Microbiological Status of Lab Animals Including Gnotobiology

1. Microbiological Status of Lab Animals Including Gnotobiology Prof. Dr. Khaled Al-Amry

2. Germ-free Life biological condition characterized by the complete absence of living microorganisms.

3. Timeline of germfree animals research Subsequent advances in methodology  during the 1920s and ’30s led to the routine raising of germfree animals. experiments were continued with chicks for more than a decade with no success. . 1920s & ’30s 1895 1895-1912 1912 The first attempts to grow germfree animals with guinea pigs at the Hygiene Institute of Berlin. The first successful germfree vertebrate experiments (with chicks). Shortly thereafter, germfree goats were kept alive for two months.

4. Gnotobiology Gnotobiology comprises the study of germfree plants and animals, as well as living things in which specific microorganisms, added by experimental methods, are known to be present. 1 When one or more known species of microorganisms are added experimentally to a germfree plant or animal, the host, of course, is no longer germfree. 2

5. Gnotobiology Both the host and the introduced species are gnotobiotic, however, since all added species are known to the investigator. 3 Precise comparisons between germfree and conventional animals cannot be made unless both are isolated from the environment and fed the same sterile diet. 4

6. Gnotobiotic research seeks to identify the specific causative agents in infectious diseases. explore the effects of microorganisms in physiological diseases. currently, it is directed toward studying the reactions of germfree animals after they have been inoculated with specific known microorganisms. explore the role of bacteria in protozoan and viral infections, the contribution of bacteria to nutrition, immunity, and animal physiology.

7. Applications of gnotobiotic research The addition of one or two specific microorganisms to germfree animals can clarify cause-and-effect relationships that are important in human disease. 1 The complex interactions of pathogenic (disease-causing) microorganisms with the bacteria that normally inhabit the body can be partially elucidated by infecting germfree laboratory animals with such organisms. 2

8. Applications of gnotobiotic research Its techniques have been used to clarify the causes of certain common human health problems. Gnotobiotic studies have shown, for example, that tooth decay is caused only by certain infectious bacteria. 3 Germfree animals are used in toxicology, pollution control, and vaccine tests. 4

9. Applications of gnotobiotic research The effects of an external force (e.g., radiation or a noxious gas) on a germfree animal are easy to distinguish because there is no interference from infection. 5 Patients with impaired immunological  defenses against bacteria can be placed in complete biological isolation using gnotobiotic techniques. 6

10. Applications of gnotobiotic research Babies suspected of lacking the ability to synthesize immunoglobulins have been delivered into germfree isolators and maintained there until laboratory tests have shown that they could synthesize gammaglobulins. 7 Hospital precautions that precede heart transplants may include elaborate gnotobiotic rooms and procedures to prevent an immune-suppressed patient from coming into contact with pathogenic microorganisms. 8

11. Germfree methodology Birds

12. Germfree methodology Plants

13. Germfree methodology Mammals

14. Germfree methodology Mammals

15. Environmental standards are a set of quality conditions that are adhered to or maintained for a particular environmental component and function. The different environmental activities have different concerns and therefore different standards.

16. Types of environmental standardization The three main types of environmental standards - ambient, emission, and technology - work together to help regulate the impact human activity have on the environment. Ambient Standards Emission standards Technological Standards The regulation of ambient pollutants in the air ensures that the air remains breathable and does not further deteriorate. Standards are put in place to encourage progressiveness across different industries. They are enacted to control the amount of pollutants that are released into the air in order to maintain air quality.

17. Microbiological standardization

18. Basic classifications for microbiological standardization of laboratory animals

19. Stainless Steel LED Display Laboratory SPF Chicken Isolator

20. Monitoring of microbiological standardization

21. Product control versus process control Product Control • Controlling laboratory animals. • FELASA (Federation of European Laboratory Animal Science Associations ) guidelinesare frequently used. • These guidelines where the monitoring frequency, sample size, micro-organisms to be tested - vary according to the microbiological quality of the animals, and test methods and are frequently updated by FELASA and shared in their web pages. Process Control • Controlling barrier systems. • Samples are taken routinely from ambient air, surfaces, base plate materials of animals, foods and waters. • Microbiological tests are carried out on these samples.

