Analysis of the Blood Type and Group among Undergraduate Physics Students of Dilla University, Ethiopia

The plasma membrane of a mature red blood cell has glycol-proteins and glycol-lipids that determine a person’s blood type. Blood types A, B, AB, and O form the ABO blood group. ABO blood type is determined by the hereditary presence or absence of antigens A and B on Red Blood Cells. Blood types and transfusion compatibility are a matter of interactions between plasma proteins and erythrocytes. A blood sample was taken from the voluntary Physics students of Dilla University. Blood type (groups) varies among each ethnic group, A, B, AB and O blood groups were analysis for these ethnic group of Physics department students found in Dilla University, Amhara state, Oromia state, South Nation Nationalities People, Tigray state and Gembella people. The distribution of blood group O was the highest with percentage frequency of 38.33, 29.44 and 28.88%, followed by blood group A and blood group B, and the least percentage frequency is that of blood group AB with 3.33% among each ethnic groups. The highest Rh-blood types were 91.66% Rh-positive blood types followed by 8.34% Rh-negative blood types. Therefore, among each ethnic groups of Ethiopian students, the blood type and groups were varies in all voluntary students of Dilla University.

All human populations share the same blood group systems; although they differ in the frequencies of specific types. The incidence of ABO and Rh groups varies markedly in different races, ethnic groups, and socio-economic groups in different part of the world [3]. The differences in human blood are due to the presence or absence of certain protein molecules called antigens and antibodies. The antigens are located on the surface of the red blood cells and the antibodies are in the blood plasma. Individuals have different types and combinations of these molecules [4]. According to ABO and Rh blood grouping systems, a person can belong to either of following 8 blood groups: A Rh+, A Rh-, B Rh+, B Rh-, AB Rh+, AB Rh-, O Rh+ and O Rh-. Our blood cells have a variety of chemicals on their surfaces called antigens. Blood cells from different people have different antigens (anti=against, opposite) on them, and which ones a person has depend on what genes that person has. External surface of RBC membrane is coated with a diverse array of glycol-proteins, complex carbohydrates, and lipoproteins, imparting antigenic structure to the membrane (Figure 1).
The blood grouping is done in laboratories by slide test which is a manual method. Most of the techniques applied are still based on the principle of interaction between antigen and antibody and subsequent agglutination of RBCs (positive result).The absence of agglutination indicates the lack of interaction (negative result) [5] (Figure 2). ABO and Rh blood groups are the most important blood groups despite the long list of several other blood groups discovered so far [6,7]. The two significant blood group systems were discovered during early experiments with blood transfusions, the ABO group in 1901 and the rhesus group in 1939 [8]. Furthermore, the discovery of ABO and Rh blood groups has contributed immensely to blood banking services and transfusion medicine in order to avoid morbidity and mortality in both adults and children.
Agglutination is the clumping of particles with antigens on their surface, such as erythrocytes by antibody molecules that form bridges between the antigenic determinants. When, antigens are situated on the red cell membrane, mixture with their specific antibodies causes clumping or agglutination of the red cells. The agglutination of red cells takes place in two stages. In the first stage-sensitization, antibodies present in the serum become attached to the corresponding antigen on the red cell surface.  [5] A red cell, which has thus coated by antibodies is said to be sensitized. In the second stage, the physical agglutination or clumping of the sensitized red cells takes place, which is caused by an antibody attaching to antigen on more than one red cell producing a net or lattice that holds the cells together. The cells form aggregates, which if large enough, are visible to the naked eye. There are also degrees of agglutination which cannot be seen without the aid of a microscope.
Agglutination is the clumping of particles with antigens on their surface, such as erythrocytes by antibody molecules that form bridges between the antigenic determinants. When, antigens are situated on the red cell membrane, mixture with their specific antibodies causes clumping or agglutination of the red cells. The agglutination of red cells takes place in two stages. In the first stage-sensitization, antibodies present in the serum become attached to the corresponding antigen on the red cell surface. A red cell, which has thus coated by antibodies is said to be sensitized. In the second stage, the physical agglutination or clumping of the sensitized red cells takes place, which is caused by an antibody attaching to antigen on more than one red cell producing a net or lattice that holds the cells together. The cells form aggregates, which if large enough, are visible to the naked eye. There are also degrees of agglutination which cannot be seen without the aid of a microscope.
The knowledge of distribution of ABO and Rh blood groups at local and regional levels are helpful in the effective management of blood banks and safe blood transfusion services. Identification of Rh system is important to prevent the erythroblastosis fetalis; which commonly arises when an Rh negative mother carries an Rh positive fetus. Knowledge of the distribution of ABO and Rh blood group is essential for effective management of blood banks inventory, be it a facility of a smaller local transfusion service or a regional or national transfusion service. It is, therefore, imperative to have information on the distribution of these blood groups in any population [13].
Knowledge of blood group distribution is also important for clinical studies, for reliable geographical information and it will help a lot in reducing the maternal mortality rate, as access to safe and sufficient supply of blood will help significantly in reducing the preventable deaths.
Knowing their blood group help to minimize the risk factor during marriage such as erythroblastosis disease occurs after marriage and miss-donating blood to the patients or miss-receiving. This study is aimed to determine frequency and distribution ABO and Rh blood group patterns among Physics student found at Dilla University, Ethiopia.

