TC GI NGUYN LN DNG V CNG TC VIN : Bi Th Vit H, Dng Vn Hp, o Th Lng, inh Thy Hng, L Hong Yn, Nguyn nh Quyn, Nguyn Vn Bc, Nguyn Hoi H, Nguyn Lin Hoa, Nguyn Kim N Tho, Phm Th Hi, Phm Vn Ty Vi hng ni u Lc s ngnh Vi sinh vt NhngcimchungcaViSinhVt Cu trc t bo Vi khun Cc nhm Vi khun ch yu X khun C khun Phng php thc nghim dng nh tn cc loi vi khun Virus Vi nm (Microfungi) Nm si (Filamentous Fungi) Nm men Vi to 1 (Microalgae) Vi to 2 Phn loi vi sinh bng Sinh Hc Phn T Gii thiu mt s k thut bo qun vi sinh vt Dinh dng ca vi sinh vt Sinh trng v pht trin ca vi sinh vt c ch vi sinh vt bng cc tc nhn vt l v ha hc Khi nim chung v trao i cht Vi sinh vt Gii phng v bo ton nng lng vi sinh vt S dng nng lng trong sinh tng hp vi sinh vt http://vietsciences.free.frvhttp://vietsciences.org

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Cng bn c.Vi sinh vt (microorganisms) l nhng sinh vt nh b n m c ch c th th y c chng di knh hin vi quang hc hay knh hin vi in t. Vi sinh vt gy ra rt nhiu bnh him ngho cho ngi, cho gia sc, gia cm. Tuy nhin s vi sinh v t gy bnh ch chim mt phn rt nh trong th gii vi sinh vt. T xa x a ng i ta bit ng dng cc vi sinh vt c ch (tuy cha h bit ti s tn ti ca chng) ch bin thc phm ( nh nu ru, lm tng, mm, nc mm, gim, sa chua, chao, mui da, mui c, ...), phn, ngm v cy ly si, xp i t, trng lun canh vi cy h u...; hoc s dng cc bin php ngn chn tc hi ca vi sinh vt (nh p mui tht, c, lm mt, phi kh c ci, tm, c...). Sau vic Leeuwenhoek pht hin ra vi sinh vt v vic Louis Pasteur pht hin ra bn cht ca cc vi sinh vt- t khai sinh ra ngnh Vi sinh v t h c (Microbiology)- th nhn loi bt u quan tm rt nhiu n lnh vc khoa h c m i m ny. Vi sinh vt hc tr thnh nn tng cho s pht trin ca Cng ngh sinh hc(CNSH). Ngi ta chia s pht trin ca CNSH ra thnh 3 giai on: * CNSH truyn thng l cc qu trnh dn d nhm ch bin , b o qun cc loi thc phm, x l t ai, phn bn phc v nng nghip... * CNSH cn i l qu trnh s dng cc ni ln men cng nghip sn xut quy m ln cc sn phm sinh hc nh m chnh (bt ngt), lizin v cc axt amin khc, cc acid hu c, cc dung mi hu c, cht khng sinh, mt s vitamin (nh vitamin B2, B12, C...), nhiu loi enzym... * CNSH hin i chia ra cc lnh vc nh CN di truyn (genetic engineering), cng ngh t bo (cell engineering), cng ngh enzym v protein (enzyme/protein engineering), CN vi sinh vt/ CN ln men (microbial engineering / fermentation), CN mi trng (environmental engineering). CNSH hin i thng gn lin vi cc c th mang gen ti t hp ( recombination gene). Vi sinh vt hc l khoa hc nghin cu v cc c th hoc cc nhn t qu nh n mc khng thy c bng mt thng, tc l cc vi sinh vt. Vi sinh v t h c ang c ging dy trong rt nhiu trng i hc, Cao ng, Trung hc chuyn nghip v cng c cp n t nhiu bc ph thng. Vi sinh vt h c l nh ng kin thc lin quan n cuc sng ca mi ngi. Bn cnh gio trnh Vi sinh vt hc c bin son t lu v c ti bn nhiu ln chng ti mu n cung c p thm cho ng o bn c, nht l cc thy c gio ang ging dy v vi sinh v t h c t p ti liu trnh by vi nhiu tranh nh v s , biu ny. C th coi y l ph n tham kho thit k cc bi ging nhm mc tiu cp nht vi cc tin b ca ngnh khoa hc lun lun i mi ny.

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gip cc bn tr tip cn c vi sch bo nc ngoi, nht l cp nht kin thc qua Internet chng ti c gng vit thm ting Anh sau cc thu t ng khoa hc v nguyn cc ch thch ting Anh trong hnh v. Thng qua vic gio vin hng dn sinh vin in ting Vit vo cc hnh v s gip ch rt nhiu cho qu trnh t hc ca sinh vin. Rt mong nhn c kin ng gp ca cc bn ng nghip v ng o bn c. V mc ch phc v phi li nhun chng ti mong c lng th vic s dng nhng hnh nh thu nhn qua Internet.

GS.TS. Nguyn Ln Dng Trung tm Cng ngh sinh hc i hc Quc gia H Ni

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Bi 1 LcsnghincuVisinhvthc

1546- Girolamo Fracastoro (1478, 1553). cho rng cc c th nh b l tc nhn gy ra bnh tt. ng vit bi th Syphilis sive de morbo gallico (1530) v t ta ca bi th , ngi ta dng t tn bnh 1590-1608- Zacharias Janssen ln u tin lp ghp knh hin vi. 1676- AntonyvanLeeuwenhoek (16321723) hon thin knh hin vi v khm ph ra th gii vi sinh vt (m ng gi l anmalcules). 1688- Nh vn vt hc ngi Francisco Redi cng b nghin cu v s pht sinh t nhin ca gii. 1765-1776- Spallanzani (1729-1799) cng kch thuyt Pht sinh t nhin 1786- Mller a ra s phn loi u tin v vi khun 1798- Edward Jenner ngh ra phng php chng m u b phong nga bnh u ma 1838-1839- Schwann v Schleiden cng b Hc thuyt t bo. 1835-1844- Basi cng b bnh ca tm do nm gy nn v nhiu bnh tt khc do vi sinh vt gy nn. 1847-1850- Semmelweis cho rng bnh st hu sn ly truyn qua th y thu c v kin ngh dng phng php v khun phng bnh. 1849- Snow nghin cu dch t ca bnh t vng London. 1857- LouisPasteur(18221895) chng minh qu trnh ln men lactic l gy nn bi vi sinh vt. 1858- Virchov tuyn b t bo c sinh ra t t bo. 1861- Pasteur chng minh vi sinh vt khng t pht sinh nh theo thuyt t sinh. 1867- Lister cng b cng trnh nghin cu v phu thut v khun. 1869- Miescher khm ph ra acid nucleic. 4

1876-1877- RobertKoch (18431910) chng minh bnh than do vi khun Bacillus anthracis gy nn. 1880- Alphonse Laveran pht hin k sinh trng Plasmodium gy ra bnh st rt. 1881- Robert Koch nui cy thun khit c vi khun trn mi tr ng c cha gelatin. Pasteur tm ra vaccin chng bnh than. 1882- Koch pht hin ra vi khun lao - Mycobacterium tuberculosis. 1884- Ln u tin cng b Nguyn l Koch. Elie Metchnikoff (18451916) miu t hin tng thc bo (phagocytosis) Trin khai ni kh trng cao p (autoclave) Trin khai phng php nhum Gram. 1885- Pasteur tm ra vaccin chng bnh di. Escherich tm ra vi khun Escherichia coli gy ra bnh tiu chy. phi. 1886- Fraenkel pht hin thy Streptococcus pneumoniae gy ra bnh vim

1887- Richard Petri phi hin ta cch dng hp lng (a Petri) nui c y vi sinh vt . 1887-1890- Winogradsky nghin cu v vi khun lu hunh v vi khun nitrat ho. 1889- Beijerink phn lp c vi khun nt sn t r u. 1890- Von Behring lm ra khng c t chng bnh un vn v bnh bch hu. 1892- Ivanowsky pht hin ra mm bnh nh hn vi khun (virus) gy ra bnh khm cy thuc l. 1894- Kitasato v Yersin khm ph ra vi khun gy bnh dch hch ( Yersina pestis). 1895- Bordet khm ph ra B th (complement) 5

1896- Van Ermengem tm ra mm bnh ng c tht (vi khun Clostridium botulinum). 1897- Buchner tch ra c cc men (ferments) t nm men (yeast). Ross chng minh k sinh trng st rt ly truyn bnh qua mui. 1899- Beijerink chng minh nhng ht virus gy nn bnh khm l thuc l. 1900- Reed chng minh bnh st vng ly truyn do mui. 1902- Landsteiner khm ph ra cc nhm mu 1903- Wright v cng s khm ph ra Khng th (antibody) trong mu c a cc ng vt min dch. 1905- Schaudinn v Hoffmann tm ra mm bnh giang mai (Treponema pallidum). 1906- Wassermann pht hin ra xt nghim c nh b th chn on giang mai. 1909- Ricketts chng minh bnh St ban ni lan truyn qua ve l do m m bnh vi khun (Rickettsia rickettsii). 1910- Rous pht hin ra ung th gia cm. 1915-1917- DHerelle v Twort pht hin ra virus ca vi khun ( thc khun th) 1921- Fleming khm ph ra lizzim (lysozyme). 1923-Xut bn ln u cun phn loi Vi khun (Bergeys Manual) 1928- Griffith khm ph ra vic bin np (transformation) vi khun. 1929- Fleming pht hin ra penicillin. 1931- Van Niel chng minh vi khun quang hp s dng cht kh nh ngu n cung cp electron v khng sn sinh xy. 1933- Ruska lm ra chic knh hin vi in t u tin. 1935- Stanley kt tinh c virus khm thuc l (TMV). Domag tm ra thuc sulfamide.

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1937- Chatton phn chia sinh vt thnh hai nhm: Nhn s (Procaryotes) v Nhn tht (Eucaryotes). 1941- Beadle v Tatum a ra gi thuyt mt gen- mt enzym. np. 1944- Avery chng minh ADN chuyn thng tin di truyn trong qu trnh bi n Waksman tm ra streptomycin. khun. 1046- Lederberg v Tatum khm ph ra qu trnh tip hp (conjugation) vi

1949- Enders, Weller v Robbins nui c virus Polio (Poliovirus) trn m ngi nui cy. 1950- Lwoff xc nh c cc thc khun th tim tan (lysogenic bacteriophages). 1952- Hershey v Chase chng minh thc khun th tim ADN ca mnh vo t bo vt ch (host). khun. Zinder v Lederberg khm ph ra qu trnh ti np (transduction) vi

1953- Frits Zernike Lm ra knh hin vi tng phn pha (phase-contrast microscope). Medawar khm ph ra hin tng nhn min dch (immune tolerance). Watson v Crick khm ph ra chui xon kp ca ADN 1955- Jacob v Monod khm ph ra yu t F l mt plasmid. Jerne v Burnet chng minh l thuyt chn lc clone (clonal selection). 1959- Yalow trin khai k thut Min dch phng x. 1961- Jacob v Monod gii thiu m hnh iu ho hot ng gen nh operon. 1961-1966- Nirenberg, Khorana v cng s gii thch m di truyn. 1962- Porter chng minh cu trc c bn ca Globulin min dch G. Tng hp c quinolone u tin c tc dng di t khun ( acid nalidixic).

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1970- Arber v Smith khm ph ra enzym gii hn (restriction endonuclease) Temin v Baltimore khm ph ra enzym phin m ngc (reverse transcriptase) 1973- Ames trin khai phng php vi sinh vt hc khm ph ra cc yu t gy t bin (mutagens). vi khun. Cohen, Boyer, Chang v Helling s dng vect plasmid tch dng gen

1975- Kohler v Milstein pht trin k thut sn xut cc khng th n dng ( monoclonal antibodies). Pht hin ra bnh Lyme. 1977- Woese v Fox tha nhn Vi khun c (Archaea) l m t nhm vi sinh v t ring bit. Gilbert v Sanger trin khai k thut gii trnh t ADN (DNA sequencing) 1979-Tng hp Insulin bng k thut ti t hp ADN. Chnh thc ngn chn c bnh u ma. 1980- Pht trin knh hin vi in t qut 1982- Pht trin vaccin ti t hp chng vim gan B. 1982-1983- Cech v Altman pht minh ra ARN xc tc. 1983-1984- Gallo v Montagnier phn lp v nh loi virus gy suy gim mi n dch ngi. Mulli trin khai k thut PCR (polymerase chain reaction). 1986- Ln u tin ng dng trn ngi vaccin c sn xu t bng k thu t di truyn (vaccin vim gan B). ngi. 1990- Bt u th nghim ln u tin liu php gen (gene-therapy) trn 1992- Th nghim u tin trn ngi liu php i ngha (antisense therapy). 1995- Hoa K chp thun s dng vaccin u g. Gii trnh t h gen ca vi khun Haemophilus influenzae.

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1996- Gii trnh t h gen ca vi khun Methanococcus jannaschii. Gii trnh t h gen nm men. 1997- Pht hin ra loi vi khun ln nht Thiomargarita namibiensis Gii trnh t h gen vi khun Escherichia coli. 2000- Pht hin ra vi khun t Vibrio cholerae c 2 nhim sc th ring bit.

Janssen

Leeuwenhoek(1632 1723)

Pasteur (18221895)

KnhhinvicaLeeuwenhoek

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Bttchmiutvisinhvtca Leeuwenhoek

Th nghim bnh c cong phnithuyttsinh (Pasteur)

RobertKoch(18431910)

Vikhunlaochpquaknh hinvi

ElieMetchnikoff(1845 1916)

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AlexanderFleming(18811955)

NmPenicilliumsn sinhpenicillin

Bi2NhngcimchungcaViSinhVt

1Visinhvtthucgiisinhvtno?

Vi sinh vt khng phi l mt nhm phn loi trong sinh gi i m l bao g m t t c cc sinh vt c kch thc hin vi, khng thy r c bng m t th ng, do phi s dng knh hin vi thng hoc knh hin vi i n t. Ngoi ra mu n nghin cu vi sinh vt ngi ta phi s dng ti phng php nui cy v khun. T trc n nay c rt nhiu h thng phn loi sinh vt. Cc n v phn loi sinh vt ni chung v vi sinh vt ni ring i t thp ln cao l Loi (Species), Chi (Genus), H (Family), B (Order), Lp (Class), Ngnh (Phylum), v Gi i (Kingdom). Hin nay trn gii cn c mt mc phn loi na gi l lnh gi i (Domain). y l cha k n cc mc phn loi trung gian nh Loi ph (Subspecies), Chi ph (Subgenus), H ph (Subfamily), B ph (Suborder),Lp ph (Subclass), Ngnh ph (Subphylum).

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John Ray

Carl Von Linnaeus

Xa kia John Ray (1627-1705) v Carl Von Linnaeus (1707-1778) ch chia ra 2 gii l Thc vt v ng vt. Nm 1866 E. H. Haeckel (1834-1919) b sung thm gii Nguyn sinh (Protista). Nm 1969 R. H. Whitaker (1921-1981) xut h thng phn loi 5 gii : Khi sinh (Monera), Nguyn sinh (Protista), Nm (Fungi), Thc vt (Plantae) v ng vt (Animalia). Khi sinh bao gm Vi khun (Bacteria) v Vi khun lam (Cyanobacteria). Nguyn sinh bao gm ng vt nguyn sinh (Protzoa), To (Algae) v cc Nm si sng trong nc (Water molds).

