09/12/2013

MICROBIOLOGY OF THERMAL
PROCESSING
Lilis Nuraida, Ph.D.
SEAFAST Center
dan
Departemen Ilmu dan Teknologi Pangan
Institut Pertanian Bogor

What causes it?

How to prevent it?

Mikrobiologi Makanan Kaleng - LN- 2013

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Outline
Microbiology of Food Processing
Ecology of Microorganism in food
Thermal inactivation of microorganism
Microbioloy of low acid thermally

processed food
Microbiology of Acidified thermally
processed food
Spoilage of themally processed food
Microbiology of Thermal Process - LN- 2013

Microbiology of Food Processing

The microorganisms of primary concern to

the food processor:

Molds
Yeast
Bacteria.
They can grow in the food and
cause spoilage and risk of
foodborne illness
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Microbiology of Food Processing

Molds
Multi-cell organisms
g
forming
g tubular filaments.
Reproduce by spores, which are borne in or on aerial
structures.
Most molds have little heat resistance and cannot
survive the thermal processes for low-acid canned
foods.
The p
presence of molds in canned meat and p
poultry
yp
products only
y as a

result of under-processing or as a post-processing contamination.

The growth of mold in canned foods indicating the food

container has an opening to the outside environment

(leakage.
Microbiology of Thermal Process - LN- 2013

Microbiology of Food Processing

Yeasts
Single cells, larger than bacteria
Widely found in nature and are particularly associated with

liquid foods containing sugars and acids.

Quite adaptive to adverse conditions such as acidity and

dehydration.
Compared to bacterial spores, yeasts possess little resistance

to heat.
Most yeast forms are destroyed on heating to 77C.
If yeast causes spoilage of canned food, under-processing or

leakage must be suspected

The growth of yeasts results in the production of alcohol and

large amounts of carbon dioxide gas, which swells the

container.

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Microbiology of Food Processing

Bacteria
Single cells, smaller than yeast
Based on cell wall composition: Gram positive and

Gram negative
The most important and troublesome of all the

microorganisms for the food processor.

Certain bacteria form endospore that remain alive

under conditions that kill other cells.

Bacterial spores are heat resitant

Formation of extracellular polysaccharides in certain

bacteria increase heat resistance

Microbiology of Thermal Process - LN- 2013

Source of microorganism
Raw materials and ingredients are the primary sources
The ultimate source for pathogens in raw meat and

p
poultry
y is their feces.
Soil or water can be a common source of food borne
microorganisms and spores.
Vegetables become contaminated during production
Those that grow close to or through the soil usually
having high numbers of bacteria and bacterial spores,
including spores of C.
C botulinum.
botulinum
The processing environment
and equipment
Employee
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BAKTERI PEMBENTUK SPORA

Clostridium dan Bacillus
Satu
S t sell vegetatif
t tif membentuk
b t k satu
t

spora

Spora merupakan bentuk dorman

Terbentuk pada kondisi lingkungan

yang tidak menguntungkan

Spora akan bergerminasi pada
lingkungan
g u ga ya
yangg sesua
sesuai,, misalnya
sa ya
terstimulasi dengan adanya nutrisi
Bakteri kokus tidak membentuk

spora

Microbiology of Thermal Process - LN- 2013

Ecology of Microorganism in Food

Faktor-faktor yang mempengaruhi pertumbuhan
mikroorganisme
Kandungan nutrisi
Air
Oksigen
Suhu
Nilai pH
Senyawa

antimikroba

Kebutuhan Nutrisi
Kebutuhan nutrisi mikroorganisme:

air, sumber energi, sumber

nitrogen, vitamin dan faktor
pertumbuhan lainnya serta mineral.
Kebutuhan nutrisi kapang
p gp
paling
g
minimal, diikuti dengan khamir,
kemudian bakteri gram negatif
Bakteri Gram positif membutuhkan
nutrisi paling lengkap.
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pH dan pertumbuhan
mikroorganisme

Ketersediaan air minimum

untuk pertumbuhan
mikroorganisme

10

11

12

13

Kapang

Khamir
Bakteri Asam Laktat

Mikroorganisme
g
Kapang

Aw minimum
untuk tumbuh

Staphylococcus
p y
aureus

Salmonella spp

0.75

Escherichia colii

pH 4.6

Khamir

0.88

C. botulinum
V. parahaemolytiicus
C. perfringens

C. botulinum

0.93

S. aureus

0.85

Salmonella

0.93

Berasam rendah (pH > 5.3) :

kacang-kacangan, daging, susu, santan,

keju
keju.