Study site and sample of population
This study was carried out at the College of Natural and Computational Science, Department of Biology, in Microbiology Laboratory at Dilla University, Ethiopia, during 2014. A total of 180 voluntary students of Physics Department were randomly selected among the Dilla University students. The blood collections were taken from the voluntary Physics students of Dilla University. The students were divided into five major ethnic groups i.e., Amhara, Southern Nations Nationalities and Peoples (SNNP), Oromia, Gambela Peoples, Tigray and ethnic groups. Out of 180 voluntary students were 142 males and 38 females. All were of ages between 19-24 years.

ABO and Rh-blood group's tests
Blood samples from each student was taken by Scrub" with "scrubbing with a piece of cotton saturated with 70% alcohol and pierce it with a sterile disposable lancet and was placed a small drop of blood on a three clean white glass microscope slide on which a few drops of antisera for blood group A and B was applied ( Figure 3).

Ethical clearance
The study protocol was reviewed and approved by Dilla University ethical clearance committee. Before data collection, an informed consent was obtained from respondents.

Date analysis
Data on the frequency of ABO and Rh blood groups was reported in simple percentages and graphic.

Results and Discussion
Knowing ABO blood type and Rh blood group is the most important antigenic system in transfusion. Blood type (groups) varies among each ethnic group, A, B, AB and O blood groups were analysis for these ethnic group of Physics department students found in Dilla University, Amhara state, Oromia state, South Nation Nationalities People, Tigray state and Gembella people. The voluntary students of Physics, males and females were taken from Amhara state, 24.32% , 24.32% 2.70%, 35.14% and 5.41%, 5.41%, O% 2.7% blood types A, B, AB and O were determined respectively. The voluntary students of Physics, males and females were taken from SNNP, 20.51% , 25.64% 2.56%, 20.51% and 5.13%, 10.25%, 0% 15.85% blood types A, B, AB and O were determined respectively.
The frequency of ABO blood groups varies from race to race. The country wise figure in Caucasians for the United States, the distribution is type O=47%, type A=41%, type B=9%, and type AB=3%. Among African American, the distribution is type O=46%, type A=27%, type B=20%, and type AB=7%. Among Western Europeans, 42% population shows blood group A, 9% blood group B, 3% blood group AB and the remaining 46% blood group O [14,15]. Many other studies have shown that blood group O was the most common blood group and blood group AB was the least common blood group in different ethnic groups [16]. For instance, in African-American ABO blood group, the distribution of type O, 46%; type A, 27%; type B, 20%; and type AB; 7%. Normally, the distribution of ABO blood groups varies from one population to another. In many other studies, blood group O has been found to be the most common blood group. In the Caucasians in the United States, the distribution is group O, 47%, group A, 41%, group B, 9% and group AB, 3% [7]. Among Western Europeans 42% are group A, 9% group B, 3% group AB and the remaining 46% group O. For blacks in United States, the distribution is group O, 46%, group A, 27%, group B, 2%, and group AB, 7% [7].
The voluntary students of Physics, males and females were taken from Oromia state, 23.07%, 13.46% 1.92%, 32. 69% and 9.62%, 9.62%, 3.85%, 5.77% blood types A, B, AB and O were determined respectively. The voluntary students of Physics, males and females were taken from Gambela Peoples, 20.00%, 40.00%, 0%, 40.00% and 0%, 0%, 0%, 0% blood types A, B, AB and O was determined respectively. The voluntary students of Physics, males and females were taken from Tigray, 30.00%, 20.00%, 0%, 40.00% and 0%, 0%, 0%, 10.00% blood types A, B, AB and O were determined respectively (Table 1). In Caucasians in the United State, the distribution is type O, 47%; type A, 41%; type B, 9%; type AB, 3%. Also, among Western Europeans, type O, 46%; type A, 42%; type B, 9%; and type AB, 3% [9,10]. Thus, the gene segregation for ABO systems always followed a particular pattern for its distribution in different ethnic group with exceptional cases.    Blood was mixed thoroughly with the antisera and rocked gently for 60 sec to observe agglutination. In case of doubt, the test was examined under a microscope, or the results was confirmed by reverse grouping using known group A and B red cells [14] (Figure 4).