Gn y hn c h thng phn loi 6 gii- nh 5 gii trn nhng thm gii C vi khun (Archaebacteria), gii Khi sinh i thnh gii Vi khun tht (Eubacteria) (P. H. Raven, G. B. Johnson, 2002). C vi khun v Vi khun tht thuc Cn

T. Cavalier-Smith (1993) th li xut h thng phn loi 8 gii: Vi khun tht (Eubacteria), C vi khun (Archaebacteria), 12

C trng (Archezoa), Sc khun (Chromista), Nm (Fungi), Thc vt (Plantae) v ng vt (Animalia).

Theo R. Cavalier-Smith th C trng (nh Giardia) bao gm cc c th n bo nguyn thu c nhn tht, c ribosom 70S, cha c b my Golgi, cha c ty th (mitochondria) ch a c th di p lc (Chloroplast), cha c peroxisome. Sc khun bao gm phn ln cc c th quang hp cha th dip lc trong cc phin (lumen) ca mng li ni cht nhn (rough endpplasmic reticulum) ch khng phi trong t bo cht (cytoplasm), chng hn nh To silic , To nu, Cryptomonas, Nm non.

Nm 1980, Carl R. Woese da trn nhng nghin cu sinh h c phn t pht hin thy C khun c s sai khc ln trong trt t nucleotid ARN c a ribosom 16S v 18S. ng a ra h thng phn loi ba lnh gii (Domain) bao gm C khun (Archae), Vi khun (Bacteria) v Sinh vt nhn thc (Eucarya). C khun l nhm vi sinh vt c ngun gc c xa. Chng bao gm cc nhm vi khun c th pht trin c trong cc mi trng cc oan (extra), ch ng h n nh nhm a mn (Halobacteriales), nhm a nhit (Thermococcales, Thermoproteus, Thermoplasmatales), nhm k kh sinh mtan ( Methanococcales, Methanobacteriales, Methanomicrobiales), nhm vi khun lu hunh a nhit (Sulfobales, Desulfurococcales). Monera trong h thng 5 gii tng ng vi Vi khun v C khun trong h thng 8 gii v trong h thng 3 lnh gii. Nguyn sinh trong h th ng 5 gi i t ng ng vi 3 gii C trng (Archaezoa), Nguyn sinh (Protista-Protozoa) v S c khun

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(Chromista) trong h thng 8 gii v tng ng vi 5 nhm sau y trong h th ng 3 lnh gii (domain): Archaezoa, Euglenozoa, Alveolata, Stramenopila v Rhodophyta. Theo h thng 3 lnh gii th Archaezoa bao gm Diplomonad, Trichomonad v Microsporidian. Euglenozoa ao gm Euglenoid v Kinetoplastid. Alveolata bao gm Dinoflagellate, Apicomplexan, v Ciliate. Strmenopila bao gm T o silic (Diatoms) , To vng (Golden algae), To nu (Brown algae) v Nm si sng trong n c (Water mold) . Rhodophyta gm cc To (Red algae). Ring To lc (Green algae) th m t phn thuc Nguyn sinh (Protista) mt phn thuc Thc vt (Plantae)

H thng phn loi 5 gii sinh vt

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H thng phn loi 6 gii sinh vt

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H thng phn loi 8 gii sinh vt

H thng 3 lnh gii (domain)

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Monera hay 2 lnh gii Vi khun v C khun thuc nhm Sinh vt nhn s (Prokaryote), cn cc sinh vt khc u thuc nhm Sinh vt nhn tht (Eukaryote). Sai khc gi a 3 lnh gi i Bacteria, Archaea v Eukarya c trnh by trn bng di y:

***- So snh ba lnh gii Bacteria, Archaea v Eukarya

c imNhn c mng nhn v hch nhn Phc hp bo quan c mng Thnh t bo Mng lipid Ti kh Khng Khng

BacteriaKhng Khng

ArchaeaC C

Eukarya

Hu ht c peptidoglycan cha acid muramic Cha lin kt este, cc acid bo mch thng C Thymine c trong phn ln tARN

Nhiu loi khc nhau, khng cha acid muramic Cha lin kt ete, cc chui aliphatic phn nhnh C

Khng cha acid muramic Cha lin kt este, cc acid bo mch thng Khng

Khng c thymine trong nhnh T C thymine hoc TyC ca tARN tARN m u cha methionine Khng C C

ARN vn chuyn tARN m u cha Nformylmethionine mARN a cistron Intron trong mARN C Khng tARN m u cha methionine C Khng Khng

Ghp ni, gn m v gn ui Khng polyA vo mARN

RibosomKch thc Yu t ko di EF2 70S Khng phn ng vi c t bch hu 70S C phn ng Khng Mn cm 80S (ribosom t bo cht) C phn ng Khng Mn cm

Mn cm vi cloramphenicol Mn cm v kanamycin Mn cm vi anisomycin Khng

ARN polymerase ph thuc ADNS lng enzym Cu trc Mn cm vi rifampicin Promoter typ Polymerase II Mt 4 tiu n v Mn cm Khng Mt s 8-12 tiu n v Khng C Ba 12-14 tiu n v Khng C

Trao i chtTng t ATPase Khng C C

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Sinh methane C nh N2 Quang hp vi dip lc Ho dng v c

Khng C C C

C C Khng C

Khng Khng C Khng

hiu c chi tit ni dung ghi trong bng ni trn gio vin c n gi i thch cho sinh vin nhng kin thc c bn thuc gio trnh T bo hc v Di truyn hc Phn ln vi sinh vt thuc v ba nhm C khun, Vi khun v Nguyn sinh. Trong gi i N m, th nm men (yeast), nm si (filamentous Fungi) v dng si (mycelia) c a m i n m l n u c coi l vi sinh vt. Nh vy l vi sinh vt khng c mt trong hai gi i ng vt v Th c v t. Ngi ta c tnh trong s 1,5 triu loi sinh vt c khong 200 000 loi vi sinh vt (100 000 loi ng v t nguyn sinh v to, 90 000 loi nm, 2500 loi vi khun lam v 1500 loi vi khu n). Tuy nhin hng nm, c thm hng nghn loi sinh vt mi c pht hin, trong c khng t loi vi sinh vt. Virus l mt dng c bit cha c cu trc c th cho nn ch a c k n trong s 200 000 loi vi sinh vt ni trn. S virus c t tn l khong 4000 loi.

Poliovirus

Virus cm g H5N1

Virus HIV/AIDS

Trong thc t, s loi vi sinh vt phi ti hng tri u loi. B o tng gi ng chu n vi sinh v t (VTCC) thuc TT Cng ngh Sinh hc, HQG H Ni hp tc vi cc nh khoa h c Nh t b n v da trn cc k thut sinh hc phn t bc u pht hi n c kh nhi u loi vi sinh v t m i c th gii cng nhn.

2-Cc c im chung ca vi sinh vt :Vi sinh vt c cc c im chung sau y :

1)-Kch thc nh b :18

Vi sinh vt thng c o kch thc bng n v micromet (1mm=1/1000mmhay1/1000 000m).viruscokchthcnvbngnanomet(1nn=1/1000000mmhay1/1000000 000m). Kchthccngbthdintchbmtcavisinhvttrong1nvthtchcngln. Chnghnngknhca1cukhun(Coccus)chc1mm,nhngnuxpychng thnh1khilpnhngcthlchl1cm3thchngcdintchbmtrngti...6m2!

Lightmicroscope:KHVquanghc Electronmicroscope:KHVint Mostbacteria:Phnlnvikhun

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Kchthcvikhunsoviukimkhu

Badngchyuvikhun:trckhun,cukhunvxonkhun.

2)Hpthunhiu,chuynhonhanh:Tuyvisinhvtckchthcrtnhbnhngchnglicnnglchpthuvchuyn 20

hovtxaccsinhvtkhc.Chnghn1vikhunlctic(Lactobacillus)trong1gicth phngiicmtlngnglactoselnhn10010000lnsovikhilngcachng. tctnghpproteincanmmencaogp1000lnsoviutngvgp100000ln sovitrub.

LactobacillusquaKHVint

3)Sinhtrngnhanh,phttrinmnh:Chnghn,1trckhunitrng(Escherichiacoli)trongcciukinthchhpch sau1220phtliphnctmtln.Nulythigianthhl20phtthmigiphnct3 ln,sau24giphnct72lnvtora4722366500000000000000000tbo(4722 366.1017),tngngvi1khilng...4722tn.Ttnhintrongtnhinkhngcc cciukintiunhvy(vthiuthcn,thiuoxy,dthaccsnphmtraoicht chi...).Trongnilnmenvicciukinnuicythchhpt1tbocthtorasau 24gikhong1000000001000000000tbo.Thigianthhcanmmendihn,v dvimenru(Saccharomycescerevisiae)l120pht.Vinhiuvisinhvtkhccndi hnna,vdvitoTiucu(Chlorella)l7gi,vivikhunlamNostocl23gi...Cth nikhngcsinhvtnoctcsinhsinynnhanhnhvisinhvt.

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VikunEscherichiacoli

Nmmen Saccharomyces cerevisiae

NmsiAlternaria VitoChlorella

4)Cnnglcthchngmnhvddngphtsinhbind:Trongqutrnhtinholudivisinhvttochomnhnhngcchiuhotrao ichtthchngcvinhngiukinsngrtkhcnhau,kcnhngiukinht scbtlimccsinhvtkhctgngkhngthtntic.Cvisinhvtsngcmi trngnngn1300C, lnhn050C,mnnnng32%muin,ngtnnng mtong,pHthpn0,5hoccaon10,7,psutcaontrn1103at.haycphng xcaon750000rad.Nhiuvisinhvtcthphttrintttrongiukintuytikkh, cnoinmsicthphttrindyctrongbngmtthivinngFormolrtcao... Visinhvtaslnbo,nbi,sinhsnnhanh,slngnhiu,tipxctrctip vimitrngsng...dortddngphtsinhbind.Tnsbindthngmc105 1010.Chsaumtthigianngncthtoramtslngrtlncccthbindcc hhsau.Nhngbindcchsalihiuqurtlntrongsnxut.Nunhkhimi phthinrapenicillinhottnhcht20nv/mldchlnmen(1943)thnayctht trn100000nv/ml.Khimiphthinraacidglutamiccht12g/lthnay tn 150g/mldchlnmen(VEDANVitNam).

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NhmyVedanVitNam

5)Phnbrng,chngloinhiu:VisinhvtcmtkhpminitrnTrit,trongkhngkh,trongt,trnnicao, dibinsu,trncth,ngi,ngvt,thcvt,trongthcphm,trnmivt... Visinhvtthamgiatchccvovicthchinccvngtunhonsinhahohc (biogeochemicalcycles)nhvngtunhonC,vngtunhonn,vngtunhonP,vng tunhonS,vngtunhonFe... Trong nc vi sinh vt c nhiu vng duyn hi (littoral zone), vng nc nng (limneticzone)vngaycvngncsu(profundalzone),vngyaoh(benthiczone). Trongkhngkhthcnglncaoslngvisinhvtcngt.Slngvisinhvttrong khngkhcckhudncngccaohnrtnhiusovikhngkhtrnmtbinvnht ltrongkhngkhBccc,Namcc... Hunhkhngchpchtcarbonno(trkimcng,grapht...)mkhnglthc ncanhngnhmvisinhvtno(kcdum,khthinnhin,formol.dioxin...).Visinh vtcrtphongphcckiudinhdngkhcnhau:quangtdng(photoautotrophy), quang d dng (photoheterotrophy), ho t dng (chemoautotrophy), ho d dng (chemoheterotrophy).t dng cht sinh trng (auxoautotroph), d dng cht sinh trng (auxoheterotroph)...

23

6)Lsinhvtxuthinutintrntrit:Trithnhthnhcchy4,6tnmnhngchonnaymichtmthyduvt cassngtcchy3,5tnm.lccvisinhvthothchcnlivttchtrong cctngc.Visinhvthothchcanhtcphthinlnhngdngrtgingvi Vikhunlamngynay.ChngcJ.WilliamSchopftmthyticctngcminTy Australia.Chngcdngabongin,nithnhsidinvichcmmvingknh khong12mmvcthnhtbokhdy.Trcccnhkhoahccngtmthyvt tchcachiGloeodiniopsiscninicchy1,5tnmvvttchcachiPalaeolyngbya cninicchy950triunm.

Vttchvikhunlam cchy3,5tnm

VttchGloeodiniopsiscchy VttchPalaeolyngbyacchy 1,5tnm 950triunm

24

Bi 3 Cutrctbovikhun1. Thnhtbo:

Thnh t bo (cell wall) gip duy tr hnh thi ca t bo, h tr s chuyn ng ca tin mao (flagellum) , gip t bo khng vi p sut thm thu, h tr qu trnh phn ct t bo , cn tr s xm nhp ca mt s cht c phn t ln, lin quan n tnh khng nguyn , tnh gy bnh, tnh mn cm vi Thc khun th (bacteriophage). Nm 1884 H.Christian Gram ngh ra phng php nhum phn bi t phn chia vi khun thnh 2 nhm khc nhau : vi khun Gram dng (G+) v vi khun Gram m (G-). Phng php nhum Gram v sau c s dng rng ri khi nh loi vi sinh vt. Thnh phn ho hc ca 2 nhm ny khc nhau ch yu nh sau :

GramdngThnhphn Peptidoglycan Acidteicoic(Teichoicacid) Lipid Protein 3095 Cao Hunhkhngc Khngchocct

Gram m520 0 20 Cao

Tl%ivikhilng khcathnhtbo

Mngsinhcht(plasmamembrane);Mngngoi(outermembrane);Chucht (Periplasmicspace)

25

Peptidoglycan l loi polyme xp, kh bn vng, cu to bi 3 thnh phn: -N-Acetylglucosamin ( N-Acetylglucosamine, NAG) -Acid N-Acetylmuramic (N-Acetylmuramic acid, NAM) -Tetrapeptid cha c D- v L- acid amin

26

27

ThnhtbovikhunGramdng

28

ThnhtbovikhunGramm

2Mngsinhcht: MngsinhchthayMngtbocht(Cytoplasmicmembrane,CM)vi khun cng tng t nh cc sinh vt khc. Chng cu to bi 2 lp phospholipid (PL), chim 3040% khi lng ca mng, v cc protein (nm trong, ngoi hay xen gia mng), chim 6070% khi lng ca mng. u phosphatcaPLtchin,phncc,anc;uihydrocarbonkhngtch in,khngphncc,knc. CMcccchcnngchyusauy: Khngchsqualicaccchtdinhdng,ccsn phmtraoicht Duytrpsutthmthubnhthngtrongtbo. Lnisinhtnghpccthnhphncathnhtbov ccpolymecabaonhy(capsule). Lnitinhnhqutrnhphosphoryloxyhovqutrnh phosphorylquanghp(vikhunquangtdng) Lnitnghpnhiuenzym,ccproteincachuih hp. Cungcpnnglngchoshotngcatinmao

29

Sinhvininchthchtheohngdncagiovin

Cutrccauvuicaphospholipid

2.