Berasam sedang (pH 4.5 5.3) :

alpokat, nangka.

Asam (pH 3.7 4.5) :

tomat, pear, nenas.

Asam tinggi (pH < 3.7) :

sawi asin, yoghurt, strawberry.

Mikrobiologi Makanan Kaleng - LN- 2012

Oksigen
Mengapa Clostridium, Bacillus,
BAL dapat tumbuh pada
makanan yang dikemas
hermetis?

BAL: mikroaerofilik
Clostridium bersifat
anaerob obligat
Bacillus besifat fakultatif
anaerob

Suhu

Psikrofilik: toleran terhadap dingin,

tumbuh pada suhu < 4 oC
Psikrotrofik:
pertumbuhan optimum pada 14 - 20
oC
C, dapat tumbuh pada 4 oC
Contoh:
Clostridium botulinum tipe E
strain non-proteolitik tipe B dan F

Mesofilik:
suhu optimum 30 -37 oC
Contoh:

Senyawa antimikroba:
Bahan pengawet: nitrat/nitrit,
bakteriosin, asam organik,
benzoat

C.
C botulinum

Termofilik:
suhu optimum 45 - 60 oC
sporanya resisten terhadap pemanasan,

tidak memproduksi toksin

Contoh: Bacillus stearothermophilus

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Thermal Inactivation of microorganism:

Pasteurization
Sterilization
Sterilization

Why 2 methodes of thermal

process?
Yeast,
Yeast molds and non
non-spore
spore former bacteria are

inactivated by boiling
C. botulinum forms spores and need higher temperature
to inactivate them
Microbiology of Thermal Process - LN- 2013

Pasteurisasi
Pemanasan untuk membunuh mikroorganisme penyebab

penyakit
Misalnya pasteurisasi susu: 63 C; 30 min., 72 C; 16 sec.,atau

ekivalen
Membunuh mikroorganisme yang tidak membentuk spora
Dua kelompok mikroorganisme tahan pasteurisasi susu:

termodurik (Streptococcus dan Lactobacillus) dan

termofilik (Bacillus, Clostridium, Alicyclobacillus, Geobacillus,
Thermoanaerobacter)
Produk pasteurisasi tidak steril

Perlu dibantu dengan teknik pengawetan lain: pendinginan, pengasaman

Pangan dengan pH tinggi (>4.6) yang dipasteurisasi tidak

bisa disimpan pada suhu kamar, harus didinginkan

Umur simpan pendek tergantung suhu dan waktu pemanasan (biasanya

2 minggu)

Contoh: susu pasteurisasi

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Sterilisasi
Commercial sterility means the absence of microorganisms capable of growing in
the food at normal non-refrigerated conditions at which the food is likely to be held
during manufacture, distribution and storage.
(Ref: Codex Alimentarius Commission (WHO/ FAO) CAC/RCP 40-1993 )
Pemanasan di atas suhu 100 C untuk menginaktivasi spora
Harus dijamin bahwa proses telah membunuh/menginaktivasi

semua mikroba berbahaya bagi kesehatan

Paling berbahaya Clostridium botulinum menghasilkan racun
botulin
Pendekatan keamanan dan komersial : Konsep 12 D
Dua hal kritis : suhu dan waktu
Produk dapat disimpan pada suhu kamar dalam waktu lama (1-2
tahun)
Contoh: Makanan kaleng (kornet, sardin dll)

Makanan yang disterilisasi jika telah dibuka kemasannya harus

disimpan pada suhu dingin (refrigerator)
Microbiology of Thermal Process - LN- 2013

Faktor-faktor yang mempengaruhi

resistensi mikroorganisme terhadap
pemanasan
Komposisi
o pos s pa
pangan:
ga
Air, Lemak, Garam, Karbohidrat, Nilai pH,

Protein dan senyawa lainnya

Jumlah Mikroorganisme pada bahan
Suhu pertumbuhan
Senyawa penghambat
Suhu dan waktu pemanasan
P l k
Perlakuan
pendahuluan
d h l
sebelum
b l
pemanasan

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Bakteri patogen tdk tumbuh

Spora bakteri tidak tumbuh
Cukup proses pasteurisasi
Target: sel vegetatif, kapang, khamir

Terdapat peluang bakteri patogen

Spora bakteri tumbuh
Sterilisasi komersial
Target: spora bakteri

Mikrobiologi Makanan Kaleng - LN- 2013

Thermal inactivation of microorganism

Low acid foods commercially sterile

products will require high temperatures

to inactivate the organisms
g
of concern.
Acidified meat and poultry products can
be processed at lower temperatures,
since only vegetative cells must be
inactivated;
The pH prevents germination and

outgrowth of the spores.

