BACILLACEAE

YUWONO, MD

 1872, Ferdinand Cohn murid Robert Koch menemukan Bacillus subtilis

Genus Bacillus, family Bacillaceae
Ciri khas famili ini endospora
Ciri genus: Gram-positif, batang, aerobik atau fakultatif, membentuk endospora

Ubiquitous in nature
Endospora resisten zat kimia dan fisik
Siklus pembentukan endospora
Produsen antibiotik
Spora dan kristal proteinnya toksik untuk beberapa serangga
Patogen Bacillus anthracis

Bacillus coagulans
Great diversity in physiology:
Mendegradasi hampir semua substrat tumbuhan dan hewan termasuk selulosa,
starch, pectin, protein, agar, hidrokarbon
Produksi antibiotik
Nitrifikasi, denitrifikasi, fiksasi nitrogen, facultative lithotrophy; autotrophy;
acidophily; alkaliphily; psychrophily; thermophily dan parasitism.
Spora merupakan strategi bertahan di tanah dimana banyak sekali bakteri. Spora
juga dapat ditemukan di udara

Nutrisi dan Pertumbuhan

Hampir semua Bacillus versatile
chemoheterotrophs yaitu mampu
respirasi menggunakan berbagai
simple organic compounds (sugars,
amino acids, organic acids).
pada beberapa kasus kadang
memfermentasi karbohidrat
memproduksi glycerol dan
butanediol.
Bacillus megaterium memerlukan
non organic growth factors
Mayoritas mesofil optimum 3045C,tapi ada juga yang termofilik
pada 65C

Mucoid-type colonies of an
encapsulated Bacillus species

Generation time di laboratorium 25 menit

Bacillus mudah diisolasi. Teknik sederhana dengan pasteurize a diluted soil
sample pada 80C 15 menit plate pada nutrient agar dan inkubasi 37C 24
jam-beberapa hari. Setelah 24 jam ada koloni basilus yang khas: catalasepositive, Gram-positive, endospore-forming rods

Surface Structure of Bacillus

Seperti permuakaan Gram-positif lainnya kompleks berhubungan dengan adherence,
resistance dan tactical responses
Vegetative cell surface terdiri dari: capsule, proteinaceous surface layer (S-layer),
several layers of peptidoglycan sheeting dan protein pada outer surface membran
plasma

Surface of a Bacillus. Transmission E.M.

C=Capsule; S=S-layer; P=Peptidoglycan
S-layers: Crystalline protein atau glycoprotein subunit yang disebut S-layers, fungsi
belum diketahui mungkin berhubungan dengan interaksi bakteri-metal, bermuatan
negatif dan dapat mencegah autoaglutinasi pada B. stearothermophilus

Kapsul
B. anthracis, B. subtilis, B. megaterium dan B. licheniformis: poly-D- or L-glutamic acid.
B. circulans, B. megaterium, B. mycoides dan B. pumilus: karbohidrat.
Polisakarida beberapa Bacillus bereaksi silang dengan genus patogen lain seperti B.
mycoides dengan Streptococcus pneumoniae type III; B. pumilus dengan Neisseria
meningitidis group A; B.alveli dengan Haemophilus influenzae type B.
Di bawah TEM beberapa kapsul polipeptida dan polisakarida tampak fibriler. Kapsul
mudah dilihat dengan mikroskop cahaya. Media berkapsul bersifat mukoid atau slimy
colony

Kapsul Bacillus anthracis FA stain (kiri), negative stain (india ink) (kanan)

Cell Walls
Tidak bervariasi seperti pada Gram-positif umumnya. Dinding sel vegetatif
terbuat dari meso-diaminopimelic acid (DAP) kecuali pada B. sphaericus, B.
pasteurii dan B. globisporus dari lysine
Flagella
Mayoritas Bacillus motil yaitu memiliki peritrichous flagella. Flagel tetap stabil
hingga pH 11.