Tbocht:

T bo cht (TBC-Cytoplasm) l phn vt cht dng keo n m bn trong mng sinh cht, cha ti 80% l nc. Trong t bo cht c protein, acid nucleic, hydrat carbon, lipid, cc ion v c v nhiu nhiu cht khc c khi l ng phn t thp. Bo quan ng lu trong TBC l ribosom (ribosome). Ribosom n m t do trong t bo cht v chim ti 70% trng lng kh c a TBC. Ribosom gm 2 tiu phn (50S v 30S), hai tiu phn ny kt hp vi nhau to thnh ribosom 70S. S l n v Svedberg- i lng o tc lng khi ly tm cao tc. Cu trc ca ribosom vi khun so vi ribosom 80S cc sinh vt nhn th t

30

(nm, thc vt, ng vt) c trnh by trong bng sau y (Gio vin gi ng sinh vin ch thch vo hnh bng ting Vit)

Ribosomvikhun

31

SosnhRibosomVikhunvccSinhvtnhntht(Eukaryoticribosome) Trongtbochtcavikhuncncthgpccchtdtrnhccht glycogen,htPHB(Polyhydroxybutyrat),Cyanophycin,Phycocyanin,ccht dnhimsc(metachromaticbody),ccgitluhunh... loivikhunditcntrngBacillusthuringiensisvBacillussphaericus cngptinhthc (parasoralbody)hnhqutrm,cbnchtproteinv chanhngctcthgithitrn100loisuhi(tinhthcchgii phngcttrongmitrngkimdoccvikhunnyhontonvhivi 32

ngi,giasc,giacm,thuhisnchiivitm).Bacillussphaericusc thditcungqungcaccloimui.

Bot(spore)vTinhthc(Crystal)Bacillusthuringiensis(tri)vBacillus sphaericus(phi).

3.

Thnhn:

Th nhn ( Nuclear body) vi khun l dng nhn nguyn thu, cha c mng nhn nn khng c hnh dng c nh, v v vy cn c gi l vng nhn. Khi nhum mu t bo bng thuc nhum Feulgen c th thy th nhn hin mu tm. l 1 nhim sc th (NST, chromosome) duy nht dng vng cha 1 si ADN xo n kp ( X khun Streptomyces c th gp nhim sc th dng thng). NST vi khun Escherichia coli di ti 1mm (!), c khi lng phn t l 3.109, cha 4,6.106 cp base nit. Th nhn l b phn cha ng thng tin di truyn ca vi khun.

33

*

ThnhntrongtbovikhunEscherichiacoli. NgoiNST,trongtbonhiuvikhuncngpnhngADNngoiNST. lnhngADNxonkpcdngvngkhpkn,ckhnngsaochpc lp,chngctnlPlasmid.

4.

Baonhy:

BaonhyhayGipmc(Capsule)gpmtsloivikhunviccmc khcnhau: Baonhymng(Vigipmc,Microcapssule) Baonhy(Gipmc,Capsule) Khinhy(Zooglea) Munquanstbaonhythnglntiubnvimctu,baonhyc mutrnghinlntrnnnti. Thnh phn ch yu ca bao nhy l polysaccarid, ngoi ra cng c polypeptid v protein. Trong thnh phn polysaccarid ngoi glucose cn c glucozamin,ramnose,acid2keto3deoxygalacturonic,aciduronic,acidpyruvic, acidaxetic... nghasinhhccabaonhyl:

34

Bovvikhuntrongiukinkhhn,bovvikhuntrnhbthc bo(trnghpPhcukhunDiplococcuspneumoniae) Cungcpchtdinhdngchovikhunkhithiuthcn Lnitchlumtssnphmtraoicht(dextran,xantan...) Gipvikhunbmvogith(trnghpccvikhungysurng nhStreptococcussalivarrius,Streptococcusmutans...)

VikhunAcetobacterxylinumcbaonhycutobicellulose.Ngitadng vikhunnynuicytrnncdachtoraThchda(Natadecoco).

da(Natadecoco)

VikhunLeuconostocmesenteroidescbaonhydychahpchtpolymel Dextranctcdngthayhuyttngkhicpcumthiuhuyttng.Sn 35

phmnyrtquantrngkhicchintranh.Vikhunnythnggpccnh myngvgytnthtngtrongccbchancpma.Nhenzym dextransuccrasemngsaccarosebchuynthnhdextranvfructose.

VikhunLeuconostocmesenteroides Mt s bao nhy ca vi khun cn c dng sn xut Xantan (Xanthane)dnglmchtphgiatrongcngnghipdum.

5.

Tinmaovkhunmao:

Tin mao (Lng roi, flagella) khng phi c m t mi vi khun, chng quyt nh kh nng v phng thc di ng ca vi khun. Tin mao l nhng s i lng di, di knh hin vi quang hc ch c th thy r khi nhum theo ph ng php ring. Di knh hin vi in t c th thy rt r cu trc c a tng si tin mao. xc nh xem vi khun c tin mao hay khng cn c cch th gin ti p nh m bi t kh nng di ng ca chng. Cy bng que cy nhn u vo mi tr ng th ch ng cha 0.4% thch (agar-agar), cn gi l mi trng thch mm. Nu thy vt c y lan nhanh ra xung quanh th chng t l vi khun c tin mao, c kh nng di ng. Tin mao c th gc (basal body), gm 1 tr nh c gn vi 4 a trn (vi khun G - ) c dng vng nhn (ring), k hiu l cc vng L,P,S v M. Vng L n m ngoi cng, tng ng vi lp liposaccarid ca mng ngoi ; vng P tng ng vi lp peptidoglycan, vng S tng ng vi lp khng gian chu cht ; vng M nm trong cng. Vi khun G+ ch c 2 vng : 1 vng nm ngoi tng ng vi thnh t bo v 1 vng trong tng ng vi mng sinh cht. Xuyn gi a cc vng l 1 tr nh (rod) c ng knh 7nm. Bao bc tin mao phn pha ngoi l mt bao ngn c hnh mc 36

(hook). Si tin mao (filament) di khong 10-20 m v c ng knh khong 1320nm. ng knh ca bao hnh mc l 17nm. Khong cch gi a vng S v vng M l 3mm, gia vng P v vng L l 9nm, gia vng P v vng S l 12nm. ng knh c a cc vng l 22nm, ng knh cc l cc vng l 10nm. Kho ng cch t m t ngoi ca vng L n mt trong ca vng M l 27nm. Si tin mao c u t o b i lo i protein c tn l flagellin, c trng lng phn t l 30 000-60 000. Mt s vi khu n c bao lng (sheath) bao bc sut chiu di si, nh trng hp chi Bdellovibrio hay vi khun t Vibrio cholera.

Tinmaovkhunmaovikhun

TinmaoVKGramdngTinmaoVKGramm

37

TinmaovikhunG+

38

Tin mao vi khun G Cc tiu phn (subunit) ca flagellin c tng hp t cc ht ribosom nm gn mng sinh chy tng hp nn v i qua li m to dn thnh si tin mao

39

Tinmaocavikhuncccloikhcnhaututngloi: Khngctinmao(vmao,atrichia) C1tinmaomccc(nmao,monotricha) C1chmtinmaomccc(chmmao,lophotricha) C2chmtinmaomc2cc(songchmmao,amphitricha) Cnhiutinmaomckhpquanhtbo(chumao,peritricha) CloitinmaomcgiatbonhtrnghpvikhunSelenomonas ruminantium.

40

Cc loi tin mao vi khun Kiu sp xp tin mao lin quan n hnh thc di ng c a vi khu n. Tin mao mc cc gip vi khun di ng theo kiu tin- li. Chng o ngc h ng bng cch o ngc hng quay ca tin mao. Vi khun chu mao di ng theo h ng no th cc tin mao chuyn ng theo hng ngc li. Khi tin mao khng t l i v m t hng th vi khun chuyn ng theo kiu nho ln. Tc di chuyn c a vi khu n c tin mao thng vo khong 20-80m/giy, ngha l trong 1 giy chuyn ng c mt khong cch ln hn gp 20-80 ln so vi chiu di ca c th chng. Cc chi vi khun thng c tin mao l Vibrio, Spirillum, Pseudomonas, Escherichia, Shigella, Salmonella, Proteus... cc chi Clostridium, Bacterium,Bacillus, ...c loi c tin mao c loi khng. cu khun ch c 1 chi (Planococcus) l c tin mao Xon th c mt dng tin mao c bit gi l tin mao chu cht (periplasmic flagella), hay cn gi l si trc ( axial fibrils), xut pht t cc t bo v qu n quang c th. Chng gip xon th chuyn ng c nh s un vn t bo theo kiu vn nt chai.

Xonth(Spirochete)quanstdiknhhinvinnen.

41

AF:Sitrc PC:ngnguynsinhcht OS:Vngoi IP:Lni

CchchuynngunvntboXonth(OS,AF,PCxemchthch hnhtrn). 42

6.

KhunmaovKhunmaogii:

Khunmao(hayTimmao,Nhungmao,Fimbriae)lnhngsilngrt mnh,rtngnmcquanhbmttbonhiuvikhunGramm.Chngc ngknhkhong79nm,rngrut(ngknhtrongl22,5nm),slng khong250300si/vikhun.Ktcucakhunmaoginnhnnhiusovi tinmao.Chngctcdnggipvikhunbmvogith(nhiuvikhungy bnhdngkhunmaobmchtvomngnhycanghhp,ng tiuho,ngtitniucangivngvt).

KhunmaovikhunE.coli CmtloikhunmaotbitgilKhunmaogii(Sexpili,Sexpiluss nhiu)cthgpmtsvikhunvislngchc110/vikhun.Nc cutogingkhunmao,ngknhkhong910nmnhngcthrtdi. Chngcthnilingiahaivikhunvlmcunichuynvtchtdi truyn(ADN)tthcho(donor)sangthnhn(recipient).Qutrnhnyc gilqutrnhgiaophi(mating)haytiphp(conjugation).Mtsthckhun th(bacteriophage)bmvoccthth(receptors)khunmaogiivbtu chutrnhphttrincachng.

43

Bi4CcnhmvikhunchyuTheo quan im hin i (NCBI- National Center for Biotechnology Information, 2005) th vi khun bao gm cc ngnh sau y : 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. -Aquificae -Thermotogae -Thermodesulfobacteria -Deinococcus-Thermus -Chrysiogenetes -Chloroflexi -Nitrospirae -Defferribacteres -Cyanobacteria -Proteobacteria -Firmicutes -Actinobacteria -Planctomycetes -Chlamydiae/Nhm Verrucomicrobia -Spirochaetes -Fibrobacteres /Nhm Acidobacteria -Bacteroidetes/Nhm Chlorobia -Fusobacteria -Dictyoglomi

Vic phn ngnh da trn cc c im hnh thi, sinh l, sinh ha, sinh thi...

Cn c vo t l G + C trong ADN ngi ta xy dng c cy pht sinh chng loi (Phylogenetic tree) v chia vi khun thnh 11 nhm sau y : 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. -Nhm Oxy ho Hydrogen -Nhm Chu nhit -Nhm Vi khun khng lu hunh mu lc -Nhm Deinococcus -Nhm Vi khun lam -Nhm Proteobacteria -Nhm Chlamydia -Nhm Planctomyces -Nhm Spirochaetes (Xon th) -Nhm Vi khun lu hunh mu lc -Nhm Cytophaga --Nhm Vi khun Gram dng

44

45

46

Svccloiinhnh

47

ckhinimvccchivikhunthnggpchngtalmquenvimtskhophnloi ngin,datrncccimvhnhthi,sinhl,sinhho.Trongthctinviccloivi khungybnhngitathngchnonthmbngphnnghuytthanh(vicckhngth chiu)

1.Vikhunquanghp(Phototrophicbacteria): 1.1.Vikhunluhunhmuta(Purplesulfurbacteria):Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang t d ng v c (photolithoautotroph), t bo c cha chlorophyll a hoc b , h th ng quang h p ch a cc mng hnh cu hay hnh phin (lamellar) gn vi mng sinh cht. dng lm ngun cho in t (electron donors) trong quang hp thng s d ng H 2, H2S hay S . C kh nng di ng vi tin mao mc cc, c loi chu mao, t l G+C l 45-70%.

aHChromatiaceae:1.1.1- Chi Thiospirium 1.1.2. Chi Chromatium 1.1.3. Chi Thiocapsa 1.1.4. Chi Thiocystis 1.1.5. Chi Thiospirillum 1.1.6. Chi Thiorhodovibrio 1.1.7. Chi Amoebobacter 1.1.8. Chi Lamprobacter 1.1.9. Chi Lamprocystis 1.1.10.Chi Thiodyction 1.1.11.Chi Thiopedia 1.1.12. Chi Rhabdochromatium 1.1.13. Chi Thiorhodococcus

48

Chromatium

Thiocapsa

Thiocystis

Thiospirillum

49

Lamprocystis

Thiopedia

bHEctothiorhodospiraceae: 1.1.14ChiEctothiorhodospirace 1.1.15ChiHalorhodospira 1.2Vi khun khng lu hunh mu ta (Nonsulfure purple bacteria)Vi khun khng lu hunh mu ta l nhm vi khun quang d dng h u c (photoorganoheterotrophs) thng k kh bt buc, mt s loi l quang t d ng v c khng bt buc (trong ti l ho d dng hu c - chemoorganoheterotrophs). T bo cha chlorophyl a hoc b, h thng quang hp cha cc mng hnh c u hay hnh phin (lamellar) gn vi mng sinh cht. dng lm ngun cho i n t (electron donors) trong quang hp thng s dng cht hu c, i khi s d ng h p cht l u hunh dng kh hoc H2. C kh nng di ng vi tin mao mc cc, hoc khng di ng, mt s loi c ti kh (gas vesicles), t l G+C l 61-72%. 1.2.1- Chi Blastochloris 1.2.2- Chi Phaeospirillum 1.2.3- Chi Rhodobacter 1.2.4- Chi Rhodobium 50

1.2.5- Chi Rhodocista 1.2.6- Chi Rhodocyclus 1.2.7- Chi Rhooferax 1.2.8- Chi Rhodomicrobium 1.2.9- Chi Rhodoplanes 1.2.10-Chi Rhodopila 1.2.11- Chi Rhodopseudomonas 1.2.12- Chi Rhodospira 1.2.13- Chi Rhodospirillum 1.2.14- Chi Rhodothalassium 1.2.15- Chi Rhodovibrio 1.2.16-Chi Rhodovulum 1.2.17- Chi Rosespira 1.2.18- Chi Rubiviva

Rhodospirillum

RhodospirillumdiKHVint

51

Rhodopseudomonas

RhodopseudomonasdiKHVint

Rhodobacter

Rhodopila

52

Rhodocycluspurpureus

Rhomicrobium

1.3.Vikhunluhunhmulc(Greensulfurebacteria)Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang t dng v c (photolithoautotroph), t bo c cha chlorophyll a cng vi b , c ho c e, ch a caroten nhm 5, h thng quang hp lin quan n cc lc th (chlorosom) v c lp i vi mng sinh cht. dng lm ngun cho i n t (electron donors) trong quang hp thng s dng H2, H2S hay S . Ht lu hunh tch lu bn ngoi t bo Khng c kh nng di ng , mt s loi c ti kh; t l G+C l 48-58%. 1.3.1- Chi Chlorobium 1.3.2- Chi Prosthecochloris 1.3.3- Chi Pelodictyon 1.3.4- Chi Ancalichliris 1.3.5- Chi Chloroherpeton

53

Chlorobium

Pelodictyon

Prosthecochloris

1.4.Vi khun khng lu hunh mu lc (Green nonsulfur bacteria)Thuc nhm ny l cc vi khun a bo, dng si,thng k kh khng b t buc ,thng l quang d dng (photoheterotroph), c loi quang t d ng ho c ho d dng. T bo c cha chlorophyll a v c, trong iu kin k kh thy c chlorosom. dng lm ngun cho in t (electron donors) trong quang d d ng l glucose, axit

54

amin, axit hu c; trong quang t dng l H 2, H2S. Di ng bng phng thc trn (gliding) , t l G+C l 53-55%. Chi in hnh l Chloroflexus., Chloronema

