1. 1 FORENSIC SCIENCE Serology

2. 2 What is Serology? Serology is the study of serums such as blood, saliva, semen and sweat. A forensic serologist applies this information to criminal activity and the law.

3. 3 What is Blood? Blood is a very complex mixture of cells, enzymes, proteins and inorganic substances. The fluid portion is the plasma. Solids are suspended in the plasma.

4. 4 Characteristics of Blood Blood plasma is made primarily of water and accounts for 55% of the total amount of blood content. Three different types of cells are suspended in the plasma. These cells are: Erythrocytes are the red blood cells and are responsible for carrying oxygen. Leukocytes are the white blood cells and are responsible for “cleaning” the system of foreign invaders. Thrombocytes are the platelets and are responsible for blood clotting. Blood serum is the liquid that separates from the blood when a clot is formed.

5. 5 Blood Questions When blood is found at a crime scene, several questions must be answered: 1. Is it blood? 2. If so, is it human or animal? 3. If human, what type of blood is it? 4. How did the blood fall or spatter?

6. 6 1 st Question – Is it blood? A crime scene investigator cannot presume that every red stain or liquid is blood. Ketchup, spaghetti sauce and red jello may all be mistaken for blood. The investigator will perform presumptive tests at the scene to determine if the substance is blood.

7. 7 Two of the common Presumptive Tests are color tests. 1.Kastle-Meyer -phenolophthalein and hydrogen peroxide are mixed together and the blood’s hemoglobin will cause the formation of a deep pink color. Several household items will yield a false positive for a color test. For example, potatoes and horseradish yield a false positive. However, since these are unusual in a crime scene setting the Kastle-Meyer is a reasonable indication of blood. The Presumptive Blood Tests

8. 8 2. Hemastix – the strip is moistened with distilled water and then dipped into the substance. A green color indicates blood. A third option is luminol. 3. Luminol reacts with blood to produce light instead of color. Luminol detects blood even if the area has been cleaned.

9. 9 It is blood, but is it human blood? At the lab, the technician will need to determine if the blood is human or animal. First, the technician will complete a microscopic examination. The red blood cells of mammals do not have a nucleus. This is an important distinction.

10. 10 Non-Mammalian Animal Blood Frog Blood Larger nucleic red blood cells that undergo cell division No platelets

11. 11 Human Blood Numerous non-nucleic red blood cells--5 to 6 million per mm3 Larger but less numerous white blood cells 5 to 10,000 per mm3 Tiny, cellular fragments 350 to 500,00 per mm3 called platelets

12. 12 It is blood, but is it human blood? The next test that the technician will conduct is the precipitin test. In this test, the blood is injected into a rabbit; antibodies are formed; rabbit’s blood is extracted as an antiserum; the antiserum is placed on sample blood. It will negatively react with human proteins. This test is very sensitive and requires only a small amount of blood.

13. 13 It is human blood, but what type? Around 1900, Karl Landsteiner discovered that there are four different kinds of human blood based on the presence or absence of specific antigens found on the surface of the red blood cells. In 1940 Landsteiner and Weiner reported the discovery of the Rh factor by studying the blood of the Rhesus monkey. 85% of Caucasians, 94% of Black Americans and 99% of all Asians are Rh positive.

14. 14 Blood Type Terminology ABO blood groups-- based on having an A, B, both or none of the factors on the red blood cell Rh factor-- may be present on the red blood cell; positive if present and negative if not Antigen-- a substance found on a red blood cell Antibody-- a substance that reacts with an antigen Agglutination-- clumping of red blood cells; will result if blood types with different antigens are mixed

15. 15 Blood Genetic Terminology Genotype--letters that represent the traits; ie, AA, AO, BO, BB, AB and OO Phenotype--words that describe the traits Heterozygous--different alleles for the same trait; ie: AO, BO, AB Homozygous--the same alleles for the trait; ie: AA, BB, OO

16. 16 Blood Typing A blood type has antigen A and will agglutinate with B. B blood type has antigen B and will agglutinate with A. AB blood type has antigen A and B and will not agglutinate with either A or B. O blood type has neither antigen A or B and will agglutinate with either.

17. 17 Blood Reactions to Antiserum REACTION BLOOD TYPE Anti-A SerumAnti-B Serum Agglutination No agglutination Agglutination No agglutination Agglutination No agglutination Type A Type B Type AB Type O

18. 18 Blood Typing Reactions to Anti-Serum

19. Blood Groups Type AntigenAntibody Can Give Blood To: Can Get Blood From A B AB O A B A and B Neither A nor B B A Neither A nor B A and B A, ABO, A B, ABO, B AB A, B, O, AB O Only certain blood types can donate or receive blood from other blood types. If the wrong blood is transfused, the person may die.

