Bakteri & Archaea

SITOLOGI bakteri
Ukuran bakteri sangat
kecil berkisar antara 0,5 – 1,0 X
2,0 – 5,0
μm.
Bakteri terbesar yang pernah
ditemukan adalah
Thiomargarita
dengan lebar mencapai
750μm
(0,75 mm) yang membuatnya
bisa terlihat dengan mata
telanjang.
bakteri
 Bhs latin: bacterium kelompok raksasa dari organisme hidup.
Mereka sangatlah kecil (mikroskopik) dan kebanyakan
uniseluler (bersel tunggal), dengan struktur sel yang relatif
sederhana tanpa nukleus/inti sel, cytoskeleton, dan organel
lain seperti mitokondria dan kloroplas
 Merupakan sel prokaryot, berbeda dengan eukaryot.
 What is different ?
Bentuk:
 Bola/kokus

 Batang/basilus

 Spiral/spirilum

 Koma/vibrio
Pembagian kokus : konfigurasinya
 Monokokus
 Diplokokus
 Streptokokus
 Stafilokokus
 Tetrad
 Kubus/sarsina
Penataan Basilus
 Monobasilus
 Diplobasilus
 Sterptobasilus
 Bentuk pagar
 Bentuk roset
 BentukV, X, Y
 Bentuk endospora
Penataan koma

Koma
 Penataan spiral

 Spiroketa

 Spirilum
Struktur utama bakteri
 Dinding sel
 Membran plasma
 Sitoplasma
 Badan nukleus/badan inti
 Mesosom
 Ribosom
 Granula
 Flagela
 Pili
Struktur Bakteri : 3
 A. Struktur luar.
a) Flagela : atrik, monotrik, lofotrik, ampitrik, peritrik
b) Pili &Kapsul ( glikokaliks= selubung gula
c) Selongsong
d) Tangkai/apendiks

 B. Struktur dalam:
o Sitoplasma
o Badan nukleus/badan inti
o Mesosom
o Ribosom
o Granula
 C. Struktur dinding sel : terdiri dari peptidoglikan
Struktur luar.
FLAGEL
ANATOMI FLAGEL PD BAKTERI GRAM
(+) & (-)
 Bakteri berdasarkan Sporanya
Spora : Suatu badan yg refraktil yg terdapat dlm induk sel,
merupakan stadium istirahat dr sel tsb, tingkat metab.
Rendah.
Media sesuaispora mengalami germinasi => sel vegetatif

1. Eksospora :
Spora yang dihasilkan di luar sel vegetatif

2. Endospora:
Spora yang dihasilkan di dalam sel vegetatif ex: genus bacillus &
clostridium
Sifat spora
 Tidak mudah di warnai
 Tahan pemanasan
 Tahan pengeringan
 Tahan bahan kimia yang beracun

 Pewarnaan thd spora : bila sel biakan telah tua. Kenapa?

 Pewarnaan schaeffer-Fulton ( malachite green sbg zat warna
utama
 Letak endospora

 Sentral/tengah

 Terminal/ujung

 Subterminal/dekat ujung
 Bentuk endospora
 Spora eliptikal

 Spora berbentuk bola

 Spora ovoid/oval
 Dinding sel bakteri
 Penentu bentuk sel
 Pelindung sel & isi sel
 Letak di bawah kapsul, di luar membran sitoplasma.
 Tebal 10-23 nµ, berat ± 20% berat kering
 Fungsi →
 penutup dan permeabilitas( mengatur pertukaran zat dalam
&luar sel
 Mempengaruhi kegiatan metabolisme
 Berperan dlm reproduksi sel
Komposisi kimiawi :

 Peptidoglikan (murein)  dinding sel Kaku

 Polimer mole. Besar pengulangan dari
 Monosakarida ( NAG dan NAM)
 NAG ( N-acetylglucamin) & NAM( N-acetyl muramic acid)
melekat pd 4 atau 5 as. Amino : L-alanin, D-alanin, D-
glutamat& lisin
 asam tekoat, mgdg alkohol & gliserol
 polisakarida,
 lipid,
 protein.
Berdasarkan perbedaan
komposisi dinding sel, bakteri :2
1. Bakteri Gram? positif.