Chloronema

Chloroflexus

1.5- Vi khun lam (Ngnh Cyanobacteria)Trc y thng nhm ln l To lam (Cyanophyta). Thc ra y l nhng c th nhn nguyn thu, khng lin quan g n to , ngoi kh nng quang h p hi u kh (quang t dng v c) v dng H2O lm cht cho in t trong qu trnh quang hp. Vi khun lam cha chlorophyll a v phycocyanin- phycobiliprotein. M t s loi c s c t phycoerythrin. Chng phi hp vi sc t lc to nn mu nu. Mng lin k t vi phycobilisom. n bo hoc a bo dng si. Khng di ng ho c di ng bng cch trn (gliding), mt s loi c ti kh (gas vesicles).Nhi u lo i c d t bo (heterocysts) v c kh nng c nh nit. Vi khun lam c mt khp m i n i, trong t, trn , trong sui nc nng, trong n c ngt v n c mn. Chng c nng l c chng chu cao hn so vi thc vt i vi cc iu ki n bt li nh nhi t cao, pH thp. Mt s loi c kh nng sng cng sinh vi cc c th khc nh Ru, D ng x , Tu...Nhiu loi cng sinh vi nm to ra a y (Lichen). Vi khun lam c th l sinh vt xut hin sm nht trn Tri t Vi khun lam c chia thnh 5 nhm (subsection) nh sau:

55

a-

Nhm I (c tc gi gi l b Chroococcales):

Hnh que hoc hnh cu n bo, khng c dng si hay dng k t khi (aggregate); phn i hoc ny chi; khng c d t bo (heterocytes). H u h t khng di ng. T l G+C l 31-71% . Cc chi tiu biu l: -Chamaesiphon -Chroococcus -Gloeothece -Gleocapsa -Prochloron

Chamaesiphon

Chroococcus

Glooeothece

Gleocapsa

56

Prochloron

b-Nhm II (c tc gi gi l b Pleurocapsales):Hnh que hoc hnh cu n bo. c th to dng kt kh i (aggregate); phn ct nhiu ln to ra cc baeocytes; khng c d t bo.Ch c cc baeocytes l c di ng. T l G+C l 40-46% . Cc chi tiu biu l:

-Pleurocapsa -Dermocapsa -Chroococcidiopsis

Pleurocapsa

Dermocapsa

Chroococcidiopsis

c-Nhm III (c tc gi gi l b Oscillatorriales):

57

Dng si (filamentous) ; dng lng (trichome) khng phn nhnh ch c cc t bo dinh dng; phn i trn mt phng, c kiu t on (fragmentation); khng c d t bo; thng di ng. T l G+C l 34-67%. Cc chi tiu biu l:

-Lyngbya -Osscillatoria -Prochlorothrix -Spirulina -Pseudanabaena

Lyngbya

Oscillatoria

Prochlorothrix

Spirulina

Pseudanabaena

d-Nhm IV (c tc gi gi l b Nostocales) : Dng si ; dng lng (trichome) khng phn nhnh c th cha cc t bo bi t ho (specialized cell) ; phn i trn mt phng, c kiu t on to thnh o n sinh sn (hormogonia) ; c t bo d hnh ; thng di ng c th sn sinh bo t mng dy (akinetes). T l G+C l 38-47%. Cc chi tiu biu l :

58

-Anabaena -Cylindrospermum -Aphanizomenon -Nostoc -Scytonema -Calothrix

Anabaena

Anabaena trong Bo hoa du

59

Cylindrospermum

Calothrix

Nostoc

Scytonema

e-Nhm V (c tc gi gi l b Stigonematales) : Lng (trichome) dng si, phn nhnh hoc do cc t bo nhiu h n m t chu i to thnh ; phn i theo nhiu mt phng, hnh thnh on sinh sn ( hormogonia) ; c t bo d hnh ; c th sn sinh bo t mng dy ( alkinetes), c hnh thi phc tp v bit ha (differentiation). T l G+C l 42-44%. Cc chi tiu biu l :

-Fischerella -Stigonema -Geitlerinema

60

Fischerella

Geitlerinema

Theo NCBT (2005) th Vi khun lam bao gm nhng b sau y:

-Chlorococcales -Gloeobacteria -Nostocales -Oscillatoriales -Pleurocapsales -Prochlorales

2- Vi khun sinh ni bo t (Endospore-forming bacteria):

A-Vi khun hnh cu

61

2.1- Chi Sporosarcina

AA-Vi khun hnh queB-K kh bt buc C-Sinh trng c nng 3-12% NaCl 2,2- Chi Sporohalobacter CC-Khng sinh trng c nng 3-12% NaCl D-Kh sulfat 2.3- Chi Desulfotomaculum DD-Khng kh sulfat E-Phn gii Axit 3-hydroxybenzoic 2.4- Chi Sporomaculum EE- Khng phn gii Axit 3-hydroxybenzoic F- Chiu rng ca t bo > 2,5mm 2.5- Chi Oscillosporia FF- Chiu rng ca t bo < 2,5mm G-S dng axit bo bo ho v axit butyric 2.6- Chi Syntrophospora GG- Khng s dng axit bo bo ho 2.7- Clostridium BB- Vi hiu kh 2.8-Sporolactobacillus BBB- Hiu kh v k kh khng bt buc C- Phn gii lignin trn mi trng kim 2.9- Amphibacillus CC- Khng phn gii lignin trn mi trng kim D-Sinh trng trn mi trng > 10% NaCl 2.10- Halobacillus DD-Khng sinh trng trn mi trng > 10% NaCl E-C th phn gii Thiamin

62

2.11- Aneurinibacillus EE- Khng c th phn gii Thiamin F-C cha acid bo vng w trong lipid 2.12- Alicycolobacillus FF- Khng cha acid bo vng w G- C th tp giao vi 515F (chy PCR) 2.13- Paenibacillus GG- Khng th tp giao vi 515F H- C th tp giao vi 1741F (chy PCR) 2.14- Brevibacillus HH- Khng th to giao vi 1741F I-Phn gii qung pyrit 2.15- Sulfidobacillus II- Khng phn gii qung pyrit 1.16- Bacillus

Sporosarcina

Desulfotomaculum

Clostridium

63

Paenibacillus

Bacillus

3TrckhunGramm,lnmen,hiukhhockkh khngbtbuc

ACatalasedngtnh,khngdinghaydingnhtinmaocc BThngdingnhtinmaocc,khngksinhngvtcxngsng

H Vibrionaceae 3.1- Chi Aeromonas 3.2- Chi Enhydrobacter 3.3- Chi Photobacterium 3.4- Chi Pleisiomonas 3.5- Chi Vibrio BB- Khng di ng, k sinh ng vt c xng sng H Pasteurellaceae 3.6- Chi Actinobacillus 3.7- Chi Haemophilus 3.8-Chi Pasteurella

64

AA- Catalase m tnh, khng di ng hay di ng nh chu mao H Enterobacteriaceae 3.9- Chi Proteus 3.10-Chi Enterobacter 3.11-Chi Pantoea 3.12-Chi Rhanella ` 3.13-Chi Providencia 3.14-Chi Morganella 3.15-Chi Tatumella 3.16-Chi Salmonella 3.17-Chi Edwardisiella 3.18-Chi Citrobacter 3.19-Chi Budvicia 3.20-Chi Pragia 3.21-Chi Leminorella 3.22-Chi Serratia 3.23-Chi Xenorhabdus 3.24-Chi Klebsiella 3.25-Chi Kluyvera 3.26-Chi Yersinia 3.27-Chi Cedecea 3.28-Chi Ewingella 3.29-Chi Buttiauxella

65

3.30-Chi Moellerlla 3.31-Chi Leclecia 3.32-Chi Escherichia 3.33-Chi Yokenlla 3.34-Chi Hafnia 3.35-Chi Tatumella

Aeromonas

Photobacterium

Vibrio

Vibrio

Actinobacillus

Proteus

Pasteurella

Pasteurella

Enterobacter

66

Morganella

Salmonella

Salmonella

Yersinia

Citrobacter

Serratia

Serratia

Klebsiella

Klebsiella

Escherichia

4TrckhunGramm,khnglnmen,hiukhhockkh khngbtbuc

67

ASinhtrngc60C

B-T bo ln, chiu rng 1,3-1,8mm 4.1- Chi Thermomicrobium BB-T bo nh, chiu rng 0,4-0,8mm C-C th dng glucose lm ngun carbon D-Sinh trng c pH 4,5 4.2- Chi Acidothermus DD-Khng sinh trng c pH 4,5 4.3- Chi Thermus CC-Khng th dng glucose lm ngun carbon duy nht 4.4- Chi Thermoleophilum AA- Khng sinh trng c 60C B-C th oxy ho etanol thnh axit axetic C-C th oxy ho etanol ti CO2 v H2O D-C th oxy ho acid DL-lactic ti CO2 v H2O 4.5- Chi Acetobacter DD-Khng oxy ho acid DL-lactic ti CO2 v H2O 4.6- Chi Acidomonas CC-Khng th oxy ho ethanol ti CO2 v H2O D-C th oxy ho acid DL-lactic ti CO2 v H2O 4.7- Chi Frateuria DD-Khng oxy ho acid DL-lactic ti CO2 v H2O 4.8- Chi Gluconobacter

68

BB-Khng oxy ho etanol thnh acid acetic C-C th c nh Nit ngoi c th D-C th c nh Nit trong iu kin hiu kh E-C th sinh bo xc (cysts) 4.9-Chi Azotobacter EE-Khng sinh bo xc F-C lipoid trong phn cc ca t bo 4.10-Chi Beijerinckia FF-Khng c lipoid trong phn cc t bo G-T bo rng > 2,0mm 4.11- Chi Azomonas GG- T bo rng < 2,0mm 4.12- Chi Derxia DD- C th c nh nit trong iu kin vi hiu kh E-Tin mao thng mc cc F-Trong t, c th trao i carbohydrat 4.13- Chi Agromonas FF-Trong nc, khng trao i carbohydrat 4.14-Loi Aquaspirillum fasciculus EE-Chu mao F-Sinh trng v c nh nit pH < 3,0 4.5- Loi Acetobacter diazotrophicus FF-Khng sinh trng v c nh nit pH < 3,0

69

4.15- Chi Xanthobacter CC- Khng th c nh nit ngoi c th D-Hnh thnh nt sn trong r, trong thn thc vt, c th nh nit t khng kh. c

E-Sinh acid trn mi trng Thch-Cao nm men-Mannit F-C th dng DL-Arginin, L-Histidin lm ngun nit duy nht. 4.16- Chi Rhizobium FF- Khng th dng DL-Arginin , L-Histidin lm ngun nit duy nht 4.17- Chi Sinorhizobium EE-Khng sinh acid trn mi trng Thch- Cao nm men- Mannit F-C Dihydrolase arginin v Decarbocylase lysin 4.18- Chi Azorhizobium FF-Khng c Dihydrolase arginin v Decarboxylase lysin 4.19-Chi Bradyrhizobium DD- Khng sinh nt sn trong c th c nh nit E- Sinh khi u r thc vt 4.20- Chi Agrobacterium EE- Khng sinh khi u r thc vt F-C nh nit mt l 4.21- Chi Phyllobacterium FF-Khng c nh nit mt l

70

G-C th dng hp cht 1 carbon lm duy nht H-C tch lu PHB trong t bo I-Gy bnh thc vt 4.22-Chi Rhizobacter II- Khng gy bnh thc vt J-Khng di ng 4.23- Loi Paracoccus Paracoccus alcaliphalus JJ-Di ng K-Nhum Gram dng tnh hoc kh bin 4.15- Chi Xanthobacter KK- Khng nh trn 4.24- Chi Methylobacter

ngun carbon

denitrificans/

HH-Khng tch lu PHB trong t bo I-C th dng methane lm ngun carbon duy nht K-Di ng 4.25- Chi Methylomonas KK-Khng di ng 4.26-Chi Methylococcus II- Khng th dng methane lm ngun C duy nht J-Catalase dng tnh

71

K-Sinh trng cn NaCl 4.27- Chi Methylophaga KK- Sinh trng khng cn NaCl 4.28- Chi Methylophilus JJ-Catalase m tnh 4.29- Chi Methylobacillus nht GG- Khng th dng hp cht 1 carbon lm ngun carbon duy H-Sinh trng cn NaCl hay nc bin I- Khng di ng 4.30- Chi Mesophilobacter II- Di ng J-C gelatinase 4.31- Chi Alteromonas JJ-Khng c gelatinase K-C tch lu PHB L-S dng glucose M-C esterase 4.32- Chi Deleya MM-Khng c esterase 4.33- Loi Oceanospirillum kriegii LL- Khng s dng glucose 4.33- Loi Oceanospirillum jannaschii KK- Khng tch lu PHB

72

L- Sinh axt t mannit 4.34- Chi Marinomonas LL- Khng sinh axt t mannit 4.36- Chi Pseudomonas (P.stanieri, P.perfectomarina, P.doudoroffii, P.nautica) HH- Sinh trng khng cn NaCl hay nc bin I- C t dng hydrogen J-C tin mao cc hay gn cc K-Khun lc mu vng L-Khng sinh trng 520C 4.35- Chi Hydrogenophaga LL- Sinh trng 520C 4.36- Loi Ps. hydrogenothermophila KK- Khun lc khng mu vng 4.36- Chi Pseudomonas (P. saccharophila, P.facillis, P.hydrogenovora) JJ- C chu mao tha 4.50- Chi Alcaligenes (A.paradoxus, biovar, A.eutrophus, A.latus, A.denitrificans subsp. xylosoxidans) II- Khng c t dng hydrogen J- Di ng K- Tin mao mc cc L-Catalase dng tnh 4.36~47 Chi Pseudomonas LL-Catalase m tnh

73

4.48- Chi Xanthomonas KK- Chu mao L-Sinh acid t ng 4.49- Chi Ochrobactrum LL-Khng sinh acid t ng 4.50- Chi Alcaligenes JJ- Khng di ng K-Khun lc mu vng L-Phospholipid cha sphingosin 4.51- Chi Sphingobacterium LL-Phospholipid khng cha sphingosin 4.52- Flavobacterium KK- Khun lc khng c mu vng L- Yu cu cht ch v dinh dng M-Lng cha G+C cao (66-70mol%) 4.53- Chi Bordetella MM- Lng cha G+C thp (40-47mol%) N-Sinh trng mnh 4.54- Chi Moraxella NN- Sinh trng yu 4.55- Chi Oligella LL-Khng yu cu cht ch v dinh dng M- Khun lc mu tm

74

4.56- Chi Chromobacterium MM- Khun lc khng c mu tm N-Catalase dng tnh O-Hnh que c lc bin hnh cu 4.57- Chi Acinetobacter OO- Khng bin thnh hnh cu 4.58- Chi Weeksella NN-Catalase m tnh O- Sinh indol 4.59- Chi Suttonella OO- Khng sinh indol 4.60- Chi Kingella

Thermomicrobium

Thermoleophilum

Acetobacter

75

Thermus

Gluconobacter

Azotobacter

Methylobacterium

76

Bordetella

Rhizobium

vntsntrnr

Sinorhizobium

vntsntrnr

77

Methylococcus

Bradyrhizobium

vntsntrnr

Agrobacterium

vntsn

Pseudomonas

Alcaligenes

Flavobacterium

vkhunlc

78

Moraxella

Chromobacterium

Acinetobacter

5CukhunGramm,kkhbtbucA- ng knh t bo > 1,3m 5.1- Chi Megasphaera AA- ng knh t bo < 1,3m B- Khun lc di tia t ngoi (360nm) c mu hunh quang, ch sinh acid bo 2 carbon 5.2- Chi Syntrophococcus BB- Khun lc di tia t ngoi (360nm) khng c mu hunh quang, c th sinh c cc acid bo khc C- Aminoacid l ngun nng lng ch yu, khng ln men lactic 5.3- Chi Acidaminococcus CC- Aminoacid khng phi l ngun nng lng ch yu, ln men lactic 5.4- Chi Veillonella

