Group no 9 Biologics presentation.pptx….

Pharmacognosy

1. Biologics:

2. Biologics: Definition: Biologics is a broad term. It includes immunizing biologics that are derivatives of animals (serums, antitoxins, globulins) or of microscopic plant organisms (vaccines, toxins, toxoids), which either directly or indirectly confer a state of protection against pathogenic micro-organisms . Immunity. Vaccines. Toxins and anti toxins Venom, antivenoms and antisera.

3. Immunity: Definition: It is the ability of an organism to resist a particular infection or toxin by the action of specific antibodies or sensitized white blood cells . Significance: Our immune system, a network of intricate stages and pathways in the body, protects us against harmful microbes . It recognizes foreign invaders like bacteria, viruses and parasites and take immediate action .

4. Classification: Immunity is classified into two major types: Natural / innate immunity. Acquired/adaptive immunity.

5. Innate immunity: It means the defense mechanisms that are present in body because of race, species specificity and multitude of other factors, but it does not include any mechanisms especially developed during lifetime of individual. Innate immunity is endowed at birth. physical barriers: skin, mucous membrane. Cells involved: phagocytes and natural killer cells. Proteins: complement, acute phase proteins and interferons.

6. Acquired immunity: It is the type of immunity that develops when a person’s immune system responds to a foreign substance , or that occurs after a person receives antibodies from another source.

7. Active immunity: It is the specific immunity developed by an individual in response to introduction of antigenic substances into the body. It is further classified as: Naturally acquired active. Artificially acquired active. Naturally acquired active: Antigens received by body in a natural manner. Developed slowly and long lasting. Example: recovery from infection such as measles or scarlet fever .

8. Artificially acquired active: Received by body through administration of vaccine or toxoid. Immunity produced as a result of series of injections. This administration of vaccine stimulates the body cells to make their own antibiotics and produce immunity that is acquired artificially Developed gradually, long lasting. Example: typhoid vaccine.

9. Passive immunity: It is the type developed by the introduction of pre-formed antibodies (not antigens) into the body. In this type, body cells are not stimulated to produce their own antibodies. The term passive is applied because immunity acquired by individual is not self developed but is passed from one individual to another.

10. Classification of passive immunity: Naturally acquired passive: Immunity developed in newborn infant through transmission of antibodies from blood of mother is naturally acquired passive immunity. This type of immunity is produced quickly but not long lasting. Artificially acquired passive: The injection of immunizing biologics containing preformed antibodies in forms such as diphtheria antitoxin produces artificially acquired passive immunity. This type of immunity is produced quickly but not long lasting.

11. Defensive mechanism of body: The body has developed defense mechanisms to control and to cope with the constant attack of micro-organisms. The body has three lines of defense: 1 st line of defense. 2 nd line of defense. 3 rd line of defense.

12. 1 st line of defense: 1 st line of defense is a combination of physical and chemical barriers that prevent all types of foreign agents from penetrating the outer layer of body. No specific foreign agent is targeted at this level. Physical barriers: skin: Cells filled with keratin, making skin impenetrable, waterproof and resistant to disruptive toxins and moist invaders. Dead cells are shed and replaced (1 million every 40 min), taking microbes with them. Mucous membranes: Line respiratory, digestive, urinary and reproductive system and protect internal lining. Mucous membranes are more vulnerable than skin.

13. 1 st line of defense : Hair: Hairs present in the nose also act as a coarse filter. Chemical barriers: Sweat produced by glands. Sticky mucous produced by mucous membranes. Saliva and tears. Cerumen(ear wax).

14. 2 nd line of defense: The second line of defense consists of defensive cells. If a pathogen penetrates the 1 st line of defense these cells play a role in destroying the pathogen before it harms the body. Phagocytes: engulf pathogens damaged tissues or dead cells. Eosinophils: discharge destructive enzymes to destroy pathogens too big for phagocytes. Natural killer cells: seek out abnormal cells ( e.g , cancer cells)

15. 2 nd line of defense : Inflammation: Redness: caused by increased blood flow. Heat: increased blood flow increases temperature in area of injury. Swelling: histamine makes capillaries more permeable than usual. Pain: causes person to protect the area. Fever: A fever is abnormally high body temperature caused by pyrogens. A mild or moderate fever helps the body to fight bacterial infections by slowing the growth of bacteria and stimulating body defense responses.

16. 3 rd line of defense: The third line of defense against pathogenic invasion is the adaptive immune response, which has two key qualities: It is specific. It is adaptive.

