2546

(Food Microbiology)

2546

 (Food Microbiology)

. . 65000

ISBN

974-7195-23-2

Essential of Food Microbiology : John Garbutt, 1997

2546

. .

ii

2
4
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iii

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6
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aw ........................................................................................74
aw .............................78
pH ..............................................................79
pH...84
.......................................86
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7
96
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D.....................................................................98
..........100
z.................................................................................................100
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iv

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8
122
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................128

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9
140
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10
157
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11
175
..175

..................................................................175
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.........................................................................190
................................................196

198

(viroids) (prion)

.. 1677 Antony van Leeuwenhoek

(amoeba)
500
20


(aseptic techniques)

(bacteriology) (mycology) (protozoology)
(phycology algology) (virology)

(medical microbiology)

(agricultural
microbiology)
(industrial microbiology) (biotechnology)

(food microbiology)

.. 1800 .. 1837 Shwann

.. 1857 1876 Pasteur

0.2 3.0 0.5 10.0


(methylene blue)
(ribose nucleic acid)
1
(Gram stain)

(light microscope) .. 1984 Christian Gram

2
(Gram positive) (crystal violet iodine complex) (decolorizing agent)
(Gram negative)
(safranin)

1
: Garbutt (1997)

18
Micrococcus spp.

Gram variable

(flagella) 1

hanging drop motility

hot spore stain

: Garbutt (1997)

(electron microscope) (bright field microscope)
resolving power (
) resolving power
1,000 0.2
0.2 0.2

1 2.5

(transmission electron microscope)

(scanning electron microscope)
2

2
: Garbutt (1997)

(cell wall)
(cell membrane) (cytoplasm) (single
chromosome) (ribosomes) (capsules)
(glycocalyx) (fimbriae) (pili) (mesosomes)
(inclusion granules)

N-acetyl glucosamine Nacetyl muramic acid (chitin)
amino sugar

3 (peptidoglycan) (murein)
(mucopeptide) (mucocomplex)

teichoic acid

( 1 10)
teichoic acid

(lipoprotein) (phospholipid)
(lipopolysaccharide)
outer envelope outer membrane



3

3
: Garbutt (1997)
(plasma cytoplasmic membrane)

7 2

(pores) permeases

(respiratory enzyme)

(exoenzymes extracellar enzymes)

10

(cytoplasm)
75
DNA
(bacterial genome)
(plasmids)
(episomes) DNA 2
( 4,000 ) DNA
0.2
15-25

5-10



(ribosomes) 25
RNA

70s (s Svedberg units
)
(capsules) (slime layers)

11

(phagocytes)

(cation) (carboxyl)

(bacteriophage)

(cottage cheese)
(starter)
(glycocalyx)

(pili, fimbriae)
0.2 - 20 0.02
Enterobacteriaceae
Pseudomonadaceae
sex pili

12

(mesosomes)


penicillinase Bacillus
(flagella) 10 - 20
0.1 - 0.7

basal body flagellin

peritrichous polar

Bacillaceae Enterobacteriaceae
peritrichous Pseudomonadaceae polar
0.1 - 0.7
hanging drop motility agars
(storage granules)
polyphosphate (volutin) granules
RNA DNA polyhydroxybutyrate (PHB)
glycogen granules
(endospores) (mother cell)
Clostridium Bacillus
()
pH
aw

13

Clostridium botulinum (low acid
foods) 4

4
: Garbutt (1997)
4
(cysteine)
(S-S bridges) (cortex)
dipicolinic

15

(malachite) 5

14

(nucleus)
(eucaryotes)

2
(unicellular) (hyphae)
Saccharomyces spp. Candida spp. Torula spp.
Schizosaccharomyces spp. 5

5 (budding)
: Garbutt (1997)

15

(filaments)
(mycelium)
6

6
: Garbutt (1997)

(granules) (globules)
(vacuoles) phase contrast
7
Rhizopus Mucor
(non-septate)

Penicillium Aspergillus
(ascomycetes)
(basidiomycetes) clamp

16

7
: Garbutt (1997)
Mucor Rhizopus

Penicillium
(septal pore)
(fruit bodies)

17

8
: Garbutt (1997)

(chitin)
(cellulase) (mannose)

(cytoplasmic membrane) 2
(sterols)

(cilia) (flagella)

18

(cytoplasm)

(nucleus) (
5 )
RNA
(DNA) (nucleoli)
ribosomal RNA
(genome)
DNA
(chromosomes)

(mitochondria)
ATP
(ribosomes) 80s

(endoplasmic reticulum)


(storage granules)
(glycogen)
(endoplasmic reticulum) (golgi
membranes) rough

19

endoplasmic reticulum (smooth)

(vacuoles)

(spores) asexual
(ascomycetes) sexual spores
(ascospores)
Byssochlamys fulva

(chlamydospores)
9
(viruses)

20-30

(nucleic acid) (capsids)

(capsomeres) DNA RNA

20

9 (A- Alternaria, B- Aspergillus, C- Botrytis,

D- Cephalosporium, E- Cladosporium, F- Fusarium, G- Geotrichum, J- Monilia, K- Mucor)
: James (1992)

(host)


10

21

10
: Garbutt (1997)

22

3

(taxonomy)
3 (nomenclature)
(classification) (identification)
18 Linnaeus
(Carls von Linne) (binomial)
- (species)
-
- (generic name)
(specific name)
Escherichia ( Professor
Escherich) coli ()

(italics)
2


Pseudomonas
putrefaciens Shewanella putrefaciens

23

Staphylococcus aureus
Pseudomonas fluorescens
Escherichia coli
Clostridium botulinum
Bacillus cereus
Lactobacillus acidophilus
Streptococcus salivarius subsp thermophilus
Penicillium citrinum
Saccharomyces cerevisiae
: Garbutt (1997)

Staph. aureus
Ps. fluorescens
E. coli
C. botulinum
B. cereus
Lb. acidophilus
Strep. salivarius subsp thermophilus
P. citrinum
Sacc. cerevisiae

ribosomal RNA
ribosomal RNA
streptococci lactobacilli 3
3 lactobacilli streptococci

Streptococcus lactis
Lactococcus lactis subsp lactis
Streptococcus cremoris
Lactococcus lactis subsp cremoris
Streptococcus diacetylactis
Lactococcus lactis subsp lactis var diacetylactis
Streptococcus thermophilus
Streptococcus salivarius subsp thermophilus
Lactobacillus bulgaricus
Lactobacillus delbreukii subsp bulgaricus
: Garbutt (1997)

24

(plasmids) DNA
(conjugation) (transformation)

( E. coli)

ribosomal RNA
DNA DNA hybridization

hierarchical 2
artificial
(utilitarian)

natural
(phylogenetic)
kingdom phylum division class order family genus species

(subgroup)

(nuclear membrane)
(chloroplast)
ribosomal RNA 3
kingdom
- (eukaryotes) (protista) (

25

)
- (eubacteria) (bluegreen algae)
- (archaebacteria) (halophiles)
(methanogens) (thermophiles) RNA

(viroids) RNA
(prion protein)
BSE

(genetic code) (translation)

Bergeys Manual of Systematic Bacteriology

species genera families
orders classes genera
Genera of uncertain affinity

26

2
differential selective
(confirmatory test) (serology)
phage typing general purpose