79

Megasphaera

Acidaminococcus

Veillonella

6Trckhun,phykhun,xonkhunGramm,kkh:

I-

Sinh trng nhit 550 C A-Khng ln men v ng ho carbohydrat , s dng peptid, aminoacid, cao nm men... B- T bo c bao, khng kh Fe3+. 6.1- Chi Thermosipho BB- T bo khng c bao C-S dng cc loi acid hu c v H2 kh Fe3+ 6.2- Chi Deferribacter CC-Khng kh Fe3+ 6.3- Chi Thermosyntropha AA-Ln men v ng ho carbohydrat B-a mn. sinh trng nng NaCl 4% C-Di ng nh chu mao, sn phm ln men ch yu l acid acetic, ethanol, H2/CO2

80

6.4- Chi Halothermothrix CC- Di ng nh tin mao mc bn cnh hay gn c c t bo, sn phm ln men ch yu l acid acetc, acid lactic, acid succinic, H2/CO2 6.5- Chi Thermotoga BB- Khng a mn, khng sinh trng nng NaCl 4% C-Sn phm ln men ch l acid acetic D-Thnh t bo in hnh Gram m, cn vitamin B12 6.6- Chi Acetothermus DD-Thnh t bo khng in hnh Gram m, cn H2S hoc Cystein 6.7- Chi Acetogenium CC-Sn phm ln men l hn hp acid acetic v cc acid hu c khc D-Di ng E-Phy khun, di ng nh tin mao mc bn hay gn cc 6.8- Chi Acetomicrobium EE-Hnh que hoc bin i,khng di ng hoc di ng nh chu mao 6.9- Chi Fervidobacterium DD-Khng di ng E-Sn phm ln men gm H2/CO2 v hn hp acid F-Sn phm ln men gm acid acetc, ethanol, H2/CO2 6.10- Chi Thermohydrogenum FF-Sn phm ln men gm acid acetic v ethanol, hoc acid acetic v H2/CO2

81

6.11- Chi Coprothermobacter EE-Khng sinh kh, sn phm ln men gm acid acetic, acid propionic, acid isopentanoic 6.12- Chi AnaerobaculumIIKhngsinhtrngnhit550C,am

A- Khng ng ha v ln men carbohydrat B-Khng nui dng c thun khit, phi nui chung vi mt vi khun khc, phn gii cc on axt bo ngn C-T bo hnh que hay hnh si di, sn phm phn gii acid bo l acid acetic v H2 /CO2 6.13- Chi Syntrophobacter CC-Phy khun, sn phm phn gii acid bo l acid acetic, acid propionic, acid isobutiric, acid isopentanoic v H2//CO2 6.14- Chi Syntrophomonas BB-C th nui cy thun khit C-Cha sc t t bo, vi hiu kh 6.15- Chi Wolinella CC- Khng cha sc t t bo, k kh bt buc D-Ch s dng acid succinic v ch sn sinh acid propionic E-Phy khun, di ng nh tin mao mc bn 6.16- Chi Schuartzia EE- Trc khun, khng di ng 6.17- Chi Succinoclastium DD- ng ho cc loi protein E-Di ng nh chu mao, khng ln men hoc ln men

82

yu glucose, sn phm l ethanol v acid butyric 6.18- Chi Tissierella EE- Khng di ng F- Trn a thch mu khun lc c mu t nu n en 6.19- Chi Porphyromonas FF- Trn a thch mu khun lc khng c mu t nu n en, ln men pyridin v arginin 6.20- Chi Synergistes AA-C th ng ho v ln men hydrat carbon, sn sinh cc loi axit hi c

1.Snphmchyulnmenhydratcarbonlacidacetichoc acidaceticviethanolvH2/CO2B-Acid acetic l sn phm ln men duy nht C- Di ng nh chu mao 6.21- Chi Acetoanaerobium CC- Khng di ng D-C th ln men aminoacid, sinh indol t tryptophan 6.22- Chi Acidaminobacter DD- Khng ln men aminoacid, ln men carbohydrat v acid hu c ESn phm ln men ngoi acid acetic cn c mt t acid formic v acid lactic 6.23- Chi Catonella EE-Sn phm ln men ngoi acid acetic cn c m t t acid isopentanoic, acid lactic, acid succinic, acid isobutyric v acid butyric 6.24- Chi Johngonella BB- Sn phm ln men gm c acid acetic, ethanol v H2/CO2

83

C-Trc khun hoc phy khun, di ng D- Di ng nh tin mao mc cc, khng a mn 6.25- Chi Acetovibrio DD- Di ng nh chu mao, a mn 6.26- Chi Haloanaerobacter CC- T bo dng si, khng di ng 6.27- Chi Acetofilamentum

2Snphmchyulnmencarbohydratlacidlactichoc acidlactic,acidaceticvccacidkhcB-Di ng, a mn 6.28- Chi Halocella BB- Khng di ng, khng a mn C-Sn phm ln men ch c acid lactic 6.29- Chi Leptotrichia CC- Sn phm ln men gm acid lactic v cc acid khc D-Sn phm ln men l acid lactic v acid acetic E- T bo phnh to gia, khng sinh kh 6.30- Chi Sebaldella EE- T bo hnh cong, sinh kh 6.31- Chi Lachnospira DD- Sn phm ln men l acid lactic, acid acetic v acid succinic ET bo ln, ng knh ti 3mm 6.32- Chi Megamonas

84

EE- T bo < 1,5mm 6.33-Chi Mitsuokella 6.34- Chi Halella

3Snphmchyulnmencarbohydratlacidpropionichoc acidpropionicvccacidkhcB-Sn sinh acid hu c v H2/CO2 C-Khng di ng, a mn, cn NaCl 1% D- Ch sn sinh acid propionic v CO2 6.35- Chi Propionigenium DD- Ngoi acid propionic v CO2 cn c cc acid khc E- Sn sinh acid propionic, CO2 ,acid acetic v ethanol 6.36- Chi Pelobacter EE- Sn sinh acid propionic, CO2 , acid aceic v acid butyric 6.37- Chi Haloanaerobium CC- Di ng, khng a mn D-T bo hnh cong, di ng nh tin mao cc hay bn E- ng knh t bo 0.9mm 6.38- Chi Selenomonas EE- ng knh t bo 0,5mm 6.39- Chi Anaerovibrio DD-T bo hnh cong hay hnh xon, di ng nh chu mao 6.40- Chi Propionispira BB- Sn sinh hn hp acid, acid propionic l chnh, khng sinh kh

85

C-Ch yu sn sinh acid propionic D-T bo hnh que 6.41- Chi Oribaculum DD-T bo cong hnh thn rn 6.42- Chi Centipeda CC-Sn sinh acid propionic, acid acetic v acid succinic D-Di ng E- Tin mao dng lc mc bn, t bo hnh cong 6.43- Chi Pectinatus EE-T bo dng cong hoc dng xon 6.44- Chi Zymophilus DD- Khng di ng E-ng knh t bo 3mm 6.32- Chi Megamonas EE- ng knh t bo < 3m F-T bo hnh que, ch sn sinh acid propionic v acid acetic 6.45 Chi Anaerorhabdus FF- T bo hnh bin i, sn sinh acid propionic v cc acid khc G-T bo nh, u nhn, sn sinh acid propionic v acid succinic 6.46- Chi Rikenella GG- Kch thc t bo thay i, sn sinh acid propionic, acid formic, acid acetic, acid lactic, v acid succinic

86

6.47- Chi Bacteroides

4Snphmchyulnmencarbohydratlacidbutyrichoc acidbutyricvccacidkhcB-Di ng nh tin mao mc cc hay gn cc, t bo hnh cong C- Sn phm ln men ch l acid butyric D- Di ng 6.48- Chi Pseudobutyrivibrio DD- Khng di ng 6.49- Chi Fusobacterium CC-Ngoi acid butyric cn c cc sn phm ln men khc D-Sn sinh acid butyric v CO2 6.50- Chi Roseburia DD-Sn sinh acid butyric, acid acetic hoc acid lactic, khng sinh kh 6.51- Chi Butyrivibrio BB-Di ng nh chu mao hoc khng di ng C- Sn sinh acid butyric, acid acetic v ethanol, khng sinh kh, cn 1% NaCl 6.52- Chi Iliobacter CC-Sn sinh acid butyric, acid acetic...v c sinh kh, cn 13% NaCl 6.37- Chi Haloanaerobium

5Snphmlnmencarbohydratlhnhpacidcchaacid succinichocacidformicB- Sn sinh hn hp acid c cha acid formic C-Sn sinh axit formic v CO2

87

6.53- Chi Oxalobacter CC-Sn sinh acid formic v cc acid khc nhng khng sinh CO2 D-T bo hnh que, sn sinh acid formic, acid acetic v acid succinic 6.54- Chi Ruminobacter DD- T bo hnh dng thay i, sn sinh acid formic, acid acetic, acid succinic, acid lactic v acid propionic 6.47- Chi Bacteroides BB-Sn sinh hn hp acid c cha acid succinic C- Di ng D-T bo hnh que ngn, ch sn sinh acid succinic 6.55- Chi Succinimonas DD- T bo hnh cong, sn sinh hn hp acid c cha acid succinic E-Sn sinh acid succinic, acid acetic v c sinh CO2 6.56- Chi Malonomonas EE- Sn sinh acid succinic, acid acetic, khng sinh CO2 6.57- Chi Succinovibrio CC- Khng di ng D-Sn sinh acid acetic, acid lactic v acid succinic 6.33- Chi Mitsuokella DD-Trong hn hp acid khng c acid lactic E-Sn sinh acid propionic v acid succinic 6.46- Chi Rikenella EE- Sn sinh acid acetic v acid succinic FT bo hnh xon, di ng

88

6.58- Chi Anaerobiospirrillum FF- Khng di ng G-T bo hnh que ngn, ch yu sng d c v manh trng 6.59- Chi Fibrobacter GG- T bo a hnh thi, ch yu sng khoang ming 6.60- Prevotella

Thermosipho

Syntrophobacternuicychungvi Methanobrevibacte

89

Wolinella

Porphyromonas gingivalis

vtcytrnthchm

Leptotrichia

Bacteroides

Bacteroides

Fusobacterium

Fusobacterium

90

Mitsuokella

7CukhunGramdnghiukhvkkhkhngbtbuc AHiukhB-Di ng C-a mn bt buc, cn NaCl 7,5% 7.1- Chi Marinococcus CC-Khng nh trn 7.2- Chi Planococcus BB-Khng di ng C-a mn, cn NaCl 7,5% D- Lng cha GC > 50mol % 7.3- Chi Salinicoccus DD- Lng cha GC < 50mol % 91

7.1- Chi Marinococcus CC-Khng nh trn D-Sinh acid t glucose 7.4- Chi Micrococcus DD-Khng sinh hay sinh t acid t glucose 7.5- Chi Deinococcus

AAKkhkhngbtbucB-T l G + C trong ADN < 30mol % 7.6- Chi Melissococcus BB- T l G + C trong ADN > 50mol % 7.7- Chi Stomatococcus BBB- T l G + C trong ADN > 30 mol % , nhng < 50mol % C- Catalase (+) D- Thnh t bo cha acid teichoic 7.8- Chi Staphylococcus DD- Thnh t bo khngch acid teichoic 7.9- Chi Saccharococcus CC- Catalase (-) D- Ngoi xp thnh i cn c dng xp thnh bn E- khng vi Vancomycin (30mg) 7.10- Chi Pediococcus EE- Khng khng vi Vancomycin 7.11- Chi Aerococcus

92

DD- Khng c dng xp thnh bn E-Xp thnh chui rt di 7.12- Chi Trichococcus EE- Khng xp thnh chui di F-Sinh trng 10C G- Sinh kh t glucose 7.13- Chi Leuconostoc GG- Khng sinh kh t glucose H- Sinh trng 45C 7.14- Chi Enterococcus HH- Khng sinh trng 45C I-Di ng, sng trong nc 7.15- Chi Vagococcus II- Khng di ng 7.16- Chi Lactococcus FF- Khng sinh trng 10C G-Lng cha G + C ca ADN>35mol % 7.17- Chi Streptococcus GG-Lng cha G + C ca ADN< 35mol % 7.18- Chi Gemella

93

Micrococcus

Deinococcus

Deinococcus

Melissococcus

Melissococcus

KhunlcStomatococcus

94

Staphylococcus

Staphylococcus

Pediococcus

Aerococcus

Leuconostoc

Leuconostoc

95

Enterococcus

Lactococcus

Lactococcus

Streptococcus

Streptococcus

Gemella

96

8- Trc khun Gram dng hiu kh hoc k kh khng bt buc

A-Hiu khB-C chu k bin ho hnh cu, hnh que C-Thnh t bo c cha DAP (acid diaminopimelic) D- C cha LL-DAP E-Di ng 8.1- Chi Pimelobacter EE- Khng di ng 8.2- Chi Terrabacter DD- Khng cha dng LL- m cha dng Meso-DAP E-Loi quinone l MK-8 (H2), MK-7 (H2) 8.3- Chi Brevibacterium EE-Khng nh trn 8.4- Chi Brachybacterium CC- Peptidoglycan khng cha DAP D-Hnh que khng quy tc 8.5- Chi Arthrobacter DD-Hnh que c quy tc 8.6- Chi Kurthia BB- Khng c bin ho hnh cu, hnh que C-C cha acid mycolic 8.7- Chi Caseobacter CC-Khng cha axt mycolic 97

9Trckhunkhngquytc,khngbot AGram(+) Banhit CTbocphnnhnh Dakim 9.1ChiAnaerobranca DDKhngakim 9.2ChiThermobrachicum CCTbokhngphnnhnh DDing 9.3ChiThermoanaerobacter DDKhngding 9.4ChiThermoanaerobium BBam CTbohnhbdc,hnhquengnhayhnhcong DTbonhnhnhnhcon,spxptngihnhlilim 9.5ChiFalcivibrio DDTbohnhbudc,hnhquengn,khngspxpthnhhnhlilim ELnmenhydratcarbonchsnsinhacidacetic,cncthtnghpraacid acetictH2vCO2 FThchhpsinhtrng30C 9.6ChiAcetobacterium

98

FFThchhpsinhtrng38C 9.7ChiAcetitomaculum EELnmenhydratcarbonsinhacidlactic,khngtnghpraacidacetictH 2 vCO2 9.8ChiAtopobium CCTbohnhquekhngquytc DTbocphnthnhhnhna(inhba),lnmenhydrat carbonkhngsinhkh 9.9ChiBifidobacterium DDTbokhngphnthnhhnhna,lnmenhydratcarbomc sinhkh ESnphmlnmenchyulacidbutyric,acidacetichocacidformic. 9.10ChiEubacterium EESnphmlnmenchyulacidacetic,acidlacticvetanol 9.11 AAGram() A Tbohnhque,anhit,khngding 9.12 BBTbodngconghaydngsi,ding CLnmenhydratcarbonsnphmchyulacidbutyric 9.13ChiButyrivibrio CCLnmenhydratcarbonsnphmchyukhnglacidbutyric ChiAcetogenium ChiCoribacterium

