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Evidence Guide: MSL975001A - Perform microbiological tests

Student: __________________________________________________

Signature: _________________________________________________

Tips for gathering evidence to demonstrate your skills

The important thing to remember when gathering evidence is that the more evidence the better - that is, the more evidence you gather to demonstrate your skills, the more confident an assessor can be that you have learned the skills not just at one point in time, but are continuing to apply and develop those skills (as opposed to just learning for the test!). Furthermore, one piece of evidence that you collect will not usualy demonstrate all the required criteria for a unit of competency, whereas multiple overlapping pieces of evidence will usually do the trick!

From the Wiki University

 

MSL975001A - Perform microbiological tests

What evidence can you provide to prove your understanding of each of the following citeria?

Receive samples and process associated request forms

  1. Check samples and request form details before they are accepted
  2. Return samples and request forms that do not comply with requirements to source with reasons for non-acceptance
  3. Log samples, recording details that allow accurate tracking and chain of custody
  4. Distribute samples for local testing or dispatch samples to other testing facilities
  5. Store samples appropriately where testing or transport is to be delayed
Check samples and request form details before they are accepted

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Return samples and request forms that do not comply with requirements to source with reasons for non-acceptance

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Log samples, recording details that allow accurate tracking and chain of custody

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Distribute samples for local testing or dispatch samples to other testing facilities

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Store samples appropriately where testing or transport is to be delayed

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare for safe microbiological work and aseptic applications

  1. Select work area and equipment required for the safe handling of materials that may contain micro-organisms of specified risk groups
  2. Wear protective apparel, replacing it when contamination is suspected
  3. Apply correct disinfection procedures to work areas before and after use
  4. Locate relevant emergency equipment for timely response to microbiological accidents
  5. Apply standard precautions when handling biological materials
  6. Minimise the production and release of aerosols, using biological safety cabinets where necessary
  7. Clean spills, and report all spills and suspected incidents to supervisor
  8. Wash hands before and after laboratory work and when contamination is suspected
  9. Ensure the safe disposal of biohazardous materials and other laboratory wastes in accordance with enterprise procedures
Select work area and equipment required for the safe handling of materials that may contain micro-organisms of specified risk groups

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Wear protective apparel, replacing it when contamination is suspected

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Apply correct disinfection procedures to work areas before and after use

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Locate relevant emergency equipment for timely response to microbiological accidents

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Apply standard precautions when handling biological materials

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Minimise the production and release of aerosols, using biological safety cabinets where necessary

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Clean spills, and report all spills and suspected incidents to supervisor

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Wash hands before and after laboratory work and when contamination is suspected

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure the safe disposal of biohazardous materials and other laboratory wastes in accordance with enterprise procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Process samples for direct examination

  1. Prepare thin smears of samples for subsequent staining to enable microscopic identification of cells
  2. Prepare liquid films of specimens for direct observation for motility or cell structure
  3. Prepare samples to concentrate material for subsequent staining or microscopy
Prepare thin smears of samples for subsequent staining to enable microscopic identification of cells

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare liquid films of specimens for direct observation for motility or cell structure

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare samples to concentrate material for subsequent staining or microscopy

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare pure cultures for microbiological work and aseptic applications

  1. Select culture media to maximise growth of micro-organisms and cells
  2. Inoculate media aseptically, applying techniques suitable for purpose of culture
  3. Incubate inoculated media in conditions to optimise growth of organisms and cells
  4. Subculture on suitable media to optimise production of pure cultures
Select culture media to maximise growth of micro-organisms and cells

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Inoculate media aseptically, applying techniques suitable for purpose of culture

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Incubate inoculated media in conditions to optimise growth of organisms and cells

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Subculture on suitable media to optimise production of pure cultures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Perform procedures that can assist in the identification of micro-organisms

  1. Select staining techniques to demonstrate required cellular characteristics
  2. Stain prepared films to demonstrate diagnostically useful characteristics
  3. Inoculate and incubate media with pure cultures to assist in the biochemical and immunological identification of micro-organisms
  4. Perform tests on pure cultures to assist in the biochemical and immunological identification of micro-organisms
Select staining techniques to demonstrate required cellular characteristics

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Stain prepared films to demonstrate diagnostically useful characteristics

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Inoculate and incubate media with pure cultures to assist in the biochemical and immunological identification of micro-organisms