17. Humoral immunity: Humoral immunity describes the production of antibodies by B lymphocytes (B cells) B cells are antibody-producing cells that develop in the bone marrow to produce a highly specific antibody that recognize one type of antigen Each B lymphocyte has a specific antibody on its surface that is capable of recognizing a specific antigen When antigens are presented to B cells (and T H  cells) by macrophages, only the B cell with the appropriate antibody will become activated and clone The majority of B cell clones will differentiate into antibody-producing plasma cells, a minority will become memory B cells (B M  cells) Plasma cells produce massive quantities of specific antibody for a limited time (~2,000 molecules per second for ~4 - 5 days)

18. Cell mediated immunity: Cell-mediated immunity describes the activation of humoral immunity by T helper cells and the targeted destruction of infected cells by cytotoxic T cells It is important to recognize that humoral immunity will not occur without activation by cell-mediated immunity When a pathogen invades the body, it is engulfed by wandering macrophages which present the antigenic fragments on its surface This macrophage becomes an antigen-presenting cell, and presents the antigen to helper T cells (T H  cells) The T H  cells bind to the antigen and become activated, and in turn activate the B cell with the specific antibody for the antigen This B cell clones and differentiates into plasma cells and memory cells

19. Summary:

20. Summary:

21. VACCINATION

22. OBJECTIVES INTODUCTION TO VACCINES History Components of vaccine Differences between vaccine and drugs

23. VACCINATION “ Vaccines are chemical compounds contain living ,attenuated, or killed viruses ,killed rickettsiae,or attenuated or killed bacteria, and they are used as inoculations to stimulate the production of antibodies, thus protecting the person from a particular disease.” The process of vaccine administration is termed as vaccination .

24. CHARACTERISTICS Important characteristics of vaccine include: They have prophylactic action ( exception rabies vaccination) Nonliving vaccines provide protection for only limited period of time thus , repeated vaccination is required as for typhoid, cholera, typhus. multiple immunization is required as in case of polio. Active immunization with living agents is generally more preferable over killed vaccines because of a superior and more long lived immune response as for measles ,rubella , mumps.

25. PRECAUTIONS Precautionary measures should be followed to ensure optimum effectiveness with minimum adverse reactions. Use of vaccines is contraindicated under conditions in which the immune response may he depressed, such as during therapy involving corticosteroids, antineoplastic agents, immunosuppressive agents, or radiation; in patients with immunoglobulin deficiency in patients with latent or active infections Active immunization may cause fever, malaise, and soreness at injection sites.

26. PRECAUTIONS Some reactions are relatively specific to a particular vaccine, such as arthralgia and arthritis following rubella vaccine convulsions following pertussis vaccine. Allergic- reactions may result either from the organism constituting the vaccine or from a protein incorporated into the vaccine during manufacture, e.g., egg protein from chick embryo tissue cultures. Consequently , a careful history of the patient should be taken before vaccination to detect possible hypersensitivity to the protein.

27. PRECAUTIONS During the 1976 "swine flu" immunization program in the United States, there was an 8-fold increase in post immunization Guillain-Barre syndrome (acute febrile polyneuritis) in comparison with unvaccinated controls. This complication arises within 8 weeks of immunization and has resulted in a 5 1/c mortality rate among patients who developed the syndrome.Therefore,a long clinical trial must be ensured for any vaccine before its availability to population.

28. HISTORY The  word  " vaccine " was created by Edward Jenner. The  word  comes from the Latin  word  vacca, meaning cow. A virus that mainly affects cows (Cowpox) was used in the first scientific demonstration that giving a person one virus could protect against a related and more dangerous one.

29. COMPONENTS OF VACCINE All types of vaccines contain two main components . The antigen (protein) –“active ingredient” Other chemicals used to make the vaccine product

30. OTHER CHEMICALS USED TO MAKE THE VACCINE PRODUCT Suspending fluid (sterile water, saline, or fluids containing protein) Preservatives and stabilizers Monosodium glutamate (MSG) and 2-phenoxy-ethanol Used as stabilizers in a few vaccines to help the vaccine remain unchanged when the vaccine is exposed to heat, light, acidity, or humidity. Thimerosal A mercury-containing preservative that is added to vials of vaccine that contain more than one dose to prevent contamination and growth of potentially harmful bacteria

31. OTHER CHEMICALS USED TO MAKE THE VACCINE PRODUCT Formaldehyde Used to inactivate bacterial products for toxoid vaccines. Also used to kill unwanted viruses and bacteria that might contaminate the vaccine during production . Most formaldehyde is removed from the vaccine before it is packaged. Antibiotics Added to some vaccines to prevent the growth of bacteria during production and storage of the vaccine. Penicillin is usually NOT USED in vaccines. other examples include albumin, phenols, and glycine

32. OTHER CHEMICALS USED TO MAKE THE VACCINE PRODUCT Adjuvants or enhancers Help improve the vaccine's effectiveness. Adjuvants help promote an earlier, more potent response, and more persistent immune response to vaccines Example include Aluminum gels or salts of aluminum Culture media Very small amounts of the culture material used to grow the virus or bacteria used in the vaccine, such as chicken egg protein. Egg protein Found in vaccines prepared using chicken eggs e.g. influenza and yellow fever vaccines Could cause “egg allergy”