(catalase) (oxidase)
genera families species

Zygomycetes
(zygospores) (sporangiospores)
(non-septate)
Rhizopus Mucor Thamnidium
Ascomycetes (ascopores)
(asci) ( 1 (ascus) 8 )
Byssochlamys Saccharomyces cerevisiae
4 Schizosaccharomyces
Eurotium Xeromyces

27

Basidiomycetes (fruiting bodies)

(basidiocarp) (basidiospores)
Deuteromycetes (imperfect fungi)
(conidia) Penicillium Aspergillus Botrytis Alternaria Fusarium
Monilia Wallemia Cladosporium
Rhodotorula Candida
ascomycetes Eurotium echinulatum Aspergillus echinulatus Neurospora
intermedia Monilia sitophila 11

11
: Garbutt (1997)

28

Moulds

zygomycetes, fungi imperfecti
ascomycetes
12

12
: Garbutt (1997)

29

(budding) (fission)
ascomycetes imperfect fungi

(

(
)

(hosts)

(
DNA RNA )
13

30

13
: Garbutt (1997)

31

(growth)

(macromolecule)

(unicellular) binary fission

(dry weight)
binary fission 14
Saccharomyces spp.
(budding) 15
(unicellular)

batch culture

32

14 binary fission
: Garbutt (1997)
-

ATP

binary fission
4

33

15
: Garbutt (1997)
4 (generation)
generation time population doubling
time mean generation time
2 - 3 generation time
30 Clostridium perfringens
8 - 10
pH 7.0 45

34

4 binary fission

0
1
2
3
4
5
6
7
8
9
10
: Garbutt (1997)

1
2
4
8
16
32
64
128
256
512
1024

generation time
aw pH (redox potential)
generation time
16
16 (exponential)
(logarithm)
binary fission

17

35

16
: Garbutt (1997)

17
: Garbutt (1997)
10 (log10)
4
5

36

0
1
20
1
2
21
2
4
22
3
8
23
4
16
24
5
32
25
6
64
26
7
128
27
8
256
28
9
512
29
10
1024
210
: Garbutt (1997)
( 0) NO 1
2 NO 2NO 2 22NO 4NO
N t Nt
1

Nt
=
2n NO

log Nt =

n log 2 + log NO

log Nt log NO = log Nt log NO

log 2
0.3010

37

mean generation time (g)

t/n t NO Nt E. coli generation time
20 103
24 ?
g E. coli 20 n 24
24 x 60 = 72 Nt 24 NO
103

n
=
log Nt log NO = log Nt log NO
log 2
0.3010
3
72
=
log Nt log 10
0.3010
(72 x 0.3010) + 3 =
log Nt
log Nt =
24.672
Nt =
antilog 24.672 = 4.7 x 1024
E. coli
( 1 10-12 ) 4.7 x 1024 x
10-12 = 4.7 x 1012
batch
1012
1010
109 106
18

38

18 batch
: Garbutt (1997)
18

lag (lag phase)

log (log/exponential phase)

generation time
aw pH

39

(stationary phase)
pH
(lactic acid)
(ethanol)
Bacillus spp.
Clostridium spp.
(death phase)

(cell lysis) pH

spreaders

19

20

40

19
: Garbutt (1997)
Rhizopus spp.
Penicillium spp.

staling effect

41

20
: Garbutt (1997)

(replicate)

DNA
bacteriophage viruse
(pinocytosis)
() (DNA RNA)


(lysozyme)

42

43

2

(nutrients)

pH / (reduction/oxidation potential)


(culture medium)

70 90 (cytoplasm)
10 - 30
(dry weight)


6

44

()
50

20

14

0.5

0.5

0.5

0.2

0.3


: Garbutt ( 1997)
6

(trace elements)

(cofactor)

45

7
7

DNA RNA

DNA RNA

trace elements
: Garbutt (1997)

21

46

21
: Garbutt (1997)
21 (photoautotroph)
(photosynthetic)
(photoheterotroph)

non-sulfer purple (chemoautotroph)

(chemolithotroph)
nitrifying
(chemoheterotroph)

ATP

22

47

22
: Garbutt (1997)
-(chemoorganotroph)
(chemorganoheterotroph)

(adenosine triphosphate, ATP)

23

48

23
: Garbutt (1997)
(Krebscycle)


sugar alcohols

49

(deamination)
(keto acid)
deamination 24
(serine)

24
: Garbutt (1997)

50

Pseudomonas cepacia 90
(starch)

DNA RNA

Rhizobium spp. Azotobactor

spp. 8
8

: Garbutt (1997)

51

(SO42-)
(SO32-)
(cysteine)
(methionine)

(H2PO-4)
(nucleotides)

K+
Mg2+

(plasma membrane)

(golgi membrane) extracellular exoenzymes
25

52

25 exoenzyme
: Garbutt (1997)
exoenzymes
(pectinase) proteinase

(lipase)
exoenzyme
9 exoenzyme

9 exoenzyme
exoenzyme

Bacillus cereus Aspergillus niger

(amylase)
Staphylococcus aureus Bacillus cereus
(proteinase)
Pseudomonas fluorescens Aspergillus oryzae
Penicillium citrinum Aspergillus spp. Erwinia carotovora
(pectinase)
Pseudomonas fluorescens
(lipase)
Staphylococcus aureus
(DNase)
Staphylococcus aureus Bacillus cereus
(lecithinase)
: Garbutt (1997)

53

E. coli
(ammonium chloride) (potassium phosphate)
(magnesium sulfate) (ferrous sulfate)
(calcium chloride) Aspergillus niger

growth factors
growth factors 3
1)
2) (purines) (pyrimidines)
DNA RNA
3) (vitamins) (coenzymes)

Salmonella typhi growth factor
(tryptophane)

growth factors
10

54

10 growth factor

E. coli
Salmonella typhi

Proteus vulgaris

Staph. aureus

Leuconostoc mesenteroids
19 10
Aspergillus niger

Mucor hiemalis

Sac. cerevisiae

: Garbutt (1997)

10 Leuconostoc mesenteroids 19
10 () 4
growth factor fastidious

bioassays Lactobacillus
plantarum

fastidious
Leuconostoc spp. Lactobacillus spp. 11

55

11

Pseudomonas fluorescens

P. fluorescens

Klebsiella spp.

Proteus

P. fluorescens

trace elements

: Garbutt (1997)

pH

56

(culture media)

() broths agar
agar agar-agar ( agar)
agar (sulphonated
mucopolysaccharide) 1.5
agar

agar pour plate streak plate

agar
-
aw
- agar

- agar

- agar 1.5 32 - 39
85
- agar 40 - 45

- agar

57

agar
(peptones) (yeast extract) (beef extract)

(peptides)
(proteose) ()
(gelatine) (soybean)
(trypsin) (pepsin)

growth factor

58

General purpose media (non-selective)



nutrient agar (NA) plate count agar (PCA) tryptone dextrose agar malt extract agar
broth potato dextrose agar (PDA)
Selective media

- pH (citric acid)
- aw (halophiles)
(osmophilic organisms)
-

- (anaerobes)
(metabisulphite)
-
(sodium azide) (thallous acetate) (lithium chloride)
(potassium tellurite) crystal violet bile salts
surfactant
- oxytetracycline
polymyxin

59

Enrichment media

selenite
broth Salmonella spp.

