LECTURE NOTES BACTERIOLOGY

7/25/2019 LECTURE NOTES BACTERIOLOGY

1/138

OCDEN, Junalyn B.Microbiology Notes (2015)

EC!"#ENO!E$

M%C#OB%OO&'

OCDEN, J"N'N B. (2015)

1


2/138


Microbiology- study of microorganismMicro- minute, smallBio-lifeLogus- study of

Branches:1. Bacteriology

- Study of bacteria- General Branch, involves pathogennon-pathogen-!athogen: disease causing bacteria to Man-study of medically important bacteria: !athogens"#ample:- Salmonella $yphi- "schirichia coli- bacteria that causes Gastroenteritis- %eisseria gonorrhae- $reponema pallidum- spirochete& subspecie: pallidum- causes syphilis

- 'ibrio cholera- cholera , indigestion of contaminated (ater). !arasitology- study of parasites

"#ample:- "ntamoeba histolytica- causes *moebiasis- !roto+oaingestion of contaminated (ater

- !lasmodium falciparum, malaria, viva#, ovaleMalaria- !roto+oa/0oly 2 because if you have one, you (ill have all

- *scaris Lumbricoides- contaminates (ater and food, )3,333 egss per day- $richiuris $richura- 0oo4(orms

. 'irology- study of virus"#ample:

- 5olds- caused by 6hinovirus rhinitis- *78S- caused by 07' , retrovirus- 0epa *- caused by picorna6%*& 0epa B- 0epadna8%*- S*6SSevere *cute 6espiratory Syndrome- "bola

9. Mycology- study of fungi"#ample:- 5andida albicans, *spergillus fumigates, 0istoplasma capsulatum

. !hycology *lcology- study of algae

2


3/138


S5;!" ;< M756;B7;L;G=:1. *gricultural Microbiology- use of microorganism to fertile soil.

>night soil- e#. ?rine, feces used as a fertili+er

). 6ectal S(ab- no collection of stool

07S$;6=:

1. 'arro: postulated principle of contagion.

5ontagion: - pertained as e#ternal forces- ;rganism that are bred, minute organisms- !resent in air, (ater and e#ternal forces

). 6oger Bacon: according to him, invisible creatures are present in the environment and itcauses diseases.

. Girolamo


4/138


!utrid meat e#posing it for one (ee4DD formation of maggots-came up (ith biogenesis

E. Fohn %eedham:

"#periment:?sed chic4en brothheatboil sealed in a container5L"*6

H.


5/138


-8iscovered and came up (ith pure culture techniKues for handling in the lab.-8escribed and discovered Bacillus *nthracis- anthray-Mycobacterium tuberculosis

9 !ostulates:

1. Microorganism or causative agent must be present in every case of disease e#cept carriers.DDan individual that carries the bacteria but doesnIt have the signs and symptoms.

8ifferent types of carriers:

a. *ctive carrier- an individual (ill acKuires the causative agentdisease process

recoverindividual (ill carry the causative agent indefinitelyb. !assive carrie- an individual acKuires the organism and does not e#perience the

manifestation of the disease but can help in the transmission. "#ample: Malaria

c. 7ncubatory- transmit disease during the incubation period. "#ample: 0epa *d. 5onvalescence 5arrier- transmits during the recovery period. "#ample: 5hic4en po#

). $he microorganism must be isolated and gro(n and culture outside the body.

"#ception: %ot all microorganisms gro( inside the body."#ample: Mycobacterium leprae- animal armadillo-treponema pallidum-subspecie: pallidumcausevenereal syphilis.

. Microorganisms must be introduced into a healthy susceptible lab animal. "#ception: $o those

(ho are resistant9. Microorganism must be isolated again and gro(n in pure culture inside the body. !urpose& for

comparison.

1E. @alter and


6/138


>differentiate organism

)1. !aul "rlich- discovered Salvarsan A5A- treatment for syphilis.- 7ntroduced 0M7 0umoral Mediated 7mmunity

>B-cellsproduced by anto-bodies

)). *le#ander


7/138


b. 5ellular ;rgani+ation:- ?nicellular or multicellularc. %utritional type:

1.


8/138


Jingdom: made up of t(o or more phylumSpecie: interbreeding organism that can produce fertile off springs& principal natural ta#onomicunit

>Strains- lo(er than a specie& organism that appear and behave differently from other organism(ithin the same specie.5lassified as:

o BiovarBiotype: Biochemical propertieso Morphovar: based on morphologyo !athovar !athotype: !ollogenecityo Serovar- antigenenic characteristics

8;M*7%S ;< L7


9/138


5yanobacteriablue-green algae, similar to pro4aryotic cells, have similar shape butnot totally:8ifferences in shape: flat, sKuare shape>*Kuatic photosynthetic- manufacture their o(n food

c. *rchaea- cross bet(een eu4aryotes and pro4aryotes"u4aryotes: Genes, metabolic path(ays- spend more energy as compared pro4aryotes.

$*P;%;M= ;< B*5$"67*

1. 8omain- Bacteria). Jingdom- Monera. 8ivision!hylum- ends (ith /es29. 5lass. ;rder- ends (ith /ales2

A.


