????? ?????

1
????? ?????ß?a?? ?????daHeterotrophic Plate
Count

• ??. ??????? ?et??????

2
???sµ?? ?a? S??p????s?µ? ?e??

• ? ?.?.?. Te??e? ?t? p?s?µ? ?e?? p??pe? ?a e??a?
• ?at?????? ??a a????p??? ?ata????s? ?a? ??a
• ????? t??? s????e?? ????a???? s??p???
• pe???aµß????ta? ?a? t?? p??s?p??? ???e???.

3

• ?a p?s?µa ?e?? ?a p??pe? ?a e??a? asfa?? ??a
  ???s? ?at? t?? d????e?a t?? ???? t?? a????p??,
  ?aµß????ta? ?p??? t?? d?af??et???? e?a?s??s?e?
  p?? s?µßa????? µ?sa ap? ta st?d?a t?? ???? t??,
  a??? de? e??a? ?at a??????, ??a ta ?e??
  ?at?????a ??a ta ?t?µa p?? ta?a?p?????ta? ap?
  s???e???µ??e? d?ata?a??? t?? a??s?p???t????.

4
???t?? pa????? p?s?µ?? ?e???

• ??p??? eµpe????e? t?? p??? ??t??s??, epe?e??as?a?
  
• ?a? d?a??µ??.
• ?? te?e?ta?? µp??e? ?a pe???aµß??e? ß????t????
• s?s?e??? se ????a?? ? ?d??µat??? ep?ped?, ?p??
• ap?s??????t??, s?s?e?? e?e???? ????a?a,
• a?t?µata µ??a??µata s?s?e??? (ae?????) ?t?.

5
Heterotrophic plate count (HPC)(?????? a???µ??
ete??t??f???? ßa?t?????)

• ?a ete??t??fa p??sd???????ta? ?? µ???????a??sµ??
  p?? apa?t??? ???a???? ????a?a ??a t?? a??pt???
  t???. ??t? pe???aµß?????, ßa?t???a, ??µ?µ???te?
  (yeasts) ?a? µ????a (mould).

6

• Heterotrophic plate count (HPC) (????µ?s?
  ete??t??f?? ßa?t?????) pa?a??t??a ?ta? ???st? ??
  Standard plate count (p??t?p? a???µ?s?
  ete??t??f?? ßa?t?????) e??a? µ?a d?ad??as?a ??a
  t?? e?t?µ?s? t?? ???ta??? ete??t??f?? ßa?t?????
  st? ?e??.
• ? µ???d?? µp??e? ?a p??sf??e? ???s?µe?
  p????f???e? ??a t?? p???t??? ?at?stas? t?? ?e???.
• ? ?e?µ???as?a p?? ???s?µ?p??e?ta? ??µa??eta?
  pe??p?? 20?C µ???? 40oC.
• ?????? ep?as?? ??µa??eta? ap? ???e? ??e? se 7
  ?µ??e? ? ?a? µe????? eßd?µ?de?.
• T?ept??? ????? pt??? ? p???s?? se pe??e?t???t?ta
  se s?stat???.
• ? ?d?a ? µ???d?? de? ?a?????e? t??? ???a??sµ???
  p?? a????e?e?.

7
?????ß?a?? a??pt??? st? ?e??

• ????????a??sµ?? a?apt?ss??ta? ?a?????? st? ?e??
  ?a? st?? ep?f??e?e? µe t?? epaf? t??? st? ?e?? ??
  ?µ????.
• ?et? t?? epe?e??as?a t?? p?s?µ?? ?e??? ? a??pt???
  p?? a??????e?ta? a?af??eta? ?? epa??pt???.
• ???pt??? a?t??at??p??eta? s?????? st?? ?????te?e?
  t?µ?? HPC se de??µata ?e???.

8

• ????µ??a ep?peda HPC pa???s?????ta? e?d??? se
  st?s?µa tµ?µata t?? s?????se?? s?st?µ?t??
  d?a??µ??
• ?d?a?????? e??atast?se?? sp?t???
• ?µf?a??µ??a ?e??
• F??t?a ????a???
• ??t?µata µ??a??µata
• ?? ?????? ?a????st???? pa?????te? t?? epa??pt????
  e??a?
• Te?µ???as?a
• ??a?es?µ?t?ta ??ept???? ??????
• ???e??? ?p??e?µµat???? ap???µa?t????
• T?ept??? s?stat??? µp??e? ?a p???????? ap? t?
  ?d?t??? pe??ß????? ?a? ? ta ????? p?? ?????ta? se
  epaf? µe t? ?e??.

9
???s? t?? HPC st?? d?a?e???s? t?? ?e???

• ?? ?et??se?? t?? HPC ???s?µ?p?????ta?
• G?a ?a de?????? t?? ap?te?esµat???t?ta t??
  d?ad??as?a ?ate??as?a? t?? ?e???, ?ts? ?? ?µes?
  ??de??? ap?µ?????s? pa??????? µ???????a??sµ??.
• O? µet?? t?? a???µ?? epa??pt???? ???a??sµ?? p??
  µp????? ? de? µp????? ?a ????? ???e??? s?µas?a
  ?a?
• O? µet?? p??a??? pa??µßas?? µe µ?t??s??
  ????ßa?t???d??? st?? µe??d??? ?a??????e?a se
  Lactose- base . ? efa?µ??? a?t? ??e? µe??µ???
  a??a ?a??? ? ?a??????e?a se ????? Lactose-base
  a?t??atast????e ap? e?a??a?t???? µe??d??? p??
  e??a? e?e??e?? ?a?t???.