Differential media

-
- pH
-

- (substrates) extracellular enzyme
(lecithinase) (proteinase) (haemolysin) (DNase)
- pH
(bromocresol purple) (methyl red)
-

-
(tellurium)
(triphenyl tetrazolium chloride) (formazan)

60

- (iron salts) (hydrogen sulfide)

(iron sulfide)
Selective / differential media

Macconkey agar
(bile salts) Enterobacteriaceae
( E.coli Enterobacter spp. Salmonella spp. Proteus spp.)

E. coli Salmonella spp.

Chemically defined media

general purpose selective
differential media minerals modified glutamate medium

Elective media

selective medium
Tomato juice agar
lactobacilli peptonized milk pH
6.1
lactobacilli
Living media (embryo)
(host)

61

agar pH
121 15 pH 115
10 pH

62

100
26
E. coli

26
: Garbutt (1997)
26
10
( A B) B
C A

63

(minimum temperature)
B
(optimum temperature) C
(maximum temperature)

Clostridium perfringens (optimum temperature) 37 45 pH aw

Staphylococcus aureus 6.7 48

pH 7.0 aw 0.99 pH 5.0 aw
3.0 48
30

5 12
12

Minimum ()
Optimum ()
Obligate psychrophile
-10
10-15
Psychrotroph
-10
20-30
Mesophile
5
28-43
Thermophile
30
50-65
Extreme thermophile
65
80-90
: Garbutt (1997)

Maximum ()
20
42
52
70
100

64

(mesophiles)

(-1 5)
37
13
13

Minimum ()
Optimum ()
Maximum ()
Salmonella spp.
5.3
37
45 - 47
Staphylococcus aureus
6.7
37
45
Clostridium perfringens
20
37 - 45
50
Clostridium botulinum A/B
12.5
37 - 40
50
Campylobacter jejuni
30
42 - 45
47
Vibrio parahaemolyticus
10
30 - 37
42
Bacillus cereus
10
28 - 35
48
: Garbutt (1997)

Staphylococcus aureus

14

65

14 Staphylococcus aureus

()
()
Minimum
6.7
10
Optimum
37
40 - 45
Maximum
45.6 ( 48)
45.6 ( 48)
: Garbutt (1997)
Obligate psychrophiles
( 0 - 5)
-10
(psychrotrophs)
Pseudomonas fragii
Debariomyces hansenii 6.5 10 12.5
-12
25 - 30 30 - 42
(
1 5 7) 15

16

66

15

Pseudomonas
Candida
Alteromonas
Torulopsis
Shewanella
Saccharomyces
Bacillus
Debariomyces
Clostridium
Rhodotorula
Lactobacillus
Brochothrix
: Garbutt (1997)

Penicillium
Aspergillus
Cladosporium
Botrytis
Alternaria
Trichosporon

16

Minimum ()
Optimum ()
Maximum ()
Yersinia enterocolitica
- 1.3
28 - 29
44
Listeria monocytogenes
- 0.4
30 - 37
45
Aeromonas hydrophila
0-4
37
45
Clostridium botulinum E
3.3
35
45
: Garbutt (1997)
(thermophiles)
(silage) 70 extreme thermophiles
100
Bacillus stearothermophilus Clostridium thermosaccharolyticum
Desulfotomaculum nigrificans

67

lag
minimum temperature lag
lag 27

27 lag
: Garbutt (1997)

68

27 25 lag 1
10 0 lag 18 60
1 5
lag

chilling injury 2
1. cold shock direct chilling injury


ATP
(stationary) log
2. indirect chilling injury
()
ATP

chilling injury mesophiles

Salmonella spp.

69

0.5 3.0
aw aw
1.0 0 0.907 -10 0.841 -18

-5
aw aw
-20
-10 Debaromyces 12.5

40 - 90
1.

2.

3. chilling injury
4.
5.

70

6.
cryoprotectants
7.

8.
(blanching)
9. (thawing)

1. (slow freezing)

(plasmolysis)

2. (fast freezing)
DNA
pH
(ionic strength)
(gas bubbles)

3. (Ultra fast freezing)

(liquid nitrogen) -196

71

(lipase)
(proteinase)

(water activity : aw)

75

15

1.
2.
3. crystallization hydration
4. (matrix effects)

(osmotic
pressure)

72

(aw)

(vapour pressure) (mmHg) 0
4.679 mmHg 25 23.8 mmHg 28
aw
28 0

4.579 mmHg
100 (aw = 1.0) 10

4.551 mmHg 99.4 (aw =
4.551/4.579 = 0.994)
aw 0 - 1.0 (equilibrium
relative humidity, ERH) ERH =
aw x 100% aw Raoult

aw

n=
w=
m=
M=

N = ()
W =

n / (N + n) = (w/m) / (w/m + W/M)

73

28 aw
: Garbutt (1997)
5 (w/v) aw
= 18 = 342

74

aw

= (100 / 18) / ( (5 / 342) + (100 / 18) ) = 0.997

Raoult aw
aw

aw
75 aw 0.98

aw
aw (salted butter)
(water in oil emulsion) 2-3 aw
0.993 - 0.989 (polar molecule)
18 aw
0.86
aw
aw aw
aw 1.0

aw 17

75

17 aw

Proteolytic Clostridium botulinum A B

Non-proteolytic Clostridium botulinum B E F
E. coli
Salmonella
Listeria monocytogenes
Clostridium perfringens
Vibrio parahaemolyticus
Staph. aureus
Yersinia enterocolitica
Campylobacter jejuni
Bacillus cereus

aw
0.94
0.975
0.93
0.93
0.94
0.93
0.94
0.86
0.98
0.98
0.91
0.8 0.61

: Garbutt (1997)
aw
aw 0.61

(halophiles)
(Na+) (xerophiles)
aw aw 0.96 Xeromyces bisporus
2 moderate halophiles
1 10 Vibrio parahaemolyticus
extreme halophiles

76

Halobacterium salinarum
12 - 36 ( aw 0.928 - 0.760)
(halotolerant) (haloduric)

Staphylococcus aureus Pediococcus halophilus
20 ( aw 0.83)
(osmophilic)
aw Saccharomyces rouxii (Zygosaccharomyces rouxii)
70 aw 0.62
20 ( aw 0.980)
(osmotolerant) aw
Saccharomyces cerevisiae
60 aw
29

29 aw
: Garbutt (1997)

77

29 aw lag

(death)
aw

aw 0.1

(hypertonic)

turgor pressure

(K+) (polyols)
(glycerol) (arabitol) (trehalose) (mannitol)

aw
humectant aw
humectant aw

78

sugar alcohols

aw
aw
aw aw
humectant Halobacterium salinarum

aw
aw
1. (high moisture foods) (aw 0.99 - 0.95)
(cheese
spread) (cottage cheese)

2. (aw 0.95 - 0.90)

3. (intermediate moisture foods) (aw 0.90 - 0.61)

(salami) ( )

4. (low moisture foods) (aw 0.61)

79

aw
aw 0.93 ( 48
10) Staphylococcus aureus Bacillus cereus

Salmonella spp.