33. COMPARISION BETWEEN DRUGS AMD VACCINES VACCINES SIMILARILITES DRUGS Preventive (usually once and done) Both are medical products Therapeutic ( usually several weeks to years) Biologics (proteins , live or attenuated viruses) Both can cause adverse events Small molecules (organic compounds) Process patents( meaning complex manufacturing processes) Vaccines and drugs all contain multiple ingredients Product patents ( simpler manufacturing) Given mostly parenterally, orally or intranasally . Both have the potential for interaction with disease, drugs and other vaccines May be given orally ,or other routes like IM,IV Longer clinical trials Shorter clinical trials Vaccines are supposed to protect whole populations (“herd immunity”) Vaccines and drugs all have to comply with standards of safety, quality and “efficacy” Use of drugs is individualized Low public tolerance for adverse events following immunization Efficacy for medicines and protective efficacy for vaccines Adverse drug reactions are more common, less reported and more tolerated

34. Learning outcomes: At the end of this session,participants are expected to:
– Know the different types of vaccines. – Know the advantages and disadvantages of different types of vaccines.
– Appreciate the need to take the advantages and disadvantages into
consideration when undertaking vaccine
pharmacovigilance

35. Types of vaccines Whole pathogen vaccines: Live attenuated vaccine Inactivated vaccine Subunit vaccines: Recombinant protein vaccines Toxoid vaccines Conjugate vaccines Nucleic acid vaccines: DNA and RNA vaccines Viral vectored vaccines: Replicating and non replicating vaccines

36. Whole-virus Composed by the whole virus,e.g.RABIPUR Split Vaccine Subvirion vaccine, split with solvents or detergents, containing surface antigens, nucleocapsides and matrix Sub-Unit Vaccine Purified-surface-antigen vaccine composed by surface antigens Polysacharide Vaccine Purified polysaccharide from bacteria,
e.g.plainPolysacch.Meningitis A

37. Live attenuated vaccine , whole pathogen vaccine Live attenuated vaccines: “Live attenuated vaccines contain a version of the living microbe that has been weakened in the lab. So it can’t cause disease” Attenuated vaccines are made by passing the disease-causing virus through a series of cell cultures.When it is given to a human, it will be unable to replicate enough to cause illness, but will still provoke an immune response that can protect against future infection.

38. Advantages and disadvantages Advantages: These are relatively easy to create for viruses. They elicit strong cellular and antibody responses and often confer immunity with only one or two doses. Protection from a live, attenuated vaccine typically outlasts that provided by a killed or inactivated vaccine. Disadvantages :
The nature of the living things is to change, or mutate.So,mutations that can occur when the vaccine virus replicates in the body may result in a more virulent strain.
Oral polio vaccine (OPV), a live vaccine that is ingested instead of injected.This vaccine’s virus can mutate into a virulent form and result in rare cases of paralytic polio.
People who have damaged or weakened immune systems cannot be given live vaccines.

39. Inactivated or killed vaccines Whole pathogen vaccine Inactivated vaccines : Killed or inactivated vaccine are created by inactivating a pathogen with Chemicals,Heat, or Radiation.This destroys the pathogen’s ability to replicate, but keeps it “intact” so that the immune system can still recognize it. “Inactivated” is generally used rather than killed to refer to viral vaccines of this type.

40. Advantages and disadvantages Advantages: Stable and safer because killed or inactivated pathogens can’t replicate at all, they can’t revert to a virulent form capable of causing disease. Don’t require refrigeration, and they can be easily stored and transported in a freeze-dried form۔ Disadvantages: Stimulate a weaker immune system response.They provide a shorter length of protection than live vaccines. Take several additional doses, or booster shots, to maintain a person’s immunity or to create long-term immunity.

41. Toxoids Subunit vaccines Toxoids: Some bacterial diseases are not directly caused by a bacterium itself, but by a toxin produced by the bacterium. Toxins are bacterial waste products that are considered poisonous to the animal body. They act as antigens owing to their power of stimulating certain cells of the body to produce antibodies called antitoxins. Example is tetanus

42. Inactivated by treating them with formalin, a solution of formaldehyde and sterilized water. Such detoxified toxins, called toxoids are safe for use in vaccines. When the immune system receives a vaccine containing a harmless toxoid, it learns how to fight off the natural toxin.The immune system produces antibodies that lock onto and block the toxin.

43. Recombinant protein vaccine Subunit vaccine Recombinant Protein Vaccines Recombinant vaccines are made using bacterial or yeast cells to manufacture the vaccine. A small piece of DNA is taken from the virus or bacterium against which we want to protect and inserted into the manufacturing cells. For example, to make the hepatitis B vaccine, part of the DNA from the hepatitis B virus is inserted into the DNA of yeast cells. These yeast cells are then able to produce one of the surface proteins from the hepatitis B virus, and this is purified and used as the active ingredient in the vaccine.