20. 20 Population Distribution of Blood Types in the U. S. TypePercent O A B AB 45 39 12 4

21. 21 Typing of Dried Blood Stains Absorption-elution technique l Antiserum is placed on the blood stain. Antibodies combine with the specific antigens. l Unreacted serum is washed off the bloodstain. l Stained material is heated to 56 degrees C, breaking the antibody-antigen bond. This process is known as elution. l Known red blood cells are added. Agglutination will occur if antigens present on the added RBC’s were also originally on the stained material.

22. 22 Differentiating Within Blood Types The determination that the blood type is O + eliminates 55% of the population. In order to narrow down the possibilities within a specific blood type, the blood is tested for certain enzymes. Enzymes are proteins that speed up or slow down chemical reactions. Enzymes that exist in different forms are called polymorphic and can be broken down into their separate proteins called iso-enzymes.

23. 23 Commonly Used Blood Enzymes and Proteins  Adenosine deaminase AD  Adenylate kinaseAK  Carbonic anhydrase IICA II  Erythrocyte acid phosphataseEAP  Esterase DEsD  Glucose-6-Phosphate dehydrogenaseG6PD  Glyoxylase UGLO I  Group-specific componentGe  HaptoglobinHp  Peptidase APep A  PhosphoglucomutasePGM  6-Phosphogluconate dehydrogenase6PGD  TransferrinTf

24. 24 PGM--phenotypes Electrophoresis can separate the protein components into these 10 iso-enzymes. 1- 1+ 2- 2+ 2-1- 2+1- 2-1+ 2+1+ 2+2- 1+1- - Three of these are most common PGM 1+, PGM 2-1+ and PGM 2 +

25. 25 EAP--phenotypes Determined by the intensity of the fluorescence. The darker the band the more it fluoresced. BCABCA A BA B CB C CA

26. 26 The final question, how did the blood fall or spatter? A group of blood spatter specialists deal with the physical properties of blood and the patterns produced under different conditions as a result of various forces being applied to the blood.

27. 27 The final question, how did the blood fall or spatter? Blood, as a fluid, follows the laws of physics. It is not influenced nor affected by race, gender, or age of the one bleeding.

28. 28 BLOOD DROPLET Characteristics A blood droplet will remain spherical in space until it drops onto a surface Once a blood droplet impacts a surface, a bloodstain is formed. A droplet falling from the same height, hitting the same surface at the same angle, will produce a stain with the same basic shape.

29. 29 BLOOD DROPLET Volume Is approximately 0.05 cc Is not the same for all blood droplets--from 0.03 cc to 0.15 cc Is directly dependent upon the surface or orifice from which it originates The impact area is called the target.

30. 30 CONDITIONS EFFECTING BLOODSTAIN SHAPE Height of the droplet Size of the droplet Angle of impact Velocity at which the blood droplet left the original surface Texture of the target surface –On clean glass or plastic--droplet will have smooth outside edges –On a rough surface--will produce scalloping on the edges

31. 31 Questions Answered by Blood Spatter Interpretation The distance between the target surface and the origin of blood at the time of blood shed The point(s) of origin of the blood Movement and direction of a person or an object The number of blows, shots, etc. causing the bloodshed and/or the dispersal of blood.

32. 32 Questions Answered by Blood Spatter Interpretation Type and direction of impact that produced the bloodshed The position of the victim and/or object during bloodshed Movement of the victim and/or object after bloodshed

33. 33 Bloodstain Terminology Angle of impact-- angle at which blood strikes a target surface. Bloodstain transfer-- When a bloody object comes into contact with a surface and leaves a patterned blood image on the surface. Backspatter-- blood that is directed back toward its source of energy. Cast-off-- blood that is thrown from an object in motion

34. 34 Bloodstain Terminology Contact stain-- general term referring to bloodstains caused by contact between a wet, blood-bearing surface and a second surface which may or may not have blood on it –Transfer-- image is recognizable and may be identifiable with a particular object –Swipe-- wet blood is transferred to a surface which did not first have blood on it –Wipe-- a non-blood bearing object moves through a wet bloodstain, altering the appearance of the original stain

35. 35 Bloodstain Terminology Directionality-- relates to the direction a drop of blood traveled in space from its point of origin Terminal velocity-- the greatest speed to which a free falling drop of blood can accelerate in air. It is dependent upon the acceleration of gravity and the friction of the air against the blood--approximately 25 feet/second. –High velocity-- greater than 100 feet/second; gives a fine mist appearance –Low velocity-- 5 feet/second or less –Medium velocity-- 5 to 25 feet/second.