pH
pH
pH -
H+ OH-
1 x 10-7
H2O H+ + OH[H+] = [OH-] = 1 x 10-7
H+ OH- H+
(acid) OH- (alkaline) pH
H+ H+ 10-7
10-5 pH 7 5 log
10 pH 4.0 pH 7.0 1,000

pH
pH Saccharomyces
cerevisiae pH 2.35 - 8.60 pH
4.5 pH 30

80

30 pH
: Garbutt (1997)
30 pH (minimum pH)
4.0 - 4.5 pH (optimum pH) 6.8 - 7.2 pH
(maximum pH) 8.0 - 9.0 Lactobacillus spp.
pH 3.8 - 7.2 optimum pH 5.0 Acinetobacter spp. pH
2.8 - 4.3 pH 3.0
pH pH
Fusarium spp. pH 1.8 - 11.0 pH
4.0 - 4.5 pH 3.0 - 3.5 pH
31

81

31 pH minimum
: Garbutt (1997)
31 pH pH
lag
pH
aw
pH
pH 7.0 ( 5.8) pH
H+ OH H+ pH
pH (permeases)

ATP pH
H+ OH-

82

pH
R-COOH

RCOO- + H+

H+
pH pK pH 50 pKa



H+
pH
(hypochlorous acid)

(propionic) (acetic) (lactic)
(citric) (phosphoric) (hydrochloric)
pH
pH 18
pH Salmonella spp. pH

pH
pH
pH
pH 19

83

18 pH Salmonella spp.

Minimum pH

4.05

4.05

4.30

4.40

5.40

5.40
: Garbutt (1997)
19 pH

: Garbutt (1997)

pH
2.2 - 2.4
3.1 - 3.9
3.9 - 4.6
3.7 - 4.7
4.5 - 4.7
5.0 - 6.0
5.4 - 6.9
6.0
6.2 - 6.6
6.3 - 6.6
8.6 - 9.6

(tartaric acid)

84

aw
pH
streptococci lactobacilli
Pseudomonas
spp.

pH


pH
pH


pH 4.2 pH
20
pH pH

lag
Salmonella 72

85

20 pH

Minimum pH
Optimum pH
Staph. aureus
4.0
6.0 - 7.0
Clostridium perfringens
5.5
7.0
Listeria monocytogenes
4.1
6.0 - 8.0
Salmonella spp.
4.05
7.0
Vibrio parahaemolyticus
4.8
7.0
Bacillus cereus
4.9
7.0
Campylobacter
4.9
7.0
Yersinia
4.6
7.0 - 8.0
Clostridium botulinum
4.2
7.0
: Garbutt (1997)

Maximum pH
9.8
8.0
9.6
9.0
11.0
9.3
9.0
9.0
9.0

(pH 4.5 4.6)

botulinum cook pH 4.5
pH

pH
pH
pH Rappaport Vasiliadis pH
5.2 Salmonella pH
(pH drift)

86

pH

pH

pH pH
lysine iron agar
pH 6.7
(lysine decarboxylase)
(amine)

(redox)

aerobic anaerobic
ATP

Obligate aerobes
(glycolysis)

ATP 38

Pseudomonas fluorescens Penicillium spp.
Pichia spp. Hansenula spp.

87

Microaerophiles
20 Campylobacter spp.
1.0 10.0 6.0

Facultative anaerobes


Saccharomyces
cerevisiae 1 ATP 38
(ethanol) ATP 2 1

Enterobacteriaceae E. coli Salmonella ssp. Staphylococcus aureus
Obligate anaerobes

Clostridium Cl. botulinum
(aerotolerant) Cl. perfringens
Oxygen-dependent organisms
byproducts
facultative anaerobes microaerophiles

(fermentative organisms)

byproducts
byproducts

88

facultative anaerobes oxygen-independent obligate anaerobes

clostridia

(superoxide) (hydrogen
peroxide)
O2 + e-
O2- (superoxide)
O2 + 2e- + 2H+
H2O2 (hydrogen peroxide)



obligate aerobes facultative anaerobes microaerophiles
(superoxide dismutase)

2 O2- + 2 H+

sup eroxide

dismutase

2 H2O + O2

(catalase)
catalase
2 H2O2

2 H2O + O2
obligate aerobes
facultative anaerobes
microaerophiles aerotolerant anaerobes

89

(peroxidase)
obligate anaerobes
microaerophiles

- (oxidation-reduction potential)
- (OR) (redox potential ; Eh)
() ()
/

()
(
) (mV)
0
(redox indicators)

( Eh +11 ) ( Eh
10 )

MeB + 2 H+ + 2 eOxidized ()

MeBH2
Reduced ()

90

(triphenyl
tetrazolium chloride) (resazurin)

1)

2)
3)

faecal streptococci
4)
dye reduction

21
21

()

Pseudomonas fluoresecens
Obligate aerobe
+500 +100
Staphylococcus aureus
Facultative anaerobe
+180 -230
Proteus vulgaris
Facultative anaerobe
+150 -600
Clostridium spp.
Obligate anaerobe
-30 -550
Clostridium perfringens
Aerotolerant anaerobe
+216 -230
: Garbutt (1997)

91

(sulphydryl group)



(
poising capacity) pH
pH 1 Eh +58

+200 150

22

92

22

: Garbutt (1997)

()
+500
+400
+200
+200
+200
-150
-200
-150
-200

- obligate aerobes
facultative anaerobes oxygen independent
-
obligate anaerobes facultative anaerobe oxygen
independent
- microaerophiles
pH aw

Byssochlamys fulva
Rhizopus spp.

93

obligate aerobes/facultative anaerobes

oxygen independent

obligate anaerobes
anaerobic jar
(sulfites)
(thioglycolate) (cysteine)
( 200 )
microaerophiles
( 6.0) (
10.0) anaerobic jar

Clostridium botulinum

94

10.0 optimum pH 7.2 optimum temperature 35

pH 5.2
5.0

aw pH
facultative anaerobes
minimum aw Staphylococcus aureus 0.86
0.90
(optimum)
(minimum)

(hurdle)
lag

(pure cultures)
pH




Brochothrix thermosphacta

95

(nisin)
Lactococcus lactis
(Kefir)

Streptococcus salivarius
spp. thermophilus Lactobacillus delbreukii spp. bulgaricus

96

(wet
heat) (dry heat) (disinfectants)
(preservatives) (antibiotics) pH (
) aw (
cold shock)

RNA DNA

(broth)

97

(thermal death)

106
maximum temperature
5 23
23

log10
()

0
0
106
6
5
900,000
105
5
4
10
990,000
10
4
3
15
999,000
10
3
20
999,900
102
2
1
25
999,990
10
1
0
30
999,999
10
0
-1
35
999,999.9
10
-1
40
999,999.99
10-2
-2
: Garbutt (1997)
23 35
1 (0.1 )
1 10 1
1 10

98

(semi-log) x
y log 101 102 103
log 32

32
: Garbutt (1997)
32 1 (log cycle)
90 10
D (D values)
D (decimal reduction time) 90
1 33

99

33 D
: Garbutt (1997)
D D121
90 121 D

D
Bacillus stearothermophilus D121 4 - 5
D121 0.20 - 0.01
D121
0.5 - 3.0

100

D
1. (stationary phase)
log D55 Salmonella 4.8 log
D55 14.6
2.
Salmonella 44 D55 42
35 D55 14.6
3. pH
(optimum pH)
optimum pH 7.0 Streptococcus faecalis D60 13 pH
8.0 6.0 D60 2.5 2.2
4.