10
S??d?a asf??e?a? t?? ?e???

• ?ate????t???e? ??aµµ?? ??a t?? p???t?ta t??
  p?s?µ?? ?e??? (???)
• ?? ßas???? st???? t?? ??e?a? ßas????ta? st??
  p??stas?a t?? ??µ?s?a? ??e?a? ?a? t?? p???????
  t?? as?e?e???
• ????????s? t?? s?st?µat?? ??a ?a ?a????ste? a? ?
  a??s?da ef?d?asµ?? ?e??? (µ???? s?µe???
  ?ata????s??) st? s????? t?? µp??e? ?a pa???e?
  ?e?? p???t?ta? p?? p???? t??? ?a????sµ?????
  st?????.
• ?a?a???????s? t?? stad??? t?? a??s?da? p?? ?????
  ?d?a?te?? s?µas?a ??a t?? e?asf???s? afa????
  p?s?µ?? ?e???.

11

• S??d?a d?a?e???s?? ??a t?? te?µ????s? t??
  s?st?µat?? a???????s?? ?a? pa?a???????s??, ?a?
  pe????af? e?e??e??? p?? p??pe? ?a a?a??f???? ap?
  ?a??????? s?????e? se a??a?e? pe??pt?se??
  s?µpe???aµßa??µ???? t?? te?µ????s?? ?a? t??
  ep????????a.
• S?st?µat??? a?e???t?t? ep?pte?a p?? ep?ßeßa???e?
  ?t? ta a??t??? ?e?t??????? s?st?.

12
?p??e?? ??µ?s?a? ??e?a? t?? ????? t?? HPC

• ?st?????? ap??e??
• Robert Koch 1883
• ??af??? µe t?t?? ??? µ???d?? a????e?s??
  µ???????a??sµ?? st? ?daf??, a??a ?a? ?e??
• ? a?af??? ???e se ????? p?? ? ßa?t???a??
  p????e?s? t?? ?????a? de? ?ta? a??µ? ???st?.
• ? e?sa???? µe??d?? ap?te???µe?? ap? 1ml ?e??? se
  ??ept??? ?????, ?a???e??e?ta? se p??t?µa se
  ?e?µ???as?a 22?C ??a 48h. ?a? µet???µe t?? a???µ?
  t?? ap?????
• ? ???d???? t?? ??e?a? s??d?eta? µe t?? pa???s?a
  µ???????a??sµ?? st? ?e??.
• ? Koch p??e?d?p???se ?t? ?p???e? ???d???? ?e?d???
  ap?te??sµat?? ßas???µe??? µ??? st? µ?t??µa t??
  ap??????.

13

• ????? a?t? t?? st??µ? de? ?p???e? e??ast???a??
  µ???d? p?? µp??e? ?a ep?t???e? t?? a????e?s? t??
  ßa?t?????, ?a? ? Koch de? µp????se ?a a?a?????se?
  t?? pa???s?a t?? pa??????? µeta?? t??
  p??????µ??f?a? t?? ?dat?????? ßa?t?????.
• Te??e? ded?µ???? ?t? t? ep?ped? t?? pa??????? st?
  ?e?? ?ta? ?aµ??? s?et??? µe ta ep?peda t?? ?????
  ßa?t?????.
• ?? 1893 ed?µ?s?e?se e??as?a d????s? ?e??? ?a?
  ?????a ?p?? ep??e ?e?? ap? t? µ???sµ??? p?taµ?
  Elbe ?a? t? ?p?ßa??e se epe?e??as?a ap? t?
  d????st???? a???? ?µµ??, te?µ???????ta? p?s?
  ?aµ??????ta? t? ep?ped? t?? ?dat??e??? ßa?t?????,
  ??t? t?? 100 ap?????? / ml µp????? ?a
  p???aµß????? t?? ep?d?µ?a ?????a? ?a? t?f? e?d??
  p??et??. ?ta? a?t? t? ep?ped? a????eta? p??? ap?
  1000 p??????e? / ml ?fe??eta? se a?epa??? /
  p??µµe?? d????s? t?? µ???sµ???? ?e???, µ?a
  ep?d?µ?a ?????a? p????pte?.

14

• Colony count apede?????e ??a? e?a??et???? de??t??
  t?? ?e?t??????? t?? d????s??.
• S?st?µat???? pa?at???se?? µet? t? 1983 ?de??a?
  ?t? ? d????s? ap? ?µµ? µp??e? ?a p????ße?
  ?dat????e?? ???µ??e?? e?? 1 ml ?e??? de?
  ep?t??peta? ?a pe????e? pe??ss?te?? ap? 100 ta
  ?p??a a?apt?ss??ta? se ??ept??? ????? st??? 20?C
  ??a 48h
• St?? Ge?µ???a, ? pe?????sµ?? t?? 100 ßa?t????? /
  ml te???? ????e p??t?p? ??a t? ß?ad????st????
  ?µµ?? ?a??? ??a a??? d????se?? ?a? µ???d??
  ap?ste???s?? (Garter 1915)
• ?e?????sµ?? t?? ?e????? ßa?t????? µ??? ?a? de?
  a?af??eta? ??a pa?????a ßa?t???a ??
  Salmonellatyphi Vibrio ?a? cholera.

