vi-sinh-vat-hoc- nguyen-lân-dũng

Vi sinh vat hoc Nguyen Lan Dung

VI SINH VT HCGS Nguyn Ln Dng ch bin Vi hng ni u Lc s ngnh Vi sinh vt Nhng c im chung ca Vi Sinh Vt 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 Nm m Blazei - sn xut v ng dng Phn loi vi sinh bng Sinh Hc Phn T Gii thiu mt s k thut bo qun vi sinh vt http://vietsciences.free.fr v http://vietsciences.or

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Lc s nghin cu Vi sinh vt hc1546- 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- Antony van Leeuwenhoek (1632-1723) 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 thy thuc v kin ngh dng phng php v khun phng bnh. 1849- Snow nghin cu dch t ca bnh t vng London. 1857- Louis Pasteur (1822-1895) 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. 1876-1877- Robert Koch (1843-1910) 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 trng 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. 2

Vi sinh vat hoc Nguyen Lan Dung Elie Metchnikoff (1845-1916) 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. 1886- Fraenkel pht hin thy Streptococcus pneumoniae gy ra bnh vim phi. 1887- Richard Petri phi hin ta cch dng hp lng (a Petri) nui cy 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) 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 ca 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 mm 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). 3

Vi sinh vat hoc Nguyen Lan Dung 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 ngun 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. 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. 1944- Avery chng minh ADN chuyn thng tin di truyn trong qu trnh bin np. Waksman tm ra streptomycin. 1046- Lederberg v Tatum khm ph ra qu trnh tip hp (conjugation) vi khun. 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). Zinder v Lederberg khm ph ra qu trnh ti np (transduction) vi khun. 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 dit khun ( acid nalidixic). 4

Vi sinh vat hoc Nguyen Lan Dung 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). Cohen, Boyer, Chang v Helling s dng vect plasmid tch dng gen vi khun. 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 mt nhm vi sinh vt 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 min dch ngi. Mulli trin khai k thut PCR (polymerase chain reaction). 1986- Ln u tin ng dng trn ngi vaccin c sn xut bng k thut di truyn (vaccin vim gan B). 1990- Bt u th nghim ln u tin liu php gen (gene-therapy) trn ngi. 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. 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.

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Janssen

Leeuwenhoek (1632-1723)

Pasteur (1822-1895)

Knh hin vi ca Leeuwenhoek

Bt tch miu t vi sinh vt ca Leeuwenhoek

Th nghim bnh c cong phn i thuyt t sinh (Pasteur)

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Nhng c im chung ca Vi Sinh Vt1-Vi sinh vt thuc gii sinh vt no? Vi sinh vt khng phi l mt nhm phn loi trong sinh gii m l bao gm tt c cc sinh vt c kch thc hin vi, khng thy r c bng mt thng, do phi s dng knh hin vi thng hoc knh hin vi in t. Ngoi ra mun 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 Gii (Kingdom). Hin nay trn gii cn c mt mc phn loi na gi l lnh gii (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).

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), 7

Vi sinh vat hoc Nguyen Lan Dung 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), 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) cha c th dip 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 hc phn t pht hin thy C khun c s sai khc ln trong trt t nucleotid ARN ca 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), chng hn 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 thng 5 gii tng ng vi 3 gii C trng (Archaezoa), Nguyn 8

Vi sinh vat hoc Nguyen Lan Dung sinh (Protista-Protozoa) v Sc khun (Chromista) trong h thng 8 gii v tng ng vi 5 nhm sau y trong h thng 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 To silic (Diatoms) , To vng (Golden algae), To nu (Brown algae) v Nm si sng trong nc (Water mold) . Rhodophyta gm cc To (Red algae). Ring To lc (Green algae) th mt 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 3 lnh gii (domain) 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 gia 3 lnh gii Bacteria, Archaea v Eukarya c trnh by trn bng di y:

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

c im Nhn c mng nhn v

Bacteria Khng Khng 11

Archaea C

Eukarya

Vi sinh vat hoc Nguyen Lan Dung hch nhn Phc hp bo quan c Khng mng Hu ht c peptidoglycan Thnh t bo cha acid muramic Cha lin kt este, cc acid Mng lipid bo mch thng Ti kh C Thymine c trong phn ln tARN ARN vn chuyn tARN m u cha Nformylmethionine mARN a cistron Intron trong mARN Ghp ni, gn m v gn ui polyA vo mARN Kch thc Yu t ko di EF2 Mn cm vi cloramphenicol v kanamycin Mn cm vi anisomycin C Khng Khng

Khng Nhiu loi khc nhau, khng cha acid muramic Cha lin kt ete, cc chui aliphatic phn nhnh C Khng c thymine trong nhnh T hoc TyC ca tARN tARN m u cha methionine C Khng Khng

C Khng cha acid muramic Cha lin kt este, cc acid bo mch thng Khng C thymine tARN m u cha methionine

Khng C C

Ribosom 70S 70S Khng phn ng vi c t C phn ng bch hu Mn cm Khng Khng Mn cm ARN polymerase ph thuc ADN S lng enzym Mt Cu trc 4 tiu n v Mn cm vi rifampicin Mn cm Promoter typ Polymerase II Khng Tng t ATPase Sinh methane C nh N2 Quang hp vi dip lc Ho dng v c Khng Khng C C C Mt s 8-12 tiu n v Khng C Trao i cht C C C Khng C C Khng Khng C Khng Ba 12-14 tiu n v Khng C 80S (ribosom t bo cht) C phn ng Khng Mn cm

hiu c chi tit ni dung ghi trong bng ni trn gio vin cn gii thch cho sinh vin nhng kin thc c bn thuc gio trnh T bo hc v Di truyn hc 12

Vi sinh vat hoc Nguyen Lan Dung Phn ln vi sinh vt thuc v ba nhm C khun, Vi khun v Nguyn sinh. Trong gii Nm, th nm men (yeast), nm si (filamentous Fungi) v dng si (mycelia) ca mi nm ln u c coi l vi sinh vt. Nh vy l vi sinh vt khng c mt trong hai gii ng vt v Thc vt. Ngi ta c tnh trong s 1,5 triu loi sinh vt c khong 200 000 loi vi sinh vt (100 000 loi ng vt nguyn sinh v to, 90 000 loi nm, 2500 loi vi khun lam v 1500 loi vi khun). 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 cha 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 triu loi. Bo tng ging chun vi sinh vt (VTCC) thuc TT Cng ngh Sinh hc, HQG H Ni hp tc vi cc nh khoa hc Nht bn v da trn cc k thut sinh hc phn t bc u pht hin c kh nhiu loi vi sinh vt mi 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 : Vi sinh vt thng c o kch thc bng n v micromet (1mm= 1/1000mm hay 1/1000 000m). virus c o kch thc n v bng nanomet (1nn=1/1000 000mm hay 1/1000 000 000m). Kch thc cng b th din tch b mt ca vi sinh vt trong 1 n v th tch cng ln. Chng hn ng knh ca 1 cu khun (Coccus) ch c 1mm, nhng nu xp y chng thnh 1 khi lp nhng c th lch l 1cm3 th chng c din tch b mt rng ti ...6 m2 !

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Light microscope : KHV quang hc Electron microscope : KHV in t Most bacteria: Phn ln vi khun

Kch thc vi khun so vi u kim khu

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Ba dng ch yu vi khun : trc khun, cu khun v xon khun.

2)-Hp thu nhiu, chuyn ho nhanh : Tuy vi sinh vt c kch thc rt nh b nhng chng li c nng lc hp thu v chuyn ho vt xa cc sinh vt khc. Chng hn 1 vi khun lctic (Lactobacillus) trong 1 gi c th phn gii c mt lng ng lactose ln hn 100-10 000 ln so vi khi lng ca chng. tc tng hp protein ca nm men cao gp 1000 ln so vi u tng v gp 100 000 ln so vi tru b.

Lactobacillus qua KHV in t

3) Sinh trng nhanh, pht trin mnh : Chng hn, 1 trc khun i trng (Escherichia coli ) trong cc iu kin thch hp ch sau 12-20 pht li phn ct mt ln. Nu ly thi gian th h l 20 pht th mi gi phn ct 3 ln, sau 24 gi phn ct 72 ln 15

Vi sinh vat hoc Nguyen Lan Dung v to ra 4 722 366 500 000 000 000 000 000 t bo (4 722 366. 10 17), tng ng vi 1 khi lng ... 4722 tn. Tt nhin trong t nhin khng c c cc iu kin ti u nh vy ( v thiu thc n, thiu oxy, d tha cc sn phm trao i cht c hi...). Trong ni ln men vi cc iu kin nui cy thch hp t 1 t bo c th to ra sau 24 gi khong 100 000 000- 1 000 000 000 t bo. Thi gian th h ca nm men di hn, v d vi men ru (Saccharomyces cerevisiae) l 120 pht. Vi nhiu vi sinh vt khc cn di hn na, v d vi to Tiu cu ( Chlorella ) l 7 gi, vi vi khun lam Nostoc l 23 gi...C th ni khng c sinh vt no c tc sinh si ny n nhanh nh vi sinh vt.