5. aw

Salmonella typhimurium D90 72 - 78
D 0.0008 (
90,000 )
z (z values)

w > x > y 34

101

34 3
: Garbutt (1997)
34

D 3 D
y log D x
35
thermal death time
curve 1 z
D 10 z 5 - 8
6 16 z D
z 10 D121 5
D131 D111 0.5 50 D z

102

35 thermal death time

: Garbutt (1997)
D1 / D2

10(T2-T1)/z

Listeria monocytogenes D60 8.3 z 5

D 71.7

D1 / D2
= 10(T2-T1)/z
8.3 / D2
= 10(71.7-60)/5.0
D2
= 0.0379

(cooking) (blanching)

103

Appertization (pasteurization)

Appertization 100
(pH 4.5) UHT
Clostridium botulinum

C. botulinum 1012 (botulinum cook) D121

0.21 12 x 0.21 = 2.52
1 1012
121 25.2 1
(commercial sterile)

C. botulinum C. sporogenes D
1.5 Bacillus stearothermophilus D 5.0
botulinum cook


105 - 106
1 1
105 106 Bacillus stearothermophilus
35

104

(nisin)
UHT
138-142 2-3 (aseptic
packaging)

(pasteurization)

60 - 80 (liquided egg) 64.4 2.5
Salmonella S. senftenberg 775W
65.6 30 71.1 10
79.4 15 ( high temperature short
time, HTST) 71.7 15
Mycobacterium tuberculosis
Salmonella Shigella

105

contact time 1
D
90

- (phenols) (phenolic compounds)

-
- (halogens)
- (alcohols)
- (aldehydes)
- (anionic detergents)
- (quaternary ammonium compounds)
- alkylating gases
- (peroxides)
- (organic acids)
- (alkalis)
- (inorganic acids)
- (ozone)
- (antibiotics)
- (synthetic antimicrobial drugs)
(toxicity)

106

Disinfectants sanitizer

(mucous membrane)

sanitizer
disinfectant
disinfectant

Antiseptic

Chemotherapeutic agents
(antibiotics) (sulpha drugs)

Preservatives
()

cidal static
germicide bacteriocide

107

sporicide fungicide algicide

(algae) viricide fungistat

(phenolic compounds)
(alkyl) (hydroxyl group)

chloroxylenol

(sulphydryl group)
(halogens)
(oxidizing agents)

disinfectant sanitizer

108

(undissociated hypochlorus acid)

(residual chlorine)
0.4
5.0
100.0 - 300.0
(hypochlorite) 10.0
disinfectant surfactant
nonionic (iodophores)
pH 4.0 - 5.0
(hypoiodous acid)
(tyrosine)
(alcohols) (ethanol) (isopropanol)
(benzyl alcohol) 50 - 70

109

(aldehydes)
DNA

(detergents) anionic amphoteric

anionic

(quarternary ammonium compounds)

cationic
(benzylkonium chloride)

Pseudomonas spp.

(biguanides)

110

(alkylating gases)

(ethylene oxide)

(peroxides)

UHT

(peracetic acid) disinfectant

(organic acids)
(alkalis)

5 100
(inorganic acids)


(ozone)

111

(antibiotics)



(nisin) (pymaricin)

sanitizer
1.

2. (contact time)

3. pH sanitizer sanitizer
(hypochlorites)
(hypochlorous) pH

(acidity)
D Bacillus cereus

25 2.5 pH 6.0 20 pH 9.0

4. (hardness of water) sanitizer

112

5.
sanitizer

6. sanitizer



7. sanitizer

(residual chlorine)
8. sanitizer
sanitizer ()

(preservatives)
()


aw

-
() (mineral acids)

(pimaricin natamycin) (smoke)

113

(sodium orthyl phenyl phenate)

(propionic acid)
Propionibacterium spp. (benzoic acid)
(cranberries)
24
24

99
2.19
50
4.19
1
6.19
: Garbutt (1997)

pH

2.87
4.87
7.17

2.75
4.75
7.05

24 pH


(ropiness)
Bacillus subtilis

114

(mineral acids)

(inorganic anions)
(metabisulphite)
(sulphite ions)
(free radicals)

DNA

lactobacilli 450
Enterobacteriaceae

(nitrate) (nitrite) cured meat

cured meat ( saltpetre)

115

Micrococcus spp.
curing salts (inject)
Clostridium spp. ( Clostridium botulinum ) Bacillus spp.
Staphylococcus aureus lactobacilli
Enterobacteriaceae Salmonella Staphylococcus aureus

pH

(fermented meats) 100

Staph. aureus C.
botulinum 100
pH 6.2 5.0
(non-proteolytic strains) 100 pH 6.2
3.0 Listeria monocytogenes
100 3.0

(nitrous acid) (nitrous oxide)




(carbon dioxide)
(modified atmosphere)
pseudomonads

116

pH

(sodium chloride) aw

(smoke)
(formaldehyde)
(phenols) (methanol)
(antibiotics) (nisin) Lactococcus lactis
Clostridium botulinum Listeria spp. Bacillus
spp. Staphylococcus aureus
Clostridium spp. Bacillus spp. Streptomyces
natalensis

(hurdle)

(Hurdle technology)

117

36

36
: Garbutt (1997)

(Storage-life (shelf-life) testing)

(ideal) (acceptable)

Challenge Testing

118

Predictive modelling

(pork slurry)

pH aw (ihhibitors)
(growth curve)
pH

(electromagnetic spectrum)
(wave length) (nanometer)
(frequency) (Hertz)

(microwaves) ()
(oscillate)

(visible light) (intensity)

119

(cytochrome) (flavins)

(ultraviolet light)

(purine) (pyrimidine) RNA DNA

260
(tryptophan) (tyrosine) (phenylalanine)
280 DNA
(thymine) DNA
DNA
DNA
(mutant)




(melanins)

(ionizing radiation)
60 (Cobalt 60)

120

OH + OH H2O2 ()
H + O2 HO2 ()
DNA

(Gray, Gy) 1 Gy 1 (joule)
1 1 kiloGy = 1,000 Gy rad 1 Gy = 100 rads



Deinococcus radiadurans D 5 kGy

-60 137
(commercial sterile)
Clostridium botulinum 1012 D

121

3.5 kGy 12 x 3.5 = 42 kGy

radappertization

Salmonella
3 - 10
2 - 5
10 radicidation
Salmonella radurization
( pasteurization )

aw aw

aw Salmonella typhimurium D
0.5 D 1.7
Clostridium botulinum D 1.6
D 3.5
(high pressure)

400 5
1
1,200

122

8




(rancidity)

(ripening)

( Bulgarian yoghurt
)

Clostridium botulinum

123

(epidermis)
(corky layers)





(epithelium)
(intestinal mucosa)
(lysozyme)
(vomiting)

124

30
1. coccobacilli Acinetobacter Aeromonas Alcaligenes
Citrobacter Enterobacter Escherichia Flavobacterium Moraxella Proteus Pseudomonas
Salmonella Shewanella Yersinia
2. Bacillus Brochothrix Clostridium Corynebacterium
Lactobacillus Listeria
3. Enterococcus Lactococcus Micrococcus
Pediococcus Staphylococcus
Mucor Rhizopus Thamnidium
imperfect fungi Cladosporium Geotrichum Sporotrichum Candida
spp.

