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Name: ______________________________ Date: ____________ Roll: _______ Score: ________ Biology: Blood Lab or “Everything you wanted to know about blood and then some!!!!” INTRODUCTION/BACKGROUND: BLOOD, a remarkable “life-giving" liquid tissue, will be examined genetically. The blood types, A, B, AB, and 0, plus the Rh factor trait will be investigated in a lab simulation set-up. Statistical data will be collected and compared with the United States population as well as with ethnic groups. GENETICS (Chapter 14) The inherited blood proteins were identified by Karl Landsteiner, (1868-1943), and Philip Levine. The proteins consist of, what are commonly called the blood types, ____, ____, ____, and ____, (first identified in 1901). (See p. 344) Finally, the mother-fetus condition, called the Rh factor was first identified in 1940. This lab will consider the commonly known blood types and the Rh factor, both of which are well known inherited traits and of great significance in blood transfusions and/or certain pregnancy conditions. The blood types, A, B, AB, and 0 are determined by the protein that is ON the red blood cells themselves. These blood cell proteins are called ANTIGENS. (See pg 1038 & notes) For example, "A" type blood has antigen A, and blood type AB has both A and B antigens. The plasma may contain proteins called ANTIBODIES, and are referred to as "ANTI-A" and/or "ANTI-B." (See pg 1038 & notes) For example, a person with "B" blood type has 'B' antigens on their red blood cells AND antibodies (ANTI) A in their plasma. While “O" people have NO antigens on their red blood cells, they have both ANTI-A and ANTI-B in their plasma. The four blood types, (A, B, AB, and 0), are inherited as the result of MULTIPLE ALLELES, ("many forms of a gene"): in this case, three alleles (forms), A, B, and 0. Since a person normally inherits only two genes for each trait, various combinations of the three alleles can exits, (i.e., AO, AB, BB, etc.). Complete this chart based upon the above information and the text, p. 344-345 & notes. Table 1 Human blood types (groups) Type "A" Type “B” Type "AB" Type "O" Possible genotypes Red blood cell antigen Plasma antibodies Reaction** with anti-a serum Reaction** with anti-b serum (**Indicate with the word "YES" or the word "NO") 1. In emergencies, blood transfusions primarily consist of BLOOD CELLS (and very little plasma). Why is this so? (THINK: WHAT DO THE CELLS DO?) __________________________________________________________________ (Incidentally, during routine surgery, when a blood transfusion is needed, the patient's blood type is known beforehand and, therefore, whole blood [cells and plasma] is given.) To prevent "CLUMPING", an antigen-antibody reaction, (which can be fatal), blood types should be known. Examine the separate, attached illustration page, "BLOOD TYPING & THE Rh FACTOR TESTS". The "dots" indicate clumping; the dark, filled-in figures indicate NO clumping; and, the clear figures, NO test performed. Notice that "ANTI-" serum is used, either ANTI-A, B, or O. The "ANTI-" sera is extracted from blood plasma. Since "anti" literally means "against," ANTI-B serum, placed on a drop of blood, mixed, with the result of clumping, may indicate the "B" blood type, (or possibly AB). 2. Complete this table by using a "+" to denote clumping and a "-" to indicate that no blood clumping will occur. See above chart, the illustration page, and the classroom chart for assistance. Blood Lab02 Page 2 of 8 Table 2 RECIPENTS' ANTIBODIES TYPE A TYPE B TYPE AB (ANTI-B) (ANTI-A) (NONE) TYPE 0 (ANTI-A & ANTI-B) A B DONORS’ ANTIGENS AB O 3. From reading the table above, name the blood type that is called the UNIVERSAL DONOR, (receives from self; donates (cells) to all): ________ 4. Name the blood type that is called the UNIVERSAL RECIPIENT, (receives [cells] from all; donates to self only): _________ The "Rh" factor: People with "Rh+" (positive) blood have a protein substance in their plasma that acts like an ANTIGEN. "Rh-" (negative) people do not have the protein in their plasma. If an Rh- person is given Rh+ blood, the Rh- person's blood will react and build-up ANTIBODIES (against) the Rh+ protein. Red blood cell damage, (erythroblastosis), being a possible result. This situation rarely occurs since blood typing and the Rh factor is easily determined with simple tests before blood transfusions are given, hence preventing incompatible bloods from being mixed. The major concern about the Rh factor involves pregnancy. Example, an Rh- mother carrying an Rh+ fetus. The fetus' Rh+ antigens CROSS the placental barrier causing the mother's blood to make antibodies which, in turn, CROSS the placenta back into the fetus' blood and destroy its red blood cells. Because this takes time, a first pregnancy child will not be affected. If it is, it is only slightly affected. Consequent pregnancies, (with the same set-up: mom -, baby +), can result in death of the fetus due to severe red blood cell damage. Some medications, (i.e. RHogam, a globulin blood extract), given to the pregnant women, can Blood Lab02 Page 3 of 8 sometimes prevent her from making the antibodies. But, RHogam, like many medications, cannot be taken by some sensitive women. Instead, long periods of time (4-5 years) between pregnancies are recommended. Such time allows the women's antibodies (to Rh+) to decrease to safer levels. Fortunately, such pregnancies as described here are rare because the Rh factor condition is inherited as a simple Mendelian dominant/recessive trait. True/False: Rh+ is dominant over Rh- ______. Table 3 Blood Type Positive Negative Genotype(s) Antigens (yes or no) Agglutinins (antibodies) 5. Using a "+" for the dominant allele and a "-" for the recessive allele, diagram a monohybrid cross for the Rh factor. a. Parent's genotypes: ______X______ b. Possible sperm: _____ _____ c. Possible eggs: _____ ______ d. Complete the Punnett