2. Bakteri Gram negatif.

Gram stain
 The Gram stain (named after Christian Gram Danish
scientist and physician, 1853–1938) is the most useful and
widely employed differential stain in bacteriology.

 It divides bacteria into two groups, gram negative and gram

positive.
DINDING SEL
BAKTERI
Ciri – Ciri bakteri Gram positif
a. Dinding sel byk mengandung peptidoglikan (murein)
 struktur kaku & tebal (15 – 80 nm) dan berlapis
tunggal.
b. Komposisi kimiawi : kandungan lipid rendah (1 - 4
%), peptidoglikan lapis tunggal (>50%), asam tekoat.
c. Kerentanan terhadap penisilin→ lebih rentan (peka).
d. Pertumbuhan dihambat oleh zat-zat warna dasar
(misal ungu kristal)
 Ciri - ciri bakteri Gram positif
e. Persyaratan nutrisi → relatif rumit pada banyak
spesies.
f. Resistensi terhadap gangguan fisik→ lebih
resisten (tahan).
g. Reaksi terhadap pewarna primer atau ungu kristal
iodium → dapat menahan sampai akhir prosedur
(sel tampak biru gelap/ungu).
Ciri-ciri bakteri Gram negatif
a. Dinding sel tipis (10-15 nm) berlapis tiga (multi).
b. Kandungan lipid tinggi : peptidoglikan (10% berat
kering), tidak ada asam tekoat.
c. Kerentanan terhadap penisilin kurang rentan.
d. Pertumbuhan tidak begitu dihambat oleh zat warna
dasar.
 Ciri-ciri bakteri Gram negatif

e. Persyaratan nutrisi → relatif sederhana.

f. Resistensi terhadap gangguan fisik→ kurang resisten
g. Kehilangan kompleks warna ungu kristal pada waktu dicuci
alkohol → terwarnai pewarna tandingan safranin (sel tampak
merah muda).
Pewarnaan Gram

Gram negatif Gram positif

The gram stain
 The gram stain is the most widely used staining procedure in
bacteriology.
 It is called a differential stain since it differentiates
between gram-positive and gram-negative bacteria.
 Bacteria that stain purple with the gram staining procedu
re are termed gram-positive;
 those that stain pink are said to be gram-negative.
 The terms positive and negative have nothing to do with
electrical charge, but simply designate two distinct
morphological groups of bacteria.
 Gram-positive and gram-negative bacteria stain differently
because of fundamental differences in the structure of their
cell walls.
 The bacterial cell wall serves to give the organism its size and
shape as well as to prevent osmotic lysis.
 The material in the bacterial cell wall that confers rigidity is
peptidoglycan.
 The gram-positive cell wall appears thick and consists of
numerous interconnecting layers of peptidoglycan.
 Also interwoven in the cell wall of gram-positive bacteria are
teichoic acids.
 Generally, 60% -90% of the gram-positive cell wall is
peptidoglycan.
 The gram-negative cell wall, on the other hand, contains a
much thinner, single layer of peptidoglycan only two or three
layers thick.
 This is surrounded by an outer membrane composed of
phospholipids, lipopolysaccharide, and lipoprotein.
 Only 10% - 20% of the gram-negative cell wall is
peptidoglycan.
The gram staining procedure involves four basic
steps:

 1. The bacteria are first stained with the basic dye crystal
violet.
 Both gram-positive and gram-negative bacteria become
directly stained and appear purple after this step.
 2. The bacteria are then treated with gram's iodine
solution.
 This allows the stain to be retained better by forming an
insoluble crystal violet-iodine complex.
 Both gram-positive and gram-negative bacteria remain
purple after this step.
 3.Gram's decolorizer, a mixture of ethyl alcohol and
acetone, is then added.
 This is the differential step.
 Gram-positive bacteria retain the crystal violet-iodine
complex while gram-negative are decolorized.
 4. Finally, the counterstain safranin (also a basic dye) is
applied.
 Since the gram-positive bacteria are already stained purple,
they are not affected by the counterstain.
 Gram-negative bacteria, that are now colorless, become
directly stained by th e safranin.
 Thus, gram-positive appear purple, and gram-negative
appear pink.
 With the current theory behind gram staining, it is thought
that in gram-positive bacteria, the crystal violet and iodine
combine to form a larger molecule that precipitates out
within the cell.