Vi kun Escherichia coli

Nm men Saccharomyces cerevisiae

Nm si Alternaria Vi to Chlorella

4) C nng lc thch ng mnh v d dng pht sinh bin d : Trong qu trnh tin ho lu di vi sinh vt to cho mnh nhng c ch iu ho trao i cht thch ng c vi nhng iu kin sng rt khc nhau, k c nhng iu kin ht sc bt li m cc sinh vt khc tgng khng th tn ti c. C vi sinh vt sng c mi trng nng n 1300C, lnh n 0-50C, mn n nng 32% mui n, ngt n nng mt ong, pH thp n 0,5 hoc cao n 10,7, p sut cao n trn 1103 at. hay c phng x cao n 750 000 rad. Nhiu vi sinh vt c th pht trin tt trong iu kin tuyt i k kh, c noi nm si c th pht trin dy c trong b ngm t thi vi nng Formol rt cao... Vi sinh vt a s l n bo, n bi, sinh sn nhanh, s lng nhiu, tip xc trc tip vi mi trng sng ... do rt d dng pht sinh bin d. Tn s bin d thng mc 10 -5-10-10. Ch sau mt thi gian ngn c th to ra mt s lng rt ln cc c th bin d cc h h sau. Nhng bin d c ch s a li hiu qu rt ln trong sn xut. Nu nh khi mi pht hin ra penicillin hot tnh ch t 20 n v/ml dch ln men (1943) th nay c th t trn 100 000 n v/ml. Khi mi pht hin ra acid glutamic ch t 1-2g/l th nay t n 150g/ml dch ln men (VEDAN-Vit Nam).

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Vi sinh vat hoc Nguyen Lan Dung Nh my Vedan-Vit Nam

5) Phn b rng, chng loi nhiu : Vi sinh vt c mt khp mi ni trn Tri t, trong khng kh, trong t, trn ni cao, di bin su, trn c th, ngi, ng vt, thc vt, trong thc phm, trn mi vt... Vi sinh vt tham gia tch cc vo vic thc hin cc vng tun hon sinh-a-ho hc (biogeochemical cycles) nh vng tun hon C, vng tun hon n, vng tun hon P, vng tun hon S, vng tun hon Fe... Trong nc vi sinh vt c nhiu vng duyn hi (littoral zone), vng nc nng (limnetic zone) v ngay c vng nc su (profundal zone), vng y ao h (benthic zone). Trong khng kh th cng ln cao s lng vi sinh vt cng t. S lng vi sinh vt trong khng kh cc khu dn c ng c cao hn rt nhiu so vi khng kh trn mt bin v nht l trong khng kh Bc cc, Nam cc... Hu nh khng c hp cht carbon no (tr kim cng, grapht...) m khng l thc n ca nhng nhm vi sinh vt no (k c du m, kh thin nhin, formol. dioxin...). Vi sinh vt c rt phong ph cc kiu dinh dng khc nhau : quang t dng (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)...

6)- L sinh vt xut hin u tin trn tri t : Tri t hnh thnh cch y 4,6 t nm nhng cho n nay mi ch tm thy du vt ca s sng t cch y 3,5 t nm. l cc vi sinh vt ho thch cn li vt tch trong cc tng c. Vi sinh vt ho thch ca nht c pht hin l nhngdng rt ging vi Vi khun lam ngy nay. Chng c J.William Schopf tm thy ti cc tng c min Ty Australia. Chng c dng a bo n gin, ni thnh si di n vi chc mm vi ng knh khong 1-2 mm v c thnh t bo kh dy. Trc cc nh khoa hc cng tm thy vt tch ca chi Gloeodiniopsis c nin i cch y 1,5 t nm v vt tch ca chi Palaeolyngbya c nin i cch y 950 triu nm.

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Vi sinh vat hoc Nguyen Lan Dung Vt tch vi khun lam cch Vt tch Gloeodiniopsis cch y 1,5 Vt tch Palaeolyngbya cch y 950 y 3,5 t nm t nm triu nm

Cu trc t bo vi khun1. Thnh t bo :

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 bit 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 :

Thnh phn Peptidoglycan Acid teicoic (Teichoic acid) Lipid Protein

Gram dng Gram m T l % i vi khi lng kh ca thnh t bo 30-95 5-20 Cao 0 Hu nh khng c 20 Khng c hoc c t Cao

Mng sinh cht (plasma membrane); Mng ngoi (outer membrane); Chu cht (Periplasmic space)

Peptidoglycan l loi polyme xp, kh bn vng, cu to bi 3 thnh phn: -N-Acetylglucosamin ( N-Acetylglucosamine, NAG) 18

Vi sinh vat hoc Nguyen Lan Dung -Acid N-Acetylmuramic (N-Acetylmuramic acid, NAM) -Tetrapeptid cha c D- v L- acid amin

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Thnh t bo vi khun Gram dng

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Thnh t bo vi khun Gram m 2- Mng sinh cht: Mng sinh cht hay Mng t bo cht (Cytoplasmic membrane, CM) vi khun cng tng t nh cc sinh vt khc. Chng cu to bi 2 lp phospholipid (PL), chim 30-40% khi lng ca mng, v cc protein (nm trong, ngoi hay xen gia mng), chim 60-70% khi lng ca mng. u phosphat ca PL tch in, phn cc, a nc ; ui hydrocarbon khng tch in, khng phn cc, k nc. CM c cc chc nng ch yu sau y: Khng ch s qua li ca cc cht dinh dng, cc sn phm trao i cht Duy tr p sut thm thu bnh thng trong t bo. L ni sinh tng hp cc thnh phn ca thnh t bo v cc polyme ca bao nhy (capsule). L ni tin hnh qu trnh phosphoryl oxy ho v qu trnh phosphoryl quang hp ( vi khun quang t dng) L ni tng hp nhiu enzym, cc protein ca chui h hp. Cung cp nng lng cho s hot ng ca tin mao

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Sinh vin in ch thch theo hng dn ca gio vin

Cu trc ca u v ui ca phospholipid

2.

T bo cht :

T bo cht (TBC-Cytoplasm) l phn vt cht dng keo nm 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 lng phn t thp. Bo quan ng lu trong TBC l ribosom (ribosome). Ribosom nm t do trong t bo cht v chim ti 70% trng lng kh ca TBC. Ribosom gm 2 tiu phn (50S v 22

Vi sinh vat hoc Nguyen Lan Dung 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 tht (nm, thc vt, ng vt) c trnh by trong bng sau y (Gio vin ging sinh vin ch thch vo hnh bng ting Vit)

Ribosom vi khun

So snh Ribosom Vi khun v cc Sinh vt nhn tht (Eukaryotic ribosome) 23

Vi sinh vat hoc Nguyen Lan Dung Trong t bo cht ca vi khun cn c th gp cc cht d tr nh cc ht glycogen, ht PHB (Poly-hydroxybutyrat), Cyanophycin, Phycocyanin, cc ht d nhim sc (metachromatic body), cc git lu hunh... loi vi khun dit cn trng Bacillus thuringiensis v Bacillus sphaericus cn gp tinh th c (parasoral body) hnh qu trm, c bn cht protein v cha nhng c t c th git hi trn 100 loi su hi (tinh th c ch gii phng c t trong mi trng kim do cc vi khun ny hon ton v hi vi ngi, gia sc, gia cm, thu hi sn- c hi i vi tm). Bacillus sphaericus c th dit cung qung ca cc loi mui.

Bo t (spore) v Tinh th c (Crystal) Bacillus thuringiensis (tri) v Bacillus sphaericus (phi).

3.

Th nhn:

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 xon 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.

24


Th nhn trong t bo vi khun Escherichia coli.

Ngoi NST, trong t bo nhiu vi khun cn gp nhng ADN ngoi NST. l nhng ADN xon kp c dng vng khp kn, c kh nng sao chp c lp, chng c tn l Plasmid. 4. Bao nhy:

Bao nhy hay Gip mc (Capsule) gp mt s loi vi khun vi cc mc khc nhau: -Bao nhy mng ( Vi gip mc, Microcapssule) -Bao nhy (Gip mc, Capsule) -Khi nhy ( Zooglea) Mun quan st bao nhy thng ln tiu bn vi mc tu, bao nhy c mu trng hin ln trn nn ti. 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, acid 2-keto-3-deoxygalacturonic, acid uronic, acid pyruvic, acid axetic... ngha sinh hc ca bao nhy l: -Bo v vi khun trong iu kin kh hn, bo v vi khun trnh b thc bo (trng hp Ph cu khun-Diplococcus pneumoniae) -Cung cp cht dinh dng cho vi khun khi thiu thc n -L ni tch lu mt s sn phm trao i cht (dextran, xantan...) -Gip vi khun bm vo gi th ( trng hp cc vi khun gy su rng nh Streptococcus salivarrius, Streptococcus mutans...)