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Perform tests on pure cultures to assist in the biochemical and immunological identification of micro-organisms

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Estimate the number and/or size of micro-organisms in samples

  1. Count cells in undiluted samples to indicate the dilution necessary to reliably count organisms in culture
  2. Prepare serial dilutions of samples aseptically for culture and colony counting
  3. Count colonies for calculating number of viable organisms per unit volume
  4. Count micro-organisms in samples and cultures using spectrometric and electronic methodologies, where relevant
  5. Estimate and document uncertainty of measurement in accordance with enterprise procedures, where relevant
Count cells in undiluted samples to indicate the dilution necessary to reliably count organisms in culture

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare serial dilutions of samples aseptically for culture and colony counting

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Count colonies for calculating number of viable organisms per unit volume

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Count micro-organisms in samples and cultures using spectrometric and electronic methodologies, where relevant

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Estimate and document uncertainty of measurement in accordance with enterprise procedures, where relevant

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Contribute to antibiotic sensitivity testing where required

  1. Prepare inoculum suitable for antibiotic sensitivity testing
  2. Dispense or position antibiotic discs as indicated by enterprise protocol
  3. Incubate inoculated media under conditions to maximise growth of cultured organism
  4. Read and record sensitivity reactions, noting phenomena that can assist in the correct interpretation of results
Prepare inoculum suitable for antibiotic sensitivity testing

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Dispense or position antibiotic discs as indicated by enterprise protocol

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Incubate inoculated media under conditions to maximise growth of cultured organism

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Read and record sensitivity reactions, noting phenomena that can assist in the correct interpretation of results

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain records of laboratory work

  1. Make entries on report forms or into computer systems, accurately calculating, recording or transcribing data as required
  2. Maintain instrument logs as required by accreditation checklists
  3. Maintain security and confidentiality of all clinical information, laboratory data and records
Make entries on report forms or into computer systems, accurately calculating, recording or transcribing data as required

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain instrument logs as required by accreditation checklists

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain security and confidentiality of all clinical information, laboratory data and records

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Assessed

Teacher: ___________________________________ Date: _________

Signature: ________________________________________________

Comments:

 

 

 

 

 

 

 

 

Instructions to Assessors

Evidence Guide

The Evidence Guide provides advice on assessment and must be read in conjunction with the performance criteria, required skills and knowledge, range statement and the Assessment Guidelines for the Training Package.

Overview of assessment

Critical aspects for assessment and evidence required to demonstrate competency in this unit

Assessors should ensure that candidates can:

safely perform tasks for the culture, isolation, identification and use of micro-organisms

not contaminate him/herself, other people, the work area, equipment or the samples or materials under test

not contaminate media or reagents during manipulations involving transfer of cultures

identify artefacts or image aberrations attributable to misalignment or obstruction of light paths or condensers used in bright field, dark ground, phase and fluorescent microscopy, or with other steps in microscopic examinations

be consistently accurate in the identification of Gram reactions

be consistently accurate in the description of bacterial colony forms on common media used in bacteriological investigations

preparedata and documentation that is accurate, concise and in accordance with enterprise requirements

report all incidents or accidents

disinfectany spillage and safely dispose of all contaminated materials

decontaminate the work area upon completion of work.

Context of and specific resources for assessment

This unit of competency is to be assessed in the workplace or simulated workplace environment.

This unit of competency may be assessed with:

MSL934002A Apply quality system and continuous improvement processes.

Resources may include:

a standard microbiology laboratory with relevant equipment, samples and reagents

enterprise procedures, test methods and equipment manuals

under duty of care requirements, off-the-job training providers will only use samples and organisms of a risk category compatible with their laboratory as defined in AS/NZS 2243.3.

Method of assessment

The following assessment methods are suggested:

review of results/data/records generated by the candidate

feedback from peers and supervisors to confirm that enterprise procedures are consistently followed and those results meet workplace requirements

oral and/or written questions associated with laboratory determinations and record keeping

integrated assessment with a case study focus, such as the isolation and identification of bacterial species in a specimen containing two or more species, by relating sample, cultural, morphological and biochemical data, and such from other relevant tests and procedures.

In all cases, practical assessment should be supported by questions to assess underpinning knowledge and those aspects of competency which are difficult to assess directly.

Where applicable, reasonable adjustment must be made to work environments and training situations to accommodate ethnicity, age, gender, demographics and disability.