Square to the right. e. Out of 200 people, how many are Rh-? _____ Homozygous dominant? ______ Rh+? _______ Heterozygous? ________ 6. Based upon the above information, what percent of the human population is Rh+ _____? ; Rh- _____? (It is now known that Rh inheritance is controlled by at least 8 pairs of alleles. For the purpose of simplification in this lab, consider only one pair of alleles: Rh+ and Rh-.) Blood Lab02 Page 4 of 8 LAB ACITIVITY: DETERMINING BLOOD TYPES AND THE Rh FACTOR TESTS SPECIAL NOTE: This alternative blood typing activity does NOT use real blood or blood sera. You will follow the exact procedure used to type actual human blood and obtain results that closely approximate real blood typing. OBJECTIVES: • To determine the ABO type of four unknown simulated blood samples • To determine the Rh of four unknown blood samples MATERIALS: (as indicated by the instructor) 1 blood typing plastic slide; toothpicks; marking pencil 4 unknown simulated blood samples (located in the classroom) • Ms. Jones • Ms. Brown • Mr. Green • Mr. Smith Anti-A simulated typing serum (located in the classroom) Anti-B simulated typing serum Anti-Rh simulated typing serum PROCEDURE: Each team will determine the blood type of one of the four unknown blood samples as assigned by the instructor. 1. With your pencil, circle your the number assigned to your team: • #1: Ms. Jones • #2: Ms. Brown • #3: Mr. Green • #4: Mr. Smith Blood Lab02 Page 5 of 8 2. SHAKE THE SIMULATED BLOOD CONTAINERS WELL BEFORE USING. Place 2 drops of Mr. Smith's or Ms. Jones's or Mr. Green's or Ms. Brown's blood (depending on your assigned number) in each of the A, B and Rh wells of your slide. 3. Place 2 drops of the simulated Anti-A serum into sample A well. 4. Place 2 drops of the simulated Anti-B serum into sample B well. 5. Place 2 drops of the simulated Anti-Rh serum into the Rh well. 6. Use three separate toothpicks to stir each sample of serum and blood mixture. Observe and look for any evidence of clumping. RECORD your team's results and report such information to the Instructor when called upon to do so. Table 4 DATA TABLE OF CLUMPING REACTIONS Anti-A Serum Anti-B Serum Anti-Rh Serum Blood Type Comments Slide #1: Ms. Jones Slide #2: Ms. Brown Slide #3:Mr. Green Slide #4: Mr. Smith DISCUSSION: 1. Give Ms. Jones's complete blood type/Rh factor: _________ 2. Give Ms. Brown's complete blood type/Rh factor: _________ 3. Give Mr. Green's complete blood type/Rh factor: _________ 4. Give Mr. Smith's complete blood type/Rh factor: _________ 5. What ABO antigens are present on the red blood cells of Mr. Green's blood? __________________________ Blood Lab02 Page 6 of 8 6. What ABO antibodies are present in the plasma of Mr. Green? __________________________ 7. If Mr. Jones needed a transfusion, what ABO type(s) of blood could she safely receive? ________________________ 8. If Ms. Brown were serving as a donor, what ABO blood type(s) could receive her blood safely? ___________________________ 9. What is the difference between antigen and antibodies? _______________________________________________________________________ _____________________________________________________________ 10. Could a man with an AB blood type be the father of an 0 child? ________________ 11. Could a man with an 0 blood type be the father of an AB child? ________________ 12. Could a Type B child with a Type A mother have a Type A father? _____________ 13. What are all of the possible genotypes of offspring from such parents: one is A, the other is B? ______________________________________________________ 14. Suppose Mr. Smith marries Ms. Brown. What are the chances for an Rh+ child? _________; an Rh- child? ____________(Assume Mr. Smith is heterozygous.) 15. Under what conditions might a person with Rh- blood develop Rh antibodies? __________________________________________________________________ 16. What is likely to happen to a donor's cells If an Rh- person who is sensitive to Rh+ blood receives a transfusion of Rh+ blood? _______________________________________________________________________ _____________________________________________________________ 17. If you have "0+" blood, could your parents have been "0-"? _________ Could they have been "A+”? _________ 18. Can the parent-child Identification ALWAYS be determined by blood types/Rh? ________; Give examples to support your answer. _______________________________________________________________________ _____________________________________________________________ Blood Lab02 Page 7 of 8 Conclusions: There are ______________ major blood types! They are: ____; ____; ____; and ____! A person with blood type "A" has a protein on the surface of his/her blood cells called antigen _____ and an antibody protein called antibody ____ in the liquid part of his/her blood, which will destroy the foreign protein of another blood type ______. Similarly, a person with blood type "B" has antigen ______and antibody ________to destroy the foreign protein ________. A person with blood type ______ has antigen "A” and antigen "B" and has ________________ antibodies. This individual is referred to as the universal ___________ because they can receive limited amounts of any of the 4 major blood types’ during a transfusion. A person with blood type “O" has _____________ antigens and antibodies ______ and _____ which will destroy the foreign blood proteins _____. and _____. Because these individuals have blood that does not contain either of the two blood proteins and will, therefore, not stimulate a response during transfusions, they are called the universal ______________________. In addition to blood type proteins, another blood factor considered in this lab is the _________________________. Persons with this plasma protein are _______, and, those without it are designated ______________. Basically, and for simplification, ___________________ is dominant and ____________ is recessive. Blood Lab02 Page 8 of 8