PHAR2812 prac manual 2019 PDF

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PHAR2812 prac manual 2019...

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PHAR2812 Microbiology and Infection

THE UNIVERSITY OF SYDNEY

School of Pharmacy Name: ........................ Group: .......................

Practical Manual 2019

Microbiology and Infection

Contents PRACTICAL SCHEDULE 2019 ................................................................................ 3 INTRODUCTION........................................................................................................ 5 PRACTICAL COURSE OBJECTIVES .................................................................... 5 COURSE ORIENTATION ........................................................................................ 5 LABORATORY RULES ........................................................................................... 8 ASSESSMENT ........................................................................................................ 10 PENALTIES ............................................................................................................ 13 SPECIAL CONSIDERATION ................................................................................ 13 PLAGIARISM ......................................................................................................... 13 SELF-HELP TUTORIALS ...................................................................................... 13 EQUIPMENT REQUIREMENTS ALL PRACTICAL CLASSES ......................... 14 EQUIPMENT REQUIREMENTS PRACTICAL CLASSES 6 – 10 ONLY............ 14 PRACTICAL 6 ........................................................................................................... 16 TOPIC 5: ASEPTIC PRODUCTION .............................................................................. 16 Exercise 6.1: Aseptic transfer ............................................................................. 18 PRACTICAL 7 ........................................................................................................... 24 TOPIC 6: FILTRATION, DISINFECTION AND PRESERVATION ..................................... 24 Exercise 7.1: Testing efficiency of filtration sterilisation with the microorganism E. coli (bacterial challenge test) .......................................................... 26 Exercise 7.2: Evaluation of preservatives ........................................................... 28 Exercise 7.3: Determination of MICs of antimicrobial agents ............................ 30 Exercise 7.4: Formulation compatibility and preservatives ................................ 32 PRACTICAL 8 ........................................................................................................... 34 TOPIC 7: PREPARATION OF PHARMACEUTICAL PRODUCTS ...................................... 34 Exercise 8.1: Preparation of zinc sulphate eye drops APF (Australian Pharmaceutical Formulary) ................................................................................ 36 Exercise 8.2: Preparation of ampoules of potassium chloride ........................... 38 PRACTICAL REPORT ............................................................................................ 41 APPENDIX 1: SELF-HELP TUTORIALS ............................................................. 42 APPENDIX 2: STANDARD OPERATING PROCEDURE (SOP) ...................... 53

PRACTICAL SCHEDULE 2019 Week Prac. Exercises

Follow Discussion up

Post-labs completed *

Self-help Tut

Microbiology component, Location: PC2 Lab 403, LEES Blg (F22)

2

3

4

5

6

1

2

3

4

5

Ex. 1.1 Macroscopic Examination of Microorganisms Ex. 1.2 Aseptic Techniques Ex. 1.3 Cultivation of Microbes from Air and Dust Ex. 1.4 Isolation and Identification of Normal Human Flora

1.1

Ex. 2.1 Preparation & Examination of Material 1.2 using Microscopy 1.3 Ex. 2.2 Bactericidal Effect of Ultra-Violet Radiation 1.4 Ex. 3.1 Isolation of Antibiotic-resistant Mutants of Bacteria Ex. 3.2 Antibiotic Sensitivity Testing Ex. 3.3 Bioassay of Antibiotics Ex. 4.1 Direct Contact Transmission of Microorganisms Ex. 4.2 Defences of the Skin

Ex. 5.1 Effect of Physical Factors on the Growth of Bacteria and Fungi Revision

1.2, 1.3, 1.4, 2.1

1.1

1.4 2.2

2.2

1.2, 1.3, Appendix 1 1.4 (Prac 2), Topic 2 2.1

1.4 3.1 3.2 3.3

3.1, 3.2, 3.3 2.2 1.4

4.1 4.2

4.1, 4.2, 5.1

Appendix 1 Topic 1

Appendix 1 Topic 3

1.4 (Prac 4), 3.1, 3.2, 3.3 To be Appendix 1 completed in Topic 4 Prac 5 4.1, 4.2, 5.1

Pharmacy component (First Group) Location: X-Lab, CPC Blg (D17) 7

8

6

7

Ex. 6.1 Aseptic Transfer (3 protocols)