The presence of nitrite and salt reduces

the amount of heat needed to achieve

commercial sterility
heat kills the vegetative cells of

pathogens; salt and nitrite prevent

germination and outgrowth of spores

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Jumllah mikroorganisme

Jumlah mikroorganisme vs
pemanasan
Jumlah mikroorganisme
yyang
g tinggi
gg p
pada bahan
pangan memerlukan
pemanasan yang lebih
lama

Xt

Waktu pemanasan
Mikrobiologi Makanan Kaleng - LN- 2013

Microorganisms vs thermal process

100-----

yang diperlukan untuk membunuh

sejumlah mikroorganisme pada suhu
tertentu
Nilai D: waktu yang diperlukan untuk
membunuh sebanyak 90%
mikroorganisme atau 1 log10 pada
suhu tertentu.
Sterilisasi makanan tidak asam

Survivors (%)

Thermal Death Time (TDT): waktu

menggunakan konsep 12 D untuk C.

10------------------1--

botulinum

The result of a heat sterilization p

process
depends on:
the initial number of microorganisms;
the decimal reduction time (heatresistance) of microorganisms;
the time and temperature of heat exposure.

D value

Waktu (min.)

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Dr value of thermophilic and mesophilic

Microorganisms
Geobacillus stearothermophillus
Thermoanaerobacterium (Clostridium)
thermosaccharolyticum
h
h l i
Clostridium nigrificans
C. botulinum (tipe A dan B)
C. sporogenes (PA 3679)
B. coagulans

Dr (menit)
4.0-5.0
3.0-4.0
2.0-3.0
0.10-0.20
0.10-1.5
0.01-0.07

In most canned products it is not the pathogen of concern that determines

the amount of heat needed but rather the spoilage microorganisms

Spoilage is more heat resistant than the pathogens.

As the spoilage is more resistant than pathogen, the heat applied provides a

wide margin of safety with respect to survival of pathogens in the product.

Microbiology of Thermal Process - LN- 2013

D value of C. botulinum 62A in various

foods
Effect of pH on D value at 115 C

pH

Spaghetti, tomattoes Spanish Rice

sauce and cheese

4.0
4.6

0.128
0.223

0.117
0.210

5.0
6.0
7.0

0.260
0.491
0.515

0.266
0.469
0.550

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Mikrorganisme vs proses thermal

Nilai z
Suhu yang

diperlukan untuk
menurunkan atau
meningkatkan 1
siklus log pada kurva
TDT (1 log nilai D)

Log Nilai D, menit

100-----

10------------------1--

Nilai z

Suhu (oF)
Microbiology of Thermal Process - LN- 2013

Fo dan konsep 12 D
Fo adalah waktu yang diperlukan (dalam menit) pada suhu 250 oF

untuk membunuh sejumlah mikroorganisme

g
Fo = Dr (log a log b)

Konsep 12 D:
Pemanasan yang harus dilakukan untuk mengurangi
survival C. botulinum sampai 10 -12
Fo = Dr (log a log b)
12)
Fo = 0.21
0 21 (l
(log 1 log
l 10 -12
Fo = 0.21 x 12 = 2.52

Pemanasan selama 2.52 menit pada suhu 250 oF

mengurangi spora C. botulinum menjadi 1 dalam 1
kaleng dari 1012 kaleng.
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The Microbiology of Low-Acid Canned

Foods
Clostridium botulinum
Great concern to home and commercial canners
Mesophilic,
M
hili G
Gram positive,
iti
rods
d
Produce toxin:
Although the spores are heat resistant, the toxin is not.
The toxin can be inactivated by boiling temperatures (100C).
Present in soil or water: easily contaminates foods
Produce heat resistant spores
Canning foods provides an anaerobic environment favorable

to growth of the organism if it has not been destroyed by the

process.
Strains:
Putrefactive: producing odor during their growth.
Non-proteolytic: more dependent on carbohydrates, such as sugars

and starch, for growth and do not produce putrefactive odors.