15

• ? pe?????sµ?? t?? 100 ßa?t????? / ml ?ta? t?
  p??t? ep?d?µ???????? ßas??? ???? ??a t?? d????s?
  t?? ?e??? ap? µ???sµ??e? p??e?, a?t? ep?t??pe?
  st? ??e??? t?? ?ast?e?te????? ???µ??e??, ?p?? ?
  ?????a, t?f?e?d?? p??et?? shigellosis.
• ???a? s?µa?t??? ? pe?????sµ?? t?? 100 colony
  forming units (cfu) / ml de? ??e? ?µes? s??s? µe
  p??a?? ???d??? ??e?a?, µ?????, a?ta?a??? st??
  ap?te?esµat???t?ta t?? µe??d?? d????s??, µe a?t?
  t?? ??????, µ??? ?µµes? s??s? e??a? a?t? p??
  ?aµ????e? t?? ???d??? t?? ???µ????, ?????? ??a
  t?? ?ast?e?te????? ???µ??e?? ?? ?p??e? ap??t??ta?
  µe ?at?p?s?.

16
Te?µ???as?a ep?as??

• G?a t?? ?ataµ?t??s? t?? ete??t??f?? ßa?t?????
  ???s?µ?p????µe 2 ?e?µ???as?e?
• 20?C (?e?µ???as?a d?µat???) ?a? t?? 37 ?C
  (?e?µ???as?a s?µat??).
• ? ?e?µ???as?a t?? 37 ?C d??e? t?? e??a???a sta
  ete??t??fa (a???????a) ßa?t???a ?a
  p???ap?as?????ta? ta????.
• St?? ?e?µ???as?a t?? 20?C p???ap?as?????ta? ta
  ß?ad??? a?apt?ss?µe?a (a?t?????a) ßa?t???a.
•  
• ? Ge?µa????? ?a????sµ?? apa?te? ? a???µ?s? t??
  ap?????? (colony count) ?a e?t?µ?te µet? ap? 48
  hr a??pt???? se ?e?µ???as?a 20 ?C ?a? / ? 37 ?C
  ?a? ? a???µ?? t?? ?a e??a? µ???? 100 cfu / ml st?
  ?e?? t?? ß??s?? (German DrinKing Water
  Regulation)

17

• ? a???µ?s? t?? ap?????? µ??e? t??? de?
  ep?t??p??? ?a e???e? ?p???d?p?te s?µpe??sµata
  s?et??? p??? t?? ???d??? t?? ???µ????. ??????,
  a?t? e??a? ??a µet?? s?????s?? ??a
• ??? ap?te?esµat???t?ta t?? d?ad??as?a? d????s??,
  ?p?? t?? d????s?? µ?s? ß?ad??? ?µµ?? ?a?
  s?µe?a??? d????s?? (sp?t???? f??t???)
• ??? ap?te?esµat???t?ta t?? ap???µa?s??.
• ?? ep?ped? ßa?t???d??? se pe????? µe µ?a a???µ???
  p??a?? µ????s?.
• ???sta?e??t?ta st? ?d?a????? s?st?µa t?? sp?t???.

18
???sfate? e?e???e??

• ??e?? pa?????te? ????? ep??e?se? t?? p??sfate?
  e?e???e?? st? ped?? t?? a???µ?s?? t?? ap??????
  ??a t??? s??p??? t?? e?t?µ?s?? t?? ???e????, t??
  p???t?ta? t?? p?s?µ?? ?e???
• ? ße?t??µ??? ??ept??? s???es? t?? ??a?, ? ?p??a
  ?p?st????e? t?? a??pt??? e??? e???te??? f?sµat??
  ?dat??e??? ßa?t?????
• ? a?a?????? sta t??? t?? de?aet?a? t?? 1960, t??
  ß??µ?ß?a??? sta ?p??a ? p?e????t?ta t?? pa???t??
  µ???????a??sµ?? e?a????????? ?a d?at?????ta? ?a?
  ta ?p??a ?a???????? t? ep?ped? t?? e?e??e???
  d?a??µa?s?? st? ?e??.
•  
• ??e? d?ad??as?e? p?? ep?t??p??? t?? ta?t?p???s?
  e??? e???te??? f?sµat?? ?dat??e??? ßa?t?????.

19
???? ????d?? ??a p??sd????sµ? a???µ?s?? t??
ap??????

• ???? ??a µ???? ???sµa (pe??p?? 0,1) t??
  ?dat??e??? µ???????a??sµ?? a?????? st?? ?µ?da t??
  ?a???e???s?µ?? ete??t??f??? ßa?t????? ?a? pe??p??
  1 ap? ta ß??s?µa ßa?t???a de? e??a?
  ?a???e???s?µa.
• ???s?µ?p???s? ???? µe??d?? a????e?s?? ßa?t?????
  ta ?p??a µp????? ?a ?a???e??????? ap? t? p?s?µ?
  ?e?? d?µ?????????e t? ??ept??? ????? R2A
  (Reasoner and Geldreich 1985)

20

• To R2? ???? ?aµ???? pe??e?t???t?ta? se ??ept???
  s?stat??? (a?tap?????eta? ?a??t??a st?? a????e?
  t?? ?d?t???? µ?????????da?) ta ßa?t???a
  a?apt?ss??ta? ß?ad?te?a, s??d?a??µe?a de µe t??
  ?aµ??? ?e?µ???as?a 20?C ?a? µe t?? pa?atetaµ???
  ep?as? ap?d?se t? µe?a??te?? a???µ? ap?µ????µe???
  µ???????a??sµ?? (Reasoner and Geldreich 1985)
• ??a µe?????t?µa t?? µe??d?? e??a? ? pa?atetaµ????
  ?????? ?a??????e?a? (5-7 ?µ??e? st?? 28?C) ?p??
  µe???e? t?? a??a t?? pa?aµ?t??? ??a t?? µ?t??s?
  t?? ap?te?esµat???t?ta? t?? d?ad??as?a?.