Distribution of ABO blood types among physics students at Dilla University
The distribution of ABO blood groups varies regionally, ethnically and from one population to another. As indicated in the ( Figure 5), the frequency distribution of blood group O was the highest with percentage frequency of 38.33, 29.44 and 28.88%, followed by blood group A and blood group B, and the least percentage frequency is that of blood group AB with 3.33% among each ethnic groups as observed in previous studies [16][17][18]. Similarly, Hussain [19] had reported at 2013 the distribution of blood group O was the highest with percentage frequency of 29.97% followed by blood groupA1 26.52%, B 20.03%, A1B 19.34%, and A2 2.90% and the least percentage frequency is that of blood group A2B which is 1.24%. Also, it has been reported in several studies that there are variation in ABO blood group among different ethnic groups [16][17][18]. Patel [20] had reported that looking at the rhesus grouping, on female sex, 7.50% were Rh positive AB blood type and remaining 0.42% were Rh negative AB blood type.

Distribution of Rh-blood group based on sex among Physics students at Dilla University
Amhara state, students had the Rh-positive blood types were 78.37% and 12.16% males and females respectively while Rh-negative blood types were 8.11% and 1.35% males and females respectively. Whereas, SNNP, students had the Rh-positive blood types were 66.66% and 2.56% males and females respectively while Rh-negative blood types were 25.64% and 5.13% males and females respectively ( Table 2). Patel [20] had reported that looking at the rhesus grouping, on male sex, 21.07% were Rh positive A blood type and remaining 0.84% were Rh negative A blood type.  Oromia state, students had the Rh-positive blood types were 67.31% and 3.85% males and females respectively while Rh-negative blood types were 25.64% and 3.85% males and females respectively. On the other hand, Tigray ethnic groups of students had the Rh-positive blood types were 80.00% and 10.00% males and females respectively while Rh-negative blood types were 10.00% and 0.00% males and females respectively. And also, Gambela Peoples ethnic groups of students had the Rh-positive blood types were 100.00% and 0% males and females respectively while Rh-negative blood types were 0.00% and 0.00% males and females respectively (Table 2). Patel [20] had reported that looking at the rhesus grouping, on male sex, 37.70% were Rh positive B blood type and remaining 1.55% were Rh-negative B blood type.