2 (intrinsic factors)
(extrinsic factors)
pH
pH (buffering capacity) (poising
capacity) aw

125

aw pH 5.0
pH
4.2 aw



succession
( 25) aw pH
aerobes facultative anaerobes lactobacilli
streptococci
pH lactobacilli
pH 3.8

pH
7.0
putrefactive anaerobes
putrefaction

Shewanella putrefaciens
20

Koch (Kochs postulates)

126

1.

2.
3.

2


Halobacterium salinarum
Pseudomonas fluorescens

(green rot) Pseudomonas fluorescens
Rhizopus spp.

Proteus spp.

lactobacilli

127

Sclerotinia fructigena (polyphenol oxidase)

(pectinase)
(calcium pectate) (parenchyma cells)
Erwinia carotovora
Penicillium citrinum
Pseudomonas fluorescens

(amines)
(indole)

(diacetyl)
(microbial
rancidity)


37

128

37
: Garbutt (1997)
37 109
()
(off odour/flavor)
106

129

(minced beef)

UHT

Micrococcus spp.

pH 6.8 3.8 - 4.2
pH

(putrid)

130

pseudomonads
Enterobacteriaceae lactobacilli Brochothrix thermosphacta Shewanella
putrefaciens

aw pH

3.5 0.01 0.35

18.0 3.0
aw 0.99
pH 7.0 pH

131

pH

pH 5.4 - 5.6

pH 6.0

(oxygen permeable film) +200

(obligate aerobes)
(facultative anaerobes)

1.5

Pseudomonas spp. Ps. fluorescens Ps. fragii


(dimethyl sulfide) (methane thiol)
pH 8.0
108
pH

( 106 )

132

20-30 1
25
Enterobacteriaceae Acinetobacter

pH
pH lactobacilli
pH

Shewanella putrefaciens Enterobacteriaceae

50.0 lactobacilli
Brochothrix thermosphacta pH
Shewanella Enterobacteriaceae
cured meat
cured meat
pH aw

133

cured meats hurdles

aw country cured ham

lactobacilli
micrococci lactobacilli pediococci leuconostocs

micrococci streptococci
corynebacteria

1. Pseudomonas Alcaligenes Acinetobacter
Flavobacterium
2. Enterobacteriaceae
3. Streptococcus Lactococcus Leuconostos
4.

( (silage)
Clostridium )

103
105

134

aw 0.98
pH 6.6 +200
lactoperoxidase/thiocyanate/hydrogen peroxide

7

2 - 4

Pseudomonas
Pseudomonas fluorescens
(metallic) (unclean)
Pseudomonas 107

2 - 4
UHT
25
streptococci
lactobacilli Enterobacteriaceae
(curd) (whey)

135

Streptococcus thermophilus
Bacillus spp.
Bacillus circulans Bacillus sphaericus

(pectinase)
rots pH 5.0 6.5
pH

Erwinia
(outer periderm) (cork layer)
(water proof substance)
(suberin)

136

Erwinia
carotovora soft rot

1.

2.
3. 3.3 - 12.8
4.
5. (polythene)

25

1.
2.
3.
4. (fungicides) (benomyl)
(biphenyl) (captan)

5.

137

25

Monilia fructigena
brown rot
(Sclerotinia fructigena)
Penicillium expansum
blue mould rot

Penicillium italicum
blue mould rot
Penicillium digitatum
green mould rot

Botrytis cinerea
grey mould rot

Alternaria tenuis
Black rot
: Garbutt (1997)

Clostridium
sporogenes

Bacillus coagulans
(pear)
Byssochlamys fulva

138

(peas)
Bacillus stearothermophilus

37


(nicin)

1.

2.
3.

139

4.

5.

Clostridium botulinum type E Salmonella typhi
Staphylococcus aureus

140

/


(pathogens)

(microflora)

Escherichia coli
Salmonella spp.



(toxins)

(enterotoxin)
Staphylococcus aureus

141

2 (exotoxin) (endotoxin)

1.
2.
3.
4.

1. (lipopolysaccharides)
2.
3.



1.
2. (pyrogens)

3.
4. (shock)

142

Staphylococcus aureus 30

1.
2.
3.
4.
5.
6.
7.

143

8.

(prions)
(bovine spongiform encephalopathy ; BSE)
(acute)

(diarrhoea)
Clostridium botulinum Listeria monocytogenes


Salmonella

Campylobacter Salmonella

Staphylococcus aureus Clostridium perfringens Clostridium botulinum Bacillus cereus

(intoxications)
(syndrome)
(enterotoxins)

(neurotoxins) (mycotoxicoses)
intoxication
(incubation periods)

144

(infections) Salmonella
spp. (Salmonellosis)

Clostridium perfringens


(mucosa)
Listeria monocytogenes Mycobacterium tuberculosis

1. Campylobacter jejuni
Campylobacteriosis
1-10 ( 3-5 )
1
Campylobacter jejuni

2. (Salmonella)

-
S. typhi S. paratyphi A B C

145

- S. gallinarum
- S. dublin

-
Salmonella
Salmonella Salmonellosis
(epithelial cells) (villi)
(connective tissue)
5 - 72 (
12 - 36 )
Salmonella typhi S. paratyphi

Salmonella 1 - 4
106
108

Salmonella
Salmonella
S. typhimurium

146

3. Shigella Shigellosis

Shigella
dysenteriae Shigella sonnei Shigella flexneri
10 Shigella dysenteriae 102 - 104
Shigella sonnei Shigella flexneri
Shigella


4. Listeria monocytogenes

Listeriosis
1 - 70

102
107

Listeria

147

5. Yersinia entercolitica Yersiniosis

24 - 36
11
5-14

Yersinia entercolitica

Yersinia

6. Escherichia coli
E. coli

- Enteropathogenic E. coli (EPEC)

- Enteroinvasive E. coli (EIEC)
- Enterotoxigenic E. coli (ETEC)
- Enterohaemorrhagic E. coli (EHEC) verocytotoxic E. coli (VTEC)
E. coli
EPEC EIEC ETEC
ETEC
enterohaemorrhagic E. coli O157 : H7
3 - 4


2 - 9

148

10 - 100
E. coli O157 : H7
7. Vibrio parahaemolyticus
12 - 24

2 - 3 105 - 107
Vibrio parahaemolyticus

103 - 104

Vibrio Vibrio cholerae

0:1
Vibrio vulnificus 40

8. Aeromonas hydrophila


9. Plesiomonas shigelloides
48

149

10. Mycobacterium bovis (Tuberculosis)

(mucosa)

11. Brucella Brucella abortus

Brucella melitensis
Brucellosis

1. Staphylococcus aureus
30 6

2-3 3
5x106
1
Staph. aureus 30 - 50

150

2. Clostridium botulinum A B C
F Botulism 2
type A (strains) type B F
type E type B F
(neurotoxin) Botulism

12 - 36 2
8

1 - 10 60
50 1

lactobacilli

Clostridium botulinum
Clostridium botulinum

151

(pH
4.6)
aw 0.93

3. Clostridium perfringens 5 type

A B C D E type type A


6 - 24
12 - 24
7 x 105

Clostridium perfringens



4. Bacillus cereus
2

8 - 16 1 - 5


105 - 108
103 - 1010

152

Bacillus cereus



Bacillus B. subtilis B. licheniformis
B. pumilis


(scombrotoxic food
poisoning)