21
???d???? ap? a????e?s?µa ßa?t???a st? ?e??

• ??a? a???µ?? µe?et?? ??e? d?s? ??s?ast??? t? ?d??
  ?a?a?t???st??? f?sµa ap? ete??t??fa ßa?t???a??
  ste????. ?a ep???at?ste?a e?d? s a?t? t? f?sµa
  e??a?
• Acinetobacter spp, Aeromonas spp, Comamonas spp,
  Enterobacter spp, Flavobacterium spp, Klebsiella
  spp, Moraxella spp, Pseudomonas spp, Sphingomonas
  spp, Stenotrophomonas spp, at?p??? Mycobacterium
  spp, Bacilluw spp, and Nocardia.

22

• ?e??te? ap? Norton and Le Chevallier (2000)
  ?de??a? ?a?a?t???st???? a??a??? st? ßa?t??????
  p????sµ? µ?s? t?? epe?e??as?a? ?a? ?ata??µ?? t??
  p?s?µ?? ?e???, ep?µ????, e??a? apa?a?t?t? ?a
  e?asfa??s??µe ?t? ? epe?e??as?a t?? ?e??? ?a? ?
  ?ata??µ? t?? ?a µ? p???a??s??? a??a?? t??
  s???es?? t?? ßa?t???a??? p????sµ?? p?? ?a
  e????s??? t?te ta e??a???a?? pa?????a.
• ?a ep???at?ste?a ???? st? sp?t?, ?????? t??
  ?est?? ?e??? st? s?st?µa ?ata??µ??, e??a?
  legionella ?a? Pseudomonas aeruginosa. ? eµf???s?
  µ???sµat???? ste?e??? µ???t??, ?p?? Fusarium spp
  ?a? Aspergillus spp st? ????a?? s?st?µa (Anaissie
  et al 2001)

23

• ?a ep???at?ste?a ???? st? sp?t?, ?????? t??
  ?est?? ?e??? st? s?st?µa ?ata??µ??, e??a?
  legionella ?a? Pseudomonas aeruginosa. ? eµf???s?
  µ???sµat???? ste?e??? µ???t??, ?p?? Fusarium spp
  ?a? Aspergillus spp st? ????a?? s?st?µa (Anaissie
  et al 2001)
• ?e? ?p???e? safe?? e?de??e?? ?t? ta ete??????a
  ßa?t???a e?????? ???d??? ??a t?? d?µ?s?a ??e?a,
  ?????? ?ta? p??s?aµß????ta? ap? ta ???? ?t?µa
  µ?s? p?s?µ?? ?e??? (Rusin etal 1997, Colford et
  al 2002)
• ????????s? ???d???? ap? t?? Rosin et al 1997 se
  e?e???t?? ap?fe?e ?t? d?se?? ap? t?? st?µat??
  d?af???? µ???????a??sµ?? e??a? apa?a?t?t? ?a
  p???a??se? µ????s?
• Pseudomonas aeruginosa, 108-109 cfu
• Aeromonas hydrophila, gt1010 cfu
• Mycobacterium avium, 104-107 cfu
• Xanthomonas maltophilia 106-109 cfu

24
??at??e?? ??a t?? p???t?ta t?? ?e??? se pe??????
?????? ???d????

• ??aµat??? a???s? t?? ???µ??e?? p?? p???a????ta?
  ap? µ???????a??sµ???, µe????? ap? a?t??
  ?fe????ta? se µe?????? ete??t??f???
  µ???????a??sµ??? (Huang et al 2002)
• ??d??? p??s??? p??pe? ?a d??e? st?? e?asf???s?
  t?? apa?a?t?t? p???t?ta ?e??? se pe?????? ??????
  ???d????, ?p?? ??s???µe?a ?a? µ??? ?p??
  a??s??atesta?µ???? as?e?e?? ?????? ?e?ape?a
• Se a?t?? t?? pe??????, t? HPC ???s?µ?p??e?ta? ap?
  µe????? a???? ??a ?p?de??? t?? ???d????, t??
  pa???s?as?? e??a???a??? pa??????? (Hargreavew et
  al 2001)
• ? p??sd????sµ?? pa??????? µe HPC se µe?????
  pe?????? (Pseudononas spp, Bur?holderia,
  Stenotrophomonas) e?f???e? ???µ??? ??????
  ???d???? ??a e?pa?? ?t?µa

