The Body Weight and Thickness in Chicken Embryo Incubated for

10 Days Induced by Turmeric Rhizome Simplisia

Herlina Pratiwi

, Analis Wisnu Wardana

, Dyah Ayu Okatvianie A. P.

, Albiruni Haryo

, Nayo Diah

Fauziah

, Rizky Pamwidya Abqariyyan

, Rifkytri Aditia

, Syadza Afra

Embryology Laboratory, Faculty of Veterinary Madicine, Brawijaya University

Anatomy and Histology Laboratory, Faculty of Veterinary Medicine, Brawijaya University

Pathology Laboratory, Faculty of Veterinary Medicine, Brawijaya University

Bachelor Student of Veterinary Medicine, Faculty of Veterinary Medicine, Brawijaya University

Keywords: Chicken Embryo, Turmeric Rhizome Simplisia, Body Wight, Body Thickness.

Abstract: The zygote develops into an embryo after going through the process of organogenesis. During

organogenesis, there is also the process of angiogenesis, the formation of blood vessels. However, the

process of angiogenesis can be hampered by several factors including the presence of foreign compounds

that enter the parent body. One compound that is believed to have an angiogenesis inhibiting effect is

curcumin from the turmeric rhizome simplisia. Inhibition of angiogenesis due to curcumin induction can

have an impact on embryonic organogenesis in terms of embryonic body weigth and thicness so this study

was conducted to determine the teratogenic effects of curcumin of turmeric rhizome simplisia on chicken

embryos. This study used fifteen unincubated chicken eggs divided into three groups, the negative control

group, the group injected with turmeric rhizome simplisia at a dose of 0.48 mg/gr and DMSO 2% as much

as 0.25 mL, and the group injected with turmeric rhizome simplisia at a dose of 0.96 mg/gr and DMSO 2%

as much as 0.25 mL. All groups were incubated for 10 days at 38 °C. The parameters measured were

embryo body weight use digital scales and measurement of the body thicness with calipers. The results

showed that the turmeric rhizome simplisia at dose of 0,48,mg/gr cause died in one of incubated chicken

egg and at dose of 0.96 mg/gr cause two of five incubated chicken egg was died. The left eggs from group

of dose 0.48 mg/gr and 0.98 mg/gr have decreased of body weight and thicness compared with negative

control group. It was showed that turmeric rhizome simplicial in this doses have effect in decrease body

weight and thickness of chicken embryos.

1 INTRODUCTION

Animals have the ability to multiply through a

reproductive process. Reproduction is the process of

breeding the living creatures, starting from the

fertilization that happen between egg-cell with the

sperm cell so that it forms a new individual called

zygote. Zigot evolved into embryos after the process

of Organogenesis and was born in the form of fetus

(Subronto and Tjahayati, 2001). In the process of

organogenesis, there is also an angiogenesis (process

of blood vessel formation). This process involves

Vascular Endothelial Growth Factor (VEGF) which

is a group of growth factor with broad function as

vascular permeability used as angiogenesis markers

during organogenesis process, one of which when

forming Heart. However organogenesis process can

be hampered by several factors including the

presence of foreign compounds that enter the parent

body. One of the compounds believed to have an

organogenesis inhibitory effect is curcumin derived

from the Simplisia rhizome of turmeric. Curcumin is

the largest part of the yellow pigment contained in

the turmeric rhizome which has several biological

benefits such as antioxidant, anti-inflammatory, and

Antineoplastic (Nurrochmad, 2004).

According to Maniago Research (2014) which

uses Chorio Allantois Membrane (CAM) Duck eggs

age 10 days, it was known that turmeric can affect

the process of organogenesis by providing a barrier

effect of vascular colateralist during process

Angiogenesis. Angiogenesis is a complex process

involving of extracellular matrices, proliferation,

endothelial cell migration, and functional maturation

Pratiwi, H., Wardana, A., A. P., D., Haryo, A., Fauziah, N., Abqariyyan, R., Aditia, R. and Afra, S.

The Body Weight and Thickness in Chicken Embryo Incubated for 10 Days Induced by Turmeric Rhizome Simplisia.

DOI: 10.5220/0009587200580061

In Proceedings of the 6th International Conference on Advanced Molecular Bioscience and Biomedical Engineering (ICAMBBE 2019) - Bio-Prospecting Natural Biological Compounds for

Seeds Vaccine and Drug Discovery, pages 58-61

ISBN: 978-989-758-483-1

of the new endothelial cells into a blood vessel.

When the process of angiogenesis interupted it will

efect the distribution of nutrition to the body part of

the embryo that can efect the development of body

weight and thickness.

2 MATERIALS AND METHOD

The tools used in this research include egg

incubator, egg tray, petri dish, Effendorf tube, vortex

mixer, scales, organ pots, dispossible 1 cc onemade,

dispossible 5 cc Onemade, object glass, cover glass,

tweezers, Scissors, egg shell punch, autoclaves,

staining jar, masking tape, glove, pipette, camera,

digital scales and calipers. The materials used in this

study include the unincubated chicken eggs,

turmeric rhizome simplisia powder, ringer's lactate

solution, NaCl 0.9% liquid and DMSO 100% liquid.

Preparation of the turmeric rhizome simplisia

with making a solution of DMSO 2% by dissolving

DMSO 100% into the solution of NaCl 0.9%.

