Biotechnology is based on one of the important branches of life sciences, which is the molecular biology that deals with facts and genetic information from a completely new and original angle. The results of biotechnology are related to human vital issues such as, health, food, environment, dwindling natural resources, animal and agricultural production, and industrial fields, which was hoped will continue to develop services to the humanity. Therefore, there should be evaluation, monitoring, and management of potential risks related to this technology through discussion and application of biosafety vocabulary to take decisions by enacting laws and may need a long period of application in agricultural development and food security . Modern biotechnology in the agricultural field has been able to move genetic materials between completely different plants and food, animals, and the kingdoms of microorganisms in ways that do not occur in nature. This is including genetically modified food, which is defined as the food resulting from manipulation of the genetic material by isolating a gene with desirable characteristics from a human, animal, plant, or microorganism, and its injection into another sex to transfer the desired trait [2-4]. Some scientists and researchers point a process of mixing where there is little control over the results that end wherever they are combined Genetic material in the host’s inheritance, and accordingly, all coordination and unexpected results can occur, such as the process of transferring genetic material that can change normal function of the host’s inheritance . The use of biotechnology for the purpose of genetic modification of animals, plants, and microorganisms receives special attention from scientists, producers, seed companies, and food production and preparation companies. The Food and Agriculture Organization of the International  has recognized that biotechnology has benefits related to several factors, among which is increased in plant and animal production. Despite these advantages, many researches and studies have indicated direct and indirect effects of GMF on human health . Generally the chicken is considered as one of the best source for food and the most enjoyed foods’ item worldwide for the human beings. This is due to its superior taste, easy availability at reasonable price as and its good nutrient quality compared to fish and red meat. Likewise, to meet the increased demand for this type of food, more chicken farms have been established to provide the large demands for them, the farmers start to think about increasing the growth rate and to fatten chickens . To achieve these aims, the breeder companies give more attention to the related nutritional requirements of these animals and they have started relaying on the GMF. Given the importance of the subjects as well as the growing interest in genetically food products at all scientific and popular levels, the idea of the current study was built . Throughout the present study, we investigated the effect of one type of GMF manufactured in Iraq by FEEDCO (Iraq), used for chicken food to speed on the chicken’s growth rate in order to meet their demands for this type of food. In the present study, the chicken was selected since it is considered as a first member in the food chain that end up with human being, as a sample of the study to look for the possible effect of the GMF feeding on the levels of anabolic steroid hormones, namely; testosterone and progesterone which are considered as important regulator and molecules that have wide biological effects in animals and human beings .
Materials and Methods
Samples of chicken have been taken from the poultry fields in Local market. They were divided into two groups according to the type of their nutrition.
A) The first group of chicken was feed with normal food for (1, 14, and 28) days to be used as a control group and the second group was feed genetic modified food (GMF) for the same period as the control. For each time period the number of the used chickens was 15.
Measurement of hormones concentration
Chicken blood have been collected from their wings after (1, 14, and 28) days of feed. The Serum was separated by centrifugation of the blood at 3000×g for 15 min. and the obtained serum was stored at -20 ᵒC.
The hormone concentrations were determined using of the commercial human Uno ELISA kits (Immuno lab GmbH, Kassel, Germany), according to the manufacturer's instructions. Standard solutions six different concentrations of each of progesterone and testosterone were prepared.
The tested samples were put in the micro wells, and then the solutions of the dissolved enzyme conjugate (peroxidase), substrate (tetramethylbenzidine-TMB) and antibodies were added, and microplates were incubated for 2 hours at room temperature in the dark. The reaction was stopped by the addition of stop solution (0.5 M sulphuric acid) and the absorbance was measured on an ELISA reader at a wavelength of 450 nm. Upon plotting the calibration curve, the serum hormone concentration was determined using of the R-Biopharma Rid soft Win software. The results were expressed in ng/mL (ppb) taking the serum dilution factors into account .
Determination of heavy metal
Determination of heavy metal (pb and Cd) were estimated in serum of chicken using an atomic absorption spectrophotometer, determination of heavy elements was based on the reference method .