Microbiology of Thermal Process - LN- 2013

Pertumbuhan C. botulinum
Untuk bergerminasi spora C. botulinum memerlukan
kondisi:

Lingkungan anaerobik
Mk
Makanan
asam rendah
dh
Suhu antara 40F sampai 120F
Kadar air relatif tinggi
Nutrisi kompleks
Tidak ada bakteri lain

C. botulinum dapat ditemukan pada:

Canned foods pH> 4.6
Vacuum packed smoked fish and sausage
Foil-wrapped baked potatoes
Vacuum packed mushrooms
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Inactivation of C. botulinum
Heat processes for low-acid canned meat and

poultry are designed to produce a product that is

safe with respect to C. botulinum.
Microbial inhibitors and pH can impact the
processes needed to inactivate C. botulinum.
Inclusion of sodium nitrite and sodium chloride in

meat and poultry products (e.g., commerciallysterile canned cured meats) can lower the thermal
sterile,
process required to produce a commercially sterile
product.

Microbiology of Thermal Process - LN- 2013

Aw vs Clostridium botulinum
Aw (water activity = aktivitas air):
air y
yang
g tersedia untuk p
pertumbuhan
mikroorganisme
Spora tidak bergerminasi pada aw < 0.93
Makanan dengan aw > 0.85:
pemanasan seperti pada makanan asam rendah
Makanan dengan
g aw < 0.85:
pemanasan untuk membunuh sel vegetatif =
makanan asam

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Aw vs Clostridium botulinum
Makanan dengan aw < 0.85
pengukuran aw harus akurat dan reguler terhadap produk jadi
kontrol
k t l harus
h
dil k k sejak
dilakukan
j k persiapan
i
i
ingredien
di sampaii

pemanasan (pencapaian suhu yang ditetapkan)

Garam vs Clostridium botulinum

Tumbuh pada kadar garam 7% tetapi tidak memproduksi toksin
Germinasi spora terhambat pada kadar garam 10% ( = aw 0.93)
Aplikasi pada produk kuring
menggunakan garam nitrat/nitrit dan NaCl

Microbiology of Thermal Process - LN- 2013

pH vs Clostridium botulinum
Spora tidak bergerminasi pada pH < 4.8
pH 4
4.6
6 merupakan batas antara

makanan asam dan asam rendah

Pada makanan dengan pH > 4.6:
pemanasan untuk membunuh spora C.
botulinum
Pada makanan dengan pH < 4.6:
4 6:
pemanasan untuk membunuh sel
vegetatif
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Pengaruh pH terhadap derajat

pemanasan
Mild heat required, since spores are
inhibited by acid

High heat required to destroy spores

Acid

Mild Heat

Mild Heat

X X

spore

spore

Bacteria with
spore

Vegetative
cells (bacteria)

Vegetative
cells (bacteria)

Bacteria with
spore

pH 4.6
High acid food

Low acid food

Mikrobiologi Makanan Kaleng - LN- 2013

Other Microorganisms Important in

Low-Acid Canned Foods
Clostridium sporogenes
Its spores have higher heat resistance than those of C. botulinum,
Processes targeted to destroy spores of C. sporogenes will also

destroy C. botulinum spores.

Bacillus stearothermophilus and Clostridium

thermosaccharolyticum,

Thermophilic bacteria
Can survive processes that achieve commercial sterility.
Not harmful to humans,
Cannot grow under normal conditions of storage.
If product is properly cooled and stored, generally the spores will not

grow.

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Microbiology of acidified canned

food
Heat processes for acidified foods target organisms other

than C. botulinum, since the pH prevents its growth.

The safety of acidified foods is achieved by controlling the
pH to 4.6 or less and applying a mild heat treatment to
inactivate the vegetative cells of pathogens
The type of organism to be destroyed by the process will
depend on the pH of the product.
If the pH is below 4.2, only vegetative cells need be

considered;
If the pH is 4
4.2
2-4
4.6,
6 processes may need to take into account

sporeforming organisms such as Bacillus coagulans and the

butyric acid anaerobes (Clostridium pasteurianum and
Clostridium butyricum).