10/138


- ) cocci- diplococci- produces a hori+ontal line in 1 plane- Series of cocci more than )- streptococci- %eisseria gonorrhea coffee bean- red Gram - ) planes : tetrad- planes: 9 infront &9 at the bac4

- Multiple !lanes: "#hibited by staphylococcus- !refi# Staphyle means grape li4e cluster- Bacilli- elongated rod shape- Bacilli in pairs- diplobacilli- Bacilli in chains: Strepto bacilli- Bacilli in 5hinese letter: e#ample: 5orynebacterium diptheriae

5ommon %ame: Jleb-LoefferIs- ability to bend to snap at the middle- !alisade !ic4et


11/138


a. 5ell membrane- semi-permeable membrane5omponents: 93O phospholipids&A3O proteins

-


12/138


%*M: %- acetyl muramic%*G: %-acetyl glucosamine

- !eptide: 9 proteins tetrapeptides


13/138


. *cid *lcohol- HO& decolori+er )3 secs& removes e#cess stain around the bacteria9. Safranin- counter stain or secondary stain

@hat causes gram positiveness of a bacteriaQ-13O-)3O peptidoglycan

. !athogenecityA. Stie of *ntibiotic *ction- penicillin destroys the cell (all& hydrolysis& cytoserine&

bacitracin& vancomysinC. Site of *ntigen determinant

-Glycocaly#- gelatinous structure outside the cell (all& not all have glycocaly#. Made upof:!olysaccharides- e#amples: Streptococcus pneumoniae 5% !reumococcus ,Jrebsiella pneumonia5%:


14/138


"u4aryotic: (hipping bacteria& countercloc4(ise:for(ard& cloc4(ise-tumbling& continuous cloc4(ise-stop.


15/138


-produce Bacteriocin protein that 4ills other bacteria9. 8egrative !lasmids- (ithstand the effects of to#ins

c. 6ibosomes- granular, function for protein synthesis0o( to determine ribosomes of:

!ro4aryotesC3 s.u. and "u4aryotesE3s.u.-site of anti-bitic actionSelective to#icity principle- certain destroy ribosomes inorder to 4ill the bacteria."#ample: "rythromycin, $etracycline, 5lindamycin, *minoglycosides

d. 7nclusion for food storage of bacteria:1. ;rganic compounds: polyhydro#y butyrate). 7norganic compounds: !olyphosphates, sulfur

e. "ndospores- dormant oval structures from vegetative bacteria- process of sporulationsporogenesis

'egetative bacteriaecposed to e#tremes and inorder to survive it , it (ill develop spores.!rocess is reversible Germination

5orte#: made up of modified peptidoglycan5oat: 4eratin li4e protein& 4eratin- rough hard (ater proof protein al(ays dry. 6esists theeffects of chemicals5rosporium: 5arbohydrates, Lipoproteins-helps the bacteria to gain the ability to form spores.


16/138


Spore location:1. 5ental spore: spore at the middle). "ccentric: a little bit distant to the middle. Sub-terminal: almost at the end

9. $erminal- present at the enda. $erminal-s(ollen:b. $erminal not s(ollen:

50*!$"6 9: B*5$"67*L !0=S7;L;G=*. 6eKuirements for Bacterial Gro(th:

1. $emperature 6eKuirement: Bacterial cells (ill al(ays have an optimum temperature best at(hich a bacteria (ill multiply rapidly

"#ample: )3335 R LLN?L)R optimum temperature)35 is the usual temperature

0igher than optimum temperatureRSlo(er rate of multiplicationa. !sychrope -135- cold-loving bacteria

"#ample: Legionella pneumophilapneumonia& can be inhaled , found in free+er and aircondition

b. !sychotroph)3935- cause food spoilage"#ample: Staphylococcus aureusR Staphylococcal


17/138


d.


18/138


!hotoautotroph Sunlight 5arbon dio#ide!hotoheterotroph Sunlight ;rganic compounds5hemolitotroph 7norganic

chemical0)S, %0,


19/138


- !urely dead cells

- 6elationship bet(een the log number cells and time

B*5$"67*L G"%"$75S

- !lasmids, bacterial chromosomes, se# piliSe#ual processes: transformation conugation transduction

*. $ransformation- $he na4ed 8%* from the environment is ta4en up by the bacterial cells- 7mportant in recombinant 8%* technology

"#ample: ". coli

treat (ith calcium

undergo heat shoc4

8%* e#tractions

placed in atest tube

B. 5onugation- 8irect transfer of 8%* bet(een bacterial cells

7nteraction of&1.


20/138


). 0fr: 0igh


21/138


) types:1. 5orrect loop out- plasmids). @rong loop out- part of plasmid (ill dissociate (ith of bacterial chromosome.

@rong loop-out:


22/138


'7. 0;S$ M756;B" 6"L*$7;%S07!!athology: !athos: suffering

- Scientific study of diseases7nfection: coloni+ation of microbes8isease: change in health status of an individual

22


23/138


!re-diseaseddiseased

>%ormal


24/138


1. 5;%$*5$ $6*%SM7SS7;%:a. 8irect 5ontact: no intermediate obect bet(een ) invidualsb. 7ndirect contact: presence of intermediate odect

7nfected7nfected ;bectanimateinanimateRcalled fomites%-77.

c. 8roplet nuclei:- Modified respiratory movements- $hrough droplet spread and travel less than 1m and doesnIt stay long in air- Snee+ing ,coughing- "#ample: !neumonia, !ertusis(hooping cough c.a.: Bordetella petussis

). 'ehicle $ransmission:a. @ater Borne $ransmission:- 7ngestion of contaminated (ater- "#ample: 5holera, *moebiasis, Gastroenteritis, $yphoid fever

b.