25


Vi khun Acetobacter xylinum c bao nhy cu to bi cellulose. Ngi ta dng vi khun ny nui cy trn nc da ch to ra Thch da (Nata de coco).

da (Nata de coco)

Vi khun Leuconostoc mesenteroides c bao nhy dy cha hp cht polyme l Dextran c tc dng thay huyt tng khi cp cu m thiu huyt tng. Sn phm ny rt quan trng khi c chin tranh. Vi khun ny thng gp cc nh my ng v gy tn tht ng trong cc b cha nc p ma. Nh enzym dextransuccrase m ng saccarose b chuyn thnh dextran v fructose.

Vi khun Leuconostoc mesenteroides

26

Vi sinh vat hoc Nguyen Lan Dung Mt s bao nhy ca vi khun cn c dng sn xut Xantan (Xanthane) dng lm cht ph gia trong cng nghip du m.

5.

Tin mao v khun mao :

Tin mao (Lng roi, flagella) khng phi c mt mi vi khun, chng quyt nh kh nng v phng thc di ng ca vi khun. Tin mao l nhng si lng di, di knh hin vi quang hc ch c th thy r khi nhum theo phng php ring. Di knh hin vi in t c th thy rt r cu trc ca tng si tin mao. xc nh xem vi khun c tin mao hay khng cn c cch th gin tip nhm bit kh nng di ng ca chng. Cy bng que cy nhn u vo mi trng thch ng cha 0.4% thch (agar-agar), cn gi l mi trng thch mm. Nu thy vt cy 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 nm 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 gia 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 (hook). Si tin mao (filament) di khong 10-20 v c ng m knh khong 13-20nm. ng knh ca bao hnh mc l 17nm. Khong cch gia 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 ca cc vng l 22nm, ng knh cc l cc vng l 10nm. Khong cch t mt ngoi ca vng L n mt trong ca vng M l 27nm. Si tin mao cu to bi loi protein c tn l flagellin, c trng lng phn t l 30 000-60 000. Mt s vi khun c bao lng (sheath) bao bc sut chiu di si, nh trng hp chi Bdellovibrio hay vi khun t Vibrio cholera.

Tin mao v khun mao vi khun

27


Tin mao VK Gram dng

Tin mao VK Gram m

28


Tin mao vi khun G +

29


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

Tin mao ca vi khun c cc loi khc nhau tu tng loi : 30

Vi sinh vat hoc Nguyen Lan Dung -Khng c tin mao (v mao, atrichia) -C 1 tin mao mc cc ( n mao, monotricha) -C 1 chm tin mao mc cc ( chm mao, lophotricha) -C 2 chm tin mao mc 2 cc ( song chm mao, amphitricha) -C nhiu tin mao mc khp quanh t bo (chu mao, peritricha) C loi tin mao mc gia t bo nh trng hp vi khun Selenomonas ruminantium.

Cc loi tin mao vi khun Kiu sp xp tin mao lin quan n hnh thc di ng ca vi khun. Tin mao mc cc gip vi khun di ng theo kiu tin- li. Chng o ngc hng bng cch o ngc hng quay ca tin mao. Vi khun chu mao di ng theo hng no th cc tin mao chuyn ng theo hng ngc li. Khi tin mao khng t li v mt hng th vi khun chuyn ng theo kiu nho ln. Tc di chuyn ca vi khun 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 qun quang c th. Chng gip xon th chuyn ng c nh s un vn t bo theo kiu vn nt chai.

31


Xon th (Spirochete) quan st di knh hin vi nn en.

AF: Si trc PC: ng nguyn sinh cht OS: V ngoi IP: L ni

32


C ch chuyn ng un vn t bo Xon th ( OS, AF, PC- xem ch thch hnh trn).

6.

Khun mao v Khun mao gii:

Khun mao (hay Tim mao, Nhung mao , Fimbriae) l nhng si lng rt mnh, rt ngn mc quanh b mt t bo nhiu vi khun Gram m. Chng c ng knh khong 7-9nm, rng rut (ng knh trong l 22,5nm), s lng khong 250-300 si/ vi khun. Kt cu ca khun mao gin n hn nhiu so vi tin mao. Chng c tc dng gip vi khun bm vo gi th ( nhiu vi khun gy bnh dng khun mao bm cht vo mng nhy ca ng h hp, ng tiu ho, ng tit niu ca ngi v ng vt).

Khun mao vi khun E.coli C mt loi khun mao t bit gi l Khun mao gii (Sex pili, Sex pilus-s nhiu) c th gp mt s vi khun vi s lng ch c 1-10/ vi khun. N c cu to ging khun mao , ng knh khong 9-10nm nhng c th rt di. Chng c th ni lin gia hai vi khun v lm cu ni chuyn vt cht di truyn (ADN) t th cho (donor) sang th nhn (recipient). Qu trnh ny c gi l qu trnh giao phi (mating) hay tip hp (conjugation). Mt s thc khun th (bacteriophage) bm vo cc th th (receptors) khun mao gii v bt u chu trnh pht trin ca chng. 33


Cc nhm vi khun ch yuTheo 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

34


35

Vi sinh vat hoc Nguyen Lan Dung 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 hp hiu 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. Mt s loi c sc t phycoerythrin. Chng phi hp vi sc t lc to nn mu nu. Mng lin kt vi phycobilisom. n bo hoc a bo dng si. Khng di ng hoc di ng bng cch trn (gliding), mt s loi c ti kh (gas vesicles).Nhiu loi c d t bo (heterocysts) v c kh nng c nh nit. Vi khun lam c mt khp mi ni, trong t, trn , trong sui nc nng, trong nc ngt v nc mn. Chng c nng lc chng chu cao hn so vi thc vt i vi cc iu kin bt li nh nhit cao, pH thp. Mt s loi c kh nng sng cng sinh vi cc c th khc nh Ru, Dng 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: aNhm I (c tc gi gi l b Chroococcal):

Hnh que hoc hnh cu n bo, khng c dng si hay dng kt khi (aggregate); phn i hoc ny chi; khng c d t bo (heterocytes). Hu ht khng di ng. T l G+C l 31-71% . Cc chi tiu biu l:

-Chamaesiphon

-Chroococcus

-Gloeothece

-Gleocapsa

-Prochloron

36


Chamaesiphon

Chroococcus

Glooeothece

Gleocapsa

37


Prochloron

b-Nhm II (c tc gi gi l b Pleurocapsales):

Hnh que hoc hnh cu n bo. c th to dng kt khi (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

38


Pleurocapsa

Dermocapsa

Chroococcidiopsis

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

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 3467%. Cc chi tiu biu l:

-Lyngbya

-Osscillatoria

-Prochlorothrix

-Spirulina

-Pseudanabaena

39


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 bit ho (specialized cell) ; phn i trn mt phng, c kiu t on to thnh on 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 :

-Anabaena

-Cylindrospermum 40


-Aphanizomenon

-Nostoc

-Scytonema

-Calothrix

Anabaena

Anabaena trong Bo hoa du

41


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 hn mt chui 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 42


-Geitlerinema

Fischerella

Geitlerinema

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

43


-Chlorococcales

-Gloeobacteria

-Nostocales

-Oscillatoriales

-Pleurocapsales

-Prochlorales

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

A-Vi khun hnh cu

2.1- Chi Sporosarcina

AA-Vi khun hnh que

44

Vi sinh vat hoc Nguyen Lan Dung B-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

45

Vi sinh vat hoc Nguyen Lan Dung 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

46

Vi sinh vat hoc Nguyen Lan Dung E-C th phn gii Thiamin

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

47

Vi sinh vat hoc Nguyen Lan Dung 2.15- Sulfidobacillus

II- Khng phn gii qung pyrit

1.16- Bacillus

Sporosarcina

Desulfotomaculum

Clostridium

Paenibacillus

Bacillus

3- Trc khun Gram m, ln men , hiu kh hoc k kh khng bt buc

AB-

Catalase dng tnh, khng di ng hay di ng nh tin mao cc Thng di ng nh tin mao cc, khng k sinh ng vt c xng sng 48

Vi sinh vat hoc Nguyen Lan Dung 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 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 49

Vi sinh vat hoc Nguyen Lan Dung 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 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

50


Pasteurella

Pasteurella

Enterobacter

Morganella

Salmonella

Salmonella

Yersinia

Citrobacter

Serratia

Serratia

Klebsiella

Klebsiella 51

Escherichia


4-Trc khun Gram m , khng ln men , hiu kh hoc k kh khng bt buc

A- Sinh trng c 60C 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 52

Vi sinh vat hoc Nguyen Lan Dung 4.8- Chi Gluconobacter 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 4.15- Chi Xanthobacter CC- Khng th c nh nit ngoi c th 53