44. Conjugate vaccines Conjugate vaccine: If a bacterium possesses an outer coating of sugar molecules called polysaccharides.This polysaccharide coatings disguise a bacterium’s antigens so that the immature immune systems of infants and younger children can’t recognize or respond to them Conjugate vaccines, a special type of subunit vaccine, get around this problem.When making a conjugate vaccine, scientists link antigens or toxoids from a microbe that an infant’s immune system can recognize to the polysaccharides.

45. Nucleic acid vaccines RNA vaccines RNA vaccines use mRNA inside a lipid membrane. This fatty cover both protects the mRNA when it first enters the body, and also helps it to get inside cells by fusing with the cell membrane. Once the mRNA is inside the cell, machinery inside the cell translates it into the antigen protein. This mRNA typically lasts a few days, but in that time sufficient antigen is made to stimulate an immune response. It is then naturally broken down and removed by the body. RNA vaccines are not capable of combining with the human genetic code (DNA)

46. Nucleic acid vaccines DNA vaccines DNA is more stable than mRNA so doesn’t require the same initial protection. DNA vaccines are typically administered along with a technique called electroporation. This uses low level electronic waves to allow the bodies’ cells to take up the DNA vaccine. DNA must be translated to mRNA within the cell nucleus before it can subsequently be translated to protein antigens which stimulate an immune response.

47. Viral vectored vaccines Replicating Replicating viral vectors retain the ability to make new viral particles alongside delivering the vaccine antigen when used as a vaccine delivery platform. As with live attenuated whole pathogen vaccines this has the inherent advantage as a replicating virus that it can provide a continuous source of vaccine antigen over an extended period of time compared to non-replicating vaccines, and so is likely to produce a stronger immune response. A single vaccine may be enough to give protection.

48. Viral vectored vaccines Non-replicating Non-replicating viral vectors do not retain the ability to make new viral particles during the process of delivering the vaccine antigen to the cell. This is because key viral genes that enable the virus to replicate have been removed in the lab. This has the advantage that the vaccine cannot cause disease and adverse events associated with viral vector replication are reduced. However, vaccine antigen can only be produced as long as the initial vaccine remains in infected cells (a few days). This means the immune response is generally weaker than with replicating viral vectors and booster doses are likely to be required.

49. Types of vaccines

50. LEARNING OBJECTIVES At the end students will have information of types of vaccines on the basis of 1. Viral vaccines 2. Bacterial vaccines Diagnostic agents which are used to identify hyper-sensitivity in individuals.

51. VIRAL VACCINES These are that vaccines which are used for prophylaxis of viral infections There are 3 types of them. TYPES VACCINES 1. Living viruses Mumps Rubella Rubeola Small Pox Yellow Fever 2. Inactivated/ Killed viruses Influenza Rabies Hepatitis B 3. Living / inactivated virus Poliomyelitis

52. SMALL POX VACCINES Smallpox vaccine is the living virus of vaccinia (cowpox) that has been grown in the skin of a vaccinated bovine calf. It is available in dried and in liquid form; the latter consists of a smooth, aqueous suspension of infected tissue that contains 40 to 60% of glycerin or of sorbitol and may contain not more than 05% of phenol as a preservative. Established by Edward Jenner Production of vaccine: A calf is taken its belly is washed and shaved, then its epidermis is cut and serum oozes out of it. The "seed virus“ is inoculated into the cut by hand rubbing (the workers are protected by rubber surgical gloves). The calf is maintained in an aseptic stall and given food and water during the growth of the virus. The vesicles that develop are removed at the time of maximum potency thoroughly triturated, and either made into a smooth suspension with an aqueous solution of glycerin or sorbitol or reduced to a dried pellet. Side effects: Cerebral or cerebellar dysfunction with headache, fever, vomiting, altered mental status, lethargy, seizures, and coma. Dose and route of administration: It is given per- cutaneously , the content of 1 capillary tube by multiple- puncture method.

53. HEPATITIS VACCINES Hepatitis B vaccine is composed of chemically inactivated hepatitis B surface antigen ( HBsAg ) particles obtained from the plasma of healthy chronic HBsAg carriers by plasmapheresis , separated from the infectious Dane particle by density gradient Centrifugation and absorbed on aluminum hydroxide. Specific antibodies develop after 2 doses or 3 doses in some individuals. Hepatitis B is disease of liver which cause cirrhosis, chronic active hepatitis, hepatocellular cancer. Side effects: Reddening of the skin, especially around the ears, sweating, swelling of the eyes, face, or inside of the nose, unusual tiredness or weakness (sudden and severe). Dose and route of administration: It is given intra muscularly as 3 doses of 1 ml first 2 doses one month apart and a booster dose after 6 months. Marketed products: Heptavax - B