1. Mucous membrane:

6espiratory System- through inhalation

G7$- ingestion

G?$-se#ual). S4in:

2*


25/138


- $hrough the openings- pores, follicles. !arental 6oute:- 8irect inoculation of the microorganism underneath the s4in- !uncture needle stic4 inuries, bites9. $ransplacental 6oute:

- Mother

fetus

8"'"L;!M"%$ ;< 87S"*S":1. 7ncubation !eriod:

- 8efinite: 13-19 days- 'ariable: rabies e#ample


26/138


$here are surface receptors

Bacteria containing adhesin:- Streptococcus mutanstooth decay

Location of adhesion: glycocaly#- "scherichia coli

Location of adhesion: fimbriae- %eisseria gonorrhea

*ttach to the G?$, pharyn#, eyes

"#tra genital gonorrhea$herefore: transmission: oro-genital

- $reponema pallidum

Location: endoflagella

). 5apsule:


27/138


d. Mycolic acid 0ydro#ymetho#y acid- *4a (a#y lipid

9. "n+ymes:

- "specially "#oen+ymesa. 5ondigest materials bet(een cellsb. !romote blood clotc. !romote digestion of blood clot

- Staphyococcus aureus:. 5oagulase: promotes the formation of blood clot

o 5onverts fibrinogen to fibrin

A. Jinase: specific: Staphylo4inase


28/138


- Mucosal membrane---- prevent attachment of bacteria to 7g*. *ntigenic 'ariation:

!athogens (ill alter the shape to resist destruction by anti bodies

A. 7nvasins- proteins that (ill rearrange *ctin

!resent in: cell membrane $o#emia- presence of to#in in blood>$o#oid- (ea4ened e#oto#in!rocess to (ea4en e#oto#in:

- 0eat-


29/138



30/138


- !rovo4e 7mmune system -it (ill release a lot of$o release $ cells $ cells


31/138



32/138


%on spore forming: 5orynebacterium, Listeria, "rysiphelothri#Spore forming: 5lostridium and Bacillus;ther: Lactobacillus

7. Gram positive cocci:- Micrococcus: resemble staphylococcal species

- !eptococcus: only anaerobic GN cocci

GEN!" "TAPHYLOCOCC!"

"TAPHYLOCOCC!"5haracteristics:

Gram N cocci in clusters

*erobic to


33/138


CHARACTERI"TIC" IR!LENCE #ACTOR" DI"EA"E" LAB TE"T"

Gram N cocci inclusters

*erobic tofacultative

anaerobic Most pathogenic GN

cocci in clusters@hyQ !resence of 1virulence factors

;nly Staphylococcusthat is coagulase N1. Bound 5oagulase

5lumping factor

*ttached to thecell (all

8irectly interacts(ith fibrogen).


34/138


SSSS-StaphylococcalScalded S4inSyndrome

*lpha Beta

8elta Gamma$o#ins-function:cytoto#ins-destroydifferent cellsin the body

!rotein *-similar to thef#n of thecoagulase,

resist theaction ofphagocytes andresist theprocessopsoni+ation

5apsule

of the heart

. TO,IGENIC DI"EA"E"+1. SSSS- due to

e#foliation&

common toneonates). $SS- common to

those (ho usetampons& causes adrop in bloodpressureR drop inblood volumeresulting to:0ypovolemicshoc4, fever,

rashes. Staphylococcal


35/138


mucus membrane ifimmunocompetent

"ndocarditis medium, grayishto (hite, non-hemolytic,translucentcolonies

"TAPHYLOCOCC!" "APROPHYTIC!"

CHARACTERI"TIC" IR!LENCE #ACTOR DI"EA"E" LABORATORY TE"T"

Gram N inclusters

5atalase N

5oagulase

;pportunisticpathogen

%ormal flora inthe Genito-?rinary $ract

%o specific

"n+yme urease-responsible for?$7 signs andsymptoms:dysuria, fever,flan4 pain-

common topantyliner use ona daily basis

1. B*M: largecolonies,entire,color(hite to yello(to orangecolonies,glistening,

smooth, opaKue,butyrous, conve#

B. "TREPTOCOCC!"+

- Streptococcaceae

5haraceristics:

- Streptococci in chains& in pairs S. pneumoniae

-


36/138


a. PYOGENIC+- C35

Streptococcus pyogenes

). IRIDAN"+- $hermophiles& 935

Streptococcus mitis: causes endocarditis, sinusitis, *bscess painful inflammation (ithpus

Streptococcus salivarius: causes dental caries

Streptococcus thermophilus: used to enhance the flavor of cheese& non- pathogenic

c. ENTEROCOCC!" #AECALI"+- 1335 *%8 935

Streptococcus faecium

Streptococcus faecalis

Streptococcus bovis Streptococcus eKuines

o *ll causes ?$7 and endocarditis

d. LACTIC+- 1335

Streptococcus lactis

Streptococcus cremoriso Both enhances the flavor of cheese

0. "MITH BRO1N/"+

- Based on hemolysis

a. Al*2a32'mol&tic a%a G(''n "t('*tococc$s i(idans- !artial destruction of 6B5- Gives of greenish color

). B'ta32'mol&tic- 5omplete destruction of 6B5 produced by pyogenes

+


37/138


Streptococcus pyogenes, Streptococcus haemolyticus, Streptococcus agalactiae

c. Gamma 2'mol&tic+- %o destruction of 6B5

Streptococcus lactis, Streptococcus cremoris, Streptococcus faecalis

d. Al*2a P(im'+) +one:

- 7nnerD *lpha& ;uterDBeta

. 6"B"55* L*%5"


38/138


-dysphagia-if left untreated for 1-) (ee4s, the bacteria release erythrogenic to#incausing:

9. Scarlet


39/138


d. G6;?! 8"%$"6;5;55?S %;%-"%$"6;5;55?S

- *lphagamma gamma- Strep. faecalis faecium - Strep. mitis bovis

- @ound, subacute bacterialendocarditis, ?$7, septisimia - subacute bacterialendocarditis, ?$7, septisimia

e. G6;?! < G- beta hemolytic8ifference in B*M:< GGray, M*$, narro( +one of betahemolysis