99

DLnmenglucose,snphmchyulacidaceticvacidlactic,khngsinh kh 9.14ChiMobiluncus DDLnmenglucose,snsinhacidformic,acidacetic,acidlacticvetanol,csinh kh 9.15ChiLachnospira

Thermoanaerobacter diknhhinviint

Bifidobacterium

Eubacterium

Mobiluncus

100

10CukhunGramdngkkhbtbuc A Tbophncttheo3mtphngthnggc,spxpthnhhnhlpth 10.1ChiSarcina AATbophncttheo1hoc2mtphngthnggc,spxpthnhtngihay tng4tbo BKhnglnmenhydratcarbon,khngsinhkh CTlG+Cmol%trongADN50 10.2ChiPeptococcus CCTlG+Cmol%trongADN 45 10.3ChiPeptostreptococcus BBLnmenhydratcarbonsinhacid CTrongsnphmlnmenngoiccacidhuckhccncacidbutyricv aceton(acetone) 10.4ChiCoprococcus CCTrongsnphmlnmenkhngcacidbutyricvaceton 10.5ChiRuminococcus

101

Peptostreptococcus

Peptostreptococcus

Ruminococcus

11PhnloixkhunXkhunlnhmvikhuncbit.Chngckhunlckhvasc dnghnhphngx(actino)nhngkhunthlicdngsiphnnhnhnh nm(myces).Vxkhunccutrcnhnnguynthunhmivikhunkhc, 102

chiungangcasicngnhnhvikhun,chonnctiliugichnglnm tialkhnghpl.Xkhunphnbrngritrongtnhin.Slng nv sinhkhunlc(CFUcolonyformingunit)xkhuntrong1gtthngtti hngtriu.Trnmitrngcasxkhunchaikoikhunty:khunty khsinh(aerialmycelium)vkhuntyccht(substratemycelium).Nhiuloi chckhuntycchtnhngcngcloi(nhchiSporichthya)lichckhun tykhsinh.Giakhunlcthngthycnhiubotmngmnggilbot trn(conidiahayconidiospores).Nubotnmtrongbonang(sporangium) thcgilnangbothaybotkn(sporangiospores).Botxkhun csinhraumtskhuntytheokiuhnhthnhccvchngn(septa). Cc chui bo t trn c th ch l 1 bo t (nh Thermoactinomyces, Saccharomonospora, Promicromonospora, Micromonospora,

Thermomonosspora...),cthc2bot(nhMicrobispora),cthlchui ngn (nh Nocardia, Pseudonocardia, Streptoverticillium, Sporichthya, Actinomadura, Microtetraspora, Streptoalloteichus, Glycomyces, Amycolata, Amycolatopsis,Catellatospora,Microellobosporia...), cthlchuidi(nh Streptomyces, Saccharopolyspora, Actinopolyspora, Kibdelosporangium, nhiu loi Nocardia, Nocardioides,

Kitasatosporia, Saccharothrix,

Pseudonocardia, Amycolatopsis, Streptoverticillium...), c th cc bo t trn nmtrnbsi(synnema),tngtbsicanm(nh Actinosynnema, Actinomadura...).Ccchuibotcththng,cthxon,cthdngln sng,cthmcnhaymcvng...Cccungsinhbot(sporophore)v cungsinhnangbot(sporangiophorres)cthringr,cthphnnhnh. Cccimhnhthinyrtquantrngkhitinhnhnhtnxkhun.

103

XkhunthucnhmvikhunGramdng,thngctlGCtrong ADNcaohn55%.Trongskhong1000chiv5000loisinhvtnhns cngbckhong100chiv1000loixkhun.Xkhunphnbchyu trongtvngvaitrrtquantrngtrongchutrnhtunhonvtchttrong tnhin.Chngsdngacidhumicvccchthuckhphngiikhctrong t.Mcdxkhunthucnhmsinhvtnhnsnhngchngthngsinh trngdidngsivthngtonhiubot.Thmchmtsloixkhun cnhnhthnhtibotnhchiStreptosporangium,Micromonosporavbot dingnhchiActinoplanes,Kineosporia. Trcy,vtrphnloicaXkhunlunlcuhigynhiutranh lungiaccnhVisinhvthc,doncnhngcimvagingVikhun vagingNm.Tuynhin,nnay,XkhuncchngminhlVikhun vinhngbngchngsauy: 1. Mtsxkhunnhccloithucchi Actinomyces v Nocardia rt gingviccloivikhunthucchiLactobacillusvCorynebacterium. 2. Xkhungingvikhunchkhngcnhntht,chngchcha nhimscchtphnbdctheoccsihoccctbo. 3.ngknhcasixkhunvbotgingvivikhun.ngthi sixkhunthngkhngchavchngn. 4.Xkhunlchtncngcaccthckhunthgingnhvikhun, trongkhi,nmkhngbtncngbithckhunth.

104

5.Xkhunthngnhycmvicckhngsinhctcdnglnvikhun, nhnglithngkhngvinhngkhngsinhtcdnglnnmnhcc polyen. 6.Xkhunkhngchachitin,chtcmttrongsivbotcanhiu nm,mkhngcvikhun.ngthigingnhphnlnvikhun,x khunkhngchacellulose. 7. Tngtvivikhun,xkhunnhycmviphnngacidcami trng,cimnykhngcnm. 8. Cc c im v si v nang bo t kn(sporangium) ca chi Actinoplaneschothycthchinylcunigiavikhunvccnm bcthp. Xkhunthucvlp Actinobacteria,b Actinomycetales,baogm10 dib,35h,110chiv1000loi.Hinnay,478loiccngbthuc chi Streptomyces vhn500loithucttcccchicnlivcxpvo nhmxkhunhim.

105

Khunlcxkhun

Khuntyxkhunvbot

106

Chuibottrncungsinhbotdngxon

Botxkhun

107

Shnhthnhhailoikhuntysaukhibotxkhunnymm

108

Sibotvchuibottrn

109

Mtsdngbotxkhun(SVtchthchtnghnh)

110

Mtsdngnangbovnangbotxkhun

111

Mtsdngchuibotxkhun(SVtchthchtnghnh) cimcakhuntykhsinhvkhuntycchtctrnhbytrongs sauy:

112

113

114

115

116

phnloixkhunngitasdngcctiuchunnhtrnhtrADN 16S,laiADN,hnhthi,sinhlsinhhavhaphnloi.Hinnay,ias ccnhkhoahcngviquannimhaichngccoilhailoiringbit nuchnggingnhaudi70%khitinhnhlaiADN.Keswanivcngs chngminhrngnustngnggiahaitrnhtrADN16Sl98.6%thxc sutmcgingnhautrongphplaiADNthphn70%sl99%.Vth gitrtngng98.6%catrnhtrADN16Sccoilngngphnbit hailoikhcnhau.Tuynhin,cngcnhiunhkhoahclygitrnyl98%. 117

cbithaphnloilrtquantrngtrongvicphnloixkhun. Chngrtcchtrongphnloimcnchi.lnhngcimsau: ng,loiacetyl,acidmycolictrongthnhtbo,menaquinone,phospholipid, acidbovtlGCtrongADN.1=Chi;2=Loithnhtbo;3=Thnhphnng;4=Loipeptidoglycan;5=Acid mycolic 6=Thnhphnacidbo;7=Menaquinonchyu;8=Loiphospholipid;9=Tlmol GCtrongADN(%)

1

2

3

4

5

6

7

8

9

HCorynebacteriaceaevmtshlinquanCorynebacterium Turicella Gordonia Williamsia Skermania Dietzia Mycobacterium Nocardia Rhodococcus Tsukamurella IV IV IV IV IV IV IV IV IV IV A A A A A A A A A A A1 A1 A1 A1 A1 A1 + + + + + + + + + 1b 1b 1a 1b 1b 1b 1b 8(H2),9(H2) 10,11 9(H2) 9(H2) 8(H4) 8(H2) 9(H2) 8(H4) 8(H2),9(H2) 9 PII PII PII PII PII PII PII PII PI 51 67 65 72 63 69 64 65 68 73 62 70 64 72 60 69 66 73

HMicromonosporaceaeMicromonospora II D A1 3b 9(H4.6), 10(H4,6) PII 71 72

118

Actinoplanes Cattelatospora Catenuloplanes Couchioplanes Dactylosporangium Pilimelia Spirilliplanes Verrucosispora

II II VI VI II II II II

D D

A1

-

3b

9(H4,6), 10(H4,6) 9(H4,6), 10(H8,6)

PII PII

72 73 71 73 71 73 69 72 69 73 71 69 70

D D D D

A3 A3 A1 A1 A1

3b 2d 2d 2b

10(H4),11(H4) PIII 9(H4) 9(H4,6,8) 9(H4,2) 10(H4) 9(H4) PII PII PII PII PII

HPropionibacteriaceaevNocardiaPropionibacterium Friedmaniella Luteococcus Microlunatus Propioniferax Tessaracoccus Nocardioides Aeromicrobium Hongia Kribbella Marmoricola

II I I I I I I I I I

A3 A3 A3 A3 A3 A3 A3 A3 A3 A3 A3

1a 3a 3a 2d 1a 2d

9(H4) 9(H4) 9(H4) 9(H4) 9(H4),7(H4) 8(H4) 9(H4) 9(H4) 9(H4) 9(H4) 8(H4) PI PI PII PI PI PI PII PI PIII PI

57 63 73 66 68 68 59 63 74 67 72 69 72 71 68 70 72

HPseudonocardiaceaePseudonocardia Actinobispora III IV

A A1 A A1 -

1

8(H4) 7(H2),9(H2)

PIII/ PII PIV

68 79 71

119

Actinopolyspora Amycolatopsis Kibdelosporangium Prauserella Saccharomonospora Saccharopolyspora Thermocrispum

IV III III IV III IV III

A A1 A A1 A A -

2c

9(H4),10(H4) PIII 9(H2,4,6) PII PII PII PII PIII PII

64 68 66 69 66 67 69 69 74 70 77 69 73

3c 3f 2a 2c

9(H4) 9(H2,4) 9(H4) 9(H4) 9(H4)

A A1 A A1 C -

HActinosynnemataceaeActinosynnema Actinokineospora Kutzneria Saccharothrix III III III III

C C C C

A1 A1

-

9(H4,6) 9(H4) 9(H4) 9(H4),10(H4)

PII PII PII PII

69 71 73 70 71 70 76

A1

-

HStreptosporangiaceaeStreptosporangium Acrocarpospora Herbidospora Microbispora Microtetraspora Nonomuraea Planobispora Planomonospora Planotetrespora III III III III III III III III III

B B B B B B B B B

A1 A1

-

3c

9(H0,2,4) 9(H2,4) 10(H4,6)

PIV PIV/ PII PIV PIV PIV PIV PIV PIV PIV

69 71 68 69 71 67 74 66 64 69 70 71 72

A1 A1 A1 A1 A1

-

3c 3c 3c 3c 3c 3c

9(H0,2,4) 9(H0,2,4) 9(H0,2,4) 9(H0,2,4) 9(H0,2,4)

120

HNocardiopsaceaeNocardiopsis Thermobifida III III C C A1

-

3d 3c

10(H2,4,6) 10(H6,8)

PIII PII

64 69

HThermomonosporaceaeThermomonospora Actinocollaria Actinomadura Spirillospora III III III III B B A1 A1 B A1

-

3e 3e 3a 3e

9(H6,8) 9(H4,6) 9(H6,8) 9(H4,6)

PI PII PI PI 73 66 70 69 71

HGlycomycetaceaeGlycomyces II

-

2c

10(H4),11(H4) PI

71 73

HStreptomycetaceaeStreptomyces Kitasatospora I I/III A3 A3

-

2c

9(H6,8) 9(H6,8)

PII PII

69 78 66 73

HFrankiaceaevmtshlinquanFrankia Geodermatophilus Microsphaera Cryptosporangium Sporichthya III III III II I A3 A1

-

12b

9(H4,6,8) 9(H4) 8(H4) 9(H6,8) 9(H6,8)

PI PII

66 71 73 76 68 70 70

PII PI

CchkhcKineosporia Thermobispora Thermoactinomyces I/III III III A3 9(H4) PIII PIV 64 79 71 53 55

-

9(H0,2) 7,9

121

Cc s liu trn c ly t sch Bergeys manual systematic of bacteriology v sch Identification manual of actinomycetes. Thnh t bo: Theo Lechevalier v Lechevalier, thnh t bo x khun c chia thnh 8 loi da trn cc c im v thnh phn acid amin, c bi t l acid diaminopimelic, lysine v thnh phn ng trong thnh t bo, c th lLoi Acid thnh Ccacidaminkhc diamino tbo

ng

I II III IV V VI VII VIII

LLA2pm Gly* meso A2pm meso A2pm meso A2pm Lys,Orn Lys Asp Asp Lys,A2bu Asp Ara,Gal*** Gly**

Tt c cc loi thnh t bo u cha alanine, acid glutamic, glucosamine v acid muramic. A2pm: Acid diamino pimelic c th gm cc ng phn quang h c LL- A 2pm, DL- A2pm (meso- A2pm), DD- A2pm. Ngoi ra cn c OH- A 2pm khi c thm nhm OH gn vo v tr C th 4. Lys: Lysine Orn: Ornitin

122

A2bu: acid diamino butyric Gly: glycin *Glycin nm trong mi lin kt gia hai mch tetrapeptid ** Glycin thay th L- alanin v tr u tin trong mch tetrapeptid Asp: Asparagin Ara: Arabinose Gal: Galactose *** Trong x khun cha acid mycolic nh Nocardia v Mycobacterium, thnh t bo cha arabinogalactan, l hp cht c tng hp t arabinose v galactose.Thnhphnng:

Thnhphn ng A B C D

Ccloingchyu Arabinose,Galactose(khngcha Xylose) Madurose(khngchaArabinosehay Xylose) Khngcngctrng Arabinose,Xylose

Peptidoglycan: Theo Schleifer v Kandler (1972), peptidoglycan x khuncchiathnhccnhmnhsau:K hiu Vtrca K milin hiu Cunigiahaimch tetrapeptid K hiu Acidamin vtrth3

123

kt

A

34

1

Trctip

LLys LOrn mesoA2pm LLys LLys LOrn mesoA2pm LLys LOrn mesoA2pm LA2bu LLys LHsr LGlu LAla LOrn LHsr LA2bu

2 3

Tiunvpeptid

Lacidaminchamtnhm carboxyl(gly)hocchuipeptid ngncaacidamin Dacidaminchahainhm carboxyl(DAsphayDGlu)

4

B

24

1

Lacidamin(GlynLLys)

2

Dacidamin(DOrnhayD A2bu)

Duphy():alaninevtrthnhttrongmchtetrapeptidcthay thbiglycine. Vd:Loipeptidoglycancachi Actinoplanes lA1cnghal:Hai mchtetrapeptidlinktvinhauvtracidamins3v4,milinktnyl trctipvacidaminvtrs3lmesoA2pm.

124

Thnhphnacidbo:K hiu Khng Bo bo ha ha

Iso-

Iso-

Anteiso-

1017 (+) ++ +++ (+)

1018 ++

14/16/18 15/17 15/17 +++ ++ +++ +++ +++ +++ ++ +++

Methyl- Methyl- Cyclopropane ++

1

a +++ +++ b +++ +++ c +++ +++

+ +++ + +++ (+) +++ + ++

(+) + +++ +++ (+) ++ + +++

+++ (+) (+) +++

2

a ++ b (+) c + d+

+ + (v) +

-

3

a +++ ++ b+ c + d++ + +

(v): t hn 5%

Thnhphnmenaquinone:X khun ch cha menaquinone, khng cha ubiquinone. K hiu 8(H 2) hay MK8(H2) ch menaquinone cha mt trong s tm n v isoprene b hydro ha.