54. CORONA VIRUS VACCINE There are four categories of vaccines in clinical trials: Whole virus: Sinopharm , Sinovac vaccine, 2 doses are given intramuscularly, it is whole virion , inactivated Corona Virus vaccine which contains an inactivated form of the virus itself (made up of killed coronaviruses , making it safe to be injected into the body). Protein subunit: Novavax , 2 doses are given intramuscularly. Viral vector: Johnson & Johnson’s Janssen COVID-19 Vaccine, 1 shot is given in the muscle of the upper arm, it does not contain eggs, preservatives and latex, Nucleic acid (RNA and DNA): Pfizer- BioNTech COVID-19 Vaccine, Moderna vaccine, 2 shots are given 21 days apart, it is mRNA type Side effects: Injection site reactions (pain, swelling, redness), headache, fever, feeling of discomfort, nausea, vomiting, rash.

55. MAIN APPROACHES TO VACCINES

56. BACTERIAL VACCINES These vaccines consist of suspensions of attenuated or killed (by moist heat )pathogenic bacteria in isotonic sodium chloride solution or other suitable diluents that activate the immune system. Antibodies are built against that particular bacteria, and prevents bacterial infection later. Bacterial strains are selected on basis of their potency and high antigenicity . Potency of vaccine = number of organisms biologic reference units Smooth S strains are more antigenic then rough R strains NAMES OF BACTERAIL VACCINES Haemophilus vaccine (polysaccharide or subunit vaccine) Pneumococcal vaccine (polyvalent polysaccharide vaccine used for 23 capsular types) Typhoid (enteric vaccine) and Cholera vaccine (killed and attenuated) Plague and Pertussis vaccine (killed and attenuated) BGG vaccine (live/ attenuated vaccine) Meningitis vaccine( capuslar polysaccharide vaccine)

57. TYPHOID VACCINES It is a sterile suspension or solid containing killed typhoid bacilli (Salmonella typhi ) of the Ty 2 strain used to treat typhoid fever. It has high antigenic efficiency 30-70% Schedule : Depending on the formulation it can be given starting at the age of two ( ViPS ), six (Ty21a), or six months (TCV) Not recommended in pregnancy . Symptoms:   Headache , poor appetite , rash, fatigue, confusion, constipation, diarrhea . Dose and route of administration : Injection (Booster), Mouth, Injection 0.5 ml dose Market available vaccines: TCV typhoid conjugate vaccine, Ty21a (a live vaccine), Vi capsular polysaccharide ViPS (subunit vaccine)

58. CHOLERA VACCINE Cholera vaccine is a sterile suspension of killed cholera vihrjos ( Vibrio cholerae ) in isotonic sodium chloride solution stored between 2 and 8 degrees expired after 18 months. It is prepared from equal portions of suspensions of cholera vibrios of the Inaba (antigenic value 35-A-3) and Ogawa (antigenic value 41)strains. Safety and adverse effects or symptoms: Types of oral vaccine are generally safe. Mild abdominal pain or diarrhea may occur. They are safe in pregnancy  and in those with poor immune function.  Nausea, vomiting, diarrhea, stomach pain, loss of appetite, headache, feeling tired Dose and route of administration: Oral, Subcutaneoulsy and Intramuscularly. Multiple dose of 0.5 ml is given after 4 weeks Marketed vaccines: Dukoral monovalent inactivated vaccine   Shanchol / mORCVAX inactivated bivalent vaccine CVD 103-HgR or Vaxchora live attenuated.

59. BCG VACCINE (active vaccine) It is a dried, living culture of the bacillus Calmette -Guerin strain of Mycobacterium tuberculosis var.bovis It is prepared from seed strain and grown in suitable medium. The expiration date of BCG vaccine is up to 1 year if it is stored at 50 C. It is immunologic protection against tuberculosis given when a person is. The vaccine is recommended when a person is exposed to tuberculosis . It should be used only with individuals who have a negative tuberculin skin test. Side Effects: Ulcers, severe skin swelling at injection site, a high fever (103 degrees F or higher), loss of appetite, weight loss, extreme tiredness, bone pain in your legs. Dose and route of adminstration : Marketed available vaccine: Intra- dermally in doses of 0.1 ml BGG vaccine (freeze-dried)

60. MENINGITIS VACCINE Meningococcal polysaccharide vaccines contain the specific bacterial capsular pol ysaccharides for Neisseria meningifidis Serogroups A, C, Y, and W-135. A bivalent vaccine with both serogroups A and C included is available, as is a vaccine with all 4 serogroups . It’s use is indicated for children over 2 years of age and for military recruits and adult populations at risk in epidemic areas. Side effects: Soreness, redness, or swelling where the shot was given, tiredness (fatigue) headache, muscle or joint pain, fever or chills, nausea or diarrhea. Dose and route of administration: Single subcutaneous injection of 0.5 ml containing 50 microgram of meningococcal polysaccharide. Marketed available vaccine : Menomune -A/C Menomune -A/C/Y/W-135

