Microbiological Status of Lab Animals Including Gnotobiology

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

2. Isolators Attributes It may be a small or large container, usually made of glass and steel or plastic, with plastic or rubber gloves.

3. Isolators Attributes Food and utensils are sterilized and taken into the isolator through an adjoining area called the sterile lock.

4. Air handling system The isolator has an air handling system

5. Positive/Negative-pressure isolator

6. Sterilization of isolators

7. Metal isolator

8. Plastic flexible pharmaceutical isolator

9. Sterilization

10. Sterility testing

11. Isolators Attributes Since the interior of the isolator is sealed off from the background cleanroom, operator access to the interior is done through glove ports or half suits. Sterile containers, stopper components, and environmental monitoring materials are brought in to and out of the isolator through air locks, mouse holes, and devices known as Rapid Transfer Ports (RTPs)

12. Isolators Attributes One of the major advantages that isolators have over RABS type systems is

13. Microbiological status of the isolator

14. Isolator

15. Aseptic filing

16. The “interface” 

17. Interface isolator

18. Plastic glove isolator

19. Integrity checks

20. The “interface”

21. Other parameters which could influence and impact the decontamination effect

22. Controlling the microbiological status of an operating isolator 

23. Air & Surface monitoring Air monitoring Surface monitoring due to the nature of the methods used for microbiological monitoring, a continuous control is not possible for surface monitoring contact plates for flat surfaces or the swab technique for irregular surfaces are used a monitoring of surfaces during production is not recommended, as additional interventions during the filling are needed for sampling and residuals of nutrient medium can endanger the asepsis of the interior of the isolator. • is performed with the semi-quantitativesettling plate method (which are exposed to the environment for a defined (and validated) time span) or the quantitative air sampling method, e.g. by the impaction method. • on a regular basis, an anaerobicmicrobiological monitoring should be performed to produce data on the anaerobic status of the filling line.

24. Environmental monitoring

25. Sampling points

26. Sampling points

27. Isolator Aseptic isolator with a, six-axis robot arm that is compatible with vapor phase hydrogen peroxide sterilization

28. Restricted Access Barrier Systems (RABS) Attributes Unlike isolators they use a combination of physical and aerodynamic barriers to prevent ingress of contaminants into the interior environment. The physical barrier is similar to machine guarding having glass or polycarbonate doors with stainless steel walls that totally enclose the machinery with an air handler supplying HEPA filtered, unidirectional airflow providing an ISO 5 environment.

29. Restricted Access Barrier Systems (RABS) Attributes RABS are typically unsealed barriers having the HEPA filtered air supplied to the RABS interior and exhausted through a gap between the RABS walls and the equipment. RABS that exhaust to the background environment are referred to as open RABS.

30. Restricted Access Barrier Systems (RABS) Attributes Passive open RABS is a barrier system that is built around equipment installed below air handlers in the background cleanroom, which provide the ISO 5 environment. Closed RABS offer another option and are by design sealed isolators that can be positive or negative pressure, but are manually cleaned and bio-decontaminated rather than utilizing an automated bio-decontamination process typical of isolators.

31. Restricted Access Barrier Systems (RABS) Attributes Glove ports and half suits are also used to further separate an operator from the sterile interior of the RABS. In the early implementation of the RABS concept by the pharmaceutical industry, many companies considered any enclosure around the process a RABS, claiming that it provided enhanced sterility protection over conventional clean room manufacturing.

32. Restricted Access Barrier Systems (RABS) Attributes But in many cases operators would continually open the RABS doors to access areas within the critical zone to perform an intervention. Thus defeating the purpose of the RABS and creating an unpredictable level of contamination Ultimately, RABS can meet the requirements of Advanced Aseptic Processing if it is used as intended; separate the operator from the process. RABS can only achieve this criterion when any and all open door interventions are prohibited.