Appendix 1 Topic 5

Ex. 7.1 Testing Efficiency of Filtration with the Micro-organisms Ex. 7.2 Evaluation of Preservatives 6.1 Ex. 7.3 Determination of MICs of Antimicrobial Agents Ex. 7.4 Formulation Compatibility and Preservatives

Mid-Semester Break

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Appendix 1 Topic 6

9

Ex. 8.1 Preparation of Zinc Sulphate Eye Drops APF Ex. 8.2 Preparation of Ampoules of Potassium Chloride

8

Mid- semester exam (all materials, including lectures and Appendix 1 practical labs Topic 7 from the first 8 weeks of semester)

7.1 7.2 7.3 7.4

Pharmacy component (Second Group) Location: X-Lab, CPC Blg (D17) 10

11

12

6

Lab Report (first group)

Ex. 6.1 Aseptic Transfer (3 protocols) Ex. 7.1 Testing Efficiency of Filtration with the 6.1 Micro-organisms Ex. 7.2 Evaluation of Preservatives Ex. 7.3 Determination of MICs of Antimicrobial Agents Ex. 7.4 Formulation Compatibility and Preservatives

7

Ex. 8.1 Preparation of Zinc Sulphate Eye Drops APF Ex. 8.2 Preparation of Ampoules of Potassium Chloride

8

7.1 7.2 7.3 7.4

13

6.1

Appendix 1 Topic 5

7.1, 7.2, 7.3 & 7.4

Appendix 1 Topic 6

8.1 & 8.2

Video Weeks Demo 10 and nstrati 11 on

1. Understanding sterile production 2. Understanding GMP

* Including exercises completely written up including all Results.

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Lab Report Appendix 1 (second group) Topic 7

INTRODUCTION PRACTICAL COURSE OBJECTIVES General Objectives: To introduce the student to the world of microorganisms and to provide familiarity and experience with the basic techniques, which are essential for the effective practice of pharmacy. Specific Objectives: 1. To train the student in the skills necessary for the handling of microorganisms in a safe and competent manner. 2.

To encourage the development of those manipulative and organisational skills that will be essential to the practice of Pharmacy. Particular emphasis is placed on aseptic handling of materials.

3.

To illustrate and reinforce the basic concepts of microbiology, which are examined in the lectures.

4.

To develop an understanding of the role of microbes in infectious disease.

5.

To be familiar with different sterilisation techniques and microbial agents

To implement these objectives the practical course is divided into seven topic areas: 1. Aseptic techniques for handling microbes and sterile objects 2. Isolation and identification procedures 3. Control of microbes: Factors influencing growth (e.g. environmental conditions, UV irradiation and antibiotics) 4. Host defence mechanisms 5. Aseptic transfer and sterilisation pharmaceutical products 6. Filtration, disinfection and preservation

COURSE ORIENTATION Practical Sessions: Week 2 to Week 6: The practical classes will run one session per week, starting in Week 2. You will have been allocated to attend either Monday 9 AM to 12 PM or Monday 2 to 5 PM or Wednesday 9 AM to 12 PM. YOU MUST ATTEND YOUR ASSIGNED PRACTICAL SESSION. Attendance details will be recorded. Week 7 to Week 12: Practical classes: Students will be divided into six groups. In weeks 7 to 9, the first three groups (A, B and C) will perform practicals 6, 7, and 8. From week 10 to 12, the remaining three groups (D, E and F) will perform practicals 6, 7, and 8. You will have been allocated to attend either Monday 9 AM to 12 PM or Monday 2 to 5 PM or Wednesday 9 AM to 12 PM.

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Video Demonstration: A video demonstration session will be on weeks 10 and 11 (Thursday 1 to 2pm). Contents will be assessed by an online quiz. All practical classes will begin with a brief introductory talk explaining: • • • • •

Specific safety warnings and precautions What work is to be completed during the practical class Rationale and theoretical considerations Location of materials Continuous assessment instructions