61. DIAGNOSTIC ANTIGENS These are the antigen containing preparations used to identify hypersensitivity is either developed or not in individuals to certain organisms. Hypersensitivity is usually the result of a previous infection caused by the specific etiologic agent Small quantities of diagnostic agents are injected intra- dermally and observations are made for 24 hours, 48 hours and 72 hours Erythema or redness develops showing positive result. Antigen containing preparations: Tuberculins Histoplasmin Coccidiodin Diphtheria toxin Mumps skin test antigen

62. Diagnostic agents Positive test with erythema (redness) on skin patch of 1cm area. Injected intradermally Dose of 0.1 ml is required. 1. Mumps Skin Test Antigen: It is formaldehyde inactivated mumps virus sterile suspension from extra embryonic fluids of mumps affected chick embryo . 3. Histoplasmin : It is a sterile liquid concentrate of soluble growth product of fungus Histoplasma capsulatum Products : Histoplasmin Diluted 2. Purified protein derivative of tuberculin: It is a sterile product obtained from tuberculi bacillus and is prepared from special media free from protein. Products: 1. Aplisol 2. Tubersol 3. Aplitest 4. Diphtheria Toxin: It is used for schick test and is sterile solution of toxic product of diphtheria bacillus 5.Coccidiodin: It is the sterile solution of antigen from byproducts of mycelial growth of fungus Coccidiodes immitis . Products: Spherulin

63. Venom introduction The term venom is derived from Latin word envenom ,meaning poison. Venoms are poisonous excretions produced by animals. such as snakes, spiders , and scorpions and typically injected into prey or aggressors by biting or stinging. Venom is nothing but a secretion of venomous animals, which are synthesized in a specific part of their of body ,called venom gland.

64. Chemistry of snake venom It is toxic saliva produced by parotid glands of snake mostly water with enzymatic proteins. CHEMICAL PROPERTIES Acidic Specific gravity 1.030-1.070 Soluble in water COMPOSITION It mainly composed of Proteinases Phospholipase A,B,C cholinesterase’s Produced from time of birth and injected in response to a prey or self defense.

65. MODE OF ACTION OF VENOMS Neurotoxic(nerve toxin) Hemotoxic(blood toxin) Neurotoxic(death of tissue) Anticoagulant(prevent the blood from clotting)

66. Neurotoxic venom This type of venom attacks nervous system This mainly include fasciculins , dendrotoxins , alpha neurotoxins. Fasciculins are toxins that attack cholinergic neurons by destroying acetylcholinesterase so Ach cannot be broken and stay in receptor. Dendrotoxins are found in the venom of black mamba snake which interferes with voltage gated k channels. Block the acetylcholine receptor or prevent the opening of ion channel . By blocking signals from nerve to muscle these toxins causes paralysis and possibly death. EXAMPLE: venom of cobra

67. HEMOTOXIC VENOM A hemotoxic venom act by lysis of erythrocytes. Venom of this type have a proteolytic action. They produce swelling , cardiovascular damage, and eventual necrosis. They also disrupt blood clotting and, in the process of destroying the bloods functionality ,severely damage internal organs. The immediate cause of death in such case is usually hypovolemic shock. EXAMPLE: rattle snake

68. CYTOTOXIC VENOM Cytotoxic venom involves phospholipase ,cardiotoxins. Phospholipase help to split cell membrane and help to digest and subdue prey Causes localized symptoms like blue spots on the site of bite due to limited blood circulation Example: venom of puff adder.

69. MYOTOXIC VENOM The venom contain peptides that destroy the muscle fiber proteins and results in myonecrosis. EXAMPLE: venom of Brazilian lancehead snake Symptoms include Dry mouth Thirst Muscular spasm Dropping eyelids

70. How does venom enter the body? To deliver  venom ,  snakes  have hollow fangs that act like hypodermic needles. When a  snake  bites, muscles in its head squeeze the  venom  glands. This pushes the liquid through its fangs muscles in its head squeeze the  venom  glands. This pushes the liquid through its fangs and into the flesh of its prey.

71. ANTIVENOM Antivenom , also known as antivenin, venom antiserum, and  antivenom  immunoglobulin, is a specific treatment for envenomation. It is composed of antibodies and used to treat certain venomous bites and stings. The only available treatment against snake bite is usage of antivenom The first antivenom for snake was developed by Albert Calmette against the Indian cobra.