25
?????? ???s? t?? HPC

• ?p?s?µ? ?d???a s?et??? µe t?? ßa?t??????????
  e??tas? t?? ?d??d?t?s?? ?a? t?? e?µ??e?a t??
  ap?te?esµ?t?? d?µ?s?e????a? ??a p??t? f??? ap? t?
  ?p????e?? ??e?a? t?? ???µ. ?as??e??? t? 1934
  G??st? ?? Report 71.
• ? p??te???µe?? µ???d?? ?a? ? d?a??µ? 1ml ?e???,
  ?a? a??µe??? µe ??ept??? a?a?. ?a? ep?????ta? ta
  t??ß??a se
• 20-22C ??a t?e?? ?µ??e?
• 37C ??a d?? ?µ??e?
• ? a???µ?? t?? ßa?t????? ? ?p???? apa???µe?ta?
  st??? 20-22C e?p????e ?t? d?de? ??p??a ??de???
• ? p?s?t?ta t??f?? p?? d?at??eta? ??a t?? d?at??f?
  t?? ßa?t?????
• ? p?s?t?ta ??µat??, s????? ?a? ????? ????? ??????
  p?? ????? ap??t?se? p??sßas? st? ?e??
  ?aµß????ta? ?p??? ?t? ? ?ataµ?t??s? se 37C
  pe????e? pe??ss?te?e? p????f???e? s?et??? µe t??
  ep????d??? ??pa?s?.

26

• ?a??? ? a??pt??? t?? ???a??sµ?? st?? ?e?µ???as?a
  t?? 37C e??a? ?????? a?t??, t?? ed?f???, t??
  ??µ?t?? ? e?te????? p????e?s?? ?a? ? a???µ??
  t???, ep?µ???? µp??e? ?a ???s?µ?p????e? ??
  de??t?? t?? ßa?µ?? ?a?a??t?ta? t?? ?e???.
• ? ???es? a?af??e? ep?s?? ?t? ? ?ataµ?t??s? t??
  ap?????? ap? ??a µ??? de??µa e??e s?et??? µ????
  s?µas?a ?a? ?t? e??a? d?s???? ?a ?a????se? ta
  ???a, e?? ?pe?ßa?????, eµpe????e? d?sµe?? s????a
  s?et??? µe t?? ???e??? p???t?ta t?? ?e???.
• ? ????? t?? a???µ?? st??? 22C se a?t??? t?? 37C
  e?p????e ?t? e??a? ???s?µ?? ??a t?? e????s? t??
  a?f??d??? d?a??µ??se??, µe ????? a?a????a
  s??d?eta? µe ßa?t???a ap? ?a?a?? ?daf?? ? ?e??
  p????e?s?? sap??f?t???? µ???????a??sµ?? ?a?, ?at?
  s???pe?a, µ????? s?µas?a?.

27
?d???a ???s?? t?? HPC

• ????? a???µ?s? ap?????? se aµf?te?e? t??
  ?e?µ???as?e? a??µ? ?a? a? ep?ßeßa??????, de?
  a?t??at?pt?????? ?at a????? ?t? t? ?e?? ap?te?e?
  ???d??? ??a t?? ??e?a.
• ?? ??e?d? t?? µet??µat?? t?? ap?????? t??
  t??ß???? ?ta? ? e?t?µ?s? t?? ap?te?esµat???t?ta?
  t?? d?ad??as?a? epe?e??as?a? t?? ?e??? pa?????ta?
  µ?a e?t?µ?s? t?? ?e????? ???e???? p???t?ta t??
  ?e???.
• ?? Report 71 a?af??e? ep?s?? ?t? ? a???s? t??
  ?????? a???µ?? ap?????? µp??e? ?a d?se? t?? p??t?
  ??de??? t?? ??pa?s??.
• ?? Report 71 (st?? 4? ?a? 5? ??d?s?) epa???aße
  ?t? ?? µet??se?? ap?????? de? e??a? s?µa?t????
  ??a t?? a???????s? t?? asf??e?a? t?? ????a???
  ?e??? ?d?e?s??.

28
??µ??e?a t?? ep?p?d?? HPC

• ? a?af??? (report) a?a??????e? ?t? µe????
  e??a???a?? pa?????a (p.?. Pseudomonas aeruginosa
  and Aeromonas sp) µp??e? ?a e??a? µ???? t??
  ?????? a???µ?? ap?????? ?a? ?? eµfa??se?? t???
  se µe?????? a???µ??? st? ?e?? de???e? ?t? ??
  s?????e? st? s?st?µa d?a??µ?? ????? ???e?
  ?at?????e? ??a t?? a??pt??? t???, se a?t??es? µe
  t?? ep?ß??s? t?? e? ???? ???a??sµ??. Ost?s?,
  ?at????e st? s?µp??asµa ?t? ????? ap?de??e??
  ??p?a??d??? µ????s??, ?????? µet??se?? ap??????
  de? p??pe? ?a a?t?µet?p????ta? µe a??s???a ?s??
  af??? t?? ??e?a t?? p????sµ?? st? s????? t??.

29

• ?a?t??? ?ataµ?t??s? t?? a???µ?? ap?????? ap? ??a
  s?st?µa d?a??µ??, ?st?s?, pa???e? ???s?µa
  st???e?a µe ta ?p??a a?????????ta? ??e? ??
  µa???p???esµe? t?se?? st? ?e???? µ????ß????????
  p???t?ta t?? p?s?µ?? ?e???.
• ?? ?a? de? ????eta? ap? t?? ????pa??? ?d???a, ?
  ?ataµ?t??s? t?? ap?????? st??? 37C, d?at??????e
  ap? t? ??µ??es?a t?? ???µ. ?as??e??? ?a? ??e?
  µe???? a??a µp??e? ?a p??sf??e? ???a??? d?????s?
  µ?a? s?µa?t???? ep?de???s?? t?? p???t?ta? t??
  ?e??? p??? ßa?t???d?a ? ????? ßa?t???a??? de??te?
  a????e?????.
• ????pa??? ?d???a ??a t? p?s?µ? ?e??
• 10/ml st??? 37C
• 100/ml st??? 22C

30
?p???e?t???? ?????? a???µ?? ap?????? st??
????p?(????et????e ap? t?? Muller 1977)
31
(No Transcript)
32
?p?d?µ??????a ?a? ?a??f?s???????? t??
a??s??ate?staµ????