Gray,M*$, (ide +one of betahemolysis

S$6"!$;5;55?S !%"?M;%7*" 87!L;5;55?S !%"?M;%7*"S$6"!$;5;55?S L*%5;"L*$?S

50*6*5$"67S$75S '76?L"%5"


40/138


bacteria tomultiplycelldivision

. !!!: !urpura!roducing

!rinciple-autolysis&caused byamidase& effect:dermalhemorrhage

. Meningitis9. Sinusitis. ;titis media

B7;50"M75*L $"S$S:1. B*57$6*57% S?S5"!$7B7L7$= $"S$:

!urpose: determine the effect of small amounts of Bacitracin

Streptococcus pyogenes- easily inhibited by Small amounts og BacitracinN result: +one of inhibition

). Bile "sculin *gar:- *gar slant

Streptococcus and "nterococcuso Gro( in 93O Bileo 0ydroly+e esculin producing: esculetin and ferric ions

5auses: color change in


41/138


Group B Strep: contains a diffusible e#tracellular protein a 5*M!


42/138


43/138


$ests:1. !yrido#al 6eKuirement $est). Satellite test. L*!


44/138


9. L!S Lipo !oly Saccharide- destroy epithelial cells, responsible for to#icity. Li!0E: for attachmentA.


45/138


$6"*$M"%$:- 8o#ycycline- 5eftria#one- Tuinolones: 5iproflo#acin, ;flo#acin, Levoflo#acin

L*B 87*G%;S7S:1. GramIs Stain: Gram diplococcic intracellular, e#tracellular). 5ulture:

a. $hayer-Martin $M

- $ype of 5*M (ith enrichment supplements:o 7sovitale#o 5olistin- inhibit gro(th of G- bacillio %ystatin- inhibit gro(th of yeasto 'ancomycin- inhibit gro(th of GN bacteria

b. Modified $hayer Martin:- *ddition of trimethoprim

c. Martin-Le(is ML8ifference:

- *ddition of anisomycin anti-fungal- 7ncreased concentration of vancomycin

d. %=5:*ppearance: transparent medium5omposition: lysed horse blood, horse plasma, yeast dialysate, colistin, vancomycin,nystatin, trimethoprim

*dvantage: permit gro(th of Mycoplasma hominis, ureaplasma urealyticum

e. 5*M:- Small, grayish (hite, conve#, translucent, shiny, smooth, irregular margins

f. F"MB"5 Fohn ". Martin Biological "nvironmental 5hamber- 5ontains 5;) generating system citric acid, %a)5;

*5


46/138


;ther tests:

1. 67* radio 7mmuno *ssay). "L7S* "n+yme Lin4 7mmunosorbent *ssay. Molecular *ssay :

- %on-amplified probe assay chemiluminescent test detect: Gonococcal6ibosomal 6%*- *mplified *ssay: for large screening

9. !article *gglutination Method:- !hadebact- Gonogen 77 test

%"7SS"67* M"%7%G7$787S5%: Meningococcus

G-diplococci, fastidious, capnophilic, %on-motile, %S>can gro( in M*5

G6*M N B*57LL7:

*


48/138


1. %on spore formers:- 5orynebacterium , Listeria,& both are catalase N and non branching& "rysipelothri# 5atalase

-

). Spore


49/138


>> .1ug4g body (eight

8isease:

1. 6espiratory 8iptheriaM;$:

- 6espiratory 8roplets: coughing, snee+ing, laughing- 7ndirect contact: (ith the use of fomites

7!: )- days

SS:1.


50/138


c. LoefflerIs medium (ith serum and egg: from beef e#tract pulp: causes coagulationconclusion: e#hibit proteolytic activity

9. !56 !olymerase 5hain 6eaction. $issue 5ulture $est

A. "le4sIs $est:

- 7mmunodiffusion test to verify if a certain strain of corynebacterium is to#igenic

Serogroups: gravis, mitis, belfanti, intermedius;$0"6 S!"57"S:

- 5.ei4eium: opportunistic- 5. urealyticum: urease ?$7- 5. pseudotuberculosis: sub-acute relapsing lymphadenitis- 5. #erosis: pharyngitis- 5. ulcerans: diphtheria li4e disease- 5. pseudodipthericum : lo( resistance to infection and endocarditis

0. List'(ia monoc&tog'n's5haracteristics:

- %on-encapsulated- 5atalase N and esculin- 'oges !ros4auer- %on-motile at 35 and motile at )35 tumbing motility

- 5hainspairs- 0abitat: G7$& females: G?$pregnant and gro(ing fetus

'


51/138


7%5?B*$7;%: ) monthsSS: lasts for )- daysS!"57M"%: placenta

5;L8 "%6750M"%$ M"$0;8:

%utrient brothincubate at 935 for (4s and mossub culture5*M: small, (hite, smooth,translucent, beta-hemolytic

>motility test:

%.Bincubate: 1-) hrs at room temperature& result: umbrella-li4e patternincubateover night and chec4 again for an umbrella-li4e pattern

. "6=S7!"L;$067P 60?S7;!*$07*"". 7%S78;S*- pleomorphicLong filamentsRrough coloniesShort filamentsR smooth coloniesV;;%;$75: pigs and cattles fishesQQ

'


52/138


). 0yaluronidase

87S"*S":*%7M*LS:

S(ine "rysipelas- rashes, dermatitis- 7nflammation of the s4in and oints

- $ransmission: direct contactM*%:

M*%:"rysipeloid:

- 7nflammation of the hands and fingers- 7tchy burning sensation s4in- 5omplication: Septicemia and endocarditis

L*B 87*G%;S7S:

