BAHAN

LABORATORIUM MIKROBIOLOGI
 Procedure to obtain microbial culture
Isolation
• the separation of a particular microorganism
from the mixed populations that exist in
nature
Cultivation
• the growth of microbial populations in
artificial environments (culture media) under
laboratory conditions.
 BAHAN

Media Pertumbuhan
Larutan Pengencer
 Media
 Medium : suatu bahan yang terdiri atas campuran nutrisi atau
zat-zat gizi yang digunakan menumbuhkan mikroorganisme di
atas atau di dalamnya.
 Medium juga dapat dipergunakan untuk :
- isolasi
- perbanyakan
- pengujian sifat fisiologis
- perhitungan jumlah mikroorganisme
 Komponen penyusun media pertumbuhan : air, sumber karbon,
sumber energi, mineral, faktor pertumbuhan, maupun
konsentrasi ion hidrogen (pH).
 Contoh ; NA/NB, PDA/PDB, PCA, MRSA, EMBA, LB
Culture media
 A culture medium is a solid or liquid preparation used to grow,
transport, and store microorganisms
 Suitable culture medium  need for microbial growth and
maintenance in laboratory
 Isolation and identification need special media  identification
microorganism in water and food, industrial microbiology, testing
for antibiotic sensitivity
 Although all microorganisms need sources of energy, carbon,
nitrogen, phosphorus, sulfur, and various minerals, the precise
composition of a satisfactory medium will depend on the species
 because nutritional requirements vary so greatly.
 Knowledge of a microorganism’s normal habitat often is useful in
selecting an appropriate culture medium  nutrient requirements
reflect it’s natural surrounding
Defined medium
 Medium that all
components are known
 Many
chemoorganotrophic
heterotrophs also can be
grown in defined media
 glucose ac carbon
source and ammonium
salt as nitrogen source
 Widely use in research
Complex medium

 Media that contain some

ingredients of unknown
chemical composition
 This media is useful as a single
complex medium 
sufficiently rich and complete
to meet nutritional
requirements of many different
microorganisms
 complex media often are
needed because the nutritional
requirements of a particular
microorganism are unknown,
and thus a defined medium
cannot be constructed
 Media
 Dari segi nutriennya/susunan kimianya, media pertumbuhan
dibedakan menjadi dua yaitu :
 media kompleks/ non sintetik
 media sintetik (defined media).
 Media semi sintetik
Media kompleks kaya akan nutrien (macam nutriennya banyak
dan konsentrasinya tinggi), akan tetapi macam dan konsentrasi
nutriennya tidak diketahui.
Media sintesis terbuat dari bahan kimia murni dengan
konsentrasi yang terukur dan dilarutkan dalam air murni.
Media semi sintetik : campuran dari media sintetik dan non
sintetik
Type of media
enriched selective
• General purpose fortified • favor the growth of
by special nutrient to particular microorganisms
encourage the fastidious • For isolation specific
heterotroph mikroorganism
• Ex: Blood agar • Ex: Endo agar, eosin
methylene blue agar, and
MacConkey agar
differential
• Media that use for
distinguish between
different group of bacteria
• permit tentative
identification of
microorganisms based on
their biological
characteristics
• MacConkey agar is both
• differential and selective.
 contains lactose and
neutral red dye, lactose-
fermenting colonies appear
pink to red in color and are
easily distinguished from
colonies of non fermenters.
Bentuk Media ; konsistensi media
Media Cair Media Padat (Media
Agar)

Nutrient Broth (NB)

Terdapat AGAR sebagai bahan
Brilliant Green Lactose Bile Broth
pemadat
(BGLBB)
Panas = cair ; dingin 43C = padat
Agar tegak / agar miring
Nutrient Agar (NA)
Plate Count Agar (PCA)
Media dan cara penyiapannya
 Media harus disimpan dengan kondisi tertutup rapat
dan pada kondisi (suhu) yang sesuai
 Perhatikan masa kadaluwarsa media
 Sendok yang digunakan untuk menimbang harus
kering dan bersih, hindari tercampurnya bahan-bahan
penyusun media
Cara Pembuatan Media
LARUTAN PENGENCER
Larutan pengencer Jenis
dibutuhkan untuk  Bufer fosfat
mengencerkan kultur  Larutan garam fisiologis
yang mempunyai (NaCl 0,85%)
konsentrasi tinggi.  Pepton dilution fluid
Mengandung bufer untuk
(pepton 0,2%)
menjaga keseimbangan
ion mikroba

9 ml (1:10); 1 ml sampel + 9 ml pengencer ;

99 ml (1:100); 1 ml sampel + 99 ml pengencer
 Perhitungan Kebutuhan Bahan
media pektin sitrus cair : de Man-Rogosa-Sharpe
yeast ekstrak 1g/L; (MRS) broth : peptone 10 g,
Na2HPO4 0,9 g/L; lab-lamco powder 8 g, yeast
KH2PO4 1g/L; extract 4 g, glukosa 20 g,
MgSO4.7H2O 0,5 g/L; sorbitol mono oleat (tween 80)
1 ml, dipotassium hydrogen
KCl 0,2 g/L; dan pektin
phosphate 2 g, sodium asetat
sitrus 1g/L)
trihidrat 5 g, magnesium sulfat
hepta hidrat 0.2 g, magnese
sulfat penta hidrat 0.05 g
dalam 1000 ml aquadest.