Morganella spp. Proteus spp. Hafnia alvei

Klebsiella pneumoniae
(histamine) (histidine
decarboxylase)
3

1 - 8
5 100 100
100
Hafnia alvei
-30 Morganella spp. -15 Klebsiella pneumoniae -7

153

(bonito) (anchovies)

100

Hepatitis A (HAV) (Hepatitis)

15 - 50 HAV 3 - 6

(secondary metabolites)

(mycotoxins)
(mushrooms) Basidiomycetes
Ascomycetes Claviceps purpurea

154

(alkaloid) Byssochlamys fulva byssochlamic acid

imperfect fungi 15
Aspergillus 20 Penicillium 15 Fusarium
6

mycotoxicoses
ergotism Claviceps purpurea

- Psilocybe
cubensis (magic mushroom) psilocybin Amanita
phalloids
- Claviceps purpurea

- Aspergillus flavus Aspergillus parasiticus
(aflatoxin)
B1 Aspergillus ochraceons ochratoxin
Penicillium expansum patulin
Fusarium trichothecenes T2 (mucous
membrane)


(oil seeds)

B1 ochratoxin
patulin
Fusarium

155

(protozoa)
20
Giardia lamblia Cryptosporidium
G. lamblia
Cryptosporidium Cryptosporidiosis
(villi) Giardia
Cryptosporidium

(algae)


Gonyaulax catanella G. tamarensis G. acatenella
(dinoflagellates)
Gonyaulax
algae bloom red tide
Gonyaulax
(paralytic
shellfish poisoning) 30 2

156

Bovine Spongiform Encephalopathy (BSE)

(prion protein)

(vacuole)

transmissible degenerative encephalopathies (TDE)

6 - 12

157

10

(fermentation)

(
)
ATP

industrial fermentations

(aerobic processes)

1) (food additives)
(antibiotics) (enzymes)
(vinegar)
2)
(single cell protein) Quorn
Fusarium graminearum
(vegetarians)
3)

158

4) (cheese) (yoghurt)
(bread)
5) (alcoholic beverages)
6) (yeast extract)

7) (mushrooms)

(biotechnology)

(stability)

(lactic acid bacteria)

(blue cheese) (soy sauce)

(salami) (peperoni) (tempeh)

159

Kaffir
(Bantus)

.. 200
2,500
1,000




(curd)


back slopping


(starter culture starter)


(genes)

160

1.
1,000
2.

3.

30 ( 0.15
5 ) Tape
2

(phytate) (glucosinolates) (lectins)

4.

2-3
5.

6.

161

(lactose
intolerance)
(lactate)
(oligosaccharides)
(stachyose)
(mono-, disaccharides)

7.

1. (acid food fermentations)
(kefir)
(sauerkraut) cerevilat

(sourdough breads)

162

2. (yeast fermentations)

(byproducts)
3. (solid state fermentations)

4.

26

(mesophiles)
Lactococcus Leuconostoc Lactobacillus Pediococcus
20 30
(thermophiles) Lactobacillus spp.
Streptococcus spp. 45

163

26

1)

2) (nutto)

3) (tempeh)

idli
4)

5) tape

: Garbutt (1997)

2 (homolactic bacteria)
(heterolactic bacteria)

38

164

38
: Garbutt (1997)


Lactococcus
var. diacetylactis Leuconostoc spp. (citrate)
(diacetyl) (cottage cheese) (quark)
(butter) (acetaldehyde)
Lactobacillus delbreukii subsp. bulgaricus
aroma bacteria
(polymers) (slime)

165

(freeze
dried)

27
27

Lactococcus lactis subsp. lactis/cremoris

Streptococcus saliverius subsp. thermophilus

Lactobacillus bulgaricus
Lactobacillus acidophilus

Lactobacillus acidophilus

Streptococcus lactis subsp. thermophilus
Lactococcus lactis subsp. cremoris (+ lactis)

Lactococcus lactis subsp. lactis/cremoris

Lactococcus lactis subsp. lactis/cremoris

Lactobacillus acidophilus Lactobacillus kefir

Lactobacillus kefiranofaciens Lactobacillus casei
: Garbutt (1997)

166

(ripening)
(cheddar cheese)
(casein)

Staphylococcus aureus
Salmonella spp.

1) (bacteriophage viruses)

2)

3)

4) (agglutinins) (antibody)
(clump)

5) (plasmid)

167

(Bio yoghurt) Bifidobacterium bifidum
(emmental)
Propionibacterium spp. (propionic acid)


Penicillium camembertii P. caseicolum P. candidum
P. rocquefortii P. gluacum Geotrichum candidum Brevibacterium linens

(buttermilk)
(acidophilus milk)
39
2 Streptococcus salivarius
ssp. thermophilus pH
Lactobacillus delbruekii ssp. bulgaricus
S. salivarius


pH 3.7 4.3 0.8 1.8 (
) 20 40

168

( )

85 30

( S. salivarius ssp. thermophilus L. delbruekii ssp. bulgaricus (1:1)

2 )

4 - 16 30 45

10 15

4.5
39
: Garbutt (1997)

Lactobacillus acidophilus
Bifidobacterium bifidum () L. acidophilus B. bifidum S. salivarius ssp.
thermophilus ()
37

169

16 pH 4.2 4.4

pH 4.4
pH






()

1)
2) (coagulation) (rennet)
3) (whey)
4) (salting)

170

5) (ripening)

(curd)

(acetate)
(hydrogen peroxide)
(bacteriocins)
( 26
50) ( 42 52) ( 48 80)
(unriped)
(surface ripened)

(50
200 )

(clostridia) Clostridium butyricum

171

(silage)
(nisin)

(wax)
(sorbate) (pymaricin)
pH
Pseudomonas ssp.

Samonella E. coli (enteropathogenic)
Listeria monocytogenes Clostridium botulinum

1)
2)

3)

172

Leuconostoc
mesenteroids Lactobacillus brevis Lb. plantarum Pediococcus pentacaseus



(heterofermentative bacteria)

(microbial succession)

3
1) Leuconostoc mesenteroids



1.1) pH
4.0 2

173

1.2)

1.3)

1.4)
()

Leuconostoc

(mannitol) (dextran)

1.5) Leuconostoc

1.6) Leuconostoc
15

1 - 2
2) pH Lactobacillus
brevis Lb. plantarum

6 8
3) 16 18 Lb. brevis
Lb. plantarum

174

pH 3.8 (
) 1.7 2.3 1 : 4

175

11

(traditional methods)
1. (Plate counts)
(clumps)

colony forming units (cfu) cfu

(microcolonies)

176

cfu

30 - 300
95 30
2 pour plate spread plate
(peptone solution)
(peptone phosphate buffer) 0.15
0.85

selective
selective differential elective
spread plate pour plate
45

37 50 - 55 18 - 25

standard plate count

(SPC) aerobic plate count (APC) general purpose tryptone
dextrose yeast extract agar (plate count agar) 30