Vi sinh vat hoc Nguyen Lan Dung D-Hnh thnh nt sn trong r, trong thn thc vt, c th c nh nit t khng kh. 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 G-C th dng hp cht 1 carbon lm 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 denitrificans/ Paracoccus alcaliphalus JJ-Di ng K-Nhum Gram dng tnh 54 ngun carbon duy nht

Vi sinh vat hoc Nguyen Lan Dung hoc kh bin 4.15- Chi Xanthobacter KK- Khng nh trn 4.24- Chi Methylobacter 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 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 GG- Khng th dng hp cht 1 carbon lm ngun carbon duy nht H-Sinh trng cn NaCl hay nc bin I- Khng di ng 4.30- Chi Mesophilobacter II- Di ng J-C gelatinase 55

Vi sinh vat hoc Nguyen Lan Dung 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 L- Sinh axt t mannit 4.34- Chi Marinomonas LL- Khng sinh axt t mannit 4.36- Chi Pseudomonas 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) 56 (P.stanieri, P.perfectomarina,

Vi sinh vat hoc Nguyen Lan Dung 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 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 57

Vi sinh vat hoc Nguyen Lan Dung 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 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

58


Thermus

Gluconobacter

Azotobacter

Methylobacterium

Bordetella

Rhizobium 59

v nt sn trn r


Sinorhizobium

v nt sn trn r

Methylococcus

Bradyrhizobium

v nt sn trn r

Agrobacterium

v nt sn

Pseudomonas

60


Alcaligenes

Flavobacterium

v khun lc

Moraxella

Chromobacterium

Acinetobacter

5- Cu khun Gram m, k kh bt buc A- 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 61

Vi sinh vat hoc Nguyen Lan Dung 5.4- Chi Veillonella

Megasphaera

Acidaminococcus

Veillonella

6-Trc khun, phy khun, xon khun Gram m, k kh :

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 6.4- Chi Halothermothrix CC- Di ng nh tin mao mc bn cnh hay gn cc t bo, sn phm ln men ch yu l acid acetc, acid lactic, acid succinic, H2/CO2 6.5- Chi Thermotoga 62

Vi sinh vat hoc Nguyen Lan Dung 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 6.11- Chi Coprothermobacter EE-Khng sinh kh, sn phm ln men gm acid acetic, acid propionic, acid isopentanoic 6.12- Chi Anaerobaculum II- Khng sinh trng nhit 550 C, a m 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 63

Vi sinh vat hoc Nguyen Lan Dung 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 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. Sn phm ch yu ln men hydrat carbon l acid acetic hoc acid acetic vi ethanol v H2/CO2 B-Acid acetic l sn phm ln men duy nht C- Di ng nh chu mao 6.21- Chi Acetoanaerobium 64

Vi sinh vat hoc Nguyen Lan Dung 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 mt 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 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 2- Sn phm ch yu ln men carbohydrat l acid lactic hoc acid lactic , acid acetic v cc acid khc B-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 65

Vi sinh vat hoc Nguyen Lan Dung 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 EE- T bo < 1,5mm 6.33-Chi Mitsuokella 6.34- Chi Halella 3-Sn phm ch yu ln men carbohydrat l acid propionic hoc acid propionic v cc acid khc B-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 66

Vi sinh vat hoc Nguyen Lan Dung BB- Sn sinh hn hp acid, acid propionic l chnh, khng sinh kh 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 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 6.47- Chi Bacteroides 4- Sn phm ch yu ln men carbohydrat l acid butyric hoc acid butyric v cc acid khc B-Di ng nh tin mao mc cc hay gn cc, t bo hnh cong C- Sn phm ln men ch l acid butyric 67 acetic

Vi sinh vat hoc Nguyen Lan Dung 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 5-Sn phm ln men carbohydrat l hn hp acid c cha acid succinic hoc acid formic B- Sn sinh hn hp acid c cha acid formic C-Sn sinh axit formic v CO2 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 68

Vi sinh vat hoc Nguyen Lan Dung 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 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

69


Thermosipho

Syntrophobacter nui cy chung vi Methanobrevibacte

Wolinella

Porphyromonas gingivalis

vt cy trn thch mu

Leptotrichia

Bacteroides 70

Bacteroides


Fusobacterium

Fusobacterium

Mitsuokella

7- Cu khun Gram dng hiu kh v k kh khng bt buc A- Hiu kh B-Di ng 71

Vi sinh vat hoc Nguyen Lan Dung 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 % 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 AA- K kh khng bt buc B-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 72

Vi sinh vat hoc Nguyen Lan Dung 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 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 73


Micrococcus

Deinococcus

Deinococcus

Melissococcus

Melissococcus

Khun lc Stomatococcus

Staphylococcus

Staphylococcus 74

Pediococcus


Aerococcus

Leuconostoc

Leuconostoc

Enterococcus

Lactococcus

Lactococcus

75

Vi sinh vat hoc Nguyen Lan Dung Streptococcus Streptococcus

Gemella

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

A-Hiu kh B-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

76

Vi sinh vat hoc Nguyen Lan Dung 8.7- Chi Caseobacter CC-Khng cha axt mycolic D-Sp xp thnh hnh ch V E- Peptidoglycan c cha D-ornitin, quinone l MK-11,MK-12 8.8- Chi Aureobacterium EE- Peptidoglycan cha L-ornitin, quinone l MK-8 8.9- Chi Sphaerobacter DD-T bo khng sp xp thnh hnh ch V EHnh que khng quy tc F-Khng c dng t bo hnh cu hay hnh que ngn G- Quinone l MK-8 8.10- Chi Rubrobacter GG- Khng nh trn H- Peptidoglycan cha D-ornitin 8.11- Chi Curtobacterium HH- Khng nh trn 8.12- Chi Clavibacter FF-C dng t bo hnh cu hay hnh que ngn G-Peptidoglycan cha DAP 8.13- Chi Aeromicrobium GG-Peptidoglycan khng cha DAP 8.14- Chi Microbacterium EE- Hnh que c quy tc 8.15- Chi Renibacterium AA- K kh khng bt buc

77

Vi sinh vat hoc Nguyen Lan Dung B-Catalase (+) C- C cha DAP D-C acid mycolic 8.16- Chi Corynebacterium DD- Khng c acid mycolic E-Sn sinh acid propionic 8.17- Chi Propionibacterium EE- Khng sn sinh acid propionic F-Hnh que khng quy tc 8.18- Chi Dermabacter FF-Hnh que c quy tc G-Chiu rng vi khun > 1,0mm 8.19- Chi Caryophanon GG- Chiu rng vi khun < 1,0mm H-Gy bnh ng vt 8.20- Chi Listeria HH- Khng gy bnh ng vt 8.21- Chi Brochothrix CC-Khng cha DAP D-Peptidoglycan cha lysin E- Di ng F- Quinone l MK-7 8.22- Chi Exiguobacterium FF- Quinone l MK-9 8.23 Chi Jonesia

78

Vi sinh vat hoc Nguyen Lan Dung EE- Khng di ng 8.24- Chi Rothia DD Peptidoglycan khng cha lysin EPhn gii cellulose 8.25- Chi Cellulomonas EE- Khng phn gii cellulose 8.26- Chi Rarobacter BB- Catalase (-) C-Peptidoglycan thuc nhm B (theo Schleiter v Kandier,1972) D- Hnh que khng quy tc 8.27- Chi Agromyces DD- Hnh que c quy tc EThng c dng hnh si 8.28- Chi Erysipelothrix EE-Khng c dng hnh si 8.29- Chi Carnobacterium CC-Peptidoglycan khng thuc nhm B m thuc nhm A D-Hnh que khng quy tc E- Peptidoglycan cha LL-DAP 8.30- Chi Arachnia EE- Peptidoglycan khng cha LL-DAP F-Khng c menaquinone 8.31- Chi Gardnerella FF- C menaquinone G-C menaquinone MK-10 (H4)

79

Vi sinh vat hoc Nguyen Lan Dung 8.32- Chi Actinomyces GG- C menaquinone MK-9 (H4) 8.33 - Chi Arcanobacterrium DD- Hnh que c quy tc 8.34- Chi Lactobacillus