5*M, 5*M, 5%*, 0B$: 0uman Blood Bilayer $(een

M / BC%% $O#E O#ME#$&enus Bacillus

BACILLUS ANTHRACISCN nt3ra4 Bacilliabitat $oilMO! 6irect contact, into 7oun6s, in3alation, ingestion

!'E$ O N!#81. Cutaneous nt3ra4 (59)

MO! s:in;s:in contactE4


53/138


; #areG causes se

; %ngestion oA 6airy


54/138


e6e@a

. et3al actor$


55/138


. $tring oA earls; DiPerentiate B.

ant3racis Aro@ B.cereus

; $usce


56/138


BACILLUS CEREUSCN rie6 #ice BacillusMO! ingestionG conta@inate6 Aoo6(Arie6 rice) soil

C#C!E#%$!%C$ F%#"ENCE C!O#$ D%$E$E B D%&NO$%$

$i@ilar 7it3 Bacillusant3racisDiPerence

; &ro7s best at *50C; Beta 3e@olytic; $e@i soli6 agarG

@otility s7ar@ing; Beta lacta@ase

1. Enteroto4in cause oAAoo6


57/138


BACILLUS SUBTILIS; Co@@on laboratory conta@inant

BM large, gray, 6ull, at, groun6 glass a


58/138


10.C%N Ce


59/138


EPEC (ENTERO PATHOGENIC ESCHERICHIA COLI)Fa. ilib. %nti@in ? cause 6estruction oA @icroHilli oA t3e s@all intestine

Disease1. %nAantile 6iarr3ea

; $oAt stool, AeHer, nausea, Ho@iting, ab6o@inal cra@


60/138


1. !$% g2. %


61/138


$3iegella 6ysenteriae CN le4ners BacillusDisease 3ili


62/138


B%OCEM%C !E$!1. !$% Q2. %MFC (,B,C) V + - -

D - + - -. % ?

SALMONELLA; Most are @otile, aerogenic (e4ce


63/138

, J yMicrobiology Notes (2015)

Mot ecalOral route%


64/138

, J yMicrobiology Notes (2015)

CITROBACTER SPECIES; Motile; Nor@al ora oA t3e colonCitrobacter Areun6eii an6 Citrobacter 6iHersus contain Hirulence Aactors

1. En6oto4in

2. Ca


65/138

J yMicrobiology Notes (2015)

EMB


66/138

Microbiology Notes (2015)

SERRATIA

$erratia @arcescens$erratia liKuiAaciens

$erratia rubi6aea ll are @otile, c3ro@ogenic, re6


67/138


roteus alcaliAaciens - +roteus stuartii V +roteus rettgeri + +roteus rustigianiis - -

; Cause "!%

"ERSINIA PESTIS; $3ort


68/138


C#C!E#%$!%C$; NON; ENC$"!ED, NON;MO!%E, $!%DO"$, E#OB%C; Contain &ranules M"C N"E$; cause bea6e6 a


69/138


. Caseous esionc3eese li:econsistency an6e@ergence oA

$$; eHer, c3ills, nig3t

s7eats,


70/138


1. !b oA t3e bones2. !b oA t3e oints. !b oA t3e s


71/138


5.ryl $ulAatase

+. !7een -0 3y6rolysis. %ron "


72/138


a.a Mantou4 !est intracutaneousa.b Fon irKuet; rub tric3oloro acetic aci6 on t3e s:in oA t3e


73/138


c. Non;


74/138


CLOSTRIDIUMabitat &%!, soil, 7ater, 6ecaying @atter%ncubation


75/138


nausea,Ho@itting ; 3ite 3alo aroun6t3e colonies

# L*,1 % *&1*:#esult Aor@ation oA stor@y clot? turbi6G 73ite to gray color

3# R&4&01& CAMP:

CO$!#%D%"M D%%C%E

CHARACTERISTICS VIRULENCE FACTORS DISEASES LABORATOR" DIAGNOSIS

; naerobic; $ub ter@inal

s


76/138


&; bacteria; CeAo4itin in3ibit

gro7t3 oA &/an6 &; bacteriaan6

; ructose; Contain

Neutral #e6 (


77/138


; !er@inal$7ollen $


78/138


COMMON %N FC""M $EED #OCE$$ OOD$B%!! $O%D%E#EN! $!#%N$ !O &G @e6ically i@


79/138


; $ub;ter@inals7ollen s


80/138

gy ( )

@ont3 %B isco@@on since@i4e6 Aee6ingis 6one

$$; Di


81/138


82/138


83/138



84/138

F%B#%O #EMO'!%C"$; Co@@a s3a


85/138

D%$E$E gastroenteritis

CAMP"LOBACTER

C69$75*&0 &*,1

C#C!E#%$!%C$; CurHe6 bacilli; $ s3a


86/138

%"#!E 'D#O'$%$

E!"$ +JEJ"N%

$"$CE!%B%%!' !Oetus Jeuni

Nali6i4ic #esistant $usce


87/138



88/138

2. "!%. %nAections


89/138

C%&ENE$ EC%$ ND C%&ENE$ ODO#N$; Motile; eritric3ous agellar arrange@ent; O


90/138


91/138



92/138

D%$E$E ertussis3oo ',>70$?8

0g&

Bro7ning oA


93/138

; Naso


94/138

%N MN; Brucella @elitensis; %ngestion oA goatss3ee


95/138



96/138

C"!"#Erie6;egg a


97/138

&ranulo@aFenereu@);73ite6isc3arge,ina@@ation oA

ly@


98/138

ll $igns an6 $y@


99/138



100/138

; %umerical *perture: pertain to the light gathering ability of obectives, determinedthrough the si+e of obectives.