Therefore, it is important that all students are present and punctual for this prelaboratory talk. Feel free to ask questions to clarify any of the procedures described. Working Units Some experiments are performed on a per group basis. Each group of students should divide the workload among themselves ensuring that each member is completely familiar with what the others are doing. REMEMBER even if you are not responsible for performing a specific task in your group work, you will still be assessed on all tasks, so it is very IMPORTANT to understand all parts of the experiment. Make sure that results are exchanged and seen by all members of the group. Demonstrators Each bench of students will be allocated a demonstrator. Consult your demonstrator if you have any difficulties understanding the method or objective of an exercise. They are there to answer your questions. Planning In order to use laboratory time effectively, it is essential that the appropriate sections of the practical notes are read PRIOR to coming to class. Pre-work To ensure that you gain maximum benefit from the laboratory work and to encourage preparation for each practical class, pre-lab tasks have been set. These must be completed (and timestamped in LabArchives) prior to coming to class. Completion of Pre-work tasks will be assessed by your demonstrator. Post-lab Tasks To maximise your benefit from the laboratory work and to facilitate your learning outcomes, post-lab tasks have been set. These are indicated in the manual and must be completed prior to coming to the practical class immediately following the

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completion of the relevant aspect of each exercise. Completion of Postwork tasks will be assessed by your demonstrator.

Please note that on-time completion of Pre-work and Post-lab Tasks contribute to your Attendance, Preparation and Application of Knowledge (APAK) mark each week.

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LABORATORY RULES As potential practising Pharmacists, it is critical that you handle pharmaceutics carefully to avoid contamination. In the following laboratory classes you will be instructed with the correct aseptic technique to use to handle such sterile products. You will also learn how to handle microbes safely. COMPLIANCE WITH ALL LABORATORY RULES CONTRIBUTES TO YOUR ATTENDANCE AND PARTICIPATION MARK The following precautions MUST be observed for the safety of everyone working in the laboratory – any student NOT COMPLYING will be ASKED TO LEAVE. Protective Clothing and Dress 1. A laboratory gown/coat MUST be worn at all times in the laboratory. It should be put on and fastened fully on entry to the laboratory and removed on leaving to reduce the risk of contaminating your clothing. 2.

SAFETY GLASSES MUST be worn when undertaking any work in the laboratory.

3.

Substantial shoes with closed in toes MUST be worn. Thongs and sandals are unsatisfactory protection and should not be worn in the laboratory.

4.

Hair, if long, MUST be tied back.

Safety Requirements 1. DO NOT EAT, DRINK OR SMOKE in the laboratory and never place pencils, pens, labels or other materials in the mouth. 2.

MOUTH PIPETTING OF LIQUIDS IS BANNED. A rubber teat or filler must be used at all times.

3.

STORE ALL PERSONAL BELONGINGS INCLUDING BAGS, COATS, UMBRELLAS ETC. IN THE LOCKERS PROVIDED

4.

CULTURES ARE NEVER TO BE TAKEN OUT OF THE LABORATORY.

5.

Inoculated media must be PROPERLY LABELLED (i.e. with name, date and the nature of the specimen) and put in the appropriate box for incubation.

6.

DO NOT SIT on the benches.

7.

Turn gas burners off when not in use.

8.

Any personal accident must be reported to a demonstrator immediately.

9.

Any spillage of culture material must be reported to a demonstrator immediately so that appropriate action may be taken.

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10. At the end of each practical session: a. Discard all used tubes, pipettes, Petri dishes etc. in the designated receptacles. b. Clear the bench top of all equipment. c. Wipe bench top with F10 disinfectant and paper towel. d. Log out of your computer! e. Discard your gloves in the yellow biological waste bin. f. Remove your glasses and place in the appropriate area. g. Remove your gown and place in the laundry baskets provided. h. Move to the EXIT SINK area and wash your hands with the disinfectant soap supplied BEFORE leaving the laboratory.

11. Ensure that you know where the nearest emergency exit is.

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ASSESSMENT SUMMARY 1. Theory Examination (2 hours) 2.

60%

Practical (including mid-term quiz)

40%

Weeks 2 - 6 a. APAK* mark b. Mid-term Quiz (Week 8)

(5%) (15%)

Weeks 7 - 12 a. Attendance and Participation b. Practical Report c. Video Demo Quiz (Weeks 10 & 11)

(3%) (13%) (4%)

* Attendance, Preparation & Application of Knowledge The assessment for this course consists of the following components: 1.

Theory Examination The theory examination will take place in the examination period at the end of the semester.

2.