Brevibacterium

Arthrobacter

Arthrobacter

Rubrobacter

Aeromicrobium

Renibacterium

80


Corynebacterium

Propionibacterium

Listeria

Cellulomonas

Cellulomonas

Agromyces

Actinomyces

Lactobacillus

Lactobacillus

9- Trc khun khng quy tc, khng bo t A- Gram (+) B- a nhit C- T bo c phn nhnh 81

Vi sinh vat hoc Nguyen Lan Dung D-a kim 9.1- Chi Anaerobranca DD- Khng a kim 9.2- Chi Thermobrachicum CC- T bo khng phn nhnh D-Di ng 9.3- Chi Thermoanaerobacter DD-Khng di ng 9.4- Chi Thermoanaerobium BB-a m C-T bo hnh b dc, hnh que ngn hay hnh cong D- T bo nh nhn hnh con, sp xp tng i hnh li lim 9.5- Chi Falcivibrio DD- T bo hnh bu dc, hnh que ngn, khng sp xp thnh hnh li lim E-Ln men hydrat carbon ch sn sinh acid acetic, cn c th CO2 F- Thch hp sinh trng 30C 9.6- Chi Acetobacterium FF-Thch hp sinh trng 38C 9.7- Chi Acetitomaculum EE- Ln men hydrat carbon sinh acid lactic, khng tng hp ra acid acetic t H2 v CO2 9.8- Chi Atopobium CC-T bo hnh que khng quy tc D-T bo c phn thnh hnh na (inh ba), ln men hydrat carbon khng sinh kh 9.9- Chi Bifidobacterium DD- T bo khng phn thnh hnh na, ln men hydrat carbom c 82 tng hp ra acid acetic t H2 v

Vi sinh vat hoc Nguyen Lan Dung sinh kh E-Sn phm ln men ch yu l acid butyric, acid acetic hoc acid formic. 9.10- Chi Eubacterium EE- Sn phm ln men ch yu l acid acetic, acid lactic v etanol 9.11AA- Gram (-) A- T bo hnh que, a nhit, khng di ng 9.12BB- T bo dng cong hay dng si, di ng C-Ln men hydrat carbon sn phm ch yu l acid butyric 9.13- Chi Butyrivibrio CC-Ln men hydrat carbon sn phm ch yu khng l acid butyric D-Ln men glucose, sn phm ch yu l acid acetic v acid lactic, khng sinh kh 9.14- Chi Mobiluncus DD- Ln men glucose, sn sinh acid formic, acid acetic, acid lactic v etanol, c sinh kh 9.15- Chi Lachnospira Chi Acetogenium Chi Coribacterium

Thermoanaerobacter di knh hin vi in t

Bifidobacterium

Eubacterium

83


Mobiluncus

10- Cu khun Gram dng k kh bt buc A- T bo phn ct theo 3 mt phng thng gc, sp xp thnh hnh lp th 10.1- Chi Sarcina AA- T bo phn ct theo 1 hoc 2 mt phng thng gc , sp xp thnh tng i hay tng 4 t bo B-Khng ln men hydrat carbon, khng sinh kh C- T l G + C mol % trong ADN 50 10.2- Chi Peptococcus CC- T l G + C mol % trong ADN 45 10.3- Chi Peptostreptococcus BB- Ln men hydrat carbon sinh acid C-Trong sn phm ln men ngoi cc acid hu c khc cn c acid butyric v aceton (acetone) 10.4- Chi Coprococcus CC- Trong sn phm ln men khng c acid butyric v aceton 10.5- Chi Ruminococcus

84


Peptostreptococcus

Peptostreptococcus

Ruminococcus

X khun11- Phn loi x khun X khun l nhm vi khun c bit. Chng c khun lc kh v a s c dng hnh phng x (actino-) nhng khun th li c dng si phn nhnh nh nm (myces). V x khun c cu trc nhn nguyn thu nh mi vi khun khc, chiu ngang ca si cng nh nh vi khun, cho nn c ti liu gi chng l nm tia l khng hp l. X khun phn b rng ri trong t nhin. S lng n v sinh khun lc (CFU- colony-forming 85

Vi sinh vat hoc Nguyen Lan Dung unit) x khun trong 1g t thng t ti hng triu. Trn mi trng c a s x khun c hai koi khun ty: khun ty kh sinh (aerial mycelium) v khun ty c cht (substrate mycelium). Nhiu loi ch c khun ty c cht nhng cng c loi (nh chi Sporichthya) li ch c khun ty kh sinh. Gia khun lc thng thy c nhiu bo t mng mng gi l bo t trn (conidia hay conidiospores). Nu bo t nm trong bo nang (sporangium) th c gi l nang bo t hay bo t kn (sporangiospores). Bo t x khun c sinh ra u mt s khun ty theo kiu hnh thnh cc vch ngn (septa). Cc chui bo t trn c th ch l 1 bo t (nh Thermoactinomyces, Saccharomonospora, Promicromonospora, Micromonospora, Thermomonosspora...),c th c 2 bo t (nh Microbispora), c th l chui ngn (nh Nocardia, Pseudonocardia, Streptoverticillium, Sporichthya, Actinomadura, Microtetraspora, Streptoalloteichus, Glycomyces, Amycolata, Amycolatopsis, Catellatospora, Microellobosporia...), c th l chui di (nh Streptomyces, Saccharopolyspora, Actinopolyspora, Kibdelosporangium, Kitasatosporia, Saccharothrix, nhiu loi Nocardia, Nocardioides, Pseudonocardia, Amycolatopsis, Streptoverticillium...), c th cc bo t trn nm trn b si (synnema), tng t b si ca nm (nh Actinosynnema, Actinomadura...). Cc chui bo t c th thng, c th xon, c th dng ln sng, c th mc n hay mc vng... Cc cung sinh bo t (sporophore) v cung sinh nang bo t (sporangiophorres) c th ring r, c th phn nhnh. Cc c im hnh thi ny rt quan trng khi tin hnh nh tn x khun. X khun thuc nhm vi khun Gram dng, thng c t l GC trong ADN cao hn 55%. Trong s khong 1000 chi v 5000 loi sinh vt nhn s cng b c khong 100 chi v 1000 loi x khun. X khun phn b ch yu trong t v ng vai tr rt quan trng trong chu trnh tun hon vt cht trong t nhin. Chng s dng acid humic v cc cht hu c kh phn gii khc trong t. Mc d x khun thuc nhm sinh vt nhn s nhng chng thng sinh trng di dng si v thng to nhiu bo t. Thm ch mt s loi x khun cn hnh thnh ti bo t nh chi Streptosporangium, Micromonospora v bo t di ng nh chi Actinoplanes, Kineosporia. Trc y, v tr phn loi ca X khun lun l cu hi gy nhiu tranh lun gia cc nh Vi sinh vt hc ,do n c nhng c im va ging Vi khun va ging Nm. Tuy nhin, n nay, X khun c chng minh l Vi khun vi nhng bng chng sau y: 1. Mt s x khun nh cc loi thuc chi Actinomyces v Nocardia rt ging vi cc loi vi khun thuc chi Lactobacillus v Corynebacterium. 2. X khun ging vi khun ch khng c nhn tht, chng ch cha nhim sc cht phn b dc theo cc si hoc cc t bo. 3. ng knh ca si x khun v bo t ging vi vi khun. ng thi si x khun thng khng cha vch ngn. 86

Vi sinh vat hoc Nguyen Lan Dung 4. X khun l ch tn cng ca cc thc khun th ging nh vi khun, trong khi , nm khng b tn cng bi thc khun th. 5. X khun thng nhy cm vi cc khng sinh c tc dng ln vi khun, nhng li thng khng vi nhng khng sinh tc dng ln nm nh cc polyen. 6. X khun khng cha chitin, cht c mt trong si v bo t ca nhiu nm, m khng c vi khun. ng thi ging nh phn ln vi khun, x khun khng cha cellulose. 7. Tng t vi vi khun, x khun nhy cm vi phn ng acid ca mi trng, c im ny khng c nm. 8. Cc c im v si v nang bo t kn(sporangium) ca chi Actinoplanes cho thy c th chi ny l cu ni gia vi khun v cc nm bc thp. X khun thuc v lp Actinobacteria, b Actinomycetales, bao gm 10 di b, 35 h, 110 chi v 1000 loi. Hin nay, 478 loi c cng b thuc chi Streptomyces v hn 500 loi thuc tt c cc chi cn li v c xp vo nhm x khun him.