Individual cells of motile Bacillus species photographed on nutrient agar, 15,000X

A. B. subtilis; B. B. polymyxa; C. B. laterosporis; D. B. alveli.

Property

Vegetative cells

Endospores

Surface coats

Typical Gram-positive
murein cell wall
polymer; crystalline Slayer

Thick spore coat, cortex, and

unique peptidoglycan core
wall; no S-layer

Microscopic appearance

Nonrefractile

Refractile

Calcium dipicolinic acid

Absent

Present in core

Cytoplasmic water activity

High

Very low

Enzymatic activity

Present

Absent

Macromolecular synthesis

Present

Absent

Heat resistance

Low

High

Resistance to chemicals and

acids

Low

High

Radiation resistance

Low

High

Sensitivity to lysozyme

Some sensitive; some

resistant

Resistant

Sensitivity to dyes and

staining

Sensitive

Resistant

Perbedaan endospora dan sel vegetatif Bacillus species

Formasi Spora

Genetik
Transforamsi pada Bacillus subtilis ditemukan pada1958
Generalized and specialized transduction juga terjadi pada B. subtilis
Genes affecting sporulation B. subtilis
Bacteriophages memediasi generalized transduction pada B subtilis, B. cereus,
B. megaterium, B. thuringiensis, B. anthracis dan B. stearothermophilus.
Conjugative plasmids (fertility plasmids capable of bringing about their own
transfer from one bacterium to another) juga terjadi pada bebrapa spesies
Insecticidal delta toxin crystal protein B. thuringiensis dikode pada large
plasmids. Plasmid ini dapat ditransfer antar spesies. Selain itu terbukti pula
bahwatransmissible plasmids of Bacillus, a conjugative transposon (Tn925)
berasal dari Enterococcus faecalis ke B. subtilis.

Ekologi
Endospora resisten terhadap stres lingkungan. Primary habitat of the aerobic
endospore-forming bacilli is the soil. Russian microbiologist Winogradsky
menyebutnya "normal flora" of the soil.

Ecophysiological groups
Acidophiles: B. acidocaldarius, B. coagulans dan B. polymyxa.
Alkaliphiles: B. alkalophilus dan B. pasteurii. Optimum pH 8 - 11
Halophiles: B. pantothenticus, B. pasteurii, beberapa tahan 10 % NaCl
Psychrophiles atau psychrotrophs: B. globisporus, B. insolitus, B. marinus,
B. macquariensis, B. megaterium, B. polymyxa, tumbuh pada 0C
Thermophiles: B. acidocaldarius, B. schlegelii dan B. stearothermophilus,
Acidophiles dan Lithoautotrophs pada kelompok ini tahan hingga 65C.
Denitrifiers: B. azotoformans, B. cereus, B. laterosporus, B. licheniformis, B.
pasteurii, B. stearothermophilus (reduce NO3 to NO2).
Nitrogen-fixers: B. macerans and B. polymyxa. B. macerans in soil and in
decaying vegetable material.

Pathogens of Insects: B. larvae, B. lentimorbis, and B. popilliae are invasive

pathogens. B. thuringiensis forms a parasporal crystal that is toxic to Lepidoptera.
B. larvae, B. lentimorbis and B. popilliae are a related cluster of Bacillus species,
being insect pathogens, with swollen sporangia, and typically catalase-negative.
Pathogens of Animals: B. anthracis, and B. cereus. B. alvei, B. megaterium, B.
coagulans, B. laterosporus, B. subtilis, B. sphaericus, B. circulans, B. brevis, B.
licheniformis, B. macerans, B. pumilus, and B. thuringiensis have been isolated
from human infections.

Bacillus
Classification

All are large Gram-positive bacilli

Are aerobic
Form endospores
Most are found in dust and soil
Bacillus anthracis is the major pathogen in the group

Morphology and Cultural Characteristics

(Bacillus anthracis)

Bacillus

G+B with square cut ends

Forms endospores
Non-motile
Capsule is made of glutamic acid (a polypeptide)
The capsule is not found in cultured bacteria
unless the bacteria are grown on bicarbonate
containing media and in the presence of increased
CO concentrations. The organism has a capsule
while inside the host which provides the conditions
necessary for capsule production.

Gram stain of Bacillus sp.