Practical Component

Weeks 2 - 6 (Coordinator - Dr Leona Campbell) a. Attendance, Preparation & Application of Knowledge (APAK) (5%) • • • • •

Arrive on time. Classes commence promptly at 5 minutes after the hour. Attendance at all practical sessions is compulsory. Attendance will be taken at each session. You must sign the attendance list to be considered present. Students are required to comply with the Laboratory Rules at all times. Attitude to work, effort and understanding will also be assessed. Pre-work tasks (for the current practical class) and Post-lab tasks (of the previous practical class) must be completed prior to attending your timetabled practical class. These will be checked by your demonstrator.

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CRITERIA FOR ATTENDANCE, PREPARATION AND APPLICATION OF KNOWLEDGE MARK Maximum possible mark for each practical session = 3 0 marks will be awarded if: Absent from practical without acceptable explanation OR present in practical BUT Pre-work and/or Post-lab tasks not attempted; student has careless, faulty or unsafe aseptic technique/biological containment; ANY safety OR laboratory rule not observed including engaging in inappropriate or unruly behaviour. 1 mark will be awarded if: Present but late to practical without acceptable explanation; Pre-work and Post-lab tasks poorly attempted ie. some answers vague or not done; Practical notes not up-to-date. 2 marks will be awarded if: Present and on time to practical; Pre-work and Post-lab tasks not fully completed or do not demonstrate thorough understanding of content; Practical notes not up-to-date. This is a Pass to Credit performance mark. 3 marks will be awarded if: Present and on time to practical; Pre-work and Post-lab tasks fully completed and demonstrate through understanding of content; Practical notes complete and up to date; observes ALL laboratory rules. This is a Distinction to High Distinction performance mark.

b. Mid-semester Exam (15%) A mid-semester exam will be held in Week 9, in your normal lecture time from 11am-12noon on Friday 3 May. This is a ‘closed book’ MCQ-style assessment. Questions in the mid-semester exam assess materials from the first 8 weeks of semester. This includes all lectures (weeks 1 to 9, including week 9) and practical lab classes (weeks 2 to 6) including theoretical background of practical classes as well as the results of each exercise. Consequently, revision will be greatly assisted by having a LabArchives practical manual that is complete and well recorded. A zero mark will be allocated to a student missing the mid-semester exam for other than a valid medical reason or as a result of serious misadventure. LabArchives Electronic Lab Notebook (ELN) Accurate and detailed results should be recorded directly into the spaces provided in your ELN. Each exercise should be completely written up by the practical class following the completion of the exercise (see Schedule) - this

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includes ALL Results, Discussion and Conclusions etc. Therefore your records of the practical work should be complete and up-to-date at all times.

Questions Question sheets are to be found with individual exercises. You should answer the questions at the time of writing up the exercise as they are designed to help you reach conclusions as well as to stimulate discussion.

Weeks 7 - 12 a. • • • • •

Attendance and Participation (3%) Classes commence promptly at 5 minutes after the hour. Attendance at all practical sessions is compulsory. Your attendance must be registered with the Instructor at each session. Students are required to comply with the Laboratory Rules at all times. Attitude to work, effort and understanding will also be assessed. Pre-lab tasks must be completed prior to coming to class. These tasks will be checked by your demonstrator during each practical session.

b. •

Practical Report (13%) The practical report must be submitted by 4:00pm, 7 days after completion of the last practical class (practical 8). A signed assignment sheet should be attached to the report. The 7-day due date is strict, and the report will not be accepted or marked once the due time has passed. The report should be carried out in group of two students.

c. •

Video Demonstration - Weeks 10 & 11 (4%) Short videos will demonstrate different aspects, requirements and techniques to prepare sterilised pharmaceutical products and Good Manufacturing Practice (GMP). After each film, the students have 4 days (by 5:00 pm on the first Monday following the Thursday workshop) to log into Canvas and complete an assessment (a series of multiple-choice questions) related to that film. The students are required to answer these questions without consulting other students. The allowed time and instructions to complete each assessment will appear on screen prior to the assessment.

•

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PENALTIES Absence from any assessable activity will result in a mark of zero being recorded for the task. If you miss an assessable activity because of certified illness or misadventure, you must apply for Special Consideration. IF YOU MISS A PRACTICAL CLASS • Inform your demonstrator of the reason for your absence and, submit a Special Consideration from the university online portal. Your demonstrator may be able to organise an alternative time for you to learn new practical skills. • It is your responsibility to obtain all information and results for the class and to fill in your Practical Manual. Where new techniques have been taught, you must take steps to learn the necessary skills. Make sure that you und...