Thnhphnphospholipid:K Thnhphnphospholipidctrng hiu PI PII PIII Phosphatidylglycerol(v) Phosphatidylethanolamine Phosphatidylcholine,phosphatidylethanolamine(v),

125

phosphatidylmethylethanolamine(v), phosphatidylglycerol(v),khngchaphospholipidcha glucosamine Phospholipidchaglucosamine, PIV phosphatidylethanolamine(v), phosphatidylmethylethanolamine(v) PV Phospholipidchaglucosamine,phosphatidylglycerol

Tt c cc loi u cha phosphatidylinositol

VtrphnloicaXkhun:LpActinobacteria DilpActinobacteridae BActinomycetales DibActinomycineae HActinomycetaceae Cc chi: Actinomyces, Arcanobacterium,Mobiluncus DibMicrococcineae HMicrococcaceae Cc chi: Micrococcus, Arthrobacter, Kocuria, Nesterenkonia,Renibacterium,Rothia HBogoriellaceae Chi:Bogoriella Actinobaculum,

126

HBrevibacteriaceae Chi:Brevibacterium HCellulomonadaceae Chi:Cellulomonas HDermabacteraceae Ccchi:Dermabacter,Brachybacterium HDermacoccaceae Ccchi:Dermacoccus,Demetria,Kytococcus HDermatophilaceae Chi:Dermatophilus HIntrasporangiaceae Cc chi: Intrasporangium, Janibacter, Terrabacter, Terracoccus,Tetraphaera HJonesiaceae Chi:Jonesia HMicrobacteriaceae Cc chi: Microbacterium, Agrococcus, Agromyces, Clavibacter, Cryobacterium, Curtobacterium, Frigoribacterium, Leifsonia, Leucobacter, Rathayibacter HPromicromonosporaceae Chi:Promicromonospora

127

HRarobacteraceae Chi:Rarobacter HSanguibacteraceae Chi:Sanguibacter Incertaesedis(chaxcnh) Ccchi:Beutenbergia,Ornithinicoccus,Kineococcus DibCorynebacterineae HCorynebacteriaceae Ccchi:Corynebacterium,Turicella HDietziaceae Chi:Dietzia HGordoniaceae Chi:Gordonia HMycobacteriaceae Chi:Mycobacterium HNocardiaceae Ccchi:Nocardia,Rhodococcus HTsukamurellaceae Chi:Tsukamurella Incertaesedis(chaxcnh)

128

Ccchi:Skermania,Williamsia DibMicromonosporineae HMicromonosporaceae Cc chi: Micromonospora, Actinoplanes, Catellatospora, Catenuloplanes, Couchioplanes, Dactylosporangium, Pilimelia, Spirilliplanes, Verrucosispora DibPropionibacterineae HPropionibacteriaceae Cc chi: Propionibacterium, Friedmaniella, Luteococcus, Microlunatus, Propioniferax, Tessaracoccus HNocardioidaceae Cc chi: Nocardioides, Aeromicrobium, Hongia, Kribbella,Marmoricola DibPseudonocardineae HPseudonocardiaceae Cc chi: Pseudonocardia, Actinobispora, Actinopolyspora,Amycolatopsis,Kibdelosporangium, Prauserella, Saccharomonospora, Saccharopolyspora,Thermocrispum HActinosynnemataceae Saccharothrix Cc chi: Actinosynnema, Actinokineospora,

Incertaesedis(chaxcnh)

129

Ccchi:Kutzneria,Streptoalloteichus DibStreptosporangineae HStreptosporangiaceae Cc chi: Streptosporangium, Acrocarpospora, Herbidospora, Microbispora, Microtetraspora, Nonomuraea,Planomonospora,Planotetraspora HNocardiopsaceae Ccchi:Nocardiopsis,Thermobifida HThermomonosporaceae Cc chi: Thermomonospora, Actinocorallia, Actinomadura,Spirillospora DibGlycomycineae HGlycomycetaceae Chi:Glycomyces DibStreptomycineae HStreptomycetaceae Ccchi:Streptomyces,Kitasatospora DibFrankineae HFrankiaceae Chi:Frankia HAcidothermaceae Chi:Acidothermus 130

HGeodermatophilaceae Ccchi:Geodermatophilus,Blastococcus HMicrosphaeraceae Chi:Microsphaera HSporichthyaceae Chi:Sporichthya Incertaesedis(chaxcnh) Chi:Cryptosporangium BBifidobacteriales HBifidobacteriaceae Ccchi:Bifidobacterium,Gardnerella DilpAcidimicrobidae BAcidimicrobiales HAcidimicrobiaceae Chi:Acidimicrobium DilpCoriobacteridae BCoriobacteriales HCoriabacteriaceae Cc chi: Coriobacter, Atopobium, Collinsella, Cryptobacterium, Denitrobacterium, Eggerthella, Slackia

131

DilpRubrobacteridae BRubrobacteriales HRubrobacteriaceae Chi:Rubrobacter DilpSphaerobacteridae BSphaerobacteriales HSphaerobacteriaceae Chi:Sphaerobacter

Mtmtschithnggp: ChiRhodococcus:Pht trin di dng que hoc khun ty c cht phn nhnh. tt c cc chng, chu trnh sng (morphogenetic) u bt ngun t giai o n hnh cu ho c que ngn. Bng cch phn on, cc t bo hnh cu s to thnh dng que ri dng si, s i phn nhnh v h si. Mt s chng cn to khun ty kh sinh phn nhnh ho c b s i (synnemata). Chng khng c kh nng chuyn ng cng nh khng hnh thnh bo t hay ni bo t. Gram dng. Acid-alcohol fast (nhum khng acid-cn) mt s giai o n pht tri n. Hiu kh. Ha d dng hu c. Catalase dng tnh. Hu ht cc chng u mc tt trn cc mi trng tiu chun 30oC, s khc cn thiamin cho sinh trng. Khun lc c th sn si hoc trn nhn, c mu vng sm, kem, vng, vng da cam, ho c khng mu. Arylsulfatase m tnh, nhy cm vi lysozyme, khng phn h y c casein, cellulose, chitin, elastin hay xylan. C th s dng c rt nhiu loi hp cht hu c lm ngun cacbon v ngun nng lng. Thnh t bo cha lng ln acid meso- diaminopimelic (meso-DAP), arabinose v galactose. Cha diphosphatidylglycerol, phosphatidylethanolamin, phosphatidylinositol mannoside. Thnh phn menaquinone chnh l MK-8(H2) v MK-9(H2). Cha lng ln acid tuberculostearic mch thng, khng bo ha v acid mycolic v i 32-66 cacbon, v i nhiu nht bn lin kt i. T l mol G+C trong ADN l 63-72%.

132

C nhiu trong t v phn gia sc. Mt s chng gy bnh cho ngi v ng vt. Loi chun: Rhodococcus rhodochrous

ChiNocardioides:Si s cp phn nhnh trn b mt, sau xm nhp vp trong th ch r i t o n thnh dng que hay cu hay khng u. Si kh sinh c th tha th t, phn nhnh ho c khng sau cng t thnh nhng mu ngn ho c dng que. Nh ng m u ny s l ngun gc ca nhng si mi. Khng c t bo di ng. Khun lc nho. Gram dng. Khng nhum khng acid ( Non-acid fast ). Catalase d ng tnh. Hi u kh b t buc. Ha d dng hu c. Mc d dng trn cc mi trng tiu chun. Nh y c m vi thc khun th c hiu. Acid amin chnh trong thnh t bo l L-DAP v glycine. Khng cha acid mycolic. Phospholipid ch yu l phosphatidylglycerol v acylphosphatidlyglycerol. Acid bo chim u th l 14-methylpentadecanoic. Thnh phn menaquinone chnh l MK-8(H4). T l mol G+C trong ADN t 66,1-72,7%. C nhiu trong t. Loi chun: Nocardioides albus

ChiPseudonocardia:Si kh sinh v si c cht u sinh bo t dng chui. Si phn on, th ng c dng zich-zac vi xu hng phng ln ngn hoc gia. Si ko di bng cch n y chi. Cc on si c chc nng ca bo t hoc bin i thnh bo t. Thnh si c hai lp. Gram dng. Khng c giai on di ng. Hiu kh. Sinh tr ng trn nhi u loi mi trng hu c hoc tng hp. a m hoc a nhit. Loi chun: Pseudonocardia thermophila

ChiSaccharopolyspora:Si c cht pht trin mnh, phn nhnh, t thnh cc on dng que v i kch th c 1x5m, ch yu nhng phn gi hn ca khun lc. Khun ty kh sinh phn o n to cc chui bo t. Gram dng. Khng nhum khng acid. Hi u kh. Khun l c mng, nh ln, hi nhn, si kh sinh t, thng to thnh chm, ch yu nh ng phn gi. C th s dng nhiu loi cht hu c nh ngun cacbon v nng l ng duy nht, c th phn gii adenine, khng nhiu lo i cht khng sinh nhng nh y c m vi lysozym. T l mol GC trong ADN l 77%. 133

Loi chun: Saccharopolyspora hirsute

ChiIntrasporangium:Si phn nhnh, c xu hng t thnh nhiu on c kch thc v hnh dng khc nhau. Khng bao gi c si kh sinh. Cc ti bo t hnh oval hay hnh qu chanh c to thnh gia hoc/v u si. Cc bo t khng di ng. Gram dng. Khng nhum khng acid. Ha d dng hu c. Catalase dng tnh. Hi u kh. M c t t nh t 28-37oC, khng mc c 45oC. Sinh trng tt hn trn cc mi trng cha peptone v cao tht. T l mol GC trong ADN l 68,2%. Thnh t bo ch a L-DAP v glycine. Thnh phn phospholipid ch yu l mt loi phospholipid ch a glucosamin cha bit. Thnh phn acid bo ch yu l cc acid bo mch thng bo ha v khng bo ha. Menaquinone chnh l MK-8. Loi chun: Intrasporangium calvum

ChiActinopolyspora:H si phn nhnh, hnh thnh rt nhiu si kh sinh c ng knh khong 1 m. S i c cht hu ht khng t on. Cung sinh bo t cha 20 hoc hn bo t d ng que ngn hoc dng cu vi v nhn hnh thnh trn si kh sinh theo chi u h ng gc. Si c cht khng sinh bo t. Gram dng. Nhum khng acid. Thnh t bo cha meso-DAP, arabinose v galactose. Khng cha acid mycolic. Hi u kh. Ha d dng hu c. T l mol GC trong ADN l 64,2%. Loi chun: Actinopolyspora halophila

ChiSaccharomonospora:Ch yu sinh bo t n trn si kh sinh. Bo t khng b n nhi t, khng c kh nng di ng, c hnh thnh trn cc cung sinh bo t n gi n, khng phn nhnh, di ngn khc nhau. Trn mi trng thch, h si dinh dng phn nhnh, bao ph b i lp si kh sinh vi cc bo t mc dy c dc trn cc si. S i kh sinh ban u c mu trng ri chuyn thnh xm xanh v xanh sm. Sc t xanh cng c s i dinh dng v khuch tn ra mi trng. Thnh t bo cha meso-DAP, arabinose v galactose. Khng cha acid mycolic. Cha lng ln iso- v anteiso- acid bo, phosphatidylethanolamin v MK-9(H4). Hiu kh. Ha d dng hu c. Nhit thch hp cho sinh trng l 35-50oC, pH 7-10. Sinh catalase, deaminase v phosphatase. Phn

134

hy c casein, gelatin, tinh bt. xylan v tyrosin. c bi t c th s d ng glycerol lm ngun cacbon. Phn lp c trong t, cht lng cn h, than bn, thy nhi u trong phn bn, phn compt, v c kh. T l mol GC trong ADN l 69-74%. Loi chun: Saccharomonospora viridis

ChiFrankia:Khng sinh si kh sinh. Ti bo t thng sinh trn cu ng sinh bo t . Bo t khng c kh nng di ng vi hnh dng khng c nh, t khng mu n mu en. Trong iu kin kh khn nh khng c qu trnh c nh m, cc ti (vesicle) c th c hnh thnh. Gram dng- hoc Gram khng c nh (variable). Hi u kh ho c vi hi u kh. Catalase dng tnh. a m. Ha d dng hu c . Th ng m c r t ch m (th i gian gia hai ln phn i t bo l 1-7 ngy). Hu ht cc chng u c kh nng c nh nit khng kh invitro v in planta. Thnh t bo cha meso-DAP, acid glutamic, alanin, acid muramic, v glucosamine. Thnh phn phospholipid gm phosphatidylinositiol mannoside, phosphatidylinositol v diphosphatidylglycerol. Acid bo dng thng, mch thng, v khng bo ha. Thnh phn ng gm xylose, madurose hoc fucose, hoc ch gm glucose hoc galactose. Nhiu chng gm 2-Omethyl-D-mannose, hu ht cc chng cha rhamnose. Hu h t c ng sinh v i m t s thc vt ht kn nht nh, to cc nt sn trn r vt ch thch h p. C th tm trong t. T l mol GC trong ADN l 66-71%. Loi chun: Frankia alni

ChiActinoplanes:Pht trin di dng si phn nhnh, khng t o n. Gram dng, nhng m t ph n si dinh dng c th l Gram m. Khng nhum khng acid. R t t s i kh sinh ho c khng c. To nhiu loi sc t c kh nng khuch tn. Bo t ch a trong ti bo t , sinh trn cung sinh bo t hoc khng cung, t khi trong th ch. D i i u ki n nh t nh, nhiu chng c h si sp xp dng que (palisade). Khi , ti bo t ch y u c sinh u cc si. Bo t hnh cu ho c que ngn, sp xp theo nhi u cch khc nhau bn trong ti bo t, c hnh thnh bng cch t on si bn trong ti bo t trc tip hoc sau vi ln phn nhnh. Sau khi ngm trong n c, bo t di ng c gii phng ra t ti bo t, trong mt s trng hp, kh nng di ng xut hin sau khi bo t c gii phng. Bo t di ng cha tin mao cc. Thnh t bo ch a meso-DAP v glycin. Thnh phn ng cha D-xylose v L-arabinose. Thnh phn phospholipid chnh l phosphatidylethanolamin. Hiu kh, ha d dng hu c , a m

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hoc a nhit va phi. Hu ht cc chng u khng cn cc nhn t sinh tr ng hu c. T l mol GC trong ADN l 72-73%. Loi chun: Actinoplanes philippinensis

ChiPilimelia:Ti bo t c sinh ra trn b mt ca c cht t cung sinh bo t . Ti bo t c hnh cu, hnh trng, hnh qu l, hnh chung hay hnh tr, c kch th c xp x 10-15 m. Ti bo t cha rt nhiu bo t xp thnh chui song song ho c cc hng cu n khng u. Gram dng. Khun ty c cht c ng knh 0,2-0,8 m, phn nhnh v c vch ngn. Khun ty kh sinh tht s khng c hnh thnh. Thnh t bo ch a meso-DAP v glycin. Thnh phn ng cha xylose v arabinose. Ch m c c trn mi trng hn hp. Khun lc nh, c hoc mm. Khun ty c ch t c mu vng chanh, vng, da cam hoc xanh xm, chuyn sang nu n nu s m khi gi. Hi u kh, ha t dng hu c, iu kin sinh trng ti u l pH 6,5-7,5 v 20-30 oC. C kh nng phn hy c cht cheratin (lng tc ca ng vt c v). T l mol GC trong ADN l 72-73%. Loi chun: Pilimelia terevasa

