*Effects of Electromagnetic Field on the Development of Chick Embryo: An In Vivo Study DOI: http://dx.doi.org/10.5772/intechopen.84704*

#### **Figure 1.**

*TriField meter measuring the electromagnetic field exposure when the phone is receiving a call (the needle moving toward the right with increasing intensity of the electromagnetic waves) (a) group 1, (b) group 2, (c) group 3, (d) group 4, and (e) during downloading, the needle is hitting extreme right.* 

 A 30-egg incubator Model EH-35, Sino-PFE Company, China was used which is equipped with temperature and humidity control and forced air ventilation; temperature was programed at 37° centigrade and humidity of 50–60% (**Figure 2a**). The experiment was run in batches of 30 eggs, due to the size of the incubator. The eggs were placed in egg holders which were programed to rotate 10 times/day. A mobile phone (power of 0.47 W/kg body and SAR 1.10 W/kg) and local service provider with 1800 MHz frequency were selected. A TriField Meter, model 100XE was used to confirm the emission of electromagnetic field of the mobile phone during the experiment (**Figure 2b**).

### *3.1.1 Exposed group*

The selected mobile phone was placed in the center of the incubator in silent mode and vibration disabled. The farthest egg was within a radius of 16 cm. This

#### **Figure 2.**

*(a) The egg incubator. (b) TriField meter showing high levels of electromagnetic waves transmitted from the mobile during call receiving (*≥*1 mW/cm2 ).* 

is important because the radius of single 1800 MHz frequency electromagnetic wavelength is approximately 16.5 cm [21].

The mobile phone placed inside the incubator was called from outside 4 times a day and each call lasted for 5 minutes. There was a gap of 4 hours in-between each call with a call free period during the night. Total exposure time/day was 20 minutes. To repeat the call on the same time every day, a ringing schedule was made, and call was recorded once the call was made. This prevented errors in making the calls. The experiment was conducted in batches of two with 30 eggs in each batch. A total of 10 embryos were scarified each at day 5 (exposure time 100 minutes), day 10 (exposure time 200 minutes), and day 15 (exposure time 300 minutes).

#### *3.1.2 Control group*

It was also conducted in batches of two. There was no mobile placed inside the incubator, and the eggs were not exposed to any electromagnetic field.

 A total of 10 randomly selected eggs were removed from the incubator in both the groups at day 5, day 10, and day 15 each, the shells were removed, and the embryos were dissected from the membranes. Mortality of the embryos was observed by observing the movements in the limbs or beating of the heart. Hamburger and Hamilton developmental stages were used to assess the embryos for gross morphological abnormalities [25]. The embryos were washed in normal saline, blotted dry with tissue paper and the dependent variables, the weight, CR-length, and eye diameter were recorded.

 A digital balance with a precision of 0.01 g (Universal Impex HA-3202) was used for measuring the weight, and a caliper (Mitutoyo Vernier calipers, Nanjing Sulang Trading Co., Ltd., China) was used to measure the length which was the length from the vertex to the tip of the coccyx and the maximum eye diameter.

For statistical analysis, student's t-test was used, and a p-value of <0.05 were considered as significant. All data were presented as the mean value ± SEM.

#### **3.2 Results**

 Mortality was not observed among both the control and the exposed groups, and all 120 eggs revealed developing embryos.

#### *3.2.1 Day 5*

 No morphological deformities were observed in both the groups. There was no significant difference between the average wet body weight in the exposed group (0.189 ± 0.035 g) and the control group (0.209 ± 0.031 g); (t = 1.67, df = 28, p < 0.11). The CR-length and eye diameter were not measured at day 5.

#### *3.2.2 Day 10*

 Gross malformation was absent in both the groups. In the exposed group, small hemorrhages were apparent under the skin with areas of paler skin indicating reduced blood flow (**Figure 3b**). In the control group, the skin was pink in color throughout (**Figure 3a**). The average weight of the exposed group (1.572 ± 0.38 g) was significantly lower than the weight of the control group (2.331 ± 0.27 g), t = 8.19, df = 48, p < 0.01 (**Figure 5a**). This trend was also observed with the C-R length and eye diameter; all showing significant differences (**Figure 5b** and **c**).

*Effects of Electromagnetic Field on the Development of Chick Embryo: An In Vivo Study DOI: http://dx.doi.org/10.5772/intechopen.84704* 

#### **Figure 3.**

*Low dose: (a) day 10: control group showing normal embryo development. (b) Experimental group: embryos were smaller in size than the control group, and marked hemorrhagic areas could be seen under the skin alternating with pale areas.* 

#### **Figure 4.**

*Low dose: (a) day 15: control group showing well developed embryo with no anomalies or deformities. Skin pink in color, covered by feathers, well-formed upper and lower extremities and normal toes and eyes completely covered by eye lids. (b) Experimental group: no anomalies or deformities were observed and embryo showing same features as revealed in the control group.* 

#### *3.2.3 Day 15*

 Gross appearance in both the groups revealed well-formed skin covered by white feathers and well-formed beak. The size of the wings and the limbs was increased, and toes can be seen well formed, middle toe the longest. The head size that was

#### **Figure 5.**

*(a) Wet body weight of the chick embryo: experimental and control groups at days 5, 10, and 15 showing significant different at day 10 (p < 0.01). (b) Eye diameter was significantly smaller in the exposed group than the control group at day 10 (p < 0.01). (c) C-R length of the chick embryo in the experimental group was significantly smaller than the control group at day 10 (p < 0.01).* 

 much bigger as compared to the body before was now become smaller, which was in accordance with the normal development of the embryo. The eyes were now fully covered by eye lids. The skin no longer shows any hemorrhage areas, and blood vessels no longer visible under the skin (**Figure 4a** and **b**). No significant differences were observed in either average wet body weight, control (14.91 ± 1.73 g) and exposed group (14.82 ± 1.57 g), eye diameter, control (1.123 ± 0.051 cm and exposed 1.14 ± 0.05) and the average C-R lengths, control (7.013 ± 0.41 cm), and the exposed groups (6.978 ± 0.348 cm) (**Figure 5a–c**).
