Assessment of Sanitation Practices

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Assessment of Sanitation Practices

• Michael S. Ramsey
• Teaching Laboratory Manager
• UCD
• mramsey_at_ucdavis.edu

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• Talk today assumes risk assessment and design and
  implementation of Winery Sanitation and Quality
  Program
• The link to my talk is still up but brutal to
  list
• Google Michael Ramsey and HACCP

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Objectives

• Discuss Quality
• Methods to Assess Sanitation Practices
• Other industries
• Non-microbiological
• Microbiological
• Cell swabbings
• ATP Bioluminescence

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What is Wine Quality?

• Quality... you know what it is, yet you dont
  know what it is.
• Robert Pirsig (1974) Zen and the Art of
  Motorcycle Maintenance pg 178

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What is Wine Quality

• All sectors of manufacturing debate the
  definition of quality
• To the ancient Greeks, quality meant truth
• They would not have understood our usage of the
  word in our modern wine making dilemma
• Aristotle said Quality (Truth) is not an act, it
  is a habit
• The Oxford English Dictionary (OED) defines
  quality
• the degree of excellence of the thing

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What is Wine Quality?

• Why isnt wine quality as easy to define as, say,
  golf ball quality?
• You cant fake quality any more than you can
  fake a good meal
• - William S. Burroughs

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What is Wine Quality?

• Example the ester, ethyl acetate
• Produced by some microbes that can be present
  during wine fermentation
• This character is found at high concentration in
  wine vinegar
• It is also a principle aroma compound of many
  nail polish removers.
• People who associate this character with nail
  polish remover tend to consider it quite
  objectionable in wines
• People who associate it with vinegar do not find
  it as unpalatable

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What is Wine Quality?

• Another example is rosemary character
• This herb character is found in soap products and
  commonly used in cooking.
• Individuals who associate the character with a
  soap find it quite offensive in foods and
  beverages
• Those who recognize it as the herb rosemary do
  not object to its presence.

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What is Wine Quality?

• As Pirsig implied, we all have different
  understandings of quality and that these
  understandings are inherently subjective

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What is Wine Quality?

• In other manufacturing sectors, quality is
  about minimizing variability and meeting clear
  specifications or zero defects
• OED definition of defect a shortcoming,
  imperfection, or lack
• Origin late Middle English (as a noun,
  influenced by Old French defect 'deficiency')
  from Latin defectus, past participle of deficere
  'desert or fail', from de- (expressing reversal)
  facere 'do'

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What is Wine Quality?

• If quality means zero defects then a
  defective wine is one that is purchased by your
  consumer in any condition other than how you
  intended for it to be in the bottle.
• Wine Quality is ultimately how you or your
  customer - define it.
• Quality means doing it right when no one is
  looking.
• Henry Ford

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• How does quality, then, impact ones assessment
  of my sanitation practices?

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Questions to Ask Yourself

• What should be the main emphasis of my definition
  of quality?
• What are the important areas of risk and
  sanitation in my winery to look into and why?
• How much risk am I willing to accept?
• Which method(s) of assessment are most suitable
  for my vision of Quality and Quality Assurance?

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• Traditionally, a combination of three methods are
  used to assess efficacy of sanitation methods
• Education and Training
• Inspections (visual assessments)
• Microbiological testing
• We are stuck with technology when what we really
  want is just stuff that works.
• Douglas Adams, The Salmon of Doubt (2002)

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Education and Training

• Requires continuous work due to employee turnover
• Which is rarely done

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Visual Assessments

• Limited
• Often, a lack of specificity (terms like
  satisfactory, acceptable, etc.) leaves the
  interpretation to the inspector, who may place
  too much emphasis on relatively unimportant
  matters and thus increase costs without reducing
  hazards.
• Not always reliable to detect sanitary conditions

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Microbiological Methods of Assessment in Food

• Molding surfaces with agar
• Staining surfaces and image analysis with
  epifluorescence microscope
• Bacterial impedance (i.e., BacTrac 4300)
• A sample is incubated and tested for changes in
  impedance in a specific medium

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Microbial Methods in Wineries

