Research article seminar

1. Andres Bejarano Agudelo
III semestre medicina
Universidad Pontificia Bolivariana

2. Introduction
Chicken coccidiosis: poultry disease that
seriously affects the poultry industry.
Chickens feel tired, eat and drink less, show
paleness and have colorless and bloody
droppings.
Developed resistance to anticoccidial drugs and
antibiotics.
Genetically engineered vaccines: tending in
coccidiology research.

3. Introduction
Eimeria tenella: it is a parasitic protozoa that
causes hemorrhagic cecal coccidiosis in young
poultry.
Acquired by fecal contamination of food and water,
and it establishes itself in the crypts of Lieberkühn
of the small intestine.
It´s zygotes are released in the chicken´s feces..

4. General objective
“This study provides a theoretical basis for further understanding of the
molecular mechanism of Eimeria tenella surface antigen gene SAG 6 and SAG
15, and it also lays a foundation for screening chicken coccidia vaccine
candidate genes”
*Taken from the article´s introduction.

5. Materials and methods
RT-PCR: reverse transcription polymerase
chain reaction
Laboratory technique combining reverse
transcription of RNA into DNA, and
amplification of specific DNA targets, using
polymerase chain reaction (PCR).
Allows real time observation of the amplified
gene.
Can be used to identify and quantify specific
target species (in this case E.tenella).

6. ELISA and plasmid transfection
ELISA: used to detect antibodies,
hormones, peptides and proteins in the
blood. Diseases: HIV, Ebola, Rotavirus,
Lyme disease, Syphilis, Toxoplasmosis,
Zika virus, among others.
Plasmid transfection: a gene of interest
delivered to a cell during division. Can be
transferred between organisms and used
to develop resistance to antibiotics,
among other uses.

7. Results
EtSAG 6 and 15
complete genes
EtSAG 6 and 15
genes from pET28a
vector
EtSAG 6 and 15 genes from
pEGFP-N1-SAG 6 and 15
plasmids
D-E: western blot analysis of
rEtSAG 6 and 15 proteins
Column M:
molecular mass
marker
Column 1: EtSAG
6 gene
Column 2: EtSAG
15 gene

8. Results pEGFP-N1-SAG 6
plasmid
pEGFP-N1-SAG
15 plasmid
pEGFP-N1
plasmid
transfected with lipofectamine
Fig A: EtSAG 6 and 15 gene
expression in 293 T cells, under
fluorescence microscopy.
RT PCR of
EtSAG 6
RT PCR of
EtSAG 15
Western blot analysis of
EtSAG 6 and 15
Column M:
molecular mass
marker
Column 1: pEGFP-
N1-EtSAG 6 and 15
plasmid
Column 2: pEGFP-
N1- SAG 6 and 15
plasmid
Column 3: empty
pEGFP-N1 plasmid.

9. Discussion
Author´s name Conclusion Does it agree with the
article?
Shirley “Eimeria tenella is a kind of
coccidia with strong pathogenicity,
and it brings about the economic
loss of the poultry industry”.
YES
Tabares “Previous studies have shown that
the SAGs of Eimeria tenella are
mainly expressed in the
sporozoite stage and the
merozoite stage”.
YES
Song “Subsequent studies have also
confirmed that as recombinant
vaccines, EtSAGs can induce the
host to produce high-level
antibodies and have a certain
effect against coccidia infection”
YES

10. Personal conclusions
-By molecular biology techniques such as RT PCR, ELISA and plasmid
transfection, it is possible to develop genetically engineered vaccines against
chicken coccidiosis.
-This vaccines will eliminate the economic losses of the poultry industry, as
well as it will provide more safety for people when they buy chicken or other
products from poultry.