Statistical significance was assessed using least significant differences–LSD (T-test) where p-value˂0.05 was considered significance, p=0.0001 as a highly significant.
Results and Discussion
Serum level of testosterone showed non- significant variation between the control group and the GMF feed group for 1 and 14 days of feeding, while a significant increase (p<0.0001) in this level was observed after 28 days of feeding (Figure 1).
The results in Figure 2 indicated that, there is a significant increase (p<0.0001) in progesterone level in the group of chicken which was feed GMF compared to that of the control group.
Whatever the source is, in the body they serve the same biological activity. Furthermore, these steroids regarding to their sources remain free in the blood stream and thereby, increasing their potential effects since they have limited bonding or linkage to human plasma proteins and any abnormality, or imbalance in their blood concentrations may cause a range of disorders in the body .
This hormone has been reported to have a predominant role in growth promoting and development of the female chicken reproductive tract .
Figure 1: Testosterone concentration in chicken group with GMF feeding compared with that of control
Figure 2: Progesterone concentration in the genetic modified feed compared with that of control
Progesterone regulates the female fertility and regulates the ovulation rate and oviduct development in chicken .
It is known that testosterone and progesterone, as well as other hormones are naturally synthesized endogenously every day in both food producing animals and in human body. In addition to that people are exposed add lesser quantiles of these hormones from consumption of animal products . The hormones residues in animal and plant are the first concern of most people and this partly because exogenous steroid hormones have been reported to affect cancer risks .
Some studies such as Canton et al.,  Zeitoun and Ahmed  indicated that even small differences in the hormone concentration may have adverse biological effects.
GM foods have both positive and negative impacts. And since there are no studies concerning the effect of this type of food feed on testosterone and progesterone concentration in chicken, in this study, we tried to investigate the effect of GMF feeding feed on the level of these hormones. This pilot study results recommended that harder regulations should be applied to stop the widespread misuse of GMF in chicken industry. Furthermore, an inspection of chickens for anabolic hormones levels is required prior to marketing these chickens. A conclusion which needs further studies to be confirmed and thus more researches are required in this area to examine the harmful effect of this type of food on human health.
Table 1 presents the Lead concentration was 0.366 p.p.m, in investigation of chicken samples, the percentage of Lead concentration was recorded in all serum chicken samples analyzed from 0.058-1.453 p.p.m, while the Cadmium concentration was 0.317 p.p.m, in the investigation of chicken samples, the percentage of Cadmium concentration was recorded in all serum chicken samples analyzed from 0.120-1.511 p.p.m.
These result was agreement with  when investigating and detecting contamination with heavy elements (Pb and Cd) in some chicken meat in the local market of the city of Hilla, and this result was in agreement with Iraqi standard and the World Health Organization (WHO) because the concentration of Cd from (0.201-0.041) p.p.m which is within the permissible limits in the Iraqi standard and WHO.Conclusion
This pilot study results recommended that harder regulations should be applied to stop the widespread misuse of GMF in chicken industry. Furthermore, an inspection of chickens for anabolic hormones levels prior to marketing these chickens should be checked, a conclusion which needs further studies to be confirmed.
The authors would like to thank all study participants of Market Research and Consumer Protection Center, University of Baghdad, Iraq. Prior to commencing this study, ethics approval was obtained from Market Research and Consumer Protection Center, University of Baghdad. (Mracpc.Un.Bagh.1486).
No potential conflict of interest was reported by the authors.
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
All authors contributed to data analysis, drafting, and revising of the paper and agreed to be responsible for all the aspects of this work.
Adil Turki Al-Musawi
Sudad Jasim Mohammed
Aliaa Saadoon Abdul- Razzaq
HOW TO CITE THIS ARTICLE
Hathama Razooki Hasan, Adil Turki Al-Musawi, Sudad Jasim Mohammed, Aliaa Saadoon Abdul- Razzaq Al-Faraji. The Effect of Genetically Modified Food on the Level of Some Steroid Hormones in Chickens: A Pilot Study. Chem. Methodol., 2023, 7(6) 483-488