72. ANTIVENOM PREPARATION Antivenom is immunoglobulins purified from the serum or plasma of horse or sheep that has been immunized with the venom of snake. Monovalent antivenom neutralize the venom of only one specie of snake. Polyvalent antivenom neutralize the venom of several different species of snake

73. Preparation of ANTIVENOM

74. ANTISRUMS A blood serum containing antibodies against specific antigens, injected to treat or protect against specific diseases. The therapeutic effectiveness of antiserums is based on their production of artificial passive immunity Each antiserum is a specific biologic employed to provide a supply of ready-made antibodies to combat the disease

75. ANTIRABIES SERUM Anti rabies serum is a sterile , non pyrogenic solution containing antiviral substance obtained from the blood serum of plasma of healthy horse that has been immunized agent against rabies. Injection of anti rabies serum provides the patient with immediate protection against rabies

76. TOXINS & ANTITOXIN

77. Objectives: At the end of the topic, students would be able to , Define toxins & antitoxin Explain types of toxins & antitoxin. Explain exotoxin & endotoxin mechanism Discuss commercial & medical application of exotoxin. Remember fluid & adsorbed toxoids

78. Toxins are bacterial waste products that are considered poisonous to the animal body they act as antigens because of their power to stimulate certain cells of the body to produce antibodies called antitoxins. Toxins can be small molecules , peptides, or proteins that are capable of causing disease on contact with or absorption by body tissues interacting with biological macromolecules such as enzymes or cellular receptors. Toxins vary greatly in their severity, ranging from usually minor to almost immediately deadly. What are toxins?

79. Types Of Toxins Exotoxins are proteins produced inside pathogenic bacteria, most commonly gram-positive bacteria, as part of their growth and metabolism. The exotoxins are then secreted into the surrounding culture medium. Endotoxins are the lipid portions of lipopolysaccharides (LPS) that are part of the outer membrane of the cell wall of gram-negative bacteria . The endotoxins are liberated when the bacteria die and the cell wall breaks apart. exotoxins endotoxins Cell wall Exotoxin Clostridium botulinum Endotoxins Salmonella typhimurium

80. Effects of Exotoxin

81. Commercial Application of Exotoxin To produce a solution of exotoxins commercially, the highly virulent organisms are cultured in beef broth medium and then killed by appropriate means. The organisms are removed by filtration through a bacterial filter, and the filtrate that contains the toxins and other products of growth is standardized on a suitable animal to determine the minimum lethal dose. This dose represents the smallest amount of the toxin that will kill a majority of a series of guinea pigs within 96 hours after subcutaneous administration. The source of "the most poisonous poison" is Clostridium botulinum , a microorganism generally unable to grow in the body of a warm-blooded animal but capable of causing death if its exotoxins are ingested. food poisoning in humans commonly is produced by types A, B, and E. When the toxins produced by this bacterium are compared with other types of protein poisons their potencies range from 10 to 1000 times higher .

82. Medical Application of Exotoxin Vaccinations This process involves inactivating the toxin, creating a toxoid that does not induce toxin-related illness and is well-tolerated. A widely used toxoid vaccine is the DPT vaccine . DPT vaccine protects against pertussis, tetanus and diphtheria infections. Cancer treatment Cancer cells can be eliminated without destroying normal cells like in chemotherapy or radiation by attaching an antibody or receptor ligand to the exotoxin, creating a recombinant toxin that is targeted to certain cells. The cancer cell is killed once the toxin is internalized; for example, Pseudomonas exotoxin disrupts protein synthesis after cellular uptake.

83. Fluid & Adsorbed Toxoids Fluid toxoids Treating exotoxins with formaldehyde reduces or eliminates the toxic Properties without affecting the antigenic properties. These products, detoxified in this manner, are called fluid toxeids . they are used to induce artificial active immunity in susceptible individuals. Adsorbed toxoid By precipitating or adsorbing the fluid toxoid with alum, aluminum hydroxide, or aluminum phosphate, an adsorbed toxoid is produced which, when administered, results in a slower release of the antigen from the site of injection and a subsequent production of higher and more prolonged antibody titers. However, the adsorbed toxoids are more prone to produce local reactions at the site of injection than are fluid toxoids. To avoid this, adsorbed toxoids should be administered by deep intramuscular injection, whereas the fluid toxoid may be administered subcutaneously. Both fluid and adsorbed toxoids are used to produce active immunity against diphtheria and tetanus. They are used alone and in combination. Repeated immunization with diphtheria and tetanus toxoids may result in increasingly severe local reactions.

84. ENDOTOXIN They are the part of the outer membrane portion of the cell wall of gram negative bacteria, e.g., Lipopolysaccharide (LPS) which is released when dead cells lyse in blood, causes macrophages to release high levels of cytokines resulting is chills, fever, weakness, aches, small blood clots, tissue necrosis, shock and death.