• ???e? µe??te? d????? st???e?a ??a t?? ep?pt?s? ?
  ep?p??asµ? ??a a??s??ate?staµ????? as?e?e??.
• US EPA 2000 (US Environmental Protection Agency)
  d??e? µ?a p??s????s? t?? ep?p??asµ?? t??
  a??s??ate?staµ???? as?e???, p?? e??a? pe??p??
  0,83 t?? ?e????? p????sµ??.
• Ta p??pe? ?a a?aµ??eta? ?t? ? ep?p??asµ?? ?a
  a????e? pe?a?t???, ap? t?? a???s? t?? ep?ß??s??
  t?? as?e??? ap? ?a????? ?at? t?? d????e?a t?? d??
  te?e?ta??? de?aet??? t?? a???s? t?? ??tas?? t??
  ??µe???e?ape?a? ?at? t?? ?d?a pe??pt?s?.
• ????s? t?? p?s?st?? µetaµ?s?e?s?? a?µ?p???t????
  ß?ast???? ??tt????.

33
???µ??e?? ap? HPC ???a??sµ??? st? p?s?µ? ?e??
µeta?? t?? a??s??atesta?µ????

• ? p??ta????? ?????a t?? e?????? t?? ???d???? ap?
  t? p?s?µ? ?e?? ??a a????p??? ???s?
  (pe???aµßa??µ???? t?? efa?µ???? t???, ?p?? p?s?µ?
  ?a? ?t???) ßas???µe?e? st?? ep?d?µ?????????
  µe??te? ?a? st?? a???????se?? t?? ???d???? p??
  af????? e?a??et??? µ???sµat????? pa?????te?,
  ?a??? ?a? ??a ?a?????? p????sµ?. ? a??a??µe???
  a???µ?? t?? a??s??atasta?µ???? as?e???, ?st?s?,
  ?a??st? a?a??a?a t?? a??pt??? ???? e?????? ??a
  t?? p??stas?a t?? as?e??? a?t?? ap? µ???sµat?????
  pa?????te? st? p?s?µ? ?e?? ?a? s??af??
  e??atast?se??.

34
???d???? ???µ???? ??a a??s??atesta?µ????? as?e?e??

• ??d???? pe??pt?se?? ???d???? ap? t? p?s?µ? ?e??,
  se a??s??atesta?µ????? as?e?e?? e??a? ap?
• ???a?a ?at?p?s? ?at? t?? d????e?a t?? ?a??µe?????
  ?d??t?at????? f???t?da?
• ???ß? t?? ß?e???????? ?at? t? d????e?a t??
  f???t?da? t?? d??t???
• ??sp??? t?? ae????µ?t?? ?at? t? d????e?a t??
  ?t???.

35
????t??se?? ap? ta s?st?µata ?e???

• Aspergilla
• Fusarium
• ?p??et???? p?e?µ?????? ???µ??e?? s?µßa????? ?ta?
  ae????µata e?sp?e?µe?a ?at? t? d????e?a t?? ?t???
  (Anaissie and Costa 2001).

36
S?µa?t???? µ???????a??sµ?? st? p?s?µ? ?e??
p???a???ta? ?dat??e?e?? ???µ??e?? se
a??s??ate?staµ????? as?e?e??
37
????????s? ???d???? t?? e??a???a??? pa???????
ßa?t?????

• ?e??ss?te?a ap? ta ete??t??fa ßa?t???a t??
  p?s?µ?? ?e??? de? e??a? pa?????a ??a t?? ?????p?.
• HPC ßa?t???a t?? p?s?µ?? ?e??? ?p?? ta a??????a
  ???? µp??e? ?a e??a? pa?????a se
  a??s??atesta?µ????? ?p?d??e??
• Pseudomonas spp.
• Acinetobacter spp.
• Moraxella spp.
• Xanthomonas spp. ?a? d??f???? µ???te?.
• ???a µ? HPC µ???????a??sµ?? pe???aµß?????
• Legionellae
• Mycobacteriae
• Cryptosporidiae

38

• ?e??t? ap? t? Rusin et al 1977
• ?????se t?? p??a??t?ta ???µ???? ap? t? p?s?µ?
  ?e??.
• e??a? ? ????a a?t?a ??s???µ?a??? ???µ??e?? ?a?
  ????? ???t?t?ta
•  
• Pseudomonas aeruginosa
  e??a? ? ????a a?t?a
• Acinetobacter
  ??s???µe?a??? ???µ??e??
  Stenotrophomonas maltophilia ?a? ?????
  ???t?t?ta
•  
• Legionella pneumophila p???a?e? t? 4-20 t??
• p?e?µ???a? st?? ?????t?ta ?a? ?atat?sseta?
• st? de?te?? ? t??t? p?? s???? a?t?a p?e?µ???a?
• sta ?t?µa p?? apa?te?ta? ??s??e?a.
• Moraxella spp. µp??e? ?a p???a??se? ???µ???
  st??? ?f?a?µ??? ?a? t?? a??te??? a?ap?e?st????.