Turmeric rhizomes simplisia powder as much as

0.48 mg/gr eggs for the treatment group of P1 and

0.96 mg/gr eggs for group treatment P2 diluted with

DMSO 2% to reach the volume of 1.25 mL then

homogenized using a vortex mixer.

The location of the egg sack was marked by a

pencil, as well as on the top of a egg sack for the

spot to punch the eggs. The next step was to

punched the egg as wide as the syringe needle in the

marked part and then injected the turmeric rhizome

simplisia as much as 0.25 mL in the P1 group and as

much as 0.25 mL in the P2 group. The hole where

was the injection done than be closed using masking

tape and the egg labeled.

The incubation of eggs was done in incubators

with a temperature of 38°C for 72 hours then carried

out the injection of the turmeric rhizome simplisia

and incubated at the temperature of 38°C until the

embryo reaches the 10th days old. The position of

the egg in the incubation was vertically with the egg

sack located above.

The egg shells are broken down by knocking

the egg sack using tweezers without cange the

position of the eggs. The embryo from the egg

caried out into the petri dish then the embryo are

taken and soaked in ringer's lactate solution while

shake several times to remove the remaining egg

yolks. A clean embryo was measured the body

weight by digital scale and the thickness by calipers.

The body weight was measured to within 0.1g

using electronic scales while the body width was

taken across the widest point of the abdomen in

embryos (Browne, 2006).

3 RESULT

The results showed that the turmeric rhizome

simplisia at dose of 0,48 mg/gr caused died in one of

incubated chicken egg and at dose of 0.96 mg/gr

caused two of five incubated chicken egg was died.

The factor that caused died in incubated egg with

inoculation of trumeric rhizome simplisia was not

known exactly, however, the embryos have been

death when we hasvest it. We suggest that apoptosis

is probably a cause of death of chicken embryos that

was inoculated with turmeric rhizome simplisia.

Piwocka et al (2001) said that curcumin a yellow

pigment from Curcuma longa, curcumin may induce

apoptosis in normal and cancer cells, it induces

nonclassical apoptosis via a still-unrecognized

mechanism, which leads to chromatin degradation

and high-molecular-weight DNA fragmentation. In

the other hand, research that conducted by Chen et al

(2010) showed that curcumin induces apoptotic

injury effects on mouse blastocysts through ROS

generation, and further promotes mitochondria-

dependent apoptotic signaling processes to impair

sequent embryonic development. The left eggs from

group of dose 0.48 mg/gr (P1) and 0.98 mg/gr (P2)

have decreased of body weight and thicness

compared with negative control group (K-) (Figure 1

and 2).

Figure 1. Body weight of Chicken Embryo Inoculated

with Turmeric Rhizome Simplisia and Incubated for 10

Days.

The Body Weight and Thickness in Chicken Embryo Incubated for 10 Days Induced by Turmeric Rhizome Simplisia

Figure 2. Body thickness of Chicken Embryo Inoculated

with Turmeric Rhizome Simplisia and Incubated for 10

Days.

4 DISCUSSION

Weight loss can be used as an indicator of

abnormalities at the time of the organogenesis phase.

The organogenesis phase involves the nutrients,

oxygen, and disposal of residual metabolic

substances. These needs can not be covered if only

from diffusion resource so that there must be a new

system for embryo development. It encourages the

occurrence of angiogenesis. The process of

angiogenesis also occurs in the process of

adipogenesis. Adipogenesis involves two processes

the formation of precursor adipocytes known as

preadipocytes of stem cells mesenchyme and

differensiation preadiposit become a mature and

functional adipocyte. The functionalization

adipocyte cells continue to develop and in its

development requires nutrients and oxygen to form

blood vessels through the process of angiogenesis

(Billon et. al., 2010).

The body weight and thickness of the chicken

embryo are closely related with grwoth of the

adipose tissue. According to the Ejaz et al (2009) the

growth of adipose tissue is similar to the growth of

cancer that requires new blood vessels. In the

adipose tissue, this angiogenesis is mediated by

adipokines which includes leptin, adiponektin,

aesistin, visfatin, TNF-α, IL-6, IL-1, and VEGF. The

curcumin in turmeric rhizome simplisia can inhibit

adipokines which induces angiogenesis. In addition

to inhibiting Adipokin, curcumin inhibits the

expression mRNA VEGF. In the process of

adipogenesis, curcumin decrease its

differentiensiation of preadipocytes so that the

amount of mature and functional adipocyte also

decline. According to Gonzalez-Castejon and

Rodriguez-Casado (2011) curcumin regulate the

genes involved in energy metabolism and the

accumulation of lipidas well as lowering

intracellular lipid levels. The ability of curcumin to

decrease the VEGF levels that affect the decrease in

blood numbers also affects angiogenesis in adipose

tissues that adipose tissue does not develop and

causes reduces body weight and thickness. We

suggest that the mechanism of curcumin that inhibit

angiogenesis and adipokines give effect to decrease

body weight and thickness of the embryo.

5 CONCLUSION

Induction of curcuma longa simplisia in chicken

embryo incubated 10 days was showed give efect to

the body weight and thicness, even it just a small

effect.

ACKNOWLEDGEMENTS

We many thanks to Faculty of Veterinary Medicine,

Universitas Brawijaya for providing research

assistance funds and supporting the completion of

this research.

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The Body Weight and Thickness in Chicken Embryo Incubated for 10 Days Induced by Turmeric Rhizome Simplisia