useful tool for the pedagogy of avian embryology.
bone and allowed the visualization of the primary
ossification center development. Prospective future works include constructing
didactic embryological models for other species and
Given multiple factors such as species, evaluating the impact that these models have upon
development stage, size, and evisceration, pedagogy. We want to stain not only the bone but
experience is needed to optimize diaphanization also the cartilage using Alcian blue in chickens and
and staining of ossification processes. Several other animal species to study the musculoskeletal
modifications of Dawson´s technique have system at earlier stages.
been proposed to optimize the diaphanization
process. Sanchez et al. (1996) used a higher KOH Acknowledgments
concentration (15%) to decrease the time spent
in diaphanization from one month to one week We wish to thank Marruecos poultry farm,
in human fetuses. Betti (2005) used 2-4% KOH owned by Super Pollo Paisa company, for supplying
to diaphanize fish larvae but only for 10 minutes, the fertilized eggs required for all the development
while Maiolino et al. (2009) used 6% KOH, under stages.
an unspecified period of time. Care must be taken
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