39
(No Transcript)
40
HPC ßa?t???a st? e???st?s?? epe?e??as?a? ?e???

• HPC ??e? µa??? ?st???a ?? de??t?? p???t?ta? ?e???
• ?et? ap? de?aet?e? e?µ??e?a? t?? ap?te?esµ?t??
  t?? HPC ??e? a????e? ?a? ap? de??t? asfa????
  ?e??? se ???? p??sd????sµ?? p???t?ta? ?e???.
• HPC ???s?µ?p??e?ta? s?µe?a ??a µ?t??s? ßa?t?????
  ?at? t?? d????e?a t?? epe?e??as?a? t?? ?e??? ?a?
  aµ?s?? µet? t?? e??at??e??? t?? ap? t? e???st?s??
  epe?e??as?a?.
• ?e???aµß??e? ?a? ???a test, ?p?? a?t?? t??
  ????ßa?t???d???, ????t?ta? ?a? t?? ?p??e?µµat????
  ???????.
• ???t?µ???ta? ta d?? te?e?ta?a test ??at? pa??????
  p????f???e? p?a?µat???? ?????? (real-time) ??a
  t?? p???t?ta t?? ?e??? ?a? t?? µe??d???
  epe?e??as?a?.
• ? s?????t??s? t?? ete??t??f?? ßa?t????? st?
  te???? ?e?? ?a p??pe? ?a e??a? 10 cfm/ml ?
  ????te?? (Fox and Reasoner 1999).
• ?aµ??? ? sta?e?? ep?ped? HPC ßa?t????? st? te????
  ?e?? p??s??te? asf??e?a st? ?t? ? µ???d??
  epe?e??as?a? ?e??? e????eta? s?st?.

41
HPC ßa?t???a st? s?st?µa ?ata??µ?? t?? ?e???

• ?ta? ß??s???ta? ????? ep?peda HPC st? te???? ?e??
  µet? t? e???st?s?? epe?e??as?a?, t? ep?ped? HPC
  st? s?st?µa ?ata??µ?? e??a? ep?s?? ?????.
• ?ta? st? te???? ?e?? ap? t? e???st?s??
  epe?e??as?a? pe????e? pa?ade?t? ep?peda HPC
  ßa?t???a, a??? ta ep?peda t?? s?st?µat??
  ?ata??µ?? e??a? p??? ap? ta s???st?µe?a ???a,
  a?t? s?????? de???e? ?t? ta ßa?t???a
  epa?apt?????a? (regrowth) st? s?st?µa d?a??µ??.
• ?a?t???a?? epa??pt??? a?af??eta? st?
  p???ap?as?asµ? (proliferation) t?? ???ta???
  ???a??sµ?? p?? e??a? pa???ta st? ?e?? µet? t??
  epe?e??as?a t?? p?s?µ?? ?e???, pe???aµß??e?
  ap??at?stas? ?a? a??pt??? t?? ???a??sµ?? p??
  p??????µ???? t?a?µat?st??a? ?at? t? d????e?a t??
  epe?e??as?a? t?? ?e???.

42

• ?p?? a?af??aµe p??????µ???? ta ete??t??fa
  ßa?t???a ap??t??? ??ept??? s?stat??? ap? t?
  pe??ß????? t??? ??a ?a ep???s??? ?a? ?a
  a?apt??????.
• ??? ?at? t? d????e?a epe?e??as?a? t?? ?e??? de?
  afa??e?e? ap? a?t? ? ???d?asp?µe?? ???a???? ???
  (?.?.?.) ?a? t? ?f?µ???s?µ? ???a???? ????a?a
  (?.?.?.) t?te pa?at??e?ta? epa??pt??? t??
  ßa?t?????.

43
HPC ßa?t???a st? S?st?µa ?ata??µ?? t?? ?e???

• ?????? s???e?t??se?? ???d?asp?µe??? ???a?????
  ???? µp??e? ?a apa?t?se? ?????te?? ??t?s?
  ap???µa?t???? p?? ?a ???s?µ?p????e?. St??
  pe??pt?s? t?? ????????, d???e?d??? t?? ???????
  ?a? ????aµ??e? a???s? t?? ??t?s?? µp??e? ?a
  µe??se? t?? ap?te?esµat??? s?????t??s? t??
  ?p??e?µµat???? ap???µa?t????.
• Se ?aµ???te?e? s???e?t??se?? ap???µa?t????
  ep??e??eta? ? ete??t??f??? ?????da, ????te?a
  a???t??? ap? t? ?p??e?µµat??? ap???µa?t??? ?a?
  µp??e? ?a??te?a ?a p???ap?as???eta? e?t?? t??
  s?st?µat?? d?a??µ??.
• ?ta? ???s?µ?p??e?ta? ?????s? ?? ap???µa?t???
  µ???d??, ta ep?peda t?? ?????? ???a????? ????a?a
  µe?????ta? a??? ?? s???e?t??se?? t?? ?f?µ???s?µ??
  ???a????? ????a?a a????eta?, p??????ta? t??
  epa??pt??? t?? ßa?t????? (Escobar et al 2001).