Distribution of Rh-blood group among Physics students at Dilla University
The highest Rh-blood types were 91.66% Rh-positive blood types followed by 8.34% Rh-negative blood types ( Figure 6). According to the Fareed [21] study on population, Khan with highest Rh(D) positive (96.03%) with lowest Rh(D) negative (3.97%) whereas Syed with lowest Rh(D) positive (87.86%) and highest Rh(D) negative (12.14%) among six populations. The Rh(D) negative frequency shows decreasing order: Syed > Mir > Malik > Mughal > Gujjar and Bakarwal > Khan. Moreover, this study further confirmed that Rh-positive has the highest percentage frequency while Rh-negative has the lowest percentage frequency as observed in previous studies among different ethnic groups [16][17][18]. Patel [20] had reported that looking at the rhesus grouping, on females sex, 35.42% were Rh positive B blood type and remaining 2.91% were Rh negative B blood type.

Distribution of Rh-blood group in each sex of Physics students at Dilla University
The 22.97%, 20.27%, 2.70%, 32.43% and 4.05%, 5.41%, 0%, 2.7% of the Physics students males and females were Rh-positive blood group A+, B+, AB+ and O+ respectively while Rh negative males and females students were 1.35%, 4.05%, 0%, 2.70% and 1.35% 0%, 0%, 0%, A-, B-, AB-and O-had this blood group, the students which came from Amhara state respectively (Table 3). Whereas, the 20.51%, 23.08%, 2.56%, 20.51% and 5.13%, 7.70%, 0%, 12.82% of the Physics students males and females were Rh-positive blood group A+, B+, AB+ and O+ respectively while Rh negative males and females students were 0%, 2.56%, 0%, 0% and 0%, 2.56%, 0%, 2.56%, A-, B-, AB-and O-had this blood group, the students which came from SNNP respectively. On the other hand, the 23.08%, 11.54%, 1.92%, 30.77% and 9.62%, 7.70%, 1.92%, 5.77% of the Physics students males and females were Rh-positive blood group A+, B+, AB+ and O+ respectively while Rh negative males and females students were 0%, 1.92%, 0%, 1.92% and 0%, 1.92%, 1.92%, 0%, A-, B-, AB-and O-had this blood group, the students which came from Oromia state respectively. According to the study of Hussain [19]    The following are the blood samples of various blood groups taken and mixed with Anti-A, Anti-B and Anti-D (Anti-Rho D) Monoclonal and observed by naked eye and under the microscope either form clumping or not. Figure 7A-7C shows the A Positive blood group sample on adding Antigen-A, Antigen-B and Antigen-D respectively. When Antibody A, Antibody B and Antibody D was added on the positive A blood type, the blood sample on slide A and C was form aggregates (agglutinate) but the blood sample found on slide B remain fluid (No aggregates). Figure 8A-8C shows the A Negative blood group sample on adding Antigen-A, Antigen-B and Antigen-D respectively. When Antibody A, Antibody B and Antibody D was added on the positive A blood type, the blood sample on slide A was form clump (agglutinates) but the two blood sample found on slide B and C remain fluid (No clump).  Figure 9A-9C shows the B Positive blood group sample on adding Antigen-A, Antigen-B and Antigen-D respectively. When Antibody A, Antibody B and Antibody D was added on the positive B blood type, the blood sample on slide B and C was form aggregates (agglutinates) but the blood sample found on slide A remain fluid (No aggregates).   Figure 11A-11C shows the AB Positive blood group sample on adding Antigen-A, Antigen-B and Antigen-D respectively. When Antibody A, Antibody B and Antibody D was added on the positive AB blood type, all blood sample found on slide A, B and C was form aggregates (agglutinates).  Figure 12A-12C shows the AB Negative blood group sample on adding Antigen-A, Antigen-B and Antigen-D respectively. When Antibody A, Antibody B and Antibody D was added on the negative AB blood type, the blood sample on slide A and B was form aggregates (agglutinates) but the blood sample found on slide C remain fluid (No aggregates).

Conclusion
The study revealed that the blood group of these five different ethnic group have different numbers of blood types that means the numbers blood type A, blood type B, blood type AB, and blood type O are various. Therefore, ABO blood type and Rh-blood groups varies among each ethnic group.