Endospores of Bacillus sp,

Bacillus
Grow well on ordinary lab media producing large
granular colonies with a coarse texture.
On BA may cause beta hemolysis.

Virulence factors
Capsule helps organism to resist phagocytosis but
antibodies are not protective.
Exotoxin is very complex and is produced only when the
bacteria is growing in animal tissues. Toxin production is
mediated by a temperature sensitive plasmid. The toxin
consists of three protein components (maximum toxicity
occurs when all three components are present).

Bacillus sp. On BA

Bacillus
Protective antigen (PA) reacts with host cell tissue receptors
where it is proteolytically activated so that it binds LF and EF
to allow entry of LF and EF into the host cells. Injection of PA
into a host will result in the production of antibody against PA
that provides short term immunity.
Lethal factor (LF) Both PA and LF are required for lethal
activity.
Edema factor (EF) Both PA and EF are required for edema
to occur. EF has adenylate cyclase activity and it becomes
active when it enters eucaryotic cells. This results in an
increase of cAMP and resulting edema.

Bacillus
Clinical significance
Anthrax which is the disease caused by B. anthracis
is essentially a disease of animals who acquire the
organism by ingestion or inhalation of spores. The
spores are extremely resistant to adverse chemicals
and physical environments. They may remain a
source of infection in soil for 20-30 years.
Man acquires anthrax usually from contact with
animal products; less commonly from working in an
agricultural setting with infected animals.

Bacillus
Man may acquire the organism through skin
abrasions, by inhalation of spores, or by ingestion.
The disease that develops depends upon the mode
of transmission:
Pulmonary (Woolsorters disease) Spores are inhaled and
germinate in the lungs where they multiply and
spread to cause a fatal septicemia or meningitis. This
is the most serious form of the disease.
Intestinal anthrax results from ingestion of spores.

Bacillus
Cutaneous anthrax when organism comes in through skin
abrasions; most common form; most damage comes from
the invasion but the toxin may play a role; spores enter
through a cut, germinate, and vegetative cells multiply;
there is an acute host inflammatory response, but the
capsule probably prevents phagocytosis; exotoxin release
and invasive damage result in rapid development of a
malignant pustule; occasionally, without treatment, the
organism will disseminate to cause septicemia and death in
a few days (rare because of B-lysin activity in the
bloodstream that kills the bacteria); vascular injury with
edema, hemorrhage, and thrombosis may occur; death
results from respiratory failure and anoxia caused by
actions of the toxin on the central nervous system (toxemic
degeneration).

Anthrax
Anthrax:primarily a disease of domesticated and wild animals terutama
herbivora.Manusia terkena karena konbtak dengan hewan-hewan tersebut:
flesh, bones, hides, hair dan excrement. Paling sering cutaneous anthrax
didapat lewat kujlit atau mukosa yang terluka,minor scratch atau abrasion
pada face or neck or arms terinokulasi spora dari tanah. Spora tumbuh
menjadi vegetatif berupa gelatinous edema kemudian papule within 12-36
jam, kemudian cepat menjadi vesicle, pustule (malignant pustule) dan
akhgirnya necrotic ulcer yang dapat menyebar menjadi septikemia.
Lymphatic swelling dalam 7 hari sebagai tanda septikemia umumnya fatal.

Anthrax
Inhalation anthrax (woolsorters' disease) karena menghirup spore-containing
dust where animal hair or hides are being handled. Dimulai dengan demam
tinggi dan nyeri dada dan perdarahan sistemik dan fatal.
Gastrointestinal anthrax analog dengan cutaneous anthrax karena tertelan
poorly cooked meat from infected animals. Gastrointestinal anthrax jarang tapi
kadang menjadi wabah
Patologi ditentukan virulence: antiphagoytic capsule, and powerful lethal toxin
ie. the anthrax toxin.