Access must be provided to appropriate learning and/or assessment support when required.

The language, literacy and numeracy demands of assessment should not be greater than those required to undertake the unit of competency in a work like environment.

This competency in practice

Industry representatives have provided the case studies below to illustrate the practical application of this unit of competency and to show its relevance in a workplace setting.

Biomedical

A patient's urine sample and request form have been brought to the laboratory for urgent testing. After preparation of the work area, the technical officer examines a cover-slipped preparation of the sample and notes the presence of pus cells and non-motile rod organisms. In a Gram stain he confirms the presence of pus cells and Gram negative bacilli. They inoculate a MacConkey's and a blood agar plate for growth and isolation of bacteria. After consultation with the supervisor they are asked to set up a direct culture for antibiotic sensitivity testing. The supervisor informs the clinician of the initial findings. The next morning the technical officer assists the supervisor to read the plates. The predominance of lactose fermenting organisms is noted. The supervisor asks the technical officer to set up a biochemical panel to assist in identifying the organism. The supervisor confirms the technical officer's reading of the direct sensitivities plate. Later in the day the team is able to confirm that the patient's urine is infected with Escherichia coli and that the organism is sensitive to a number of antibiotics, including a sulphonamide and a cephalosporin.

Food processing

A swollen can of tuna was received at the company laboratory for microbiological investigation. The technical officer recorded the details supplied with the can and prepared for the investigation. A range of media, including cooked meat media and nutrient broth were prepared and aseptic can opening equipment was sterilised. After the can was opened in the biohazard cabinet, the state of the contents was recorded, pH checked and Gram stains prepared and examined. The media was inoculated with the food samples and incubated at a range of temperatures under aerobic and anaerobic conditions. The can was then emptied for double seam tear down to determine the cause of the spoilage. The next day the technical officer examined the media and broth cultures. From all the data collected the technical officer and supervisor were able to determine that pre-processing spoilage had occurred, probably due to excessive delays in the process prior to can sterilisation. The results were reported to production personnel so that they could follow up the circumstances relating to the delays, and ensure that the SOP had been followed and sufficient product rejected.

Required Skills and Knowledge

Required skills

Required skills include:

using protective clothing and biological safety cabinets

safely performing tasks for the culture, isolation, identification and use of micro-organisms

not contaminatingoneself, other people, the work area, equipment or the samples or materials under test

not contaminating media or reagents during manipulations involving transfer of cultures

identifying artefacts or image aberrations attributable to misalignment or obstruction of light paths or condensers used in bright field, dark ground, phase and fluorescent microscopy, or with other steps in microscopic examinations

Gram reactions

describing bacterial colony forms on common media used in bacteriological investigations

preparing documentation that is accurate, concise and in accordance with enterprise requirements

reporting incidents or accidents

disinfectingspillage and safely disposing of all contaminated materials

decontaminating the work area upon completion of work

Required knowledge

Required knowledge includes:

microbiological terminology, including, where relevant, that of bacteriology, parasitology, virology and mycology

disinfection and sterilisation as applied to practical aspects of microbiology

microbial diversity

micro-organisms of importance in medicine, in production of foods and other manufactured goods, and in assessment of the natural environment

cell biology and chemistry related to laboratory phenomena, such as growth and isolation of organisms for identification

microbial genetics

rationale for sample dilution when preparing materials for enumerating organisms and other pure culture work (e.g. Most Probable Number (MPN) technique)

need for accurate identification of sample source (e.g. body, specimen, process line and field location)

relevant health, safety and environment requirements

Specific industry

Additional knowledge requirements may apply for different industry sectors. For example:

Biomedical and biotechnology:

aspects of normal and abnormal anatomy, physiology, biochemistry and immunology as these pertain to the microbiological investigation of health and disease of animals and plants

interactions of micro-organisms with hosts

issues of pathogenicity

antimicrobial agents and antibiotic susceptibility/sensitivity testing

use of polymerase chain reaction (PCR) procedures in virology testing

handling of genetically altered cells

freezing and thawing of cultured cells

in tissue culture settings, maintaining the proper growth or storage conditions for the preservation of pure cell culture lines

maintaining the proper containment and preservation of genetically altered cell lines

use of micro-organisms in enzyme, vitamin, preservative and amino acid production