177

48 total bacterial counts

(TBCS) total viable count (TVC)
Standard plate count

2. (membrane filtration)
(cellulose acetate) 0.45



differential

3. most probable number (MPN) multiple tube

E. coli
plate count MPN
3
2 1

178

MPN 40

40 most probable number 3

: Garbutt (1997)
MPN
(MPN table)
MPN
37 10-1 10-2
10-3 2 1 1 MPN 28
MPN 20

179

28 MPN

10-1
10-2
10-3
0
1
0
1
0
0
1
1
0
1
2
0
2
0
0
2
0
1
2
1
0
2
1
1
2
2
0
3
0
0
3
0
1
3
1
0
3
1
1
3
2
0
3
2
1
3
2
2
3
3
0
3
3
1
3
3
2
3
3
3

MPN /
2
4
7
11
9
14
15
20
21
23
40
40
70
90
150
210
200
500
1100
>1100

: Garbutt (1997)
MPN 3 2

/

180

4. (direct
microscopic count)
0.01
1
(methylene blue) (oil immersion lens)
25

105 Howard mould count

5. (dye reduction tests)

(resazurin)




streptococci


6. (indicators)

E. coli

181

48
37 (bile salts) surfactant
E. coli Enterobacter spp. Klebsiella spp. Citrobacter spp.
E. coli

(faecal coliforms)
44 - 45.5
E. coli Klebsiella Enterobacter
Enterobacteriaceae


E. coli
Salmonella Shigella
(Rapid methods)

182

1. ATP photometry ATP (adenosine
triphosphate)
(light)
luciferin (substrate) luciferase
ATP
luciferin + luciferase + ATP + O2

Mg2+

oxyluciferin + luciferase + AMP + light

ATP
(1 ATP 1 ) ATP
( ATP ATP)
(photometer)
ATP
ATP 1 (fg) 100 fg ( 1 fg = 10-15 g)
ATP
ATP
ATP



ATP
ATP photometry
UHT

183

2. direct epifluorescent filter


(acridine orange)
epifluorescent


epifluorescent

RNA DNA

45

surfactant
(trypsin) (somatic
cells)
3. electrical impedance
Impedance
impedance 2 capacitance
(electrode)
conductance

184

conductance

impedance



conductivity ( impedance
)
impedance (Bactometer)

impedance
41

41 impedance
: Garbutt (1997)

185

impedance
106 - 107
impedance detection time (DT)
DT
DT
lag
cfu
impedance plate count
42

42 log cfu impedance detection time

: Garbutt (1997)

186

E. coli 5

selective
differential MPN

(false positives)
E. coli E. coli
(false negatives)

plate count MPN

(injured cells) (resuscitation)
selective enrichment
selective differential
(serolgy)

187

growth factors
lag

(dyes) surfactants pH

pre-enrichment
non-selective
selective Salmonella lactose broth pre-enrichment
selective
(pyruvate) Baird-Parker Staphylococcus
aureus
non-selective
selective differential
tryptone soya agar 2
selective differential
selective differential
(selective temperature)
37 44 - 45.5

minerals modified glutamate

188

37 4
tryptone bile agar 44 18
(Kovacs)
selective enrichment

1
(loop) (streak) selective differential
(replicates)

rapid methods
(highly specific)
Enzyme-linked immunosorbent assay (ELISA)
Salmonella Listeria monocytogenes
ELISA (antibody)
(antigen) (enzyme) (peroxidase)
(alkaline phosphatase)
(substrate)

ELISA 90 pre-enrichment
enrichment

189

( 105 )
2
Salmonella (dipstick)
preenrichment
ELISA
1
ELISA
Staphylococcus aureus
(mycotoxins)

(commercially
sterile)
30 5 (streak)

impedance

190

slit sampler

2 contact plate
swabbing

.. 1950

191

(standard for
frozen prawns)
- TVC 106
- 102
- Staphylococcus aureus 10
- Salmonella

..1962
International Commission on the
Microbiological Specifications for Foods (ICMSF) .. 1974

(ingredients) .. 1975
FAO/WHO

(good manufacturing practice)

192

ICMSF
ICMSF 3
1.

2.

HACCP
3.


(sampling plans) ICMSF
(sampling plans) ICMSF
1. Two-class plan 2
(accept) (reject)

n
m
m = 0
c
m

Staphylococcus aureus
n = 5 , c = 0 m = 103 /

193

5 S. aureus
103/
c = 0 103/g
m
Salmonella
n = 10 , m = 0 c = 0
10 25 Salmonella
c = 0
Two-class plan
29
2. Three-class plan 3

0

m
m

M
M
M
m
M
m M c
aerobic plate count
n = 5 , c = 2 , m = 5 x 105 M = 107
5 aerobic
plate count 30

194

29

Clostridium botulinum
Salmonella typhi
Salmonella paratyphi
Salmonella cholerae suis
Shigella dysenteriae
Brucella melitensis
Clostridium perfringens type C
Mycobacterium tuberculosis (bovis)
Vibrio cholerae
Infections hepatitis virus A
Salmonella typhimurium
Salmonella
Shigella sonne flexnerii
Vibrio parahaemolyticus
Escherichia coli (enteropathogenic)
Beta hemolytic streptococcus

Bacillus cereus
Clostridium perfringens
Brucella abortus
Staphylococcus aureus
Listeria monocytogenes
Campylobacter jejuni
: Garbutt (1997)

195

30
(cfu/g)
1
1
4.0 x 104
m
5
2
3.2 x 10

5
3
4.2 x 10
4
9.6 x 104
5
4.9 x 105
2
1
6.3 x 106
2 m M
5
2
4.8 x 10
m c = 2
5
3
2.1 x 10

4
5.9 x 105
5
3.6 x 105
3
1
3.2 x 105
1 M
7
2
7.8 x 10

3
4.8 x 105
4
1.3 x 105
5
4.9 x 104
: Garbutt (1997)
Three class plan
Enterobacteriaceae E. coli
S. aureus
Vibrio parahaemolyticus

196

(destructive analysis)



(Hazard Analysis Critical Control Point : HACCP)
.. 1960


Hazard
Analysis Critical Control Point (HACCP)

HACCP
HACCP
1.



(pesticide residues)

197

2. (critical control points : CCPS)

ICMSF CCP 2 CCP1
CCP2
3. (Risk)
3

4. (Severity)

5. (Monitoring)
(visual inspection) (smell)

(corrective action)

6. (Verification)
HACCP

198

Adams, M. R. and M. O. Moss. 1995. Food Microbiology. The Royal Society of Chemistry,
Cambridge, England.
Frazier, W. C. and D. C. Westhoff. 1988. Food Microbiology. 4th ed. McGraw- Hill, Inc.,
USA.
Garbutt, J. 1997. Essentials of Food Microbiology. Arnold, London, England.
ICMSF. 2000. Microorganisms in Foods 6 : Microbial Ecology of Food Commodities. Aspen
Publishers. Inc., Maryland, USA.
Jay, J. M. 1992. Modern Food Microbiology. 4th ed. Van Nostrand Reinhold, London,
England.
Kraft, A. A. 1992. Psychrotrophic Bacteria in Foods : Disease and Spoilage. CRC Press, Inc.,
Florida, USA.
Montville, T. J. 1987. Food Microbiology. Vol 2. New and Emerging Technologies. CRC
Press, Inc. Florida, USA.
Ray, B. 2001. Fundamental Food Microbiology. 2nd ed. CRC Press LLC, Florida, USA.