Anthrax

Anthraks kutaneus paling sering terjadi pada manusia

Anthraks pneunonik fatal jika terjadi

ESCHAR

B. Anthracis

B. Anthracis (kanan) dan B. cereus (kiri)

Bacillus cereus

Ditemukan di lingkungan seperti tanah, debu

Patogen oportunistik pada luka, abses paru dan LCS
Keracunan makanan menyebabkan diare, mual dan muntah
terutama makanan yang dimasak kurang matang atau makanan
yang disimpan/diawetkan

Koloni: Medusa-head morphology

Bacillus cereus food poisoning

Bacillus cereus causes two types of food-borne intoxications:
1. Characterized by nausea and vomiting and abdominal cramps and has an
incubation period of 1 to 6 hours. It resembles Staphylococcus aureus food
poisoning in its symptoms and incubation period. This is the "shortincubation" or emetic form of the disease.
2. Manifested primarily by abdominal cramps and diarrhea with an incubation
period of 8 to 16 hours. Diarrhea may be a small volume or profuse and
watery. This type is referred to as the "long-incubation" or diarrheal form of
the disease, and it resembles more food poisoning caused by Clostridium
perfringens. In either type, the illness usually lasts less than 24 hours after
onset.
The short-incubation form is caused by a preformed heat-stable enterotoxin.
The mechanism and site of action of this toxin are unknown. The longincubation form is mediated by a heat-labile enterotoxin which activates
intestinal adenylate cyclase and causes intestinal fluid secretion.

Bacillus subtilis dan spesies lain

Ditemukan di lingkungan
Patogen oportunistik

GRAM POSITIVE BATANG NON SPORA

CASE
The patient was a 2,150 gram female neonate
Born at 32 weeks gestation. She had poor vital signs and she was
intubated at birth because of poor respiratory effort.
The mother presented at the time of delivery with complaints of lower
abdominal pains with a temperature of 39oC and a white blood cell
count of 25,000/mm3.
The mother was given ampicillin at the time of delivery. She
recovered.
Blood cultures were taken from the infant soon after birth. These
grew Gram-positive coccobacilli that produced beta hemolysis on
sheep blood agar.
Lumbar punctures were attempted, but they were too traumatic and
non-revealing.

Listeria monocytogenes

Gram positif, batang pendek bulat (kokobasiler)

Fakultatif anaerob, b-hemolytic on sheep blood agar
Tumbuh pada suhu 4C. Katalase positif
Commonly associated with dairy products

-hemolysis

Gejala klinis
Meningoencephalitis: Most common in neonates
Low grade septicemia during pregnancy
Granulomatosis infantiseptica-intrauterine
Early onset syndrome- before delivery characterized by
septicemia, disseminated abscesses and granulomas
Late onset syndrome- meningitis 2-3 weeks after birth
(MUST BE differentiated from Group B Streptococcal
meningitis)

Bacillus
Antibiotic susceptibility and treatment
Penicillin or tetracycline
A short-term PA vaccine is available for industrial
workers and others at high risk.

Other Bacillus species

Bacillus subtilis, and occasional other species
may occasionally cause opportunistic infections.
Bacillus cereus is a major cause of enterotoxin
food poisoning; the toxin is protein in nature and
can be destroyed by heating; food poisoning occurs
after ingestion of pre-formed toxin; vomiting occurs
1-5 hours after ingestion

Bacillus
B. cereus is also an opportunistic pathogen that
has been cultured from cases of septicemia,
endocarditis, meningitis, wound infections,
pneumonia, and fulminant eye infections; in addition
to the enterotoxin that bacteria may
produce, a
dermonecrotic and a lethal toxin, hemolysins,
lecithinase, proteases, and nucleases may be
involved in its pathogenesis; clindamycin with or
without gentamycin may be used for treatment
infections

Basil gram (+) berspora

I.Aerob
a.Bacillus cereus
b.B.subtilis
c.B.anthracis
II.Anaerob
a.Clostridium tetani
b.Cl.perfringens (welchii)
c.Cl.botulinum
d.Cl.difficile

Bacillus anthracis
Menyebabkan peny.anthrax (zoonosis).
Infeksi terjadi pd manusia melalui spora.
1.kulit : berupa papula vesikula pustula maligna
ulkus nekrotik.
kalau menjalar keseluruh tubuh septikemia.
2.gasterointestinal :melalui makanan (daging hewan)
3.pneumonia.per inhalasi paru paru
secara alamiah : woolsorter disease.
dapat dibuat senjata biologis (WMD)
bersifat sangat fatal (mematikan)