44
HPC ßa?t???a sta µ??a??µata epe?e??as?a? ?e???

• Health Canada
• US Environmental Protection Agency (EPA)
• US Consumer Safety Commission
• ?ta???? ??ß????s?
• ???te??a? t?? apa???e?s? t?? f??t??? e?e????
  ????a?a p?? ???s?µ?p?????ta? st? sp?t?
• ????s? t?? HPC ßa?t???a st? ????? t?? ????a?a
  a???????sa? ap? a???s? t?? µ?t??s?? t?? HPC st?
  d????µ??? ?e?? (Regunathan and Beauman 1994).
• ?et? ap? µe??te? ?? t?sse??? ap?f?s?sa? ?a ?????
  t?? apa???e?s?.

45
HPC ßa?t???a se eµf?a??µ??? ?e??

• St? eµf?a??µ??? ?e??, ta HPC ßa?t???a
  a?apt?ss??ta? se ????? s?????t??s? e?t?? ??????
  ?µe??? ap? t?? eµf????s?.
• Se p?s?t??? µe??t? t?? ßa?t???a??? p????sµ?? se
  µeta????? ?e??, sta HPC ßa?t???a (se ep?as? 22C)
  a??????e ap? t?? a????? 101-102 cfu/ml p??
  ?p???a? st?? p??? t?? ?e??? se 105-106 cfu/ml st?
  eµf?a??µ??? µet? ap? t?e?? ?µ??e? ap????e?s??.
• ?a ßa?t???a a?apt?ss??ta? ?a? de? staµat??? t??
  a??pt??? a??µ? ?a? ?ta? t? ?e?? ap????e??e? se
  6C (Gonzalez et al 1987).
• ?e? pa?at??????e ?a ??e? e?d????e? ep?d?µ?a
  ???µ?d??? as???e?a? s?et???µe?? µe t?? ?????
  s?????t??s? HPC ßa?t????? st? eµf?a??µ??? ?e??.

46
?a???e? ?a? ?d???e?

• ?? p??sfat?? ?a???e? ? ?d???e? ??a ta HPC
  ßa?t???a st? ?e?? t?? ß??s?? d?af??e? e????sta
  µeta?? d?af??et???? ??at??. Ge????, ? de??t?? HPC
  ???s?µ?p??e?ta? ?? e??a?e?? ??a t?? ap??t?s?
  p????f????? ??a t?? µ???d? epe?e??as?a? ?e??? ?a?
  ?e???? t?? ßa?t?????????? p???t?ta t?? ?e??? ap?
  t?? a?a????s? t?? ap? t? e???st?s?? epe?e??as?a?
  ??? st?? e?s?d? t?? µ?sa st? s?st?µa d?a??µ??.

47
?a???e? ?a???sµ?a? ??????s?? ??e?a?

• ?a???e? ??a t?? p???t?ta p?s?µ?? ?e??? (2003)
• ?p?te??sµata HPC st??? 22C pe?????f??ta? ?t?
  e??a? µ????? ??e????µ???? a??a?.
• ???a? µ?a ?a?? ??de??? ?a??? ?e?t?????a? t??
  epe?e??as?a? ?e???, s???e???µ??a t?? µe??d??
  s???????s??, d????s?? ?a? ap???µa?s??.
• S??p?? e??a? ?a d?at???s??µe HPC ?s? t? d??at?
  ?aµ???.
• ??a a???s? HPC ßa?t????? st??? 37C s???????µe??
  µ a?t? p?? f?s???????? ß??s???ta? µp??e? ?a
  e??a? ta p??ta s?µ?d?a µ????s?? ?????? e?? de?
  s???de?eta? µe t?? a?t? a???s? HPC ßa?t?????
  st??? 22C.
• ?af???? ? p???de?t??? a???s? HPC st? ?e?? t??
  s?????? µp??e? ?a de???e? eµp???t?sµ? t?? ?e???
  µe ?f?µ???s?µ? ???a???? ????a?a se µ?a ?d?e?s? ?
  µp??e? ?a ?fe??eta? st?? e?s?d? st? ?e??
  d?a??µ??.

48
????pa???? ?d???e?

• ?a ?????ß???????? p??t?pa s?µpe???aµß????? HPC
  ?aµ?a s?µa?t??? a???s? p??? ap? ta f?s????????
  ep?peda ?ta? ep?????ta? se 22C ?a? 37C ?a? ??a
  ta eµf?a??µ??a ?e?? e?t?? 12 ???? ap? t??
  eµf????s?
• 100 cfm/ml ?ta? ep?????ta? se 22C ??a 72 ??e?
• 20 cfm/ml ?ta? ep?????ta? se 37C ??a 38 ??e?
• (Barrell et al)

49
Ge?µa????? ?a????sµ??

• ? Ge?µa????? ?a????sµ?? ??s?µ?? ?e??? a?af??e?
  ?t? t? p?s?µ? ?e?? de? p??pe? ?a pe????e?
  pe??ss?te?? ap? 100 cfm/ml sta HPC ßa?t???a
  (Hambsch 1999).
• ???sd?????e? t?? ?e?µ???as?a ep?as?? 20C ?a?
  36C ??a pe???d? 40 hr.

• ?a?ad?? ??s?µ? ?e?? ????te?a ap? 500 cfm/ml
  (s???e?t?. HPC).
• ?µf?a??µ??? ????te?a ap? 100 cfm/ml.
• ??? S?????t??s? HPC ??t? t?? ep?t?ep?µe??? 500
  cfm/ml.
• ??st?a??a ?a?ade?t? HPC (ep?as? 35-37C ??a
  48hr)
• ????te?a ap? 100 cfm/ml ??a ??????µ??? ?e??.
• ????te?a t?? 500 cfm/ml a??????t? ?e??.