• Microbial Swabbing
• ATP Bioluminescence

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Microbiological Swabbing

• Apply sterile swab over 4 in. x 4 in. surface
  area
• Swab is transferred to sterile liquid vortexed
• Liquid is put on selective plates which allow
  microbes to grow

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Microbiological Swabs
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Microbiological Swabs

• Positive attributes
• Possible to quantify number and type of microbes
  present
• Can be used to trend sanitary condition of
  facility/equipment.
• Technology presumes there's just one right way to
  do things and there never is.
• Robert M. Pirsig, Zen and the Art of Motorcycle
  Maintenance (1974), p. 166

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Microbiological Swabs

• Microbiological testing also has some limitations
  
• Results not available for days
• TVC (Total Viable Counts) is not really total.
  It is only a measure of the fraction of the
  microbes able to produce colonies in the medium
  used under the conditions of incubation.
• Difficulties related to sampling
• Training, swab material, and technique

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ATP Bioluminescence

• Widespread use in dairy, food, and beer
  industries since the early 1990s
• ATP is short for adenosine triphosphate.
• This compound is the basic currency for energy in
  all living cells.

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ATP Bioluminescence

• The presence of this molecule may be assayed
  simply using an enzyme and coenzyme complex
  (luciferase/luciferin) found in the tail of the
  firefly, Photinus pyralis.

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ATP Bioluminescence

• The reaction is essentially the stoichiometric
  conversion of ATP to photons of light in the
  presence of the enzyme luciferase

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ATP Bioluminescence

• luciferin ATP O2 Mg2
• oxyluciferin AMP PPi CO2 light
• The breakthrough idea was to use ATP from all
  biological residues on a surface as an indicator
  of cleanliness, rather than try to ascertain the
  number of microorganisms present.

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ATP Bioluminescence

• The total ATP present on a swabbed surface can be
  extracted, assayed, and potentially uploaded to a
  computer database extremely rapidly (within five
  minutes).

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ATP Bioluminescence

• The light given off in this reaction is assayed
  as Relative Light Units (RLUs) a measure of the
  light produced by the ATPase assay relative to a
  control sample.
• RLU readings above the number you have chosen
  (based on the sum of your interferences) would be
  FAILevents in your cleaning/sanitation program.

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ATP Bioluminescence

• Since the results are relative, we can not
  compare results from one system to another.
• However, these assays are sensitive, direct,
  objective tests of cleaning efficiency and risk.

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ATP Bioluminescence

• Although the ATP bioluminescence assay is very
  rapid, it is not very specific.
• The results tell us nothing about the nature of
  the contaminant, whether it is juice, wine,
  random organic matter, microorganisms or
  combinations.
• There is no direct correlation between ATP
  results and microbial counts
• ATP amounts can vary widely between microbes.

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ATP Bioluminescence

• If your QA strategy is to provide quick and
  sensitive answers to your winery cleaning
  efficiency, or your bottling line sanitation
  requires zero contaminants regardless of the
  nature of that contamination, ATP Bioluminescence
  assays might just be for you.

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Issues With Both Methods

• Cell culture and ATP methods tend to under count
  remaining cells
• Training and technique
• There is a limit of detection with swabbing
• Depending on area swabbed and swab material
• All ATP-Bioluminescence systems also have a limit
  of detection
• No system can presently detect 100 of all ATP
  present.
• The lower the limit of detection, the more
  sensitive a system will be. Generally, LODs
  decrease as the price of the unit increases.
• You will need to decide how much sensitivity fits
  into your vision of quality and your budget.

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Conclusions

• A combination of three methods are used to assess
  efficacy of sanitation methods
• Education and Training
• Inspections (visual assessments)
• Microbiological testing
• For further discussion of ATP methods in the
  winery, refer to Ramsey, M. S. and Mills, D. A.  
  2013.  Winery Biofilms  A source for
  contamination throughout the winery.  Practical
  Winery and Vineyard Journal.  Winter Issue.

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Thank you

• Making an art out of your technological life is
  the way to solve the problem of technology.
• Robert Pirsig,1974