87


Khun lc x khun

Khun ty x khun v bo t

Chui bo t trn cung sinh bo t dng xon

88


Bo t x khun

S hnh thnh hai loi khun ty sau khi bo t x khun ny mm

89


Si bo t v chui bo t trn

Mt s dng bo t x khun (SV t ch thch tng hnh) 90


Mt s dng nang bo v nang bo t x khun

91


Mt s dng chui bo t x khun (SV t ch thch tng hnh)

c im ca khun ty kh sinh v khun ty c cht c trnh by trong s sau y:

92


93


94


95


96


phn loi x khun ngi ta s dng cc tiu chun nh trnh t rADN 16S, lai ADN, hnh thi, sinh l sinh ha v ha phn loi. Hin nay, i a s cc nh khoa hc ng vi quan nim hai chng c coi l hai loi ring bit nu chng ging nhau di 70% khi tin hnh lai ADN. Keswani v cng s chng minh rng nu s tng ng gia hai trnh t rADN 16S l 98.6% th xc sut mc ging nhau trong php lai ADN thp hn 70% s l 99%. V th gi tr tng ng 98.6% ca trnh t rADN 16S c coi l ngng phn bit hai loi khc nhau. Tuy nhin, cng c nhiu nh khoa hc ly gi tr ny l 98%. c bit ha phn loi l rt quan trng trong vic phn loi x khun. Chng rt c ch trong phn loi mc n chi. l nhng c im sau: ng, loi acetyl, acid mycolic trong thnh t bo, menaquinone, phospholipid, acid bo v t l GC trong ADN. 1= Chi; 2=Loi thnh t bo ; 3=Thnh phn ng ; 4=Loi peptido glycan ; 5=Acid mycolic 6=Thnh phn acid bo ; 7=Mena quinon ch yu ; 8=Loi phospho-lipid ; 9=T l mol GC trong ADN (%) 97

Vi sinh vat hoc Nguyen Lan Dung 1 2 3

4

5

6

7

8

9 5167 6572 6369 6465 68 73 6270 6472 6069 6673 7172 7273 7173 7173 6972 6973 71 69 70 5763 73 6668 68

H Corynebacteriaceae v mt s h lin quan Corynebacterium 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

H Micromonosporaceae Micromonospora Actinoplanes Cattelatospora Catenuloplanes Couchioplanes II II II VI VI D D D D D D D A3 A3 A1 A1 A1 A1 A1 3b 2d 2d 2b 3b 3b 9(H4.6), 10(H4,6) 9(H4,6), 10(H4,6) 9(H4,6), 10(H8,6) 10(H4), 11(H4) 9(H4) 9(H4,6,8) 9(H4,2) 10(H4) 9(H4) PII PII PII PIII PII PII PII PII PII

Dactylosporangium II Pilimelia Spirilliplanes Verrucosispora II II II

H Propionibacteriaceae v Nocardia Propionibacterium I Friedmaniella Luteococcus Microlunatus I I I A3 A3 A3 A3 1a 2d 9(H4) 9(H4) 9(H4) 9(H4) 98 PI PI PII

Vi sinh vat hoc Nguyen Lan Dung Propioniferax Tessaracoccus Nocardioides Aeromicrobium Hongia Kribbella Marmoricola I I I I I I I A3 A3 A3 A3 A3 A3 A3 1a 3a 3a 2d 9(H4), 7(H4) 8(H4) 9(H4) 9(H4) 9(H4) 9(H4) 8(H4) PI PI PI PII PI PIII PI PIII/ PII PIV PIII PII PII PII PII PIII PII 5963 74 6772 6972 71 6870 72 6879 71 6468 6669 66 6769 6974 7077 6973 6971 73 7071 7076

H Pseudonocardiaceae Pseudonocardia Actinobispora Actinopolyspora Amycolatopsis III IV IV III A A A A A A A A C A1 A1 A1 A1 A1 A1 3c 3f 2a 2c 2c 1 8(H4) 7(H2), 9(H2) 9(H4), 10(H4) 9(H2,4,6) 9(H4) 9(H2,4) 9(H4) 9(H4) 9(H4)

Kibdelosporangium III Prauserella IV

Saccharomonospora III Saccharopolyspora IV Thermocrispum III

H Actinosynnemataceae Actinosynnema Actinokineospora Kutzneria Saccharothrix III III III III C C C C A1 A1 A1 9(H4,6) 9(H4) 9(H4) 9(H4), 10(H4) PII PII PII PII

H Streptosporangiaceae Streptosporangium III Acrocarpospora Herbidospora Microbispora III III III B B B B A1 A1 A1 3c 3c 9(H0,2,4) 9(H2,4) 10(H4,6) 9(H0,2,4) 99 6971 68PIV/ PII 69 PIV 71 PIV 67PIV

Vi sinh vat hoc Nguyen Lan Dung Microtetraspora Nonomuraea Planobispora Planomonospora Planotetrespora III III III III III B B B B B A1 A1 A1 A1 3c 3c 3c 3c 3c 9(H0,2,4) 9(H0,2,4) 9(H0,2,4) 9(H0,2,4) PIV PIV PIV PIV PIV 74 66 6469 7071 72

H Nocardiopsaceae Nocardiopsis Thermobifida III III C C B B B A1 A1 A1 A1 3d 3c 3e 3e 3a 3e 10(H2,4,6) 10(H6,8) 9(H6,8) 9(H4,6) 9(H6,8) 9(H4,6) PIII PII PI PII PI PI 6469

H Thermomonosporaceae Thermomonospora III Actinocollaria III Actinomadura Spirillospora III III 73 6670 6971 7173 6978 6673 6671 7376 68 70 70 6479 71 5355

H Glycomycetaceae Glycomyces II 2c 10(H4), 11(H4) PI

H Streptomycetaceae Streptomyces Kitasatospora I I/III A3 A3 2c 9(H6,8) 9(H6,8) PII PII

H Frankiaceae v mt s h lin quan Frankia Geodermatophilus III III A1 A3 1 2b 9(H4,6,8) 9(H4) 8(H4) 9(H6,8) 9(H6,8) PI PII PII PI

Microsphaera III Cryptosporangium II Sporichthya I

Cc h khc Kineosporia Thermobispora I/III III A3 9(H4) 9(H0,2) 7,9 100 PIII PIV

Thermoactinomyces III


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 bit l acid diaminopimelic, lysine v thnh phn ng trong thnh t bo, c th l Loi thnh t bo I II III IV V VI VII VIII

Acid diamino Cc acid amin khc LL-A2pm meso- A2pm meso- A2pm meso- A2pm Lys, Orn Lys Lys, A2bu Gly* Gly**

ng

Ara, Gal*** Asp Asp Asp

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 hc LL- A2pm, DL- A2pm (mesoA2pm), DD- A2pm. Ngoi ra cn c OH- A2pm khi c thm nhm OH gn vo v tr C th 4. Lys: Lysine Orn: Ornitin 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

101

Vi sinh vat hoc Nguyen Lan Dung 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. Thnh phn ng: Thnh phn ng A B C D Cc loi ng ch yu Arabinose, Galactose (khng cha Xylose) Madurose (khng cha Arabinose hay Xylose) Khng c ng c trng Arabinose, Xylose

Peptidoglycan: Theo Schleifer v Kandler (1972), peptidoglycan x khun c chia thnh cc nhm nh sau: V tr ca K mi lin Cu ni gia hai mch tetrapeptid hiu kt 3-4 1 Trc tip

K hiu A

K Acid amin hiu v tr th 3 L-Lys L-Orn meso- A2pm L-Lys L-Lys L-Orn meso- A2pm L-Lys L-Orn meso- A2pm L-A2bu L-Lys L-Hsr L-Glu L-Ala L-Orn

2 3

4

B

2-4

1

2

Tiu n v peptid L-acid amin cha mt nhm carboxyl (gly) hoc chui peptid ngn ca acid amin D-acid amin cha hai nhm carboxyl (D-Asp hay D-Glu) L-acid amin (Glyn-L-Lys) D-acid amin (D-Orn hay D-A2bu) 102

Vi sinh vat hoc Nguyen Lan Dung L-Hsr L-A2bu

Du phy (): alanine v tr th nht trong mch tetrapeptid c thay th bi glycine. V d: Loi peptidoglycan ca chi Actinoplanes l A1 c ngha l: Hai mch tetrapeptid lin kt vi nhau v tr acid amin s 3 v 4, mi lin kt ny l trc tip v acid amin v tr s 3 l meso- A2pm. Thnh phn acid bo: 10Methyl- Cyclopropane 18 ++ +++ (+) (+) +++ ++ -

K hiu 1 a b c a b c d a b c d

Khng IsoIsoBo bo ha 14/16/18 15/17 ha +++ +++ +++ ++ (+) + + +++ + + + +++ +++ +++ + + (v) + ++ + + + +++ ++ +++ +++ +++ +++ ++ +++ + +++ + +++ (+) +++ + ++

Anteiso15/17 (+) + +++ +++ (+) ++ + +++

10Methyl-17 (+) ++ +++ (+)

2

3

(v): t hn 5%

Thnh phn menaquinone: X khun ch cha menaquinone, khng cha ubiquinone. K hiu 8(H2) hay MK-8(H2) ch menaquinone cha mt trong s tm n v isoprene b hydro ha. Thnh phn phospholipid: K Thnh phn phospholipid c trng hiu PI Phosphatidylglycerol(v) PII Phosphatidylethanolamine 103

Vi sinh vat hoc Nguyen Lan Dung Phosphatidylcholine, phosphatidylethanolamine(v), phosphatidylmethylPIII ethanolamine(v), phosphatidylglycerol(v), khng cha phospholipid cha glucosamine Phospholipid cha glucosamine, phosphatidylethanolamine(v), PIV phosphatidylmethylethanolamine(v) PV Phospholipid cha glucosamine, phosphatidylglycerol Tt c cc loi u cha phosphatidylinositol