). "lectron Microscope

; $"M: $ransmission "lectron Microscope; S"M: Scanning "lectron Microscope; Scanner- .13& M!;-.)&0!;-.A&;7;-1.)- (et using

; *s magnification increases , aperture increases & direct relationship

6efractive 7nde#: the refractive inde# of the glass is the same to the refractive inde#

>*voids refraction- Measure of the relative velocity of light as it passes through a medium

"#ample of Light Microscopes:

1. !hase contrast- (or4s by WWWWWWWWWWWWW the differences bet(een the refraction inde# andsurrounding medium.67- clearedenser image

). 8ar4-filled microscopes- illuminating only the specimen but not the field.

; 6esolution or 6esolving !o(er: ability to distinguish fine details or structures (ithin.; Minimum distance bet(een t(o obects (herein these obects could be seen as t(o separate

entities.

; @or4ing distance:o 8istance bet(een obective and specimeno 7n mmo Scanner- 1E. mm& M!;-13.Amm&0!;-3.Amm& ;7;-.1mm

o Magnification increases as (or4ing distance decreases& indirect relationship; 5ontrast: the number of visible shades seen in the specimen; Stain the slide before focusing

7llumination System: light source

; 5ondenser: focus the light; 7ris diaphragm: regulate light;


101/138

Support System: base, arm, stage, observative tubebody tube , revolving nose piece*dustable System: 5lostridium botulinum R canned good bacilli>Botulism- no contraction of muscles, difficulty in s(allo(ing- it cannot remove the endospores of thermophiles - most are non-pathogenic

8isinfection- reducing the number of harmful microorganisms- cannot destroy endospores- can destroy: vegetative pathogens or non-spore forming bacteria>8isinfectant- for non-living things>*ntiseptic- for living tissues

>8egerming- mechanical removal of microbes in a limited area

Sanitation- More on in the food industry- use of chemical agents to reduce microbes in food handing eKuipments.6eason:- Meet public health standards- 6educe bacterial contamination

101


M"$0;8S $; 5;%$6;L M756;B7*L G6;@$0:


102/138

M"$0;8S $; 5;%$6;L M756;B7*L G6;@$0:*. !0=S75*L denature or alter the structure of microorganism

-. !s' of 2'at+) types: Moist 0eat denature by coagulation and 8ry 0eathave an o#idi+ing effect

>$8!- $hermal 8eath !oint- pertains to the lo(est temperature at (hich microbes in a liKuidsuspension (ill be 4illed in 13 minutes.

"#ample: ". coli- C3

5

9)3

5- 13

1 mins.

>$8$- $hermal 8eath $ime- minimal length of time at (hich all microbes (ill be 4illed at a giventemperature.

>86$- 8ecimal 6eduction $ime- minimal length of time at (hich H3O of microbes (ill be 4illed at agiven temperature.

M;7S$ 0"*$:

1. Boiling- 13335 at sea level- Jills: vegetative bacteria, fungi some viruses e#cept those (ith endospores.

"#ample: 0epa *- survives boiling at 3mins. *nd bacteria (ith endospores can survive )3 hours.). *utoclave- steam under pressure. !asteuri+ation

- L$0- A-A35 or classical- ?0$- ?ltra 0igh $emperature- 19335 at sec.- cool in vacuum.

86= 0"*$:1. 8irect


103/138

1. 0"!* - 0igh "fficiency !articulate *ir


104/138

8isadvantages: capable of penetrating the s4in causing genetic mutation to an individual.

B.?ltraviolet radiation- non-ioni+ing radiation (ill produce thymine dimmers and it (ill causemicrobial mutation. But it cannot penetrate: paper, cloth and glass.@avelength:

5.Micro(ave 1mm-1m- it (ill 4ill microbesin the presence of (ater

-release heat absorbed by (ater4ills microorganisms"sp. 'egetative bacteria

5oo4ing por4 in micro(ave$richinosis& causative agent: $richinella spiralis

>*sepsis- absence of contamination>Sepsis- presence of bacterial contamination>*septic techniKue- methods in removingor avoiding contamination esp. among patients,instruments and medical personnels>Bacteriostatic- agent that (ill inhibit the gro(th of microbes.

>Germicidal- agent that acts as Bactericide, fungicide, viricidedoes not 4ill viruses but itinactivates them>Sporicide- destroys endosporesprotection fungal sporeshelp bacteria reproduce

B. 50"M75*L M"$0;8S:- Most in the form of disinfectants

1. !henols !henolics: 4ills microorganisms by destroying or disrupting the cell membrane ofmicrobes."#ample: 5arbolic *cid additive in throat spray- local anaesthetic8isadvantage& strong irritating odor s4in irritant

!henolics: chemical derivative of phenols$=!"S:1. 5resols

"#ample: Lysol- comes from coal tar effective disinfectant). Biphenols

"#ample: !hische#- 4ills staphylococcus and streptococcus bacteria*dvantage:

1. $hey are stable

10*


105/138


"#ample:


106/138

"#ample:1. Silver- 1O Silver %itrate- used to prevent the occurrence of opthalmia neonatum from:

neonate& results to purulent conunctivitis>5redeIs method- dropping of Silver %itrate in the eye.

). Mercury- merthiolate disinfect s4in (ounds Mercurochlome

. 5opper- 5opper Sulfate- anti microbial7t can destroy: algae in pools and fishtan4s9. Selenium- anti- fungal properties. Vinc- Vinc 5hloride- compound for mouth(ash- anti bacterial& +inc chloride- paints

.Tuarternary *mmonium 5ompounds *J* T?*$S- most effective disinfectant0o(Q By destroying the bacterial structures"#ample: Vephiran and 5epacol- both can be found in lab spray bottles"#ception: resistant R pseudomonas8isadvantages:

1. 7t (ill form foams). "asily inactivated by soaps . @hyQ Because soaps are anionic detergents.. "asily inactivated by organic compounds.