 The alcohol/acetone mixture then causes dehydration of

the multilayered peptidoglycan in the gram-
positive call wall, thus decreasing the space between the
molecules and causing the cell wall to trap the crystal violet-
iodine complex within the cell.
 In the case of gram-negative bacteria, the alcohol/acetone
mixture, being a lipid solvent, dissolves the outer
membrane of the gram-negative cell wall (and may
also damage the cytoplasmic membrane to which the
peptidoglycan is attached).

 The thin layer of peptidoglycan is unable to retain the crystal

violet-iodine complex and the cell is decolorized.
 It is important to note that gram-positivity (the ability to
retain the purple crystal violet-iodine complex) is not an all-
or-nothing phenomenon but a matter of degree.
Beberapa faktor yang mempengaruhi hasil Pewarnaan Gram:
 1. Teknik pengerjaan yang salah.
◦ Terlalu lama (Overheating) selama proses fiksasi.
◦ Terlalu banyak penggunaan alkohol pada tahap decolorization
(pencucian zat warna kristal violet)
◦ Terlalu banyak penggunaan air pada tahap pencucian
 Ini menyebabkan bakteri Gram positif tidak terwarnai dan tidak
membentuk kompleks crystal violet-iodine
 2. Umur kultur yang digunakan.
 Kultur yang berumur lebih dari 24 jam akan kehilangan kemampuan untuk
diwarnai dengan kompleks crystal violet-iodine
 3. Bakteri itu sendiri
 Beberapa bakteri Gram positif susah diwarnai dengan kompleks crystal violet-
iodine .
Bahan dan alat
 Bahan :
 Kultur bakteri
 Larutan kristal violet
 Gram’s iodine
 2 g potassium iodide in 300 ml distilled water plus 1 g iodine
crystals)
 95% ethanol and/or isopropanol-acetone mixture (3:1 v/v)
 safranin
 Alat :
 clean glass slides ( kaca objek)
 inoculating loop (jarum Ose)
 Bunsen burner (lampu spiritus)
 bibulous paper (kertas saring)
 microscope
 lens paper and lens cleaner
 immersion oil ( minyak immersi)
PROCEDURE

 1. Heat-fix a smear of a mixture of the bacterium as follows:

 a. Using the dropper bottle of distilled water found in your
staining rack, place a small drop of water on a clean slide
by touching the dropper to the slide
 b. Aseptically remove a small amount of Staphylococcus
epidermidis from the agar surface and mix it generously with
the water.
 Flame the loop and let it cool.
 Now aseptically remove a small amount of Escherichia coli
and sparingly add it to the water.
 Flame the loop and let it cool
 c. Using the loop, spread the mixture over the entire
slide to form a thin film
 d. Allow this thin suspension to completely air dry
 e. Pass the slide (film-side up) through the flame of the
bunsen burner 3 or 4 times to heat-fix
 2. Stain with Hucker's crystal violet for one minute.
 Gently wash with water.
 Shake off the excess water but do not blot dry between
steps.
 3. Stain with gram's iodine solution for one minute
and gently wash with water.
 . Decolorize by adding gram's decolorizer drop by drop
until the purple stops flowing.
 Wash immediately with water.
 5. Stain with safranin for one minute and wash with
water.
 6.Blot dry and observe using oil immersion microscopy
Gambar Mikroskop
ARCHAEA
Archaea
 Termasuk golongan eubakteri
 Mayoritas berasal dari lingkungan yang ekstrim
 Ekstrim (panas, dingin, asam garam tinggi)
 Kelompok : 3 :
 bakteri metanogenik,penghasil metan
 B. Halofilik  kadar garam tinggi ;3,5-5 M NaCl
 B. Termoasidofilik. panas ekstrim & keasaman ekstrim
 Materi archaeae selanjutnya agar di Baca pd
:pratiwi hal: 32-35
 Bersambung
 terimakasih