22. Microbial air sampler for continuous monitoring

23. Dips and swabs surface monitoring

24. The use of animals in research and testing Around the world it is estimated that +100 million animals are used in experiments every year CHINA EUROPEAN UNION NEW ZEALAND CANADA c.16m 4.3m 11.5m 0.24m JAPAN c.11m AMERICA c.20m AUSTRALIA c.6.7m

25. The use of animals in research and testing

26. Statistics of Scientific Procedures on Living Animals (2016) 72.8 % 13.6 % Fundamental Research 28.5% Human & Veterinary Medicine 8.6% Safety & Toxicity Testing 13.5% 6.3 % 3.9 % 3.4 % Total numbers of procedures 3,936,723

27. Animal Testing It is the use of non-human animals in experiments that seek to control the variables that affect the behavior or biological system under study. 1 This approach can be contrasted with field studies in which animals are observed in their natural environments or habitats. 2

28. Animal Testing Experimental research with animals is usually conducted in universities, medical schools, pharmaceutical companies, defense establishments and commercial facilities that provide animal-testing services to industry. 3 The focus of animal testing varies on a continuum from pure research, focusing on developing fundamental knowledge of an organism, to applied research, which may focus on answering some question of great practical importance, such as finding a cure for a disease. 4

29. Areas of animal use

30. 3 Rs and animal testing ethics Replacement using humane methods which avoid or replace the use of animals 3 Rs and animal testing ethics Reduction minimizing the numbers of animals used 3 Rs and animal testing ethics Refinement improving experiments, housing and care to reduce suffering throughout animals’ lives

31. Animal testing: pros versus cons

32. Animal testing: pros versus cons

33. Animal testing: pros versus cons

34. Animal testing: pros versus cons

35. Animal testing: pros versus cons

36. Animal testing: pros versus cons

37. Animal testing: pros versus cons

38. Lab animals 1. Rabbit Appearance The body of the rabbit is closely covered with smooth hair or fur, except at a few places (the tip of the nose, a small portion of scrotum and the inguinal spaces). The rabbit has large and erect pinnae (external ears) with readily visualised musculature. The small external opening of the mouth is bound by a cleated upper lip and an undivided lower lip.

39. Lab animals 1. Rabbit Appearance The external nostrils are ovoid in shape and are connected with the cleft in the upper lip. Prominent whiskers are present around the nose, over each eye and one or two on each cheek. The rabbit has large eyes (pink in albino rabbit) directed more laterally than those of most mammals. The hind legs are longer, muscular and powerful than forelimbs.

40. Lab animals 1. Rabbit Lab Use Rabbit was used in Friedman test of pregnancy, but with the advent of more easy and refined techniques, its use for this purpose has become limited.

41. Lab animals 1. Rabbit Rabbit as a lab animal Research areas where rabbit is experimental animal of choice include: Hypertension, cardiac surgery, infectious diseases, metabolic disorders, ophthalmology. In recent times, rabbit is considered as excellent animal model for the study of gastrointestinal diseases. It is also used in serology because its large ear vein facilitates easy withdrawal of blood.

42. Lab animals 1. Rabbit In microbiology laboratory (a) Differentiate between human and bovine types of tubercle bacilli since it is susceptible only to bovine type; (b) Prepare small quantity of antisera; (c) Prepare Rabies vaccine; (d) Isolate herpes, smallpox viruses.

43. Lab animals 1. Rabbit Blood Collection Marginal ear vein is the preferred site of blood collection; blood can also be withdrawn from the jugular vein, the orbital sinus, for large quantity (20 ml) of blood, the cardiac puncture of anaesthetized rabbit is preferred.

44. Lab animals 2. Guinea Pig Appearance The guinea pig is a short rodent, it does not have a tail. The head seen in profile is blunt and rectangular with small ears. The short thick neck emerges with the trunk.

45. Lab animals 2. Guinea Pig Appearance The limbs are unequal in length, the hind legs are longer than the forelegs. The guinea pig differs from the other laboratory rodents in many respects. It needs Vitamin C in the diet and is very susceptible to tuberculosis and to anaphylactic shock.

46. Lab animals 2. Guinea Pig Lab Use The guinea pig is very widely used in immunology (delayed hypersensitivity), immune responses, anaphylactic shock and allergic encephalomyelitis). It is also used in the field of biochemistry, toxicology, physiology, pharmacology, microbiology.

47. Lab animals 2. Guinea Pig Blood Collection Small samples of blood (less than 0.25 ml) can be obtained by simple venesection of the marginal ear vein; large quantity by the cardiac puncture or from the metatarsal veins of anaesthetized guinea pig; repeated samples of small quantities from the orbital sinus.

48. Lab animals 2. Guinea Pig In Microbiology Lab Particularly in microbiology, laboratory, guinea pig is quite useful: (a) To isolate both human and bovine types of tubercle bacilli; (b) For “Virulence” test for diphtheria bacilli and tetanus bacilli; (c) To obtain “Complement” for serological test; (d) To study rickettsial disease (typhus fever), viral diseases.

49. Lab animals 3. Rat Appearance The albino rat has a wide head, rough fur and long ears. The tail length is always less than the body length. It has long cylindrical body, long thin tail and very short legs.

50. Lab animals 3. Rat Appearance The head has a pointed snout with two silt-like nostrils, a narrow mouth with short lower jaw, and split upper lips, two small beady eyes, set so that they can look diagonally forwards and sideways, they have numerous long whiskers. It has a short neck. The trunk is slightly wider than the head.