85. Effects of Endotoxin 1 2 3 4

86. ANTITOXIN An antitoxin is an antibody with the ability to neutralize a specific toxin . Antitoxins are produced by certain animals, plants , and bacteria . Although they are most effective in neutralizing toxins, they can kill bacteria and other microorganisms. Antitoxins are made within organisms, but can be injected into other organisms, including humans. This procedure involves injecting an animal with a safe amount of a particular toxin. Then, the animal’s body makes the antitoxin needed to neutralize the toxin. Later, the blood is withdrawn from the animal. When the antitoxin is obtained from the blood, it is purified and injected into a human or other animal, inducing passive immunity . To prevent serum sickness , it is often best to use antitoxin generated from the same species .

87. ANTITOXIN antitoxin, antibody , formed in the body by the introduction of a bacterial poison, or toxin, and capable of neutralizing the toxin. People who have recovered from bacterial illnesses often develop specific antitoxins that confer immunity against recurrence. For medical use in treating human infectious diseases, antitoxins are produced by injecting an animal with toxin; the animal, most commonly a horse, is given repeated small doses of toxin until a high concentration of the antitoxin builds up in the blood. The resulting highly concentrated preparation of antitoxins is called an antiserum.

88. ANTITOXIN In the past, diphtheria antitoxin consisted of unprocessed serum which, when injected, often caused numerous cases of sensitivity to horse serum proteins. Today, depending on the manufacturer, either of 2 processing methods is employed: The first involves a series of precipitations using varying concentrations of ammonium sulfate. During this process, the euglobulin and fibrinogen fractions are initially "salted" out, followed by the pseudoglobulin fraction, which contains the antitoxin. The latter fraction is redissolved , dialyzed, and filtered. The second method utilizes a pepsin solution to digest the plasma, thus removing up to 80% of the protein; however, a loss of about 20% in antitoxin content also occurs. The digested material is then treated with ammonium sulfate solution, redissolved , dialyzed, and filtered. Antitoxins are standardized in terms of "antitoxin units.“

89. TYPES OF ANTITOXINS Diphtheria antitoxin Diphtheria antitoxin is a sterile, non-pyrogenic solution of the refined and concentrated proteins, chiefly globulins, containing antitoxic antibodies obtained from the blood serum or plasma of healthy horses that have been immunized against diphtheria toxin or toxoid. It has a potency of not less than 500 antitoxin units per ml. The expiration date with a 20% excess of potency is not later than 5 years after the date of manufacture or of issue. Diphtheria antitoxin should be stored at a temperature of between 2 and 8° C. USES AND DOSE Diphtheria antitoxin is a passive immunizing agent capable of inducing passive immunity against diphtheria. Any person with clinical symptoms of diphtheria should receive the antitoxin at once without waiting for bacteriologic confirmation. The usual prophylactic dose, intramuscularly or intravenously, is 1000 to 10,000 units; the therapeutic dose, 20,000 units to 120,000 units.

90. Botulism antitoxin Botulism antitoxin is a sterile, nonpyrogenic solution of the refined and concentrated antitoxic antibodies, chiefly globulins, obtained from the blood serum or plasma of healthy horses that have been immunized against the toxins produced by both the type A and type B and/or type F strains of Clostridiurn botulinum . This antitoxin contains not more than 20% of solids and should be stored at a temperature of between 2 and 8° C. The expiration date is not later than 5 years after the date of issue.  This multivalent antitoxin is used to treat all cases of toxemia caused by the types of botulinus bacteria Used in its preparation. A multivalent antitoxin is advantageous because the prescribing physician is not required to wait for a determination of the type of the causative organism.  USE AND DOSE. Botulism antitoxin is classed as a passive immunizing agent to be used in the treatment of botulism. The usual dose is, intravenously, 20,000 units, repeated at 2- to 4-hour intervals, as necessary.

91. Tetanus Antitoxin Tetanus a control antitoxin is a sterile, nonpyrogenic solution of the refined and. Concentrated proteins, chiefly globulins, containing antitoxic antibodies obtained from the blood serum or plasma of healthy horses that have been immunized against tetanus toxin or toxoid. It has a potency of not less than 400 antitoxin units per ml. Tetanus antitoxin should be stored at a temperature of between 2 and 8°C. The expiration date of the liquid antitoxin is not later than 5 years after the date of manufactureor issue with a 20% excess of potency. USES AND DOSE Tetanus antitoxin is employed in the treatment and prophylaxis of tetanus if tetanus immune globulin is not available. It creates passive immunity to tetanus. Like diphtheria antitoxin, it is a valuable therapeutic agent when used early in the disease. Prophylactic doses should he given to individuals who have had 2 or less injections of tetanus toxoid and who have tetanus-prone injuries that are more than 24 hours old. Tetanus toxoid should also be administered at a different site on the patient. The usual prophylactic dose, intramuscularly or subcutaneously, is 1500 to 5000 units; the therapeutic dose is 50,000 to 100,000 units or more with at least part of the dose given intravenously.