A.*ldehydes:0o(Q By forming covalent cross lin4s (ith several functional groups"#ample:

1.


107/138

E.!ero#ygens>;+one ;- highly reactive form of o#ygen e#pose to ?ltraviolet Light-very reactive in chlorine-disinfect pools-neutrali+e the taste and odor from (ater8isadvantage: less stable more e#pensive

>0ydrogen !ero#ide 0);)- effective as a disinfectant8isadvantage: not suitable for large (ounds because 0ydrogen !ero#ide is immediately acted on as

catalase@aterN;#ygen-can be used as sporadicidal not in room temperature but at high temperature.

>Ben+oyl !ero#ide>!eracetic acid- effective liKuid sporicide- 4ills bacteria and fungi in mins- it 4ills endospores viruses in 3 minutes.-used to clean food handling eKuipments because it doesnIt leave any to#ic residues.

5lean 5atch Method or Mid-Stream Method

*fter urine collection: it must be at least one hour7f process is delayed: refrigerate for 1E-)9 hours at 935

1. Grams Stain). 5ulture. 5olony 5ount: assess bacteriuria from contamination

"#ample: less than 13,333ml contaminantMore than 133,333ml- bacteriuria

a. !our !lateb.


108/138


). Lung biopsy


109/138

g p y. $ranstracheal *spiration near (indpipe9. Bronchial lavage

!repare specime:) slides methodStain:

1. Grams Stain Strep. pneumoniae and Jlebsiella5ulture>%T-%eufeld-Tuellung $est- capsular s(elling test

). *


110/138

g

?se !asteur pipette

Grams Stain5ulture

*bscess, @ounds "#udates&Syringe needles

*spiratecontainer!rocess

L*B M"$0;8S:7. 876"5$ M"$0;8:- "#amining microorganism in natural and living state

a. @et mount Saline mountb. 0anging drop->motility

77. 7%876"5$ M"$0;8:-


111/138


112/138


1. 5arbol fuschin


113/138

). 6osolic *cid in *lcohol. Meth. Blue

BaumgartenIs Method:- 8iluted alcohol fuschin- Myco. leprae 6ed& Myco. tuberculosis Blue

Special Stains:1.


114/138

- Section of environment safety that suggest safety!urpose:$o reduce ris4 of ha+ards

Levels:

1 Biosafety Level 1 BSL- microorganisms that do not consistently cause disease in healthyindividual."#ample: ". coli J1)

S. cerevisiae!olyomavirus

>Jno( basic laboratory techniKues:-


115/138


!67%57!L" ;< B7;S*


116/138

!67%57!L" ;< B7;S*


117/138

Generation $ime doubling $ime double $ime- $ime reKuired for bacterial cells to divide at regular intervals.- 5an be e#pressed as:

G$R Generation $ime

*. 876"5$ M"$0;8:1. !late Method 'iable 5ell count:

5ount the number of living cells

5


118/138


119/138


). 6eason (hy )- blood cultures:- $o reduce missing out a transient bacteria


120/138

- $o reduce missing out a transient bacteria>1ml of bloodRfe( bacteria

0o( (ill you collectQ- 1 hour interval

. 'olume*dult: 13 ml5hildren: )- ml%eonate 7nfant: 1-) ml

9. S4in disinfection:- Must be meticulously prepared

8ifferent disinfectants:

Blood culture: C3 percent alcohol, 3. O 5hlorhe#idine in C3O 6;0, 13 !orvidone$incture of 7odine: (ipe (ith C3O 6;0 to lessen s4in irritation

. *nti- coagulant:

S!S: Sodium !olyanethol Sulfate

7nhibit anti-bacterial of serum and phagocytes

5lot- trap microorganisms

%o S!S

@ith anti coagulant but no S!S

%o coagulant, %o S!S, %o broth

120


- Let it clot


121/138

A. 7ncubate:- C days, can incubate in ) (ee4s

C. 5hoice of broth:

$SB- $ryptic Soy Broth

*ll purpose liKuid media

E. Tuantity of Broth:'olume of broth:3 ml: 13P the volume of blood

H. Broth preparation:-scre(-capped conical tube

should not be tightly /scre(ed2

121


122/138


More than one hourR disintegrate13. Gram stain- most have peptidoglycans


123/138

13. Gram stain most have peptidoglycans7f $B is suspected: *


124/138

5ulture Media: anything that contains nutritional and environmental supplements$=!"S:

*. 5;%S7S$"%5=:1. Solid Media: contain )-O agar). Semi-solid media: also called as Gelatinous media& .-1O agar- used to chec4 motility of microorganisms.>motile: if there is cloudiness or ha+iness after incubation>non-motile: no ha+iness or clear. LiKuid or Broth Media:-no agar& al(ays be placed in a test tube

B. 5;M!;S7$7;%:1. Synthetic 8efined Media:

- "#act chemical composition of media is 4no(n

). %on-synthetic ?ndefined Media:

- 5hemical composition of one component is not e#actly 4no(n

. $issue 5ulture Media:- 5ontains living cells:

o @7E: normal human s4ino 0ep): 0uman epithelial cancer cellso 0ela cells: contains human cervical cancer cells

5. 0o( is it dispensed:1. !lated Media: placed on a !etri dish). $ube media: placed on test tubes

8. ?S":1. Simple *ll purpose Media

- *llo(s the gro(th of (ide variety of microbes"#ample: %utrient agar, B*M, $S* $ryptic Soy *gar

). "nrichment media:- !romote the gro(th of pathogens (hile inhibiting the gro(th of normal flora.