Microbiology of Thermal Process - LN- 2013

Tanda-tanda kebusukan makanan

kaleng karena bakteri
Pembentukan gas (kaleng kembung)
Perubahan penampakan makanan
Perubahan bau dan rasa makanan
Produk makanan hancur
Larutan yang seharusnya bening

menjadi keruh

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Bacterial decomposition of thermally

processed product
Incipient spoilage growth of bacteria before processing;
Typically incipient spoilage manifests itself as low or no vacuum in the

container
t i andd a slight
li ht change
h
iin pH
H off th
the product.
d t
Generally no viable microorganisms are recovered in subculture media.
Although the product presents no risk to public health, if there is sufficient
growth, the product may be considered to be adulterated
Contamination after processing leakage of bacteria into the container;
Inadequate heat processing: the growth of mesophilic
Growth of thermophilic bacteria in the processed food;
Spoilage by acid-tolerant sporeforming bacteria (specific to products

with reduced pH)

Spoilage due to improper curing (specific to cured products).

Microbiology of Thermal Process - LN- 2013

Tanda-tanda kebusukan makanan kaleng

karena kurang pemanasan dan kebocoran
Penampakan

Kurang
pemanasan

Bocor

Kaleng

Kaleng datar atau

gembung; seam
umumnya normal

Datar

Penampakan produk

Berlendir atau
terfermentasi

Kental dan terfermentasi

Odor

Normal, asam, bau

busuk, seragam semua
kaleng

Asam, bau busuk,

umumnya bervariasi
antara kaleng

Mikroskopik dan kultur

Pembentuk spora

Campuran

Kebusukan terjadi pada

beberapa batch atau
bagian kemasan

Kebusukan acak

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Spoilage by Acid-Tolerant Sporeformers

Acidified foods (those products with a pH 4.6 or

below)) do not require

q
a severe thermal p
process
to assure product safety.
Therefore a variety of acid-tolerant sporeformers may

survive the process.

A thermal process scheduled for acidified foods is designed to

inactivate a certain level of these sporeformers.

Their survival of those bacteria:

a result of excessive pre-processing contamination.

Underprocessing
The organisms of spoilage significance are butyric-acid
producing anaerobes and aciduric flat sour sporeformers.

Microbiology of Thermal Process - LN- 2013

Spoilage by Acid-Tolerant Sporeformers

The butyric-acid producing anaerobes:
Spores
p Clostridium butyricum
y
and Clostridium ppasteurianum are capable
p
of germination and growth at pH values as low as 4.2-4.4 and
consequently are of spoilage significance in acidified foods, particularly if
the pH is above 4.2.
Growth of these organisms in foods is characterized by a butyric
odor and the production of large quantities of gas.
Aciduric flat
flat sours
sours are facultative anaerobic sporeformers
Seldom produce gas in spoiled products.
Spoiled cans remain flat; hence the term flat sour.

Causing spoilage in acid foods such as tomato products (by Bacillus

coagulans
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Bacteria causing spoilage of thermally processed foods

Low acid Foods pH >4.6
Thermofilik flat sour

Bacillus (Geobacillus) stearothermophilus

B. coagulans

Pembusuk Sulfida

Desulfotomaculum (Clostridium nigrificans), C, bifermentans

Pembentuk gas

Thermoanaerobacterium (Clostridium) thermosaccharolyticum

Mesophilic

Putrefaktif anaerob: P.A. 3679, C. botulinum

Acid Foods 4 4.6

Thermophilic

B. Coagulans

Mesophilic

B. polymixa, B. Betanigrificans,
C. Pasteurianum, C. Butyricum (produksi asam butirat),
C. Sporogenes, C. Putrefaciens (busuk sulfida dan penggembungan
kaleng) Thermoanaerobacterium (Clostridium) thermosaccharolyticum
kaleng),
(pembentuk gas)
laktobasili, kamir,
Kapang (jika cukup oksigen): Byssochlamys fulva, Talaromyces flavus
dan Neosartorya fischeri

High Acid Food: 3.7-4

Mesophilic

Molds,Yeast and lactic acid bacteria

Microbiology of Thermal Process - LN- 2013

Terima Kasih
[email protected]; [email protected]

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