Sifat sifat B.anthracis

batang besar,ujung persegi,1 x 3-4 um, tersusun spt rantai
panjang (mirip bambu) berspora sentral- nonmotil.
Pembiakan :koloni bulat,transparan,hemolisis(-),mencairkan
gelatin.
Kapsul terdiri dari asam glutamat, sedangkan badan berupa
protein dan polisakarida.
Menghasilkan toksin :
1. A protein
2. Faktor edema
3. Faktor lethal

Diagnosis laboratorium
1.
2.
3.
4.

Bahan pemeriksaan : pus, darah,sputum, jr biopsi

Pengecatan : basil gram (+),bentuk rantai.
Pembiakan pd agar darah.
Tes Ascoli : terbentuk cincin presipitat apabila serum
imun + ekstrak jaringan terinfeksi.

Terapi : penisilin
Pencegahan : vaksin anthrax
Epidemiologi : spora tanah hewan (domba) manusia

Kontrol :
1. Bangkai hewan dibakar.
2. Produk tercemar di autoklaf
3. Gunakan sarung tangan,masker,baju
pelindung, goggle, dll.
4. Vaksinasi hewan ternak.

Clostridium tetani
Dijumpai di seluruh dunia : tanah,kotoran hewan,dll.
Basil gr(+), anaerob, spora terminal(+), motil.
Menghasilkan eksotoksin yg bekerja pd CNS.
spasme ,hiperrefleksi, kejang ok tetanospasmin.
Kuman tidak invasif : masuk melalui luka yg dalam/kotor.
(bayi :umbilikus, luka tusuk, lubang gigi, dll.)
Spora vegetatif eksotoksin ke axon/darah CNS.
Gejala : trismus (lockjaw),risus sardonicus.
Pencegahan : DPT,ATS profilaksis,booster TT.
Pengobatan : wound cleansing,penisilin,ATS & TT.

Pemeriksaan laboratorium
Spesimen : pus ,dibiakkan pd anaerobic jar .
Tampak koloni kecil spt jala halus & berjalinan.
Pada media cair (chopped cooked meat) : tumbuh pada
dasar tabung.
Pemeriksaan lab jarang diminta !
Kalau secara klinis (+) gejala tetanus :
Bayi : tetanus neonatorum, orang dewasa (luka kotor)
langsung MRS terapi .

Clostridium botulinum
Penyebaran di seluruh dunia (tanah,kotoran hewan)
Menghasilkan 7 jenis toksin, A s/d G.
A, B, dan E : peny.pd.manusia (keracunan makanan)
C
: limber neck (unggas)
D
: botulismus pd peternak.
Penyakit timbul pasca mengkonsumsi makanan yang
diawetkan : kalengan, makanan yg diasap,divakum.
Pd bayi : madu sids (sudden infant death syndrome)

Toksin rusak jika dipanaskan 100 C / 20 menit

Gejala penyakit :
18-96 jam pasca makan makanan yg tercemar
flaccid paralysis berupa gangguan penglihatan,
tak dpt menelan, kesulitan bicara, paralisis bulbar
progresif, tanpa demam.
Kematian akibat paralisis otot pernafasan dan
jantung.

Clostridium perfringens
Spora masuk melalui luka dalam/kotor (fraktur terbuka)
Menyebabkan gas gangrene (clostridial myonecrosis)
ok alfa toksin (lecithinase) yg menghidrolisis membran sel.
Gejala : nyeri hebat, menjalar pembentukan gas pd
jr.subkutan & otot .Kalau diraba ber krepitasi.
Terapi : oksigen hiperbarik dan amputasi.

Clostridium difficile
Menyebabkan kolitis pseudomembranosa
terutama pd anak.
Diare th clindamycin flora normal mati
Berkembang Cl.diff toksin kolitis ps
dapat berakibat fatal.
Terapi : vancomycin
Lab : mencari eksotoksin dr tinja pasien.

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