12*


- Most are in liKuid form"#ample:


125/138

p


126/138

g

h. *ntibiotics:

'ancomycin '5%- inhibits gro(th of GN bacteria

5olistin 5%- inhibits the gro(th of G- bacteria

%ystatin: inhibits fungal gro(th

i. p0:

6ogosasI$omato Fuice agar: Gro(th of Lactobacillus acidophilus

S8* Saborauds 8e#trose *gar- inhibit bacterial gro(th& allo(s fungal gro(th. p0R.AA. SelectiveSpecific: for the gro(th of one type or a specific type of a bacteria

"#ample:

!etragnaniIs media& for the gro(th of Mycobacterium tuberculosis.

$M: $hayer-Martin: for bacteria %eisseria

8ieudonneIs media: for the gro(th of 'ibrio 5holerae

Mac Bride: for the gro(th of Listeria monocytogens

E,PERIMENT --7noculation: 7ntroduction of microorganism in a media

$echniKues:1. LiKuidBroth:

7nsert inoculating needle at the acute angle

). *gar slant strea4:- Strea4 in +ig+ag manner from top-bottom

. *gar butt:- %eedle-DStab-D) of the culture media

9. *gar slant and butt:

12+


- Stab and strea4


127/138

>$S7- $riple Sugar 7ron

. !laced Strea4 Methods:

a. Simple Strea4:- 1 drop of the inoculums on the farthest side- Strea4 , forming parallel lines

b. 6adial Strea4:- Same 1ststep- Strea4 in a concentric fashion

c. Multiple Strea4- ?se of petri dish (ith 9 divisions- Separate strea4 per division using simple strea4

d. ;verlap:- Same 1ststep- 6otate : A3 degrees angle

- Strea4e. 7nterrupted Strea4&

- Same 1ststep- Strea4 and stop at the middle- $ilt for 1E3 degrees- Strea4

f. Multiple 7nterrupted:- 8ivide culture media into - Strea4 the 1stdivision and stop at the ape#-


128/138

E,PERIMENT -0INC!BATION

M"$0;8S:

1. *erobic incubation:- Simplest method of incubation

). 5apnophilic incubation:- 6eKuires -13O 5arbon- ?se of candle ar

. *naerobic incubation:$ypes:*. ?se of *naerobic ar:- 6emoval of ;)in the chamber (ith the reaction to 0)added to the system in the presence

of catalysts.

) techniKues:a. 8isposable Gas Generator "nvelope

- "#ample: Gas !ac4

- 5olorless plastic pac4ageblue basis for the absorbance of ;)

b. "vacuation 6eplacement $echniKue "6-$ech

- @ith the use of vacuum pump- ;)must be replaced by the follo(ing:

%itrogen- EO0)- 13O5;)- O

B. ?se of reducing Media- "#ample of culture media containing chemical component (ill combine or dissolve ;) to

remove ;).- "#amples:

12-


129/138


b. Shape:

5ircular


130/138

7rregular- no definite shape


131/138

8. 0"M;L=S7S:- *lpha- hemolysis

o 7ncompletepartial destruction of 6B5

5hange in the cell membrane colonies to greenish- Beta- hemolysis:o 5omplete destruction of 6B5o $here is clearing of colonies

". 5;L;6


132/138


133/138

a. 7ndole:Media: tryptophan tryptone broth media!rinciple: Bacteria that contain tryptophanase0o(Q) processes

b. Methyl 6ed:Media: M6-'! Methyl 6ed-'oges !ros4auerSubstances: 5lar4-LubIs 8e#trose Broth

M6 '! (ould producestrong acids p0: belo( 9.@hat are formedQ

c. 'oges !ros4auer:Media: M6 '!Substance: 5lar4-LubIs 8e#trose Broth

M6 '! producesacetoin chief end product of glucose metabolism from ButyleneGlycol !ath(ay

1


*cetoin N 93O J;0,;) *cetoin is bro4en do(n into simple products (hich is diacetyl N alpha-naphtol! iti lt d


134/138

!ositive result: red

d. 5itrate ?tili+ation $est:Media: Simmon 5itrate *gar Slantp0 indicator: bromthymol blueif color green: neutralif acidic, slant is color yello( or blue: al4aline

Bacteria that can utili+e citrate as source of carbon can also utili+e ammonium phosphateas their source of nitrogen (ith the production of ammonia to ammonium hydro#ide.

!ositive result: Green!russian Blue

A. ?rease $est :Stuarts ?rea Broth 5hristensenIs ?rea *garBacteria containing urease:

p0 indicator: phenolred!ositive result: red color%egative result: yello(

C. 5arbo#ylase $est:Media: MoeherIs decarbo#ylase*mino acids: Lysine, ;rnithine, *rginine

1*


135/138


- !ostive result: lysis clearing plaKue

E,PERIMENT -63 A"T


136/138

*ntimicrobial test: inhibiting activity of an agent to(ards the microbe

*nti microbial: microbial product in lo( concentration ugml

!urpose: guide for clinicians0elp to predict and monitor the effectiveness of a treatment

>Susceptible->6esistant->7ntermediate- some bact. *re inhibited, some are resistant$=!"S:

-. AGAR DI##!"ION METHOD KIRBY BA!ER+

INOC!L!M MEDIA ANTIBIOTIC" INC!BATION READING INTERPRETATION

*M;?%$ ;

Date post:	28-Feb-2018
Category:	Documents
Upload:	junalyn-banawe-ocden
View:	381 times
Download:	0 times

LECTURE NOTES BACTERIOLOGY

Documents