ChiDactylosporangium:Ti bo t hnh ngn tay hoc hnh chy c hnh thnh t cung sinh bo t ngn ca khun ty c cht. Cc ti bo t pht trin n l hoc t thnh tng m trn b mt c cht. Mi ti bo t cha mt dy gm ba n bn bo t . Bo t c hnh ch nht, hnh elip, hnh trng hoc hi tr, c kh nng di ng nh m t tm tin mao cc. Khng c khun ty kh sinh thc s. Khun ty c cht c ng knh 0,5-1,0 m, phn nhnh v t khi c vch ngn. Gram dng , Khng nhu m khng acid. Thnh t bo cha meso-DAP v glycin. Thnh phn ng cha xylose v arabinose. C kh nng sinh trng trn nhiu loi mi trng. Khun lc c, h i th, th ng ph ng, i khi nh ln mt b mt trn hoc hi nhn. Khun ty c cht c mu vng xanh, da cam, hay nu. Hiu kh, ha t dng hu c , i u ki n sinh tr ng t i u l pH 6,0-7,0 v 25-37oC. T l mol GC trong ADN l 71-73%. Loi chun: Dactylosporangium aurantiacum

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ChiMicromonospora:C h si sinh trng mnh, phn nhnh, c vch ngn, ng knh trung bnh 0,5 m. Bo t khng c kh nng di ng mc trc ti p hoc t cu ng sinh bo t . Khng c khun ty kh sinh. Gram dng, khng nhum khng acid. Thnh t bo bo cha meso-DAP, v/ hoc acid 3-hydroxy diaminopimelic, glycin. Thnh phn ng ch a xylose v arabinose. Thnh phn phospholipid gm phosphatidylethanolamin, phosphatidylinositol v phosphatidylinositol mannoside. Thnh phn menaquinone chnh l MK-9(H4), MK-10(H4), MK-10(H6) v MK-12(H6). Hiu kh n vi hiu kh. Ha d dng hu c. Nhy cm vi pH di 6,0. C kh nng sinh trng nhit t 2040oC. T l mol GC trong ADN l 71-73%. Loi chun: Micromonospora chalcea

ChiStreptomyces:H si sinh dng phn nhnh nhiu ln, t khi t on, ng knh 0,5-2,0 m. Khun ty kh sinh giai on trng thnh to chui t ba n nhiu bo t. M t s t loi hnh thnh chui bo t ngn trn khun ty c cht. Bo t khng c kh nng di ng. Cc khun lc ban u thng trn nhn nhng sau khun ty kh sinh s pht trin rt mnh m. Sinh nhiu loi sc t khc nhau cng nh sc t c kh nng khuch tn ra mi trng. Rt nhiu chng sn sinh ra m t ho c nhi u lo i ch t khng sinh. Hiu kh, Gram dng, khng nhum khng acid-c n, ha d d ng h u c, catalase dng tnh. Thng c kh nng kh nitrate thnh nitrit, phn hy adenine, esculin, casein, gelatin, hypoxanthine, tinh bt v L-tyrosine. C kh nng s d ng nhiu hp cht hu c lm ngun cacbon duy nht. Nhit sinh tr ng t i u l 2535 oC, pH ti u l 6,5-8,0. Thnh t bo cha L-DAP, khng ch a acid mycolic. Thnh phn acid bo gm phn ln acid bo bo ha, iso- v anteiso-. Thnh phn menaquinone chnh l MK-9(H6), MK-9(H8). Thnh phn phospholipid chnh l diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol v phosphatidylinositol mannoside. C nhiu trong t, phn compt. M t s loi gy bnh cho ngi v ng vt, mt s loi gy bnh thc vt. T l mol GC trong ADN l 69-78%. Loi chun: Streptomyces albus

ChiActinomadura:H si sinh dng phn nhnh, rt pht trin. Khun ty c cht khng t o n, c hoc khng c khun ty kh sinh. Khun ty kh sinh giai o n tr ng thnh s hnh thnh chui ngn, i khi di ca cc bo t t (arthrospore). Chu i bo t c th 137

dng thng, un cong hoc xon khng u (1-4 vng). B mt bo t nhn ho c c cc nt. Khun ty kh sinh khi hnh thnh bo t c mu tr ng, xm hay nu, vng, , xanh lc, xanh lam hay tm. Hiu kh, ha d dng hu c . Nhi t sinh tr ng t 20 n 45oC. Mt s loi a nhit. Gram dng. Thnh t bo cha meso-DAP, khng cha acid mycolic. T bo cha madurose. Thnh phn menaquinone chnh l MK-9(H4) v MK-9(H6). T l mol GC trong ADN l 65-69%. Loi chun: Actinomadura madurae

ChiMicrobispora:Khun ty kh sinh phn nhnh, hnh thnh tng cp hai bao t gn v i nhau. Bo t mc trc tip hoc trn cung sinh bo t ngn, hnh c u ho c oval, ng knh trung bnh 1,2-1,6m v khng c kh nng di ng. Thnh t bo cha acid muramic, mesoDAP nhng khng cha ng c trng. Thnh phn ng c a ton t bo ch a madurose. Thnh phn menaquinone chnh l MK-9(H 4). Thnh phn phospholipid chnh cha phosphatidylcholine v phospholipid cha glucosamine. Gram d ng, hi u kh, ha d dng hu c. a m v a nhit. Hu ht cc loi sinh tr ng u cn cc vitamin nhm B, c bit l thiamin. Trong t nhin thng tn ti trong d u. T l mol GC trong ADN l 71-73%. Loi chun: Microbispora rosea

ChiStreptosporangium:Ti bo t hnh cu ng knh khong 10m c hnh thnh trn khun ty kh sinh. Cc bo t c hnh thnh bng cch hnh thnh vch ngn c a si cu n xo n khng phn nhnh bn trong ti bo t. Bo t hnh cu, hnh oval ho c hnh que, khng c kh nng di ng. Thnh t bo cha acid m uramic, meso-DAP nhng khng cha ng c trng. Thnh phn ng ca ton t bo ch a madurose. Thnh ph n phospholipid chnh cha phosphatidylcholine v phospholipid cha glucosamine. Gram dng, hiu kh, ha d dng hu c. a m, m t s loi c kh nng ch u c nhit cao. Mt s loi cn cc vitamin nhm B cho sinh trng. T l mol GC trong ADN l 69-71%. Loi chun: Streptosporangium roseum

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ChiThermomonospora:Khun ty kh sinh sn sinh ra cc bo t n l, bn nhit, khng c kh nng di ng. Bo t c th sinh trc tip nhng thng l trn u c a cc cung sinh bo t c hoc khng phn nhnh. Bo t cng c h c sinh ra t khun ty c ch t. Gram dng. Thnh t bo cha meso-DAP nhng khng cha cc loi acid amin hay ng c trng. Hiu kh, ha d dng hu c. Cn cung c p ngu n acid amin v cc vitamin cho sinh trng nh cao nm men. Tt c cc chng u c th sinh tr ng c nhit 40-48oC, pH 7,0-9,0. pH > 8,0 thch hp cho s sn sinh khun ty kh sinh v hnh thnh bo t. Sn sinh catalase, deaminase, -glucosidase v galactosidase. C kh nng phn gii esculin, xylan, casein, gelatin v cacboxymetylcellulose. Bo t cht 90oC trong 30 pht trong nc. Tt c cc chng u nhy cm vi novobiocin (50 g/ml). C th phn lp c t t nhng t n ti nhiu trong phn bn, phn compt v c kh sy. Loi chun: Thermomonospora curvata

ChiActinosynnema:Khun ty gm hai loi l khun ty c cht hnh thnh b si (synnemata) trn b m t thch v khun ty kh sinh hnh thnh t b si. Khu n ty sinh ra cc chu i bo t . Cc bo t c kh nng hnh thnh tin mao trong mi trng n c. Thnh t bo ch a meso-DAP, acid glutamic, alanin, glucosamin v acid muramic. T bo khng ch a thnh phn ng c trng. Thnh phn phospholipid chnh l phosphatidylinositol mannoside, phosphatidylinositol, phosphatidylethanolamine v phosphatidylglycerol. Acid bo gm loi mch thng v mch nhnh. Thnh phn menaquinon chnh l MK9(H4) v MK-9(H6). Gram dng, khng acid fast, sinh catalase. Hiu kh. a m. Ha d dng hu c. Hu ht cc chng c phn lp tr c ti p t m th c v t nh l c bn b sng. T l mol GC trong ADN l 73%. Loi chun: Actinosynnema mirum

ChiNocardiopsis:Khun ty c cht pht trin mnh, phn nhnh nhiu, c th t on thnh cc th hnh cu v hnh que. Khun ty kh sinh di, phn nhnh khng u, th ng, u n ho c hnh zch zc, t on thnh cc bo t vi chi u di khc nhau. Bo t hnh oval hoc ko di, b mt nhn. Gram dng, hiu kh, ha d dng h u c , khng nhum khng axt. Thnh t bo cha meso-DAP, khng cha ng c trng, khng c acid mycolic. Thnh phn menaquinon chnh l MK-10(H2,4,6) hoc MK-9(H4,6). T l mol GC trong ADN l 64-69%. 139

Loi chun: Nocardiopsis dassonvillei Di y l hnh nh mt s chi x khun thng gp:

Frankia cng sinh r cy Phi lao

Nangbo

NocardiaKhunlcvkhuntyton

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MtsloiStreptomyces

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Mt s chi x khun: A-Microtetraspora; B- Planomonospora; C- Actinosynnema; DActinobispora; E- Saccharothrix; F- Crosiella

142

143

ChiActinomadura

Cc chi: A- Micromonospora; B- Dactylosporangium; C- Verrucosispora

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D- Pilimelia; E- Catenuplanes

Chi Actinoplanes

Loi Actinomyces viscosus

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Trn y l quan h pht sinh chng loi ca khong 90 chi x khun khc nhau

Mtsphngphpphnlpxkhun: MitrngHV(Humicacidvitaminagar)Thnhphn: Acidhumic CaCO3 FeSO4.7H2O KCl MgSO4.7H2O Na2HPO4 HnhpvitaminnhmB* Cycloheximide Thch Ncct pH 1g 0,02g 0,01g 1,71g 0,05g 0,5g 5ml 500mg 18g 1L 7,2

*Hn hp vitamin nhm BThiamineHCl Riboflavin Niacin PyridoxinHCl Inositol CaPantothenate AcidpAminobenzoic Biotin Ncct 10mg 10mg 10mg 10mg 10mg 10mg 10mg 10mg 10ml

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MitrngYS(YeastextractSolublestarchagar)Caonmmen Tinhbttan Thch Ncct pH 2g 10g 18g 1L 7,3

Thng s dng mi trng YS pha long 5 ln phn lp x khu n v pha long 2 ln nghin cu.

MitrngMaltoseBennetteCaonmmen Caotht NZamine Maltosemonohydrat Thch Ncct pH 1g 1g 2g 10g 18g 1L 7,3

MitrngISP2 Caonmmen Caomalt Glucose Thch Ncct pH 4g 10g 4g 18g 1L 7,2

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MitrngdchthYG(YeastextractGlucose)Yeast extract Glucose 1% 1%

PhngphpRC(RehydrationCentrifugationmethod):dng phnlpxkhunckhnngding mu kh t nhin trong 3-5 ngy, nghin nh mu Ly 0,5 g mu cho vo 50 ml m phosphat 10 mM- 5% cao nm men (pH 7) Loi b phn ni trn b mt Gi 28oC trong 1,5 gi Chuyn 8 ml lp dch pha trn sang ng ly tm Ly tm vi tc 3000 rpm (1500 x g) trong 20 pht Gi nhit phng trong 30 pht Chuyn 3 ml lp dch trn sang ng khc. Ly 1 ml pha long vi nc ct (10-3, 10-4) Tri trn mi trng HV vi pha long 10-2, 10-3, 10-4 Nui cy nhit 28oC trong 14-21 ngy

PhngphpSDSYE(SodiumdodecylsulfateYeastextract method):dngphnlpttcccloixkhun mu kh t nhin trong 3-5 ngy, nghin nh mu Ly 1 g mu cho vo 10 ml nc ct v trng (10-1)

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Ly 1 ml dch trn cho vo 9 ml SDS-YE trong m phosphat (10 -2) (SDS 0,05%, cao nm men 6%, m phosphat 5mM, pH 7,0) Sc nhit 40oC trong 20 pht Pha long bng nc ct v trng Tri trn mi trng HV Nui cy 28oC trong 14-21 ngy

Phng php DH (Dry heating method): dng phn lp x khunchyuthucchiStreptomyces mu kh t nhin trong 3-5 ngy, nghin nh mu Sy mu 100oC trong 30 pht Rc mt lng rt nh mu ln mi trng HV Nui cy 28oC trong 7-14 ngy

Mtsphngphphaphnloixkhun: Chunbthnhtbo1. T bo t (1-2 ml) + 5ml nc ct v trng + ht thy tinh (loi ln v nh, ng knh 0.11-0.12mm). Ph t bo bng sng siu m ca.180 trong 1- 1.5 gi cho n khi dung dch tr nn trong. Chuyn sang ng falcon 50ml. Nu ngng cng vic y, gi mu 4oC. Ly tm vi tc 3000g trong 30 pht. Chuyn dch trn sang ng ly tm tc cao (c np y bn trong). Ly tm vi tc 17000rpm trong 30 pht. B dch trn. Thm 3ml SDS 4% (SDS c tc dng loi lipid v protein). Nu dng cng vic y, gi mu nhit phng. Gi 100oC trong 40 pht, nh vn cht np.

2.

3. 4.

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5. 6. 7. 8.

Ly tm vi tc 17000 rpm trong 30 pht 30oC (nu gi nhit thp, SDS s b kt tinh). Loi b dch trn. Ra vi nc t nht 3 ln vi iu kin ging trn (17000rpm, 30 pht) Nu cn thit, x l bng RNase v DNase. ng kh qua m.

Phntchthnhphnacidamintrongthnhtbo1. 2. 3. 4. 2-3mg thnh t bo + 200l HCl 4N, vn cht np, gi 100oC qua m Lm kh bng ht chn khng (khong 1 ngy 1 m) Thm 200 l nc ct, trn u, chuyn sang ng eppendorf, ly tm vi tc 12000 rpm trong 5 pht. Lc mu. Chy sc k bn mng TLC: Dung mi: Methanol Nc ct 6N HCl Pyridine 80ml 17.5ml 4ml 10ml

Cellulose TLC Chun (1l): DAP, Ala, Gly, Glu, Lys Mu: 3-5l Nhn bit bng phun ninhydrin, gi 100oC trong 5 pht. Cc acid amin s hin ln di dng cc chm mu tm. lu, cc chm ny dn dn bin mt, nhng ring chm ca DAP s chuyn sang mu vng.

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Chun b mu chy HPLC: chun b cho c mu v chun 10-20 l (25nmol) mu, lm kh bng ht chn khng (khong 2h) 151

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Thm 20 l Ethanol/DW/Triethylamine 2/2/1, trn u, lm kh bng ht chn khng (khong 2h) Thm 20 l Ethanol/DW/Triethylamine/Phenylisothiocyanate 7/1/1/1, trn u, gi nhit phng trong 20 pht, lm kh bng ht chn khng (khong 2h) Thm 0.5ml dung dch A(dich chy HPLC), lc mu dng lm mu chy HPLC

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iukinchyHPLC hp th (absorba