36. 36 Bloodstain Pattern Terminal Velocity Directionality Angle of Impact

37. 37 Blood Stain Patterns The shape of a blood stain: l Round--if it falls straight down at a 90 degree angle. l Elliptical--Blood droplet elongates as the angle decreases from 90 to 0 degrees. The angle can be determined by the following formula: width= sine of the impact angle length

38. 38 IMPACT The more acute the angle of impact, the more elongated the stain. 90 degree angles are perfectly round with 80 degree angles taking on a more elliptical shape. At about 30 degrees the stain will begin to produce a tail. The more acute the angle, the easier it is to determine the direction of travel. The greater the height the drop falls the larger the spatter.

39. 39 Height of the Droplet and Spatter Size

40. 40 Angle of Impact and Spatter Shape

41. 41 Blood Stains The harder and less porous the surface, the less the blood drop will break apart. The softer and more porous the surface, the more a blood drop will break apart. The pointed end of the blood stain faces the direction the stain is traveling.

42. 42 Area of Intersection The location of the blood source can be determined by drawing lines the various blood droplets to the point where they intersect. This is the blood’s origin.

43. 43 Area of Convergence The area of convergence is the point of origin; the spot where the “blow” occurred. It is determined by drawing a line from the area of intersection straight up to where to where the angle of impact would intersect

44. 44 CRIME SCENE What evidence can you see in this crime scene? What story does the scene tell?

45. 45 Bring In The Dogs! The dog locates human scent. A closer look shows that the ashes have human remains and clothing. In addition, look closely at the rocks on the next slide.

47. 47 Blood Evidence Class evidence for blood would include blood type. If you can determine the DNA you would have individual evidence. Blood stain patterns are considered circumstantial evidence in a court room. Experts could argue many points including direction of stains, height of the perpetrator, position of the victim, left/right hand, whether the body was moved, etc.

48. 48 Another Body Fluid - Sperm Among the smallest and most highly specialized cells in the human body. Has a head and a tail Contains 23 chromosomes with the genetic material found in the head Males release 2.5 to 6 milliliters of seminal fluid per ejaculation with approximately 100 million sperm per milliliter. Magnified 400X

49. 49 Semen Determination of Seminal Fluid - Suspect stain may have stiff, crusty appearance - Acid phosphatase color test- presumptive test –the presence of acid phosphatase, the enzyme secreted by the prostate gland into the seminal fluid, will turn purple when sodium alpha naphthylphosphate and Fast Blue B solution are placed on it. – It will also fluoresce under UV light when it comes in contact with 4-methyl umbelliferyl phosphate.

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52. 52 Detection and Collection of Semen Sample

53. 53 Semen (cont.) Determination of Seminal Fluid Prostate Specific Antigen (PSA) or P30--unique to seminal plasma –P30 is isolated and injected into a rabbit where antibodies are produced (anti-P30) –The stain extract is placed in one well of an electrophoresis plate and the anti-P30 in the opposite well. The electric is applied and the antigens and antibodies move toward each other. The formation of a precipitation line between the wells shows the presence of P30 in the sample stain. It must be seminal fluid.

54. Microscopic Examination of Sperm A microscopic examination will also determine if the sample is semen The presence of spermatozoa confirms that the sample is semen 54

55. No spermatozoa? The male could have oligospermia – a low sperm count The male could have aspermia – no sperm due to a vasectomy 55

56. 56 Collection of Rape Evidence The Medical Examination: Collection of Physical Evidence The following types of physical evidence should be collected during the medical examination for laboratory analysis: HAIR (both pubic and head hair) –Cannot individually identify a person unless the root is present; –Does provide strong collaborative evidence individual was at the crime scene; –Can be distinguished between human and animal; –May be able to determine race; and –May be able to establish the part of the body from which the hair came.

57. 57 Collection of Rape Evidence (cont) FIBERS –Limited value due to mass production of clothing; and –In rare cases, can be used to deduce individual identification with a high degree of certainty. BLOOD –Can be distinguished as to human or animal; and –DNA analysis can provide individual identification. FINGERNAIL SCRAPINGS –Used to collect DNA evidence for individual analysis and identification. VAGINAL, RECTAL AND ORAL SWABS AND SMEAR –Used to collect DNA evidence for individual analysis and identification

58. 58 Secretors 80% of people are considered secretors. Their blood-type antigens are found in high concentration in their body fluids such as saliva, semen, vaginal secretions and gastric juice. If you are a secretor, you will have a higher concentration of A and B antigens than does your blood!! With the advent of DNA, the secretor evidence is not as important as it once was.