33. Restricted Access Barrier System Restricted Access Barrier System (RABS) encloses a vial filling, and helps to ensure the aseptic environment

34. Robotics in Aseptic Processing

35. Robotics in Aseptic Processing robot safety compliant RABS unit that protects the operator from the robot system and protects the product from contamination originating from the operator

36. The combination of Isolator-Barrier technology with robot system

37. Microbiology Lab Practices and Safety Rules Wash your hands with disinfectant soap when you arrive at the lab and again before you leave. Do not put anything in your mouth such as pencils, pens, labels, or fingers. Do not store food in areas where microorganisms are stored. Absolutely no food, drinks, chewing gum, or smoking is allowed in the laboratory

38. Microbiology Lab Practices and Safety Rules This garment must: cover your arms, with long sleeves, with buttons or snaps and be able to be removed without pulling it over your head. Leave protective clothing in the lab and do not wear it to other non-lab areas. Purchase a lab coat and safety glasses, bring them to class, and use them Avoid loose fitting items of clothing Wear appropriate shoes (sandals are not allowed) in the laboratory.

39. Microbiology Lab Practices and Safety Rules Backpacks, purses, and coats should be placed in the cubbyholes by the front door of the lab. Place needed items on the floor near your feet, but not in the aisle. Keep your workspace free of all unnecessary materials Laboratory equipment and work surfaces should be decontaminated with an appropriate disinfectant on a routine basis, and especially after spills, splashes, or other contamination. Disinfect work areas before and after use with 70% ethanol or fresh 10% bleach

40. Microbiology Lab Practices and Safety Rules This saves a lot of time and effort, and keeps you away from many accidents. Label everything clearly Replace caps on reagents, solution bottles, and bacterial cultures Do not open Petri dishes in the lab unless absolutely necessary.

41. Microbiology Lab Practices and Safety Rules You can simply impress your audience and add a unique zing and appeal to your Presentations. Sterilize Inoculating loops and needles in a Bunsen burner before you lay them down Do not leave open flames unattended . when not is Turn off Bunsen burners use

42. Microbiology Lab Practices and Safety Rules Be sure to remove all papers, notebooks, combustible materials and excess chemicals from the work area. When you flame sterilize with alcohol, be sure that you do not have any papers under you. Use appropriate care and do not take cultures out of the laboratory. Treat all microorganisms as potential pathogens

43. Microbiology Lab Practices and Safety Rules If you are working with a sample that may contain a pathogen, then be extremely careful to use good bacteriological technique. Wear disposable gloves when working with potentially infectious microbes or samples Sterilize equipment and materials Long hair must be restrained if Bunsen burners are in use.

44. Microbiology Lab Practices and Safety Rules Mouth- pipetting has been known to result in many laboratory-acquired infections. With the availability of mechanical pipetting devices, mouth pipetting is strictly prohibited. Never pipette by mouth. Use a pipetting aid or adjustable volume pipettors Do not pour anything down the sink. Autoclave liquids and broth cultures to sterilize them before discarding. Consider everything a biohazard

45. Microbiology Lab Practices and Safety Rules Dispose of all solid waste material in a biohazard bag And autoclave it before discarding in the regular trash. (e.g., eye-wash station, shower, sinks, fire extinguisher, biological safety cabinet, first aid kit, emergency gas valve). Familiarize yourself with the location of safety equipment in the lab

46. Microbiology Lab Practices and Safety Rules As with unconventional glass, wrap the fragments in paper and seal them tight if you use plastic bags.  Dispose of broken glass in the broken glass container protect waste handlers from both physical and contamination hazards that they present. Dispose of razor blades, syringe needles, and sharp metal objects in the “sharps” container

47. Microbiology Lab Practices and Safety Rules Report spills and accidents immediately to your instructor Clean small spills with care (see instructions below). Seek help for large spills. Report all injuries or accidents immediately to the instructor No matter how small they seem.

48. Laboratory safety equipment

49. Safety shower Eyewash station Safety can

50. Causes of Contamination of Laboratory Samples and their Prevention