Biological and environmental:

sampling for the microbiological testing of drinking water which should conform to the guidelines published by the National Health and Medical Research Council (NHMRC) and the Australian Water Resources Council

testing procedures for the microbiological content of water which should be guided by advice of relevant national and state/territory environment protection agencies

aspects of ecology and other biological disciplines as these pertain to the microbiological investigation of the natural environment

use of micro-organisms in waste and toxic spill recovery

use of micro-organisms in site remediation

identification of micro-organisms to assist in determining the cause, time or nature of pollution

Food processing:

sampling and test batteries which should conform to relevant food standards code

aspects of food, pharmaceutical and other relevant processing as these relate to the involvement of micro-organisms in the production process and the microbiological monitoring of the production process

use of bacteria as probiotics

multiple resistant antibiotic strains of bacteria and their relevance to the food industry

importance of hazard analysis and critical control points (HACCP) to production processes

involvement of bacteria in food spoilage and poisoning

identification procedures for determining the source of a food poisoning event

limiting bacterial growth in foods and food preservation

Range Statement

The range statement relates to the unit of competency as a whole. It allows for different work environments and situations that may affect performance. Bold italicised wording, if used in the performance criteria, is detailed below. Essential operating conditions that may be present with training and assessment (depending on the work situation, needs of the candidate, accessibility of the item, and local industry and regional contexts) may also be included.

Codes of practice

Where reference is made to industry codes of practice, and/or Australian/international standards, it is expected the latest version will be used

Standards, codes, procedures and/or enterprise requirements

Standards, codes, procedures and/or enterprise requirements may include:

Australian and international standards, such as:

AS 2252 Biological safety cabinets

AS ISO 17025-2005 General requirements for the competence of testing and calibration laboratories

AS/NZS 2243.3:2002 Safety in laboratories - Microbiological aspects and containment facilities

ISO/TS 19036:2006 Microbiology of food and animal feeding stuffs - Guidelines for the estimation of measurement of uncertainty for quantitative determinations

ISO7218:2007 Microbiology of food and animal feeding stuffs - General requirements and guidance for microbiological examinations

cleaning, hygiene, personal hygiene requirements

enterprise procedures, standard operating procedures (SOPs) and operating manuals

guidelines for small scale genetic manipulation work from the gene technology regulations

incident and accident/injury reports

instructions to comply with new legislation, standards, guidelines and codes

quality system and continued improvement processes

safety requirements for equipment, materials or products and material safety data sheets (MSDS)

sampling procedures (labelling, preparation, storage, transport and disposal)

schematics, work flows and laboratory layouts

test procedures (validated and authorised)

waste minimisation, containment, processing and disposal procedures

Equipment, materials and systems

Equipment, materials and systems may include:

protective and physical containment facilities and equipment for safe handling of micro-organisms personal protective equipment, such as gloves, gowns, masks and safety glasses and gloves for working with extremes of heat and cold

carbon dioxide cabinets and incubators

transfer equipment, such as inoculating loops, pipettes (quantitative and qualitative), flasks, tubes and spatulas

liquid nitrogen containers for cell storage

filtration membranes

microscopes with bright field and other relevant illumination systems and stereomicroscopes

counting chambers for micro-enumeration

colony counting devices

Bunsen burners and bench incinerators

Incubators and water baths

anaerobic jars, fermentation chambers, continuous culture systems and other devices for controlling growth environments of micro-organisms

laboratory information management systems (LIMS), databases, record and filing systems

stains, media, reagents and biological materials necessary for laboratory testing

laboratory glassware and measuring equipment

disinfecting and sterilising solutions and equipment, such as ultraviolet (UV) lamps

materials suitable for the safe containment, collection, processing and disposal of biological and non-biological wastes

autoclaves

Communication

Communication may involve:

supervisors and managers (laboratory, quality and customer service)

personnel in other laboratories in the enterprise or in other enterprises to which work may be referred

customers, patients and clients

external auditors and accreditation agencies (e.g. National Association of Testing Authorities (NATA))

Occupational health and safety (OHS) and environmental management requirements

OHS and environmental management requirements:

all work will assume the potential infectivity of samples and materials presented for laboratory processing

all operations must comply with enterprise OHS and environmental management requirements, which may be imposed through state/territory or federal legislation - these requirements must not be compromised at any time

all operations assume the potentially hazardous nature of samples and require standard precautions to be applied

where relevant, users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council (NHMRC) and State and Territory Departments of Health