V tr phn loi ca X khun: Lp Actinobacteria Di lp Actinobacteridae B Actinomycetales Di b Actinomycineae H Actinomycetaceae Cc chi: Actinomyces, Actinobaculum, Arcanobacterium, Mobiluncus Di b Micrococcineae H Micrococcaceae Cc chi: Micrococcus, Arthrobacter, Kocuria, Nesterenkonia, Renibacterium, Rothia H Bogoriellaceae Chi: Bogoriella H Brevibacteriaceae Chi: Brevibacterium H Cellulomonadaceae Chi: Cellulomonas H Dermabacteraceae Cc chi: Dermabacter, Brachybacterium H Dermacoccaceae 104

Vi sinh vat hoc Nguyen Lan Dung Cc chi: Dermacoccus, Demetria, Kytococcus H Dermatophilaceae Chi: Dermatophilus H Intrasporangiaceae Cc chi: Tetraphaera H Jonesiaceae Chi: Jonesia H Microbacteriaceae Cc chi: Microbacterium, Agrococcus, Agromyces, Clavibacter, Cryobacterium, Curtobacterium, Frigoribacterium, Leifsonia, Leucobacter, Rathayibacter H Promicromonosporaceae Chi: Promicromonospora H Rarobacteraceae Chi: Rarobacter H Sanguibacteraceae Chi: Sanguibacter Incertae sedis (cha xc nh) Cc chi : Beutenbergia, Ornithinicoccus, Kineococcus Di b Corynebacterineae H Corynebacteriaceae Cc chi: Corynebacterium, Turicella H Dietziaceae Chi: Dietzia H Gordoniaceae Chi: Gordonia H Mycobacteriaceae 105 Intrasporangium, Janibacter, Terrabacter, Terracoccus,

Vi sinh vat hoc Nguyen Lan Dung Chi: Mycobacterium H Nocardiaceae Cc chi: Nocardia, Rhodococcus H Tsukamurellaceae Chi: Tsukamurella Incertae sedis (cha xc nh) Cc chi : Skermania, Williamsia Di b Micromonosporineae H Micromonosporaceae Cc chi: Micromonospora, Actinoplanes, Catellatospora, Catenuloplanes, Couchioplanes, Dactylosporangium, Pilimelia, Spirilliplanes, Verrucosispora Di b Propionibacterineae H Propionibacteriaceae Cc chi: Propionibacterium, Friedmaniella, Propioniferax, Tessaracoccus H Nocardioidaceae Cc chi: Nocardioides, Aeromicrobium, Hongia, Kribbella, Marmoricola Di b Pseudonocardineae H Pseudonocardiaceae Cc chi: Pseudonocardia, Actinobispora, Actinopolyspora, Amycolatopsis, Kibdelosporangium, Prauserella, Saccharomonospora, Saccharopolyspora, Thermocrispum H Actinosynnemataceae Cc chi: Actinosynnema, Actinokineospora, Saccharothrix Incertae sedis (cha xc nh) Cc chi: Kutzneria, Streptoalloteichus Di b Streptosporangineae H Streptosporangiaceae 106 Luteococcus, Microlunatus,

Vi sinh vat hoc Nguyen Lan Dung Cc chi: Streptosporangium, Acrocarpospora, Herbidospora, Microbispora, Microtetraspora, Nonomuraea, Planomonospora, Planotetraspora H Nocardiopsaceae Cc chi: Nocardiopsis, Thermobifida H Thermomonosporaceae Cc chi: Thermomonospora, Actinocorallia, Actinomadura, Spirillospora Di b Glycomycineae H Glycomycetaceae Chi: Glycomyces Di b Streptomycineae H Streptomycetaceae Cc chi: Streptomyces, Kitasatospora Di b Frankineae H Frankiaceae Chi: Frankia H Acidothermaceae Chi: Acidothermus H Geodermatophilaceae Cc chi: Geodermatophilus, Blastococcus H Microsphaeraceae Chi: Microsphaera H Sporichthyaceae Chi: Sporichthya Incertae sedis (cha xc nh) Chi: Cryptosporangium B Bifidobacteriales H Bifidobacteriaceae 107

Vi sinh vat hoc Nguyen Lan Dung Cc chi: Bifidobacterium, Gardnerella Di lp Acidimicrobidae B Acidimicrobiales H Acidimicrobiaceae Chi: Acidimicrobium Di lp Coriobacteridae B Coriobacteriales H Coriabacteriaceae Cc chi: Coriobacter, Atopobium, Denitrobacterium, Eggerthella, Slackia Di lp Rubrobacteridae B Rubrobacteriales H Rubrobacteriaceae Chi: Rubrobacter Di lp Sphaerobacteridae B Sphaerobacteriales H Sphaerobacteriaceae Chi: Sphaerobacter M t mt s chi thng gp: Chi Rhodococcus: 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 on hnh cu hoc que ngn. Bng cch phn on, cc t bo hnh cu s to thnh dng que ri dng si, si phn nhnh v h si. Mt s chng cn to khun ty kh sinh phn nhnh hoc b si (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 on pht trin. Hiu kh. Ha d dng hu c. Catalase dng tnh. Hu ht cc chng u mc tt trn cc mi trng tiu chun 30 oC, 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, hoc khng mu. Arylsulfatase m tnh, nhy cm vi lysozyme, khng phn hy 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. Collinsella, Cryptobacterium,

108

Vi sinh vat hoc Nguyen Lan Dung 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 vi 32-66 cacbon, vi nhiu nht bn lin kt i. T l mol G+C trong ADN l 63-72%. C nhiu trong t v phn gia sc. Mt s chng gy bnh cho ngi v ng vt. Loi chun: Rhodococcus rhodochrous

Chi Nocardioides: Si s cp phn nhnh trn b mt, sau xm nhp vp trong thch ri t on thnh dng que hay cu hay khng u. Si kh sinh c th tha tht, phn nhnh hoc khng sau cng t thnh nhng mu ngn hoc dng que. Nhng mu 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 dng tnh. Hiu kh bt buc. Ha d dng hu c. Mc d dng trn cc mi trng tiu chun. Nhy cm 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

Chi Pseudonocardia: Si kh sinh v si c cht u sinh bo t dng chui. Si phn on, thng c dng zich-zac vi xu hng phng ln ngn hoc gia. Si ko di bng cch ny 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 trng trn nhiu loi mi trng hu c hoc tng hp. a m hoc a nhit. Loi chun: Pseudonocardia thermophila

Chi Saccharopolyspora: Si c cht pht trin mnh, phn nhnh, t thnh cc on dng que vi kch thc 1x5m, ch yu nhng phn gi hn ca khun lc. Khun ty kh sinh phn on to cc chui bo t. Gram dng. Khng nhum khng acid. Hiu kh. Khun lc mng, nh ln, hi nhn, si kh sinh t, thng to thnh chm, ch yu nhng phn gi. C th s dng nhiu loi cht hu c nh ngun cacbon v nng lng duy nht, c th phn gii adenine, khng nhiu loi cht khng sinh nhng nhy cm vi lysozym. T l mol GC trong ADN l 77%. Loi chun: Saccharopolyspora hirsute

Chi Intrasporangium:

109

Vi sinh vat hoc Nguyen Lan Dung 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. Hiu kh. Mc tt nht 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 cha L-DAP v glycine. Thnh phn phospholipid ch yu l mt loi phospholipid cha 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

Chi Actinopolyspora: H si phn nhnh, hnh thnh rt nhiu si kh sinh c ng knh khong 1 m. Si c cht hu ht khng t on. Cung sinh bo t cha 20 hoc hn bo t dng que ngn hoc dng cu vi v nhn hnh thnh trn si kh sinh theo chiu hng 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. Hiu kh. Ha d dng hu c. T l mol GC trong ADN l 64,2%. Loi chun: Actinopolyspora halophila

Chi Saccharomonospora: Ch yu sinh bo t n trn si kh sinh. Bo t khng bn nhit, khng c kh nng di ng, c hnh thnh trn cc cung sinh bo t n gin, khng phn nhnh, di ngn khc nhau. Trn mi trng thch, h si dinh dng phn nhnh, bao ph bi lp si kh sinh vi cc bo t mc dy c dc trn cc si. Si kh sinh ban u c mu trng ri chuyn thnh xm xanh v xanh sm. Sc t xanh cng c si 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 hy c casein, gelatin, tinh bt. xylan v tyrosin. c bit c th s dng glycerol lm ngun cacbon. Phn lp c trong t, cht lng cn h, than bn, thy nhiu trong phn bn, phn compt, v c kh. T l mol GC trong ADN l 69-74%. Loi chun: Saccharomonospora viridis

Chi Frankia: Khng sinh si kh sinh. Ti bo t thng sinh trn cung 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). Hiu kh hoc vi hiu kh. Catalase dng tnh. a m. Ha d dng hu c. Thng mc rt chm (thi 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 110

Vi sinh vat hoc Nguyen Lan Dung gm glucose hoc galactose. Nhiu chng gm 2-O-methyl-D-mannose, hu ht cc chng cha rhamnose. Hu ht cng sinh vi mt s thc vt ht kn nht nh, to cc nt sn trn r vt ch thch hp. C th tm trong t. T l mol GC trong ADN l 66-71%. Loi chun: Frankia alni

Chi Actinoplanes: Pht trin di dng si phn nhnh, khng t on. Gram dng, nhng mt phn si dinh dng c th l Gram m. Khng nhum khng acid. Rt t si kh sinh hoc khng c. To nhiu loi sc t c kh nng khuch tn. Bo t cha trong ti bo t, sinh trn cung sinh bo t hoc khng cung, t khi trong thch. Di iu kin nht nh, nhiu chng c h si sp xp dng que (palisade). Khi , ti bo t ch yu c sinh u cc si. Bo t hnh cu hoc que ngn, sp xp theo nhiu 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 nc, 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 cha 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 hoc a nhit va phi. Hu ht cc chng u khng cn cc nhn t sinh trng hu c. T l mol GC trong ADN l 72-73%. Loi chun: Actinoplanes philippinensis

Chi Pilimelia: 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 thc xp x 10-15 m. Ti bo t cha rt nhiu bo t xp thnh chui song song hoc cc hng cun 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 cha meso-DAP v glycin. Thnh phn ng cha xylose v arabinose. Ch mc c trn mi trng hn hp. Khun lc nh, c hoc mm. Khun ty c cht c mu vng chanh, vng, da cam hoc xanh xm, chuyn sang nu n nu sm khi gi. Hiu kh, ha t dng hu c, iu kin sinh trng ti u l pH 6,5-7,5 v 20-30oC. 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

Chi Dactylosporangium: 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 mt 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 nhum 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, hi th, thng phng, 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, iu kin sinh trng ti u l pH 6,0-7,0 v 25-37 oC. T l mol GC trong ADN l 71-73%. 111

Vi sinh vat hoc Nguyen Lan Dung Loi chun: Dactylosporangium aurantiacum

Chi Micromonospora: 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 tip hoc t cung 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 cha xylose v arabinose. Thnh phn phospholipid gm phosphatidylethanolamin, phosphatidylinositol v phosphatidylinositol mannoside. Thnh phn menaquinone chnh l MK-9(H4), MK10(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 20-40oC. T l mol GC trong ADN l 71-73%. Loi chun: Micromonospora chalcea

Chi Streptomyces: 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. Mt 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 mt hoc nhiu loi cht khng sinh. Hiu kh, Gram dng, khng nhum khng acid-cn, ha d dng hu 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 dng nhiu hp cht hu c lm ngun cacbon duy nht. Nhit sinh trng ti u l 25-35 oC, pH ti u l 6,5-8,0. Thnh t bo cha L-DAP, khng cha 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. Mt 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

Chi Actinomadura: H si sinh dng phn nhnh, rt pht trin. Khun ty c cht khng t on, c hoc khng c khun ty kh sinh. Khun ty kh sinh giai on trng thnh s hnh thnh chui ngn, i khi di ca cc bo t t (arthrospore). Chui bo t c th dng thng, un cong hoc xon khng u (1-4 vng). B mt bo t nhn hoc c cc nt. Khun ty kh sinh khi hnh thnh bo t c mu trng, xm hay nu, vng, , xanh lc, xanh lam hay tm. Hiu kh, ha d dng hu c. Nhit sinh trng t 20 n 45 oC. 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

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Vi sinh vat hoc Nguyen Lan Dung Chi Microbispora: Khun ty kh sinh phn nhnh, hnh thnh tng cp hai bao t gn vi nhau. Bo t mc trc tip hoc trn cung sinh bo t ngn, hnh cu hoc oval, ng knh trung bnh 1,2-1,6m v khng c kh nng di ng. Thnh t bo cha acid muramic, meso-DAP nhng khng cha ng c trng. Thnh phn ng ca ton t bo cha madurose. Thnh phn menaquinone chnh l MK-9(H4). Thnh phn phospholipid chnh cha phosphatidylcholine v phospholipid cha glucosamine. Gram dng, hiu kh, ha d dng hu c. a m v a nhit. Hu ht cc loi sinh trng u cn cc vitamin nhm B, c bit l thiamin. Trong t nhin thng tn ti trong du. T l mol GC trong ADN l 71-73%. Loi chun: Microbispora rosea

Chi Streptosporangium: 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 ca si cun xon khng phn nhnh bn trong ti bo t. Bo t hnh cu, hnh oval hoc hnh que, khng c kh nng di ng. Thnh t bo cha acid muramic, meso-DAP nhng khng cha ng c trng. Thnh phn ng ca ton t bo cha madurose. Thnh phn phospholipid chnh cha phosphatidylcholine v phospholipid cha glucosamine. Gram dng, hiu kh, ha d dng hu c. a m, mt s loi c kh nng chu 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

Chi Thermomonospora: 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 ca cc cung sinh bo t c hoc khng phn nhnh. Bo t cng c h c sinh ra t khun ty c cht. 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 cp ngun acid amin v cc vitamin cho sinh trng nh cao nm men. Tt c cc chng u c th sinh trng c nhit 40-48 oC, 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 tn ti nhiu trong phn bn, phn compt v c kh sy. Loi chun: Thermomonospora curvata

Chi Actinosynnema: Khun ty gm hai loi l khun ty c cht hnh thnh b si (synnemata) trn b mt thch v khun ty kh sinh hnh thnh t b si. Khun ty sinh ra cc chui bo t. Cc bo t c kh nng hnh thnh tin mao trong mi trng nc. Thnh t bo cha meso-DAP, acid glutamic, alanin, glucosamin v acid muramic. T bo khng cha 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 MK-9(H4) v MK-9(H6). Gram dng, khng acid fast, sinh 113

Vi sinh vat hoc Nguyen Lan Dung catalase. Hiu kh. a m. Ha d dng hu c. Hu ht cc chng c phn lp trc tip t m thc vt nh l c bn b sng. T l mol GC trong ADN l 73%. Loi chun: Actinosynnema mirum

Chi Nocardiopsis: 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, thng, un hoc hnh zch zc, t on thnh cc bo t vi chiu di khc nhau. Bo t hnh oval hoc ko di, b mt nhn. Gram dng, hiu kh, ha d dng hu 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%. Loi chun: Nocardiopsis dassonvillei Di y l hnh nh mt s chi x khun thng gp:

Frankia cng sinh r cy Phi lao

Nang bo

Nocardia- Khun lc v khun ty t on

114


Mt s loi Streptomyces

115


Mt s chi x khun: A-Microtetraspora; B- Planomonospora; C- Actinosynnema; D- Actinobispora; ESaccharothrix; F- Crosiella

116


Chi Actinomadura

117


Cc chi: A- Micromonospora; B- Dactylosporangium; C- Verrucosispora D- Pilimelia; E- Catenuplanes

118


Chi Actinoplanes

Loi Actinomyces viscosus

119


Trn y l quan h pht sinh chng loi ca khong 90 chi x khun khc nhau Mt s phng php phn lp x khun:

120

Vi sinh vat hoc Nguyen Lan Dung Mi trng HV (Humic acid-vitamin agar) Thnh phn: Acid humic CaCO3 FeSO4.7H2O KCl MgSO4.7H2O Na2HPO4 Hn hp vitamin nhm B* Cycloheximide Thch Nc ct pH 1g 0,02 g 0,01 g 1,71 g 0,05 g 0,5 g 5 ml 500 mg 18 g 1L 7,2

*Hn hp vitamin nhm B Thiamine-HCl Riboflavin Niacin Pyridoxin-HCl Inositol Ca-Pantothenate Acid p-Aminobenzoic Biotin Nc ct 10 mg 10 mg 10 mg 10 mg 10 mg 10 mg 10 mg 10 mg 10 ml

Mi trng YS (Yeast extract-Soluble starch agar) Cao nm men Tinh bt tan Thch Nc ct pH 2g 10 g 18 g 1L 7,3

Thng s dng mi trng YS pha long 5 ln phn lp x khun v pha long 2 ln nghin cu. 121

Vi sinh vat hoc Nguyen Lan Dung Mi trng Maltose-Bennette Cao nm men Cao tht NZ amine Maltose monohydrat Thch Nc ct pH 1g 1g 2g 10 g 18 g 1L 7,3

Mi trng ISP-2

Cao nm men Cao malt Glucose Thch Nc ct pH

4g 10 g 4g 18 g 1L 7,2

Mi trng dch th YG (Yeast extract-Glucose) Yeast extract Glucose 1% 1%

Phng php RC (Rehydration- Centrifugation method): dng phn lp x khun c kh nng di ng 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 122

Vi sinh